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Sample records for improves protein digestive

  1. An improved trypsin digestion method minimizes digestion-induced modifications on proteins.

    Science.gov (United States)

    Ren, Da; Pipes, Gary D; Liu, Dingjiang; Shih, Liang-Yu; Nichols, Andrew C; Treuheit, Michael J; Brems, David N; Bondarenko, Pavel V

    2009-09-01

    Trypsin digestion can induce artificial modifications such as asparagine deamidation and N-terminal glutamine cyclization on proteins due to the temperature and the alkaline pH buffers used during digestion. The amount of these artificial modifications is directly proportional to the incubation time of protein samples in the reduction/alkylation buffer and, more important, in the digestion buffer where the peptides are completely solvent exposed. To minimize these artificial modifications, we focused on minimizing the trypsin digestion time by maximizing trypsin activity. Trypsin activity was optimized by the complete removal of guanidine, which is a known trypsin inhibitor, from the digestion buffer. As a result, near complete trypsin digestion was achieved on reduced and alkylated immunoglobulin gamma molecules in 30min. The protein tryptic fragments and their modification products were analyzed and quantified by reversed-phase liquid chromatography/tandem mass spectrometry using an in-line LTQ Orbitrap mass spectrometer. The reduction and alkylation reaction time was also minimized by monitoring the completeness of the reaction using a high-resolution time-of-flight mass spectrometer. Using this 30-min in-solution trypsin digestion method, little protocol-induced deamidation or N-terminal glutamine cyclization product was observed and cleaner tryptic maps were obtained due to less trypsin self-digestion and fewer nonspecific cleavages. The throughput of trypsin digestion was also improved significantly compared with conventional trypsin digestion methods. PMID:19457431

  2. Improvement of Automatic In-Gel Digestion by in Situ Alkylation of Proteins

    OpenAIRE

    Yokono, Takeshi; Mineki, Reiko; Taka, Hikari; Kotaniguchi, Hiroto; Murayama, Kimie

    2003-01-01

    We have recently improved the automation of an in-gel digestion system, DigestPro 96, using in situ alkylation of proteins with acrylamide, conducted during one-dimensional (1D) SDS-PAGE. The improved method included the processes of destaining, dehydration, trypsin digestion, and extraction but excluded the reduction and alkylation steps following staining of proteins with CBB. The extracted peptide mixtures were directly loaded onto a micro C18 LC column of the mass spectrometer. The result...

  3. Improvement of Protein Digestibility in Jatropha curcas Seed Cakes by Gamma Irradiation

    International Nuclear Information System (INIS)

    Full text: The effect of gamma radiation on protein digestibility of Jatropha curcas press cake was investigated using in vitro digestibility technique. Six varieties of Jatropha curcas seeds were subjected to cobalt-60 gamma radiation at doses of 10-100 kGy. All treated seeds were defatted by screw press. In vitro protein digest abilities in defatted seeds were assayed using trinitrobenzene sulphonic acid (TNBS) method, by which the contents of alpha amino induced from the function of enzymes were determined using L-alanine as a reference standard. It was found that irradiation treatment at 60 kGy significantly increased the protein digestibility by 15-92%. Also, the results showed that moisture, crude protein, fat and ash contents were unchanged by irradiation, whereas fiber was significantly decreased (p < 0.05). Therefore, irradiation could serve as a possible processing method for protein utilization improvement in defatted Jatropha curcas seeds before using as a protein supplement in animal feed

  4. Improvement of automatic in-gel digestion by in situ alkylation of proteins.

    Science.gov (United States)

    Yokono, Takeshi; Mineki, Reiko; Taka, Hikari; Kotaniguchi, Hiroto; Murayama, Kimie

    2003-09-01

    We have recently improved the automation of an in-gel digestion system, DigestPro 96, using in situ alkylation of proteins with acrylamide, conducted during one-dimensional (ID) SDS-PAGE. The improved method included the processes of destaining, dehydration, trypsin digestion, and extraction but excluded the reduction and alkylation steps following staining of proteins with CBB. The extracted peptide mixtures were directly loaded onto a micro C18 LC column of the mass spectrometer. The resultant spectra were processed with "Mascot" search engine to estimate the sequence coverage of the bovine serum albumin (BSA). The original method, designed for Laemmli ID SDS gel applications, consisted of reduction and post-alkylation with iodoacetamide, which produced carboxyamidemethyl (CAM; -S-CH2CONH2) derivatives. The original method also included a desalting step essential for mass spectrometry, especially matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. We compared the original and improved methods using BSA (3 pmol loaded to the gel, one third of digested peptide mixture injected into LC-MS). The original method yielded both CAM and propionicamide (PAM;-S-CH2CH2CONH2) derivatives. The source of PAM derivatives is the unpolymerized acrylamide formed during electrophoresis. The sequence coverage of CAM derivatives of BSA by the original method was 10% with desalting and 19% without desalting. The sequence coverage of PAM derivative by the improved method was 32%. Our results clearly show the advantage of our improved automated in-gel digestion method for in situ PAM alkylated protein with respect to peptide recovery, compared with the original method with CAM post-alkylation. PMID:13678149

  5. Improvement of the digestibility of the proteins of the red alga Palmaria palmata by physical processes and fermentation.

    Science.gov (United States)

    Marrion, Olivier; Schwertz, Annie; Fleurence, Joël; Guéant, Jean Louis; Villaume, Christian

    2003-10-01

    Palmaria palmata (dulse) is an edible red alga constituting a potential protein source in human diet. However, previous studies showed that the digestibility of dulse proteins is bad because of the cell-wall encapsulating cytoplasmic proteins and the presence of fibers. The water-soluble xylan, present in high proportions in dulse, could be involved to explain the weak digestibility of proteins. To limit the influence of fibers and to improve the nutritional quality of these proteins, we have treated dulse by physical processes or by fermentation by moulds. After a 30 min predigestion by pepsin followed by a 6 h digestion into a cell dialysis containing porcine pancreatin, the corrected in vitro digestibility of crude dulse was very low (about 1.5% after correction by digestibility blank). The in vitro protein digestibility was estimated to 58% of that of casein for dulse samples obtained after washing in demineralized water and grinding in liquid nitrogen. The in vitro protein digestibility of fermented samples was 45%-65% of that of casein. After physical treatment, the digestibility improvement was related to the elimination of soluble molecules such as xylan and mineral salts. The improvement observed after fermentations seemed due to the degradation of insoluble fibers. PMID:14609091

  6. Digestibility of sorghum proteins.

    OpenAIRE

    Axtell, J. D.; Kirleis, A. W.; Hassen, M. M.; D Croz Mason, N.; Mertz, E. T.; Munck, L.

    1981-01-01

    Published information indicates that rice, maize, and wheat proteins are much more digestible in children than sorghum proteins are (66-81% compared with 46%). However, this digestibility difference cannot be demonstrated with the weanling rat, which gave digestibility values of 80% for cooked and 85% for uncooked sorghum gruels. Therefore, a search was made for a laboratory system sensitive to the digestibility differences between sorghum and other cereals. We found that porcine pepsin in vi...

  7. Improving the in vitro protein digestibility of sorghum with reducing agents

    OpenAIRE

    Hamaker, B. R.; Kirleis, A. W.; Butler, L. G.; Axtell, J. D.; Mertz, E. T.

    1987-01-01

    We have shown in previous reports that cooked sorghum protein is less digestible than other cooked cereal proteins. The pepsin-indigestible proteins in sorghum were found to be mainly prolamin proteins. Cooking sorghum in the presence of 2-mercaptoethanol increased protein digestibility (in vitro with pepsin or trypsin/chymotrypsin) to a level comparable with other cereals. At a concentration of 100 mM, other reducing agents (dithiothreitol, sodium bisulfite, and L-cysteine) were equally effe...

  8. Nanobiocatalysis for protein digestion in proteomic analysis

    OpenAIRE

    Kim, Jungbae; Kim, Byoung Chan; Lopez-ferrer, Daniel; Petritis, Konstantinos; Smith, Richard D.

    2010-01-01

    The process of protein digestion is a critical step for successful protein identification in the bottom-up proteomic analysis. To substitute the present practice of in-solution protein digestion, which is long, tedious, and difficult to automate, a lot of efforts have been dedicated for the development of a rapid, recyclable and automated digestion system. Recent advances of nanobiocatalytic approaches have improved the performance of protein digestion by using various nanomaterials such as n...

  9. Isotopic Changes During Digestion: Protein

    Science.gov (United States)

    Tuross, N.

    2013-12-01

    Nutrient and hydrological inputs traverse a complicated route of pH, enzymatic and cellular processes in digestion in higher animals. The end products of digestion are the starting products for biosynthesis that are often used to interpret past life-ways. Using an artificial gut system, the isotopic changes (dD, d18O, d13C and d15N) of protein are documented. Three separate protein sources are subjected to the conditions, chemical and enzymatic, found in the stomach and upper small intestine with only a small shift in the oxygen isotopic composition of the proteins observed. Middle to lower small intestine parameters produced both greater isotopic effects and significantly lower molecular weight products. The role of the gastric enterocyte and the likely involvement of the internal milieu of this cell in the isotopic composition of amino acids that are transported to the liver are reported.

  10. Proteinase K digestion of proteins improves detection of bacterial endotoxins by the Limulus amebocyte lysate assay: application for endotoxin removal from cationic proteins.

    Science.gov (United States)

    Petsch, D; Deckwer, W D; Anspach, F B

    1998-05-15

    Cationic proteins, such as lysozyme, ribonuclease A, and human IgG, impaired the detection of endotoxins with the Limulus amebocyte lysate assay (LAL assay) through formation of endotoxin-protein complexes, demonstrating pronounced masking of endotoxins. Methods, such as phenol extraction, dilution heating, and perchloric acid treatment failed to demask the endotoxins. Also, digestion with trypsin, chymotrypsin, or pronase recovered only 10 to 20% of the applied endotoxins. However, endotoxin recoveries up to 100% were obtained with proteinase K digestion of the samples prior to the LAL assay. This method was then applied to examine the impact of endotoxin masking on endotoxin removal from protein solutions by selective adsorption on membrane adsorbers. It was found that poly-L-lysine and poly(ethyleneimine) as endotoxin-selective ligands were able to pull endotoxins off the proteins studied, thereby guaranteeing successful decontamination. PMID:9606141

  11. Effects of Modified Digestion Schemes on the Identification of Proteins from Complex Mixtures

    OpenAIRE

    Klammer, Aaron A.; Maccoss, Michael J.

    2006-01-01

    In shotgun proteomics, a complex protein mixture is digested to peptides, separated and identified by microcapillary liquid chromatography followed by tandem mass spectrometry (LC-MS-MS). In this technology, complete protein digestion is often assumed. We show that, to the contrary, modifications to a standard digestion protocol demonstrate large, reproducible improvements in protein identification, a result consistent with digestion being a limiting factor in the efficiency of protein identi...

  12. Digestion of Protein in Premature and Term Infants

    OpenAIRE

    Dallas, David C.; Underwood, Mark A.; Zivkovic, Angela M.; German, J. Bruce

    2012-01-01

    Premature birth rates and premature infant morbidity remain discouragingly high. Improving nourishment for these infants is the key for accelerating their development and decreasing disease risk. Dietary protein is essential for growth and development of infants. Studies on protein nourishment for premature infants have focused on protein requirements for catch-up growth, nitrogen balance, and digestive protease concentrations and activities. However, little is known about the processes and p...

  13. Improvement of anaerobic digestion of sludge

    Energy Technology Data Exchange (ETDEWEB)

    Dohanyos, Michael; Zabranska, Jana; Kutil, Josef; Jenicek, Pavel

    2003-07-01

    Anaerobic digestion improvement can be accomplished by different methods. Besides optimization of process conditions is frequently used pretreatment of input sludge and increase of process temperature. Thermophilic process brings a higher solids reduction and biogas production, the high resistance to foaming, no problems with odour, the higher effect of destroying pathogens and the improvement of the energy balance of the whole treatment plant. Disintegration of excess activated sludge in lysate centrifuge was proved in full-scale conditions causing increase of biogas production. The rapid thermal conditioning of digested sludge is acceptable method of particulate matter disintegration and solubilization. (author)

  14. Digestibility of proteins and starch (in vitro) of amphidiploids (black gram x mung bean) as affected by domestic processing and cooking.

    Science.gov (United States)

    Kataria, A; Chauhan, B M; Punia, D

    1992-04-01

    The effects of different domestic processing and cooking methods on starch digestibility (in vitro) and protein digestibility (in vitro) of four strains of amphidiploids (black gram x mung bean) were investigated. An increase of 35 to 48% and 22 to 25% was observed in starch digestibility and protein digestibility, respectively, when the seed of amphidiploids were soaked for 18 h. Cooking (both of unsoaked and soaked seeds) and germination improved significantly the starch digestibility and protein digestibility of all the varieties. PMID:1374184

  15. Comparative study of in vitro digestibility of food proteins and effect of preheating on the digestion.

    Science.gov (United States)

    Takagi, Kayoko; Teshima, Reiko; Okunuki, Haruyo; Sawada, Jun-ichi

    2003-07-01

    Information on the comparative digestibility of food allergens and non-allergenic proteins is crucial when stability to digestion is to be used as a criterion to assess the allergenic potential of novel proteins. Preheating effect on in vitro digestibility has not been fully examined. In this study we investigated the preheating effect of in vitro digestibility of several proteins and their proteolytic fragments in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF). Five major food allergens, ovalbumin (OVA), ovomucoid (OVM), beta-lactoglobulin (BLG), bovine serum albumin (BSA), soybean trypsin inhibitor (STI), four proteins of unproven allergenicity, horseradish peroxidase (HRP), ribulose-1,5-bisphosphate carboxylase/oxidase (RBC), phosphinothricin acetyltransferase (PAT) and zein from corn, and plant lectin, concanavalin A (Con A) were preheated (at 100 degrees C for 5 min) or not preheated, and then digested in SGF or SIF. Food allergens were relatively stable in both SGF and SIF. Among the allergens, digestibility of OVA in both SGF and SIF was markedly decreased, and BLG and STI were relatively stable after preheating. Digestibility of ConA in SGF and SIF was markedly decreased by preheating. Digestibility of non-allergenic proteins in SGF was higher than the allergenic proteins. From these results, because of the marked increase of the digestibility in several proteins by preheating, systematic information concerning the effect of food treatment on protein digestion is necessary to assess the relationship between allergenic potential and the digestibility of food protein. PMID:12843620

  16. Comparative evaluation of fast protein enzymatic digestion.

    Czech Academy of Sciences Publication Activity Database

    Dy?ka, Filip; Bobá?, P.; Bobálová, Janette

    2011. s. 166. ISBN 978-972-99358-9-3. [International Congress on Analytical Proteomics /2./. 18.07.2011-20.07.2011, Ourense] R&D Projects: GA MŠk 1M0570; GA MŠk 1M06030; GA AV ?R IAA600040701 Institutional research plan: CEZ:AV0Z40310501 Keywords : in-gel digestion * protein Subject RIV: CB - Analytical Chemistry, Separation

  17. Improved digestibility of ?-lactoglobulin by pulsed light processing: a dilatational and shear study.

    Science.gov (United States)

    del Castillo-Santaella, Teresa; Sanmartín, Esther; Cabrerizo-Vílchez, Miguel Angel; Arboleya, Juan Carlos; Maldonado-Valderrama, Julia

    2014-12-28

    Modifying the protein conformation appears to improve the digestibility of proteins in the battle against allergies. However, it is important not to lose the protein functionality in the process. Light pulse technology has been recently tested as an efficient non-thermal process which alters the conformation of proteins while improving their functionality as stabilizers. Also, in order to rationally design emulsion based food products with specific digestion profiles, we need to understand how interfacial composition influences the digestion of coated interfaces. This study has been designed to investigate the effects of pulsed light (PL) treatment on the gastrointestinal digestion of protein covered interfaces. We have used a combination of dilatational and shear rheology which highlights inter and intra-molecular interactions providing new molecular details on protein digestibility. The in vitro digestion model analyses sequentially pepsinolysis, trypsinolysis and lipolysis of ?-lactoglobulin (BLG) and pulsed light treated ?-lactoglobulin (PL-BLG). The results show that the PL-treatment seems to facilitate digestibility of the protein network, especially regarding trypsinolysis. Firstly, PL treatment just barely enhances the enzymatic degradation of BLG by pepsin, which dilutes and weakens the interfacial layer, due to increased hydrophobicity of the protein owing to PL-treatment. Secondly, PL treatment importantly modifies the susceptibility of BLG to trypsin hydrolysis. While it dilutes the interfacial layer in all cases, it strengthens the BLG and weakens the PL-BLG interfacial layer. Finally, this weakening appears to slightly facilitate lipolysis as evidenced by the results obtained upon addition of lipase and bile salts (BS). This research allows identification of the interfacial mechanisms affecting enzymatic hydrolysis of proteins and lipolysis, which demonstrates an improved digestibility of PL-BLG. The fact that PL treatment did not affect the functionality of the protein makes it a valuable alternative for tailoring novel food matrices with improved functional properties such as decreased digestibility, controlled energy intake and low allergenicity. PMID:25358648

  18. Protein-sparing effect with high energy levels and obtaining the optimum digestible energy/digestible protein ratio in growth diets to Oreochromis niloticus(L.

    Directory of Open Access Journals (Sweden)

    Felix Walter Gutierrez

    2014-03-01

    Full Text Available The sparing effect of the dietary protein using diets with high energy levels was evaluated. A factorial design was used to examine the interaction of two levels of protein (30 and 35% and two levels of digestible energy (3.3 and 3.7 kcal/g on the weight gain (WG, food conversion FC, retained protein (PR, retained energy (ER and protein efficiency ratio (PER in “sex reversal Nile tilapia” (Oreochromis niloticus. Diets contained anchovy and soybean meal as protein sources and yellow corn and wheat bran as energy sources. The fish oil was used to adjust the energy levels. There were highly significant interactions (P<0.01 between the experimental diets. In diets with 30% proteins, the increase of digestible energy significantly enhances WG, FC, PR, ER and PER, where the level of 3.7 kcal/g had the best performance. With 35% protein, the increase of digestible energy had not significant influence on the GP, CA, PR, ER and PER, however the growth of sex reversal Nile Tilapia was improved with the level of 3.3 kcal/g. When were compared both diets, were not find significant differences. Considering the protein cost, it can be concluded that best response was obtained with a diet containing 30 % of protein and 3.7 kcal/g of digestible energy and optimum digestible energy/ digestible protein ratio of 13.75 kcal/g protein.

  19. IN VITRO GASTRO – INTESTINAL DIGESTION OF WHEAT COELIAC ACTIVE PROTEINS

    Directory of Open Access Journals (Sweden)

    Želmíra Balážová

    2012-02-01

    Full Text Available In vitro gastro-duodenal digestion of wheat coeliac active proteins was examined. Three sets of samples of wheat variety Brea were prepared from gastric and another three from duodenal digested samples. Some of proteins were found to be completely resistant to hydrolysis throughout the simulated gastric digestion and some of them throughout the simulated duodenal digestion. The most of polypeptides during subsequent incubation with enzymes resulted in gradual digestion, with only a trace of the parent proteins remaining after gastric digestion, whose majority disappeared after complete gastro-duodenal digestion. Despite the fact that the most of proteins were broken down by gastro-duodenal digestion into low molecular weight peptides, these molecules retain an allergenic potential.

  20. Digestion of Native Proteins for Proteomics Using a Thermocycler

    OpenAIRE

    Turapov, Obolbek A.; Mukamolova, Galina V.; Bottrill, Andrew R.; Pangburn, Michael K.

    2008-01-01

    Efficient protein digestion is a critical step for successful mass spectrometry analysis. Here we describe simultaneous tryptic digestion and gradual unfolding of native proteins by application of a temperature gradient using a single cycle of 5 min or less in a PCR thermocycler. Chemicals typically used for chromatographic techniques did not affect the digestion efficiency. Tryptic digestion was performed in a small volume (3 ?L) with 1.5 ?g of trypsin without denaturing agents. This rapid...

  1. Pepsin digestibility of proteins in sorghum and other major cereals

    OpenAIRE

    Mertz, Edwin T.; Hassen, Mohamed M.; Cairns-whittern, Carleen; Kirleis, Allen W.; Tu, Lichuan; Axtell, John D.

    1984-01-01

    We have shown previously that sorghum is highly digestible in the rat. However, other workers have shown that sorghum is much less digestible than wheat, maize, and rice in young children. Because the rat does not show these digestibility differences, we developed an empirical pepsin digestion method, first reported in 1981, which simulates the digestion values found in children. In this report the method has been improved and used to analyze wheat, maize, rice, millet, and sorghum and certai...

  2. Protein digestability of vegetables and field peas (Pisum sativum). Varietal differences and effect of domestic processing and cooking methods.

    Science.gov (United States)

    Bishnoi, S; Khetarpaul, N

    1994-07-01

    Protein digestibility was found to be 60.4 to 66.5 percent in raw unprocessed seeds of different pea cultivars. Protein digestibility (in vitro) was improved by the common methods of domestic processing and cooking including soaking, dehulling, ordinary cooking, pressure cooking and sprouting of legume grains. Pressure cooking had more pronounced effect on protein digestibility followed by ordinary cooking, sprouting, soaking for 18 h and (12 h) and dehulling. Pressure cooking of soaked and dehulled seeds was noticed to give most improved protein digestibility. PMID:7971789

  3. Fast On–Line Digestion System for Protein Characterization

    OpenAIRE

    Lo?pez-ferrer, Daniel; Petritis, Konstantinos; Lourette, Natacha M.; Clowers, Brian; Hixson, Kim K.; Heibeck, Tyler; Prior, David C.; Pasa-tolic?, Ljiljana; Camp, David G.; Belov, Mikhail E.; Smith, Richard D.

    2008-01-01

    An efficient on-line digestion system that reduces the number of sample manipulation steps has been demonstrated for high throughput proteomics. By incorporating a pressurized sample loop into a liquid chromatography-based separation system, both sample and enzyme (e.g., trypsin) can be simultaneously introduced to produce a complete, yet rapid digestion. Both standard proteins and a complex Shewanella oneidensis global protein extract were digested and analyzed using the automated on-line pr...

  4. Protein and starch digestibilities and mineral availability of products developed from potato, soy and corn flour.

    Science.gov (United States)

    Gahlawat, P; Sehgal, S

    1998-01-01

    A technique for development of potato flour was standardized. Five products viz. cake, biscuit, weaning food, panjiri and ladoo were prepared incorporating potato flour, defatted soy flour and corn flour. Baking and roasting were the major processing techniques employed for the development of these products. Protein, ash and fat contents of potato flour were almost similar to those of raw potatoes. Significant differences in protein, ash and fat contents of all the products were observed. Protein and starch digestibility of potato flour was significantly higher than that of raw potatoes. Protein digestibility increased by 12 to 17 percent on baking or roasting of products. Processed products had significantly higher starch digestibility and mineral availability compared to raw products. Thus, it can be concluded that roasting and baking are effective means of improving starch and protein digestibility and mineral availability of products. PMID:9839814

  5. Engineering the rabbit digestive ecosystem to improve digestive health and efficacy.

    Science.gov (United States)

    Combes, S; Fortun-Lamothe, L; Cauquil, L; Gidenne, T

    2013-09-01

    In rabbits, the bacterial and archaeal community of caecal ecosystem is composed mostly of species not yet described and very specific to that species. In mammals, the digestive ecosystem plays important physiological roles: hydrolysis and fermentation of nutrients, immune system regulation, angiogenesis, gut development and acting as a barrier against pathogens. Understanding the functioning of the digestive ecosystem and how to control its functional and specific diversity is a priority, as this could provide new strategies to improve the resistance of the young rabbit to digestive disorders and improve feed efficiency. This review first recalls some facts about the specificity of rabbit digestive microbiota composition in the main fermentation compartment, and its variability with some new insights based on recent molecular approaches. The main functions of the digestive microbiota will then be explained. Finally, some possible ways to control rabbit caecal microbiota will be proposed and a suitable timing for action will be defined. PMID:23769161

  6. Improving products of anaerobic sludge digestion by microaeration.

    Science.gov (United States)

    Jenicek, P; Celis, C A; Krayzelova, L; Anferova, N; Pokorna, D

    2014-01-01

    Biogas, digested sludge and sludge liquor are the main products of anaerobic sludge digestion. Each of the products is influenced significantly by specific conditions of the digestion process. Therefore, any upgrade of the digestion technology must be considered with regard to quality changes in all products. Microaeration is one of the methods used for the improvement of biogas quality. Recently, microaeration has been proved to be a relatively simple and highly efficient biological method of sulfide removal in the anaerobic digestion of biosolids, but little attention has been paid to comparing the quality of digested sludge and sludge liquor in the anaerobic and microaerobic digestion and that is why this paper primarily deals with this area of research. The results of the long-term monitoring of digested sludge quality and sludge liquor quality in the anaerobic and microaerobic digesters suggest that products of both technologies are comparable. However, there are several parameters in which the 'microaerobic' products have a significantly better quality such as: sulfide (68% lower) and soluble chemical oxygen demand (COD) (33% lower) concentrations in the sludge liquor and the lower foaming potential of the digested sludge. PMID:24569280

  7. Protein fragment imaging using ink jet printing digestion technique

    Science.gov (United States)

    Komatsu, Manabu; Murayama, Yohei; Hashimoto, Hiroyuki

    2008-12-01

    We have developed a capable tool for visualizing protein distribution at the sub-cell level with TOF-SIMS. This new method of visualizing a protein includes: (1) printing a digestive enzyme with ink jet (IJ) printing technology onto a tissue section, (2) measuring the two-dimensional distributions of decomposed peptide ions derived from digested protein on the tissue section by TOF-SIMS with a bismuth cluster ion (Bi 3+) source and using the aqueous ion enhancement method, and (3) analyzing the two-dimensional distributions of proteins using the results of proteome analysis. We have not yet tried this method onto a real tissue section. To show the possibility of this method, TOF-SIMS results of visualizing dot-patterned digested fragment peptide ions using IJ printing digestion technique onto a protein film are presented.

  8. Digestion

    Science.gov (United States)

    Digestion is a complex and miraculous process fundamental to our daily living. The complex nature of our digestive system, however, can also contribute to a variety of disorders. The following websites offer information and resources pertaining to the digestive system and digestive disorders. The first link leads to website for the American Gastroenterological Association which is an online resource for Association members as well as "other medical professionals with an interest in digestive diseases, patients and the general public." The site posts links to related news; and to sections for clinical resources, practice management, educational resources, and publications-to name a few (1). The second link leads to GastroLab, a long-standing online gastrointestinal resource specializing in digestive disorders. The site offers visitors a vast array of endoscopic images and video clips as well as a comprehensive dictionary of gastrointestinal terms (2). The third site presents the Gut Foundation, an organization that "provides professional and public education and promotes research into digestive disorders to improve gastrointestinal health." The Foundation website contains a variety of information about gastrointestinal conditions, diagnoses, medical research, membership, and more (3). The fourth website, from ViaHealth provides a brief overview of the digestive system; and information sections addressing common digestive disorders, rectal and colon cancer, diagnostic procedures, and online resources (4). From the ThinkQuest Library archives, the fifth site-titled La Casa de Comida-was created by a team of high school students for elementary school children. This section of the site offers a brief and basic comparison of digestive systems in birds, horses, insects, snakes, crustaceans, and mollusks (5). The sixth site contains a section of an online Biology Textbook-developed by Dr. Michael Gregory of Clinton Community College-providing a concise overview of the digestive system (6). The final link leads to a kid-friendly website from KidsHealth containing a short summary of the digestive system complete with a cartoony diagram illustrating all of the system's major parts (7).

  9. Mathematical modeling for digestible protein in animal feeds for tilapia

    Directory of Open Access Journals (Sweden)

    Luiz Vítor Oliveira Vidal

    2012-06-01

    Full Text Available The objective of this study was to formulate mathematical models to estimate digestible protein in some animal feeds for tilapia. Literature results of the proximate composition of crude protein, ether extract, and mineral matter, as well as digestible protein obtained in biological assays, were used. The data were subjected to multiple linear stepwise backward regression. Path analysis was performed to measure the direct and indirect effects of each independent variable on the dependent one. To validate the model, the experience used data from independent studies and values obtained from a digestibility trial with juvenile Nile tilapia testing five meat and bone meals, using the Guelph feces collecting system and chromium oxide (III as an indicator. The obtained model used to estimate digestible protein values (DP of animal origin is: DP(g kg-1 = -204.15+1.203xCP;R² = 0.953. The path coefficients showed a high direct positive effect (0.900 of crude protein on the digestible protein content. The mineral matter content has an indirect negative effect on protein digestibility (-0.710, reducing the crude protein content and quality.

  10. Mathematical modeling for digestible protein in animal feeds for tilapia

    Scientific Electronic Library Online (English)

    Luiz Vítor Oliveira, Vidal; Wilson Massamitu, Furuya; Elias Nunes, Martins; Tadeu Orlandi, Xavier; Mariana, Michelato; Themis Sakaguti, Graciano.

    1346-13-01

    Full Text Available The objective of this study was to formulate mathematical models to estimate digestible protein in some animal feeds for tilapia. Literature results of the proximate composition of crude protein, ether extract, and mineral matter, as well as digestible protein obtained in biological assays, were use [...] d. The data were subjected to multiple linear stepwise backward regression. Path analysis was performed to measure the direct and indirect effects of each independent variable on the dependent one. To validate the model, the experience used data from independent studies and values obtained from a digestibility trial with juvenile Nile tilapia testing five meat and bone meals, using the Guelph feces collecting system and chromium oxide (III) as an indicator. The obtained model used to estimate digestible protein values (DP) of animal origin is: DP(g kg-1) = -204.15+1.203xCP;R² = 0.953. The path coefficients showed a high direct positive effect (0.900) of crude protein on the digestible protein content. The mineral matter content has an indirect negative effect on protein digestibility (-0.710), reducing the crude protein content and quality.

  11. In Mesopore Protein Digestion: A New Forthcoming Strategy in Proteomics

    OpenAIRE

    Rosa Terracciano; Rocco Savino; Francesca Casadonte

    2011-01-01

    The conventional protocols for in solution or in gel protein digestion require many steps and long reaction times. The use of trypsin immobilized onto solid supports has recently captured the attention of many research groups, because these systems can speed-up protein digestion significantly. The utilization of new materials such as mesoporous silica as supports, in which enzyme and substrate are dramatically concentrated and confined in the nanospace, offers new opportunities to reduce the ...

  12. Effects of different industrial heating processes of milk on site-specific protein modifications and their relationship to in vitro and in vivo digestibility.

    Science.gov (United States)

    Wada, Yasuaki; Lönnerdal, Bo

    2014-05-01

    Heating processes are applied to milk and dairy products to ensure their microbiological safety and shelf lives. However, how differences in "industrial" thermal treatments affect protein digestibility is still equivocal. In this study, raw milk was subjected to pasteurization, three kinds of ultra-high-temperature (UHT) treatment, and in-can sterilization and was investigated by in vitro and in vivo digestion and proteomic methods. In-can sterilized milk, followed by UHT milk samples, showed a rapid decrease in protein bands during the course of digestion. However, protein digestibility determined by a Kjeldahl procedure showed insignificant differences. Proteomic analysis revealed that lactulosyllysine, which reflects a decrease in protein digestibility, in ?-lactalbumin, ?-lactoglobulin, and caseins was higher in in-can sterilized milk, followed by UHT milk samples. Thus, industrial heating may improve the digestibility of milk proteins by denaturation, but the improvement is likely to be offset by heat-derived modifications involved in decreased protein digestibility. PMID:24720734

  13. Enhanced Digestion Efficiency, Peptide Ionization Efficiency, and Sequence Resolution for Protein Hydrogen/Deuterium Exchange Monitored by FT-ICR Mass Spectrometry

    OpenAIRE

    Zhang, Hui-min; Kazazic, Sas?a; Schaub, Tanner M.; Tipton, Jeremiah D.; Emmett, Mark R.; Marshall, Alan G.

    2008-01-01

    Solution-phase hydrogen/deuterium exchange (HDX) monitored by high-resolution FT-ICR mass spectrometry offers a rapid method to study protein conformations and protein-protein interactions. Pepsin is usually used to digest proteins in HDX and is known as lack of cleavage specificity. To improve digestion efficiency and specificity, we have optimized digestion conditions and cleavage preferences for pepsin and protease type XIII from Aspergillus saitoi. A dilution series of the proteases was u...

  14. Phytase studies in pigs and poultry: effect on protein digestion and energy utilization

    OpenAIRE

    Kies, A. K.

    2005-01-01

    Phytase is applied for improving digestibility of phosphorus in pig and poultry diets. Indepen-dently, phytase also improves animal performance. The mechanisms to explain this effect were investigated and quantified. Protein can be complexed with phytate, especially under the acid conditions that occurs in the stomach of animals. Dietary phytase supplementation pre-vents formation of such complexes or, if such complexes are formed, helps to release protein faster and to a larger extent from p...

  15. ENZYME DIGEST AND ACID HYDROLYZED INDEX OF PROTEIN QUALITY EVALUATION

    OpenAIRE

    Mostafavi, H. Mohammadiha P.

    1984-01-01

    A pancreatopeptidase (Elastase) digest index was devised for a rapid and accurate estimation of protein quality. This index was calculated on the basis of all the amino acids released by an in-vitro Elastase digestion, acid hydrolyses of same sample and the residue of enzyme hydrolyzed. The amino acids were determined by Thin-Layer Chromatography. Samples used were cooked white kidneybeans, cooked and over-heated soybean powder, and skimmed milk powder. Good correlation was observed between e...

  16. Ostrich (Strutio camelus meat protein quality and digestibility

    Directory of Open Access Journals (Sweden)

    LS Reis

    2008-09-01

    Full Text Available The purpose of the study was to evaluate ostrich meat protein quality, as its consumption has significantly increased in the last few years in Brazil. Male Wistar rats were distributed in groupe of six elements. The standard group received a casein-based diet, the control group received a protein-free diet, and the experimental group received ostrich meat diet as protein source. The evaluated biological parameters were protein efficiency ratio (PER, net protein ratio (NPR, net protein utilization (NPU, and true digestibility (TD. There were differences (p<0.05 among treatment groups for all evaluated biological parameters. Mean true digestibility values were 92.12% and 75.77% for casein and ostrich meat, respectively.

  17. Ostrich (Strutio camelus) meat protein quality and digestibility

    Scientific Electronic Library Online (English)

    LS, Reis; TC, Oliveira.

    2008-09-01

    Full Text Available The purpose of the study was to evaluate ostrich meat protein quality, as its consumption has significantly increased in the last few years in Brazil. Male Wistar rats were distributed in groupe of six elements. The standard group received a casein-based diet, the control group received a protein-fr [...] ee diet, and the experimental group received ostrich meat diet as protein source. The evaluated biological parameters were protein efficiency ratio (PER), net protein ratio (NPR), net protein utilization (NPU), and true digestibility (TD). There were differences (p

  18. INVITRO DIGESTIBILITY OF PROTEIN FROM BARLEY AND OTHER CEREALS

    DEFF Research Database (Denmark)

    Buchmann, N. B.

    1979-01-01

    An in vitro method for measuring barley protein digestibility is presented. Samples were first incubated with pepsin in HCl; pancreatin was then added concomitantly with a bacteriostatic borate buffer. After TCA-precipitation, soluble nitrogen was measured. The digestion was unaffected by accumulated free amino acids. There were no free amino acids following pepsin treatment, but the essential ones were well liberated by pancreatin. Results for barley grown in the field or in pots, and for decortified barley fractions agreed with true digestibility values determined with rats. Of these samples, the field-grown barleys per se differed too little for the accuracy to be confirmed. The other cereals tested, oats, rye, maize, wheat, and rice, gave unsatisfactory results with pepsin/pancreatin, and also with pepsin, pancreatin, or pronase used separately. The ranking of the cereals according to in vitro digestibility depended on the type of enzyme and on the enzyme-to-substrate ratio.

  19. Towards quantitative prediction of proteasomal digestion patterns of proteins

    CERN Document Server

    Goldobin, Denis S

    2014-01-01

    We discuss the problem of proteasomal degradation of proteins. Though proteasomes are important for all aspects of the cellular metabolism, some details of the physical mechanism of the process remain unknown. We introduce a stochastic model of the proteasomal degradation of proteins, which accounts for the protein translocation and the topology of the positioning of cleavage centers of a proteasome from first principles. For this model we develop the mathematical description based on a master-equation and techniques for reconstruction of the cleavage specificity inherent to proteins and the proteasomal translocation rates, which are a property of the proteasome specie, from mass spectroscopy data on digestion patterns. With these properties determined, one can quantitatively predict digestion patterns for new experimental set-ups. Additionally we design an experimental set-up for a synthetic polypeptide with a periodic sequence of amino acids, which enables especially reliable determination of translocation ...

  20. ENZYME DIGEST AND ACID HYDROLYZED INDEX OF PROTEIN QUALITY EVALUATION

    Directory of Open Access Journals (Sweden)

    H.Mohammadiha P. Mostafavi

    1984-08-01

    Full Text Available A pancreatopeptidase (Elastase digest index was devised for a rapid and accurate estimation of protein quality. This index was calculated on the basis of all the amino acids released by an in-vitro Elastase digestion, acid hydrolyses of same sample and the residue of enzyme hydrolyzed. The amino acids were determined by Thin-Layer Chromatography. Samples used were cooked white kidneybeans, cooked and over-heated soybean powder, and skimmed milk powder. Good correlation was observed between elastase index value and their biological values reported in the literature from feeding trials. The pattern of aminoacids released by acid and by enzyme hydrolysis was about the same.

  1. Microbial Delignification with White Rot Fungi Improves Forage Digestibility

    OpenAIRE

    Akin, D. E.; Sethuraman, A.; Morrison, W. H.; Martin, S. A.; Eriksson, K. -e L.

    1993-01-01

    Three wild-type white rot fungi and two cellulase-less mutants developed from Phanerochaete chrysosporium K-3 (formerly Sporotrichum pulverulentum) were tested for their ability to delignify grass cell walls and improve biodegradation by rumen microorganisms. Fungal-treated and control stems of Bermuda grass were analyzed for their content of ester- and ether-linked aromatics by using alkali extraction and gas chromatography, for in vitro dry weight digestion and production of volatile fatty ...

  2. Following the digestion of milk proteins from mother to baby.

    Science.gov (United States)

    Holton, Thérèse A; Vijayakumar, Vaishnavi; Dallas, David C; Guerrero, Andrés; Borghese, Robyn A; Lebrilla, Carlito B; German, J Bruce; Barile, Daniela; Underwood, Mark A; Shields, Denis C; Khaldi, Nora

    2014-12-01

    Little is known about the digestive process in infants. In particular, the chronological activity of enzymes across the course of digestion in the infant remains largely unknown. To create a temporal picture of how milk proteins are digested, enzyme activity was compared between intact human milk samples from three mothers and the gastric samples from each of their 4-12 day postpartum infants, 2 h after breast milk ingestion. The activities of 7 distinct enzymes are predicted in the infant stomach based on their observed cleavage pattern in peptidomics data. We found that the same patterns of cleavage were evident in both intact human milk and gastric milk samples, demonstrating that the enzyme activities that begin in milk persist in the infant stomach. However, the extent of enzyme activity is found to vary greatly between the intact milk and gastric samples. Overall, we observe that milk-specific proteins are cleaved at higher levels in the stomach compared to human milk. Notably, the enzymes we predict here only explain 78% of the cleavages uniquely observed in the gastric samples, highlighting that further investigation of the specific enzyme activities associated with digestion in infants is warranted. PMID:25385259

  3. Protein utilization and amino acid digestibility of canola meal in response to phytase in broiler chickens.

    Science.gov (United States)

    Kong, C; Adeola, O

    2011-07-01

    The regression method was used in a 14-d broiler chicken study to determine the true ileal digestibility of amino acid (AA) and protein utilization in canola meal (CM, 388 g of CP/kg) without or with added phytase. Experimental treatments consisted of 2 factors, phytase at 2 levels (0 or 1,500 phytase units/kg) and CM at 3 levels (125, 250, or 375 g/kg). Birds received a standard starter diet from d 1 to 8 and the assay diets from d 8 to 22 posthatch. On d 8, a total of 384 birds were allocated to 6 dietary treatments in a randomized complete block design; excreta were collected from d 12 to 14 and d 19 to 21, and ileal digesta were collected on d 22 posthatch. True ileal indispensable AA digestibility of CM were derived from the regression of AA flow (mg/kg of DM intake) at the terminal ileum against the intake of AA (mg/kg of dietary DM) of birds fed diets without or with phytase. Body weight gain (BWG), protein gain, and protein intake increased linearly (P < 0.001) with increasing CM level, regardless of phytase supplementation. Effects of phytase (P < 0.05) were observed on BWG and the protein efficiency ratio from d 8 to 15, whereas effects of phytase (P < 0.05) were observed on BWG and protein gain from d 15 to 22. There was no effect of phytase on protein intake and net protein utilization from d 8 to 22. Phytase supplementation at 1,500 phytase units/kg did not affect true ileal digestibility of any AA in CM. In conclusion, the results of this study indicated that phytase supplementation improved the protein efficiency ratio of birds fed diets containing CM as the sole protein source from d 8 to 15 posthatch but did not affect the true ileal digestibility of AA in CM as determined by the regression method. PMID:21673166

  4. QTL Mapping of a High Protein Digestibility Trait in Sorghum bicolor

    OpenAIRE

    Winn, Jennifer A.; Esten Mason, R.; Robbins, Adriana L.; Rooney, William L.; Hays, Dirk B.

    2009-01-01

    Compared with other cereal grains, Sorghum bicolor shows lower protein digestibility. The low digestibility is thought to result from disulfide cross linking in the ?- and ?-kafirins. In contrast, the single recessive high digestibility/high lysine content (HD) mutation which confers greater grain digestibility exists in sorghum that is thought to result from reduced accumulation of ?-kafirin that allows greater access to the high digestible ?-kafarin fraction. In ...

  5. Toxic Trypsin Digest Fragment from the Bacillus thuringiensis Parasporal Protein

    OpenAIRE

    Aronson, John N.; Arvidson, Harold C.

    1987-01-01

    Enzymatic digestion in vitro of the Bacillus thuringiensis protoxin presumably releases and activates the toxin in a manner analogous to that which occurs when a B. thuringiensis sporulated fermentation preparation passes through the midgut of a lepidopteran larva. Therefore, a sporulated culture of B. thuringiensis subsp. kurstaki (serotype 3a3b) HD-263 was treated with trypsin to release an activated toxin soluble in bicarbonate buffer. A 63-kilodalton protein, toxic to cabbage looper larva...

  6. Rapid and efficient protein enzymatic digestion: An experimental comparison.

    Czech Academy of Sciences Publication Activity Database

    Dy?ka, Filip; Bobá?, P.; Mazanec, Karel; Bobálová, Janette

    2012-01-01

    Ro?. 33, ?. 2 (2012), s. 288-295. ISSN 0173-0835 R&D Projects: GA MŠk 1M0570; GA MŠk 1M06030; GA AV ?R IAA600040701 Institutional research plan: CEZ:AV0Z40310501 Keywords : in-gel digestion * MALDI-TOF MS * protein Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 3.261, year: 2012

  7. PHYSIOLOGICAL DIGESTION OF PEANUT PROTEINS IN THE SWINE MODEL OF FOOD ALLERGY

    Science.gov (United States)

    Rationale: The physiological digestion of peanut protein/allergens has not been critically evaluated. To compare the physiological digestion of peanut proteins/allergens to in vitro pepsin digestion, piglets were administered a peanut meal followed by periodic sampling and analysis of digesta. Me...

  8. Quantitative assessment of in-solution digestion efficiency identifies optimal protocols for unbiased protein analysis : Quantitative assessment of protein digestion protocols

    DEFF Research Database (Denmark)

    Leon, Ileana R; Schwämmle, Veit

    2013-01-01

    The majority of mass spectrometry-based protein quantification studies uses peptide-centric analytical methods and thus strongly relies on efficient and unbiased protein digestion protocols for sample preparation. We present a novel objective approach to assess protein digestion efficiency using a combination of qualitative and quantitative LC-MS/MS methods and statistical data analysis. In contrast to previous studies we employed both standard qualitative as well as data-independent quantitative workflows to systematically assess trypsin digestion efficiency and bias using mitochondrial protein fractions. We evaluated nine trypsin-based digestion protocols, based on standard in-solution or on spin filter-aided digestion, including new optimized protocols. We investigated various reagents for protein solubilization and denaturation (dodecyl sulfate, deoxycholate, urea), several trypsin digestion conditions (buffer, RapiGest, deoxycholate, urea), and two methods for removal of detergents prior to analysis of peptides (acid precipitation or phase separation with ethyl acetate). Our data-independent quantitative LC-MS/MS workflow quantified over 3700 distinct peptides with 96% completeness between all protocols and replicates, with an average 40% protein sequence coverage and an average of 11 peptides identified per protein. Systematic quantitative and statistical analysis of physicochemical parameters demonstrated that deoxycholate-assisted in-solution digestion combined with phase transfer allows for efficient, unbiased generation and recovery of peptides from all protein classes, including membrane proteins. This deoxycholate-assisted protocol was also optimal for spin filter-aided digestions as compared to existing methods.

  9. Gross and true ileal digestible amino acid contents of several animal body proteins and their hydrolysates.

    Science.gov (United States)

    Cui, J; Chong, B; Rutherfurd, S M; Wilkinson, B; Singh, H; Moughan, P J

    2013-07-01

    Amino acid compositions of ovine muscle, ovine myofibrillar protein, ovine spleen, ovine liver, bovine blood plasma, bovine blood globulins and bovine serum albumin and the amino acid compositions and in vivo (laboratory rat) true ileal amino acid digestibilities of hydrolysates (sequential hydrolysis with Neutrase, Alcalase and Flavourzyme) of these protein sources were determined. True ileal amino acid digestibility differed (P<0.05) among the seven protein hydrolysates. The ovine myofibrillar protein and liver hydrolysates were the most digestible, with a mean true ileal digestibility across all amino acids of 99%. The least digestible protein hydrolysate was bovine serum albumin with a comparable mean true ileal digestibility of 93%. When the digestible amino acid contents were expressed as proportions relative to lysine, considerable differences, across the diverse protein sources, were found in the pattern of predicted absorbed amino acids. PMID:23567135

  10. Microwave-assisted Protein Digestion on Various Locations of a Microplate

    OpenAIRE

    Jeongkwon Kim*; Jangmi Hong; Kyu Hwan Park; Hyun Sik Kim; Ji-hye Lee; Sehwan Park; Sunyoung Lee

    2011-01-01

    The effectiveness of microwave-assisted protein digestion in different well positions of a 96-well microplate wasinvestigated where microwave-assisted protein digestion of bovine serum albumin was performed in 10 different wells of a 96-wellmicroplate in a microwave oven. Similarly increased sequence coverages (~70%) were generally observed for the 10 microwaveassistedprotein digestion samples compared to conventional overnight digestion (63%), which is possibly due to higher miscleavageratio...

  11. Differential digestion of human milk proteins in a simulated stomach model.

    Science.gov (United States)

    Zhang, Qiang; Cundiff, Judy K; Maria, Sarah D; McMahon, Robert J; Wickham, Martin S J; Faulks, Richard M; van Tol, Eric A F

    2014-02-01

    A key element in understanding how human milk proteins support the health and development of the neonate is to understand how individual proteins are affected during digestion. In the present study, a dynamic gastric model was used to simulate infant gastric digestion of human milk, and a subsequent proteomic approach was applied to study the behavior of individual proteins. A total of 413 human milk proteins were quantified in this study. This approach demonstrated a high degree of variability in the susceptibility of human milk proteins to gastric digestion. Specifically this study reports that lipoproteins are among the class of slowly digested proteins during gastric processes. The levels of integral lysozyme C and partial lactadherin in milk whey increase over digestion. Mucins, ribonuclease 4, and macrophage mannose receptor 1 are also resistant to gastric digestion. The retention or enhancement in whey protein abundance can be ascribed to the digestive release of milk-fat-globule-membrane or immune-cell enclosed proteins that are not initially accessible in milk. Immunoglobulins are more resistant to digestion compared to total milk proteins, and within the immunoglobulin class IgA and IgM are more resistant to digestion compared to IgG. The gastric digestion of milk proteins becomes more apparent from this study. PMID:24694256

  12. Rapid characterization of protein chips using microwave-assisted protein tryptic digestion and MALDI mass spectrometry.

    Science.gov (United States)

    Ha, Na Young; Kim, Shin Hye; Lee, Tae Geol; Han, Sang Yun

    2011-08-16

    We demonstrate that the microwave-assisted protein enzymatic digestion (MAPED) method can be successfully applied to the mass spectrometric characterization of proteins captured on the affinity surfaces of protein chips. The microwave-assisted on-chip tryptic digestion method was developed using a domestic microwave, completing the on-chip proteolysis reaction in minutes, whereas the previous on-chip digestion methods by incubation took hours of incubation time. For the model protein chips, antibody-presenting surfaces were prepared, where anti-?-tubulin1 and antibovine serum albumin (BSA) were immobilized on self-assembled monolayers. The resulting digestion efficiency, displaying sequence coverages of 30 and 14% for ?-tubulin1 and BSA, respectively, was comparable to the previous time-consuming incubation studies. It allowed the characterization of immunosensed proteins by MASCOT search using peptide mass fingerprinting. In an example of this method for protein chip applications, BSA naturally involved in fetal bovine serum was unambiguously identified on a model protein chip by imaging mass spectrometry. This work shows that biomass spectrometry techniques can be implemented for surface mass spectrometry and biochip applications. Along with recent advances in imaging mass spectrometry, this technique will provide a new opportunity for high-speed, and thus high-throughput in the future, label-free mass spectrometric assays using protein arrays. PMID:21774472

  13. A hydrophilic immobilized trypsin reactor with N-vinyl-2-pyrrolidinone modified polymer microparticles as matrix for highly efficient protein digestion with low peptide residue.

    Science.gov (United States)

    Jiang, Hao; Yuan, Huiming; Liang, Yu; Xia, Simin; Zhao, Qun; Wu, Qi; Zhang, Lihua; Liang, Zhen; Zhang, Yukui

    2012-07-13

    In this work, a novel kind of N-vinyl-2-pyrrolidinone (NVP) modified poly acrylic ester microspheres was prepared, followed by trypsin immobilization to prepare a hydrophilic immobilized enzyme reactor (IMER), to achieve highly efficient protein digestion with low peptide residue. The nonspecific adsorption of peptides on such an IMER was evaluated by the in sequence digestion of bovine serum albumin (BSA) and myoglobin. Without NVP modification, both proteins could be identified after digestion by a 5 cm-length IMER, but 18 peptides of BSA were found in the digests of myoglobin caused by the nonspecific adsorption of the matrix. With NVP modification, the hydrophilicity of IMER was greatly improved, resulting in not only the sequence coverage of myoglobin increased from 63% to 73%, but also no residual peptides from BSA observed in myoglobin digests. Although the sequence coverages of proteins obtained by the IMER were comparable to those obtained by in-solution digestion, the digestion time was shortened from 24h to 1 min. By such an IMER, a protein mixture, containing BSA, myoglobin, and cytochrome c (100, 1 and 0.01 ?g/mL, respectively), was digested, and all proteins were unambiguously identified with improved sequence coverages than that achieved by in-solution digestion. Furthermore, the hydrophilic IMER was also off-line coupled to nano-RPLC-ESI-MS/MS for the analysis of proteins extracted from yeast. After 1.5 min digestion, 271 protein groups with at least 2 distinct peptides were identified, much more than those obtained by 24h in-solution digestion (192 protein groups), indicating the great potential of such an IMER for proteome analysis. PMID:22446077

  14. Effect of Phytase Supplementation on the Digestibility of Crude Protein, Amino Acids and Phosphorus of Cowpea (Vignia unguiculata) in Broilers

    OpenAIRE

    Iyayi, Eustace A.

    2013-01-01

    Cowpea contains phytate-P and other nutrients like amino acids which are poorly digested in broilers. Supplementation of such diets with microbial phytase helps to improve digestibility of these nutrients. In 2 experiments the effect of phytase supplementation on the digestibility of crude protein, amino acids and P of cowpea in broilers were investigated. Experiment One, had 6 diets in which 0, 150 or 300 g kg-1 cowpea replaced maize starch and 0 or 500 units of phytase enzyme (Natupho...

  15. Protein Digestibility of Different Animal and Plant Protein Sources for Labeo rohita Fingerlings

    Directory of Open Access Journals (Sweden)

    P. K. Sarker

    2000-01-01

    Full Text Available A laboratory trial was conducted on the protein digestibility of rohu (Labeo rohita fingerlings in a static indoor fish rearing water system of glass made aquaria (90 cm x 30 cm x 30 cm for a period of 75 days. Each treatment had two replicates containing 12 fingerlings. Five experimental diets A, B, C, D and E were prepared from different combination of five major sources of protein viz., fish meal, protein concentrate, meat & bone meal, mix of animal protein sources and mix of plant protein sources each containing 33% dietary protein level. The apparent protein digestibility (APD values of the above mentioned diets were 81.70, 78.66, 75.53, 72.53 and 69.96%.

  16. Effect of suppressing the synthesis of different kafirin subclasses on grain endosperm texture, protein body structure and protein nutritional quality in improved sorghum lines

    OpenAIRE

    Da Silva, Laura S.; Jung, Rudolf; Zhao, Zuo-yo; Glassman, Kimberly; Taylor, Janet; Taylor, J. R. N.

    2011-01-01

    To improve sorghum grain protein nutritional quality, improved sorghum lines were transformed to suppress the synthesis of different kafirin sub-classes, or backcrossed into transgenic lines with improved protein quality. Co-suppression of the alpha-, gamma- and delta-kafirin sub-classes and removal of the tannin trait resulted in transgenic sorghum lines with high cooked protein digestibility ( 80%), improved Amino Acid Score (0.8) and Protein Digestibility Corrected Amino Acid S...

  17. In vivo digestion of infant formula in piglets: protein digestion kinetics and release of bioactive peptides.

    Science.gov (United States)

    Bouzerzour, Karima; Morgan, François; Cuinet, Isabelle; Bonhomme, Cécile; Jardin, Julien; Le Huërou-Luron, Isabelle; Dupont, Didier

    2012-12-28

    The first months of life correspond to a key period in human life where dramatic physiological changes (establishment of microbiota, development of the immune system, etc.) occur. In order to better control these changes it is necessary to understand the behaviour of food in the gastrointestinal tract of the newborn. Infant formula is the only food for the newborn when breast-feeding is impossible. The kinetics of digestion of milk proteins and the nature of the peptides liberated in the small intestine throughout infant formula digestion have never been extensively investigated so far and were therefore studied using the piglet as a model of the newborn child. Piglets were fed infant formula by an automatic delivery system during 28 d, and slaughtered 30, 90 and 210 min after the last meal. Contents of stomach, proximal and median jejunum and ileum were collected and characterised. The extent of ?-lactoglobulin (?-lg), ?-lactalbumin (?-la) and casein proteolysis was monitored by inhibition ELISA, SDS-PAGE, immunoblotting and MS. At 30 min after the last meal, caseins were shown to be extensively hydrolysed in the stomach. Nevertheless, peptides originating mainly from ?-caseins (from 509 to 2510 Da) were identified in the jejunum and ileum of the piglets. ?-Lg partially resisted gastric digestion but completely disappeared in the stomach after 210 min. ?-La had a similar behaviour to that of ?-lg. Two large peptides (4276 and 2674 Da) generated from ?-lg were present in the ileum after 30 and 210 min and only one (2674 Da) after 90 min. PMID:22377314

  18. Susceptibility of sweet potato (Ipomoea batatas) peel proteins to digestive enzymes

    OpenAIRE

    Maloney, Katherine P.; Truong, Van-den; Allen, Jonathan C.

    2014-01-01

    Sweet potato proteins have been shown to possess antioxidant and antidiabetic properties in vivo. The ability of a protein to exhibit systemic effects is somewhat unusual as proteins are typically susceptible to digestive enzymes. This study was undertaken to better understand how digestive enzymes affect sweet potato proteins. Two fractions of industrially processed sweet potato peel, containing 6.8% and 8.5% protein and 80.5% and 83.3% carbohydrate, were used as a source of protein. Sweet p...

  19. Relação lisina digestível: proteína digestível em rações para tilápias-do-nilo / Digestible lysine/digestible protein ratio in diets for Nile tilapia

    Scientific Electronic Library Online (English)

    Giovani Sampaio, Gonçalves; Luiz Edivaldo, Pezzato; Margarida Maria, Barros; Leonardo, Tachibana; Maria Julia Santa, Rosa; Igo Gomes, Guimarães.

    2299-23-01

    Full Text Available Este estudo foi realizado para determinar a melhor relação lisina digestível:proteína digestível em rações para juvenis de tilápia-do-nilo. As tilápias (11,0 ± 0,43 g) foram distribuídas em 36 aquários (250 litros) numa densidade de seis peixes/aquário. Foram formuladas 12 rações com 22,0; 26,0 ou 3 [...] 0,0% de proteína digestível, com base na composição em aminoácidos digestíveis, e 4,5; 6,0; 7,5 ou 9,0% de lisina, em relação à proteína digestível, fornecidas à vontade durante 60 dias. Houve interação significativa dos fatores lisina digestível/proteína digestível para o ganho de peso, uma vez que o aumento dos níveis de lisina em rações com 26,0 e 30,0% de proteína digestível promoveu aumento linear desse parâmetro. Os melhores valores de conversão alimentar foram obtidos com os níveis de 26,0 e 30,0% proteína digestível, que determinaram aumento do consumo diário de proteína digestível. O nível de lisina promoveu redução linear da conversão alimentar e aumento linear da taxa de eficiência proteica. O nível de 26,0% de proteína digestível pode ser utilizado em rações para juvenis de tilápiado-nilo, entretanto essas rações devem ser formuladas com aminoácidos digestíveis e relação lisina digestível/proteína digestível de 6,0%. Quando utilizados níveis superiores a 26,0% de proteína digestível, o nível máximo de 7,5% de lisina digestível em relação à proteína digestível melhora o ganho de peso na espécie. Abstract in english This study was carried out to determine the best digestible protein/digestible lysine ratio that should be present in feed fed to Nile tilapias. Two hundred and sixteen tilapias (11.0 ± 0.43g) were distributed in 36 fish tanks (205 L) at a density of 6 fish/tank. Twelve feeds were formulated with th [...] ree different digestible protein (DP) levels 22.0; 26.0 and 30.0% (based on digestible amino acids) and four different lysine percentages of 4.5; 6.0; 7.5 or 9.0% in relation to digestible protein. The fish were fed ad libtum during a 60-day period. There was significant effect of the digestible protein and digestible lysine ratio on weight gain, because the increase in lysine levels in feeds with 26 and 30% DP promoted linear increase in this parameter. The best values for feed conversion were obtained at the levels 26 and 30% DP that increased the daily consumption of digestible protein. The lysine level caused a linear reduction in feed conversion and linear increase in the protein efficiency rate. The results suggested that the 26% DP level might be used in ration to feed Nile tilapia juveniles; however, these should contain digestible amino acids and the digestible lysine/digestible protein ratio should be 6.0%. However, for levels higher than 26% DP, a maximum digestible lysine level of 7.5% DP improved weight gain for the species.

  20. Co-digestion of municipal sewage sludge and solid waste: Modelling of carbohydrate, lipid and protein content influence.

    Science.gov (United States)

    Nielfa, A; Cano, R; Pérez, A; Fdez-Polanco, M

    2015-03-01

    Solid wastes from industrial, commercial and community activities are of growing concern as the total volume of waste produced continues to increase. The knowledge of the specific composition and characteristics of the waste is an important tool in the correct development of the anaerobic digestion process. The problems derived from the anaerobic digestion of sole substrates with high lipid, carbohydrate or protein content lead to the co-digestion of these substrates with another disposed waste, such as sewage sludge. The kinetic of the anaerobic digestion is especially difficult to explain adequately, although some mathematical models are able to represent the main aspects of a biological system, thus improving understanding of the parameters involved in the process. The aim of this work is to evaluate the experimental biochemical methane potential on the co-digestion of sewage sludge with different solid wastes (grease; spent grain and cow manure) through the implementation of four kinetic models. The co-digestion of grease waste and mixed sludge obtained the best improvements from the sole substrates, with additional positive synergistic effects. The Gompertz model fits the experimental biochemical methane potential to an accuracy of 99%, showing a correlation between the percentage of lipid in the substrates and co-digestions and the period of lag phase. PMID:25698789

  1. Growth of Nile tilapia post-larvae from broodstock fed diet with different levels of digestible protein and digestible energy

    Scientific Electronic Library Online (English)

    Silia Maria de Negreiros, Sousa; André, Freccia; Lilian Dena dos, Santos; Fábio, Meurer; Lucélia, Tessaro; Robie Allan, Bombardelli.

    2013-08-01

    Full Text Available SciELO Brazil | Language: English Abstract in english This study was carried out to evaluate the influence of different levels of digestible energy and digestible protein in diets of Nile tilapia (Oreochromis niloticus) broodstock on offspring growth during sex reversal stage. A total of 2,700 post-larvae (8.2±0.001 mg) derived from breeders fed nine d [...] iets with distinct levels of digestible protein (28, 34 and 40% DP) and digestible energy (2,800; 3,400; and 4,000 kcal DE.kg-1) were stored in twenty-seven 70 L tanks. After 30 days of growth, their average final weight (AFW, g), average weight gain (AWG, g), final standard length (SL, cm), condition factor (CF), specific growth rate (SGR, %/day), stock uniformity (UNI, %), survival (SUR, %) and sex reversal rate (SRR, %) were measured. Although female nutrition is thought to influence the performance or quality of progeny during early life stages, no influence of diets supplied to broodstock was detected on any parameter. Nonetheless, the offspring presented commercially satisfactory growth rates. Thus, diets containing 28% of digestible protein and 2,800 kcal of digestible energy.kg of diet-1 can be used to feed Nile tilapia broodstock without jeopardizing offspring performance during the sex reversal phase.

  2. Digestion and absorption of protein along ovine gastrointestinal tract

    International Nuclear Information System (INIS)

    Digestion and absorption of protein were determined in ovine gastrointestinal tract with cerium-141 as an unabsorbed reference substance. Nitrogen flows changed little in rumen and reticulum, but in the proximal small intestine flows increased because of secretion of .9 g nitrogen per day per kg body weight. This secretion included trypsin, chymotrypsin, elastase, and carboxypeptidases A and B; maximal activity was in proximal segments of the small intestine and decreased with distance from the pylorus. Activity of chymotrypsin decreased more rapidly than that of trypsin. Amino acid flows reflected the influx of protein in the duodenum; absorption was approximately 55% in the terminal ileum. No major changes of proportions of individual amino acids were observed. Overall nitrogen absorption was 72.6% of which 6% was in the large intestine. The major soluble protein fraction in the gastrointestinal tract consisted of peptides with molecular weight 7,000 to 14,000 daltons. Soluble high molecular weight protein was observed only in rumen and duodenum. Low molecular weight peptides and amino acids accumulated only in the proximal small intestine. Solubilization of protein and breakdown of peptides of 7,000 to 14,000 molecular weight appear to be rate limiting for protein absorption in sheep

  3. Proteins of Bacuri almonds: nutritional value and in vivo digestibility

    Scientific Electronic Library Online (English)

    Magalli Costa Barbosa, Lima e Silva; Priscila Aiko, Hiane; José Antônio, Braga Neto; Maria Ligia Rodrigues, Macedo.

    2014-03-01

    Full Text Available SciELO Brazil | Language: English Abstract in english Bacuri (Scheelea phalerata Mart.) is a type of palm fruit tree widely distributed in the Brazilian Cerrado. The objective of this paper was to study the almonds of bacuri, in their form in natura and processed, focusing on their nutritional value through the profile of amino acids, anti-nutritional [...] factors and in vivo digestibility. Raw and toasted samples of the almond presented a high level of proteins and fiber. Proteins of raw bacuri almond showed no limiting amino acid when compared to the ones recommended by FAO/WHO, and histidine was the most limiting essential amino acid in the toasted almonds. The almond of bacuri does not present anti- nutritional factors. In an assay with rats fed with control (casein), tests (bacuri almond flours) and aproteic diets, we verified the quantity of ration ingested and body weight gain, determining the urinary and metabolic nitrogen. Rats treated with the test diets presented inferior values of True Digestibility (DV), (82.9 and 72.3%, respectively for the raw and toasted almonds) when compared to the control group (92.3%). The raw bacuri almond presented a superior nutritional value to the one found in the toasted almond.

  4. On-Membrane Tryptic Digestion of Proteins for Mass Spectrometry Analysis

    OpenAIRE

    Luque-garcia, Jose L.; Neubert, Thomas A.

    2009-01-01

    Identification of proteins and characterization of posttranslational modifications are crucial steps for many biological, biochemical, and biomedical studies, and mass spectrometry has become the method of choice for these analyses. Here we describe two methods for the on-membrane digestion of proteins electroblotted onto nitrocellulose membranes prior to analysis by mass spectrometry. These on-membrane methods take approximately half the time of in-gel digestion and provide better digestion ...

  5. The role of protein digestibility and antacids on food allergy outcomes

    OpenAIRE

    Untersmayr, Eva; Jensen-jarolim, Erika

    2008-01-01

    Digestion assays with simulated gastric fluid have been introduced for characterization of food proteins to imitate the effect of stomach proteolysis on dietary compounds in vitro. By using these tests, dietary proteins can be categorized as digestion-resistant class 1 (true allergens triggering direct oral sensitization) or as labile class 2 allergens (nonsensitizing elicitors). Thus the results of these digestion assays mirror situations of intact gastric proteolysis. Alterations in the gas...

  6. An On-Line, High-Pressure Digestion System for Protein Characterization by Hydrogen/Deuterium Exchange and Mass Spectrometry

    OpenAIRE

    Jones, Lisa M.; Zhang, Hao; Vidavsky, Ilan; Gross, Michael L.

    2010-01-01

    The rapid and complete digestion of proteins is important when protein characterization by hydrogen deuterium exchange (HDX) is coupled with mass spectrometry. We developed a single-pump, on-line, high pressure digestion system that relies on UPLC technology to aid in the digestion of proteins. Two model proteins, A?1–42 and NBSA, were used to demonstrate the efficacy of the high pressure system. Both model proteins readily aggregate and are difficult to digest under normal conditions. Our...

  7. Evaluation of interspecimen trypsin digestion efficiency prior to multiple reaction monitoring-based absolute protein quantification with native protein calibrators.

    Science.gov (United States)

    van den Broek, Irene; Smit, Nico P M; Romijn, Fred P H T M; van der Laarse, Arnoud; Deelder, André M; van der Burgt, Yuri E M; Cobbaert, Christa M

    2013-12-01

    Implementation of quantitative clinical chemistry proteomics (qCCP) requires targeted proteomics approaches, usually involving bottom-up multiple reaction monitoring-mass spectrometry (MRM-MS) with stable-isotope labeled standard (SIS) peptides, to move toward more accurate measurements. Two aspects of qCCP that deserve special attention are (1) proper calibration and (2) the assurance of consistent digestion. Here, we describe the evaluation of tryptic digestion efficiency by monitoring various signature peptides, missed cleavages, and modifications during proteolysis of apolipoprotein A-I and B in normo- and hypertriglyceridemic specimens. Absolute quantification of apolipoprotein A-I and B was performed by LC-MRM-MS with SIS peptide internal standards at two time points (4 and 20 h), using three native protein calibrators. Comparison with an immunoturbidimetric assay revealed recoveries of 99.4 ± 6.5% for apolipoprotein A-I and 102.6 ± 7.2% for apolipoprotein B after 4 h of trypsin digestion. Protein recoveries after 20 h trypsin incubation equaled 95.9 ± 6.9% and 106.0 ± 10.0% for apolipoproteins A-I and B, respectively. In conclusion, the use of metrologically traceable, native protein calibrators looks promising for accurate quantification of apolipoprotein A-I and B. Selection of rapidly formed peptides, that is, with no or minor missed cleavages, and the use of short trypsin incubation times for these efficiently cleaved peptides are likely to further reduce the variability introduced by trypsin digestion and to improve the traceability of test results to reach the desirable analytical performance for clinical chemistry application. PMID:24168082

  8. MALDI-target integrated platform for affinity-captured protein digestion

    OpenAIRE

    Tajudin, Asilah Ahmad; Adler, Belinda; Ekstro?m, Simon; Marko-varga, Gyo?rgy; Malm, Johan; Lilja, Hans; Laurell, Thomas

    2013-01-01

    To address immunocapture of proteins in large cohorts of clinical samples high throughput sample processing is required. Here a method using the proteomic sample platform, ISET (Integrated Selective Enrichment Target) that integrates highly specific immunoaffinity capture of protein biomarker, digestion and sample cleanup with a direct interface to mass spectrometry is presented. The robustness of the on-ISET protein digestion protocol was validated by MALDI MS analysis of model proteins, ran...

  9. Adenovirus terminal protein protects single stranded DNA from digestion by a cellular exonuclease.

    OpenAIRE

    Dunsworth-browne, M.; Schell, R. E.; Berk, A. J.

    1980-01-01

    Adenovirus 5 DNA-protein complex is isolated from virions as a duplex DNA molecule covalently attached by the 5' termini of each strand to virion protein of unknown function. The DNA-protein complex can be digested with E. coli exonuclease III to generate molecules analogous to DNA replication intermediates in that they contain long single stranded regions ending in 5' termini bound to terminal protein. The infectivity of pronase digested Adenovirus 5 DNA is greatly diminished by exonuclease ...

  10. Effects of Malting and Fermentation on Anti-Nutrient Reduction and Protein Digestibility of Red Sorghum, White Sorghum and Pearl Millet

    Directory of Open Access Journals (Sweden)

    C. A. Onyango

    2012-11-01

    Full Text Available Sorghum and millet and their products require specialized treatment in order to improve their nutritive value, organoleptic properties and shelf-life. They contain anti-nutrients which are the major phytochemicals which negatively affects their nutritive values. The phytochemicals of concern include tannins and phytates, which interfere with mineral absorption, palatability and protein digestibility. Malting and fermentation treatments were applied to reduce the anti-nutrients, improve protein digestibility, and acidity to increase the products shelf life. The effects of malting and fermentation on the cereals nutritive value and anti-nutrient reduction were studied and evaluated for a period of 8 days. A treatment combining malting for 3 days and fermentation for 2 days respectively both at room temperatures (25°C was employed. Tannins and phytates were significantly reduced (p ? 0.05 by malting and fermentation. Protein digestibility was significantly (p ? 0.05 improved by malting and fermentation treatments; malted cereals digestibility ranged between 34.5-68.1% while the fermented flours protein digestibility range was 97.4-98.3%. The pH values were lowered to below 4.0, a level at which they could effectively inhibit spoilage microorganisms at the end of the fermentation period. A combination of optimum time treatments of malting and fermentation for 3 days and 2 days respectively were effective in reducing tannins and phytates and improving protein digestibility of the cereals.

  11. The protein composition of the digestive fluid from the Venus flytrap sheds light on prey digestion mechanisms

    DEFF Research Database (Denmark)

    Schulze, Waltraud X; Sanggaard, Kristian Wejse

    2012-01-01

    The Venus flytrap (Dionaea muscipula) is one of the most well-known carnivorous plants because of its unique ability to capture small animals, usually insects or spiders, through a unique snap-trapping mechanism. The animals are subsequently killed and digested to assimilate nutrients as the plants grow in mineral-deficient soils. We deep sequenced the cDNA from Dionaea traps to obtain transcript libraries, which were used in the mass spectrometry-based identification of the proteins secreted during digestion. The identified proteins consisted of peroxidases, nucleases, phosphatases, phospholipases, a glucanase, chitinases, and proteolytic enzymes, including four cysteine proteases, two aspartic proteases, and a serine carboxypeptidase. The majority of the most abundant proteins were categorized as pathogenesis-related proteins, suggesting that the plant's digestive system evolved from defense-related processes. This in-depth characterization of a highly specialized secreted fluid from a carnivorous plant provides new information about its prey digestion mechanism and the evolutionary processes driving its defense pathways and nutrient acquisition.

  12. Susceptibility of sweet potato (Ipomoea batatas) peel proteins to digestive enzymes.

    Science.gov (United States)

    Maloney, Katherine P; Truong, Van-Den; Allen, Jonathan C

    2014-07-01

    Sweet potato proteins have been shown to possess antioxidant and antidiabetic properties in vivo. The ability of a protein to exhibit systemic effects is somewhat unusual as proteins are typically susceptible to digestive enzymes. This study was undertaken to better understand how digestive enzymes affect sweet potato proteins. Two fractions of industrially processed sweet potato peel, containing 6.8% and 8.5% protein and 80.5% and 83.3% carbohydrate, were used as a source of protein. Sweet potato proteins were incubated with pepsin, trypsin, and chymotrypsin and protein breakdown was visualized with SDS-PAGE. After pepsin digestion, samples were assayed for amylase inhibitory activity. Sporamin, the major storage protein in sweet potatoes, which functions as a trypsin inhibitor as well, exhibited resistance to pepsin, trypsin, and chymotrypsin. Sporamin from blanched peel of orange sweet potatoes was less resistant to pepsin digestion than sporamin from outer peel and from extract of the white-skinned Caiapo sweet potato. Trypsin inhibitory activity remained after simulated gastric digestion, with the Caiapo potato protein and peel samples exhibiting higher inhibitory activity compared to the blanched peel sample. Amylase and chymotrypsin inhibitory activity was not present in any of the samples after digestion. PMID:25473492

  13. High efficiency and quantitatively reproducible protein digestion by trypsin-immobilized magnetic microspheres

    OpenAIRE

    Sun, Liangliang; Li, Yihan; Yang, Ping; Zhu, Guijie; Dovichi, Norman J.

    2011-01-01

    Aldehyde- and NHS-activated magnetic microspheres were used to immobilize trypsin (CHO-trypsin and NHS-trypsin), and their performance for protein digestion was evaluated by reversed phase liquid chromatography-electrospray ionization-tandem mass spectrometry using an LTQ Orbitrap Velos instrument. NHS-trypsin provided greater sequence coverage and identified more peptides for the digestion of bovine serum albumin. A one-minute digestion at room temperature using the immobilized trypsin also ...

  14. Processing technologies and cell wall degrading enzymes to improve nutritional value of dried distillers grain with solubles for animal feed: an in vitro digestion study.

    Science.gov (United States)

    de Vries, Sonja; Pustjens, Annemieke M; Kabel, Mirjam A; Salazar-Villanea, Sergio; Hendriks, Wouter H; Gerrits, Walter J J

    2013-09-18

    Currently, the use of maize dried distillers grain with solubles (DDGS) as protein source in animal feed is limited by the inferior protein quality and high levels of non-starch polysaccharides (NSP). Processing technologies and enzymes that increase NSP degradability might improve digestive utilization of DDGS, enhancing its potential as a source of nutrients for animals. The effects of various combinations of processing technologies and commercial enzyme mixtures on in vitro digestion and subsequent fermentation of DDGS were tested. Wet-milling, extrusion, and mild hydrothermal acid treatment increased in vitro protein digestion but had no effect on NSP. Severe hydrothermal acid treatments, however, effectively solubilized NSP (48-78%). Addition of enzymes did not affect NSP solubilization in unprocessed or processed DDGS. Although the cell wall structure of DDGS seems to be resistant to most milder processing technologies, in vitro digestion of DDGS can be effectively increased by severe hydrothermal acid treatments. PMID:23964718

  15. Quantitative Assessment of In-solution Digestion Efficiency Identifies Optimal Protocols for Unbiased Protein Analysis*

    OpenAIRE

    Leo?n, Ileana R.; Schwa?mmle, Veit; Jensen, Ole N.; Sprenger, Richard R.

    2013-01-01

    The majority of mass spectrometry-based protein quantification studies uses peptide-centric analytical methods and thus strongly relies on efficient and unbiased protein digestion protocols for sample preparation. We present a novel objective approach to assess protein digestion efficiency using a combination of qualitative and quantitative liquid chromatography-tandem MS methods and statistical data analysis. In contrast to previous studies we employed both standard qualitative as well as da...

  16. Host hemolymph proteins and protein digestion in larval Habrobracon hebetor (Hymenoptera: braconidae).

    Science.gov (United States)

    Baker, J E; Fabrick, J A

    2000-10-01

    Host plasma proteins and protein digestion in larval parasitoids were studied during trophic interactions of the ectoparasitoid Habrobracon hebetor Say (Hymenoptera: Braconidae), with a host, larvae of the Indianmeal moth, Plodia interpunctella Hübner (Lepidoptera: Pyralidae). We could detect no apparent differences in host hemolymph protein patterns up to 72 h after paralysation and/or parasitization by H. hebetor. A 190 kDa putative apolipophorin I present in host hemolymph could not be detected in the midguts of feeding H. hebetor larvae indicating that it is rapidly digested. The major 60 kDa storage proteins (putative hexamerins) in host hemolymph were detected in the parasitoid midgut and were completely digested 24 h after cessation of feeding and the beginning of cocoon formation. Host hemolymph had a pH of about 6.4. The pH optima of the midgut proteinases in the larval parasitoid were in the alkaline region, but midgut fluid in feeding parasitoid larvae was about pH 6. 8. Based on enzyme activity against selected artificial proteinase substrates including azocasein, N-alpha-benzoyl-L-Arg p-nitroanilide (BApNA), succinyl-Ala-Ala-Pro-Phe p-nitroanilide (SAAPFpNA), succinyl-Ala-Ala-Pro-Leu p-nitroanilide (SAAPLpNA), and inhibition by selected proteinase inhibitors, serine proteinases appear to be the predominant class of enzymes involved in protein digestion in the midguts of H. hebetor. There is also an active aminopeptidase (LpNA) associated with the microsomal fraction of midgut preparations. There was no evidence for preoral digestion or ingestion of proteinases from host hemolymph by the parasitoid larva. There was a very active BApNAase in the soluble fraction of midgut extracts. This activity increased on a per midgut basis up to 24 h after the beginning of cocoon formation but decreased rapidly by 48 h. Two major (P1 and P3) and several minor proteinases were detected in midgut extracts of H. hebetor analysed with gelatin zymograms. The apparent molecular mass of P1 varied from 95 to 49 kDa depending on protein loading. P3 had an apparent molecular mass of 39 kDa that was independent of protein loading. In summary, electrophoretic evidence indicates that host hemolymph protein patterns do not change significantly for at least 72 h after paralysation by H. hebetor. The role, if any, of envenomization in preventing breakdown of hemolymph proteins during this time remains to be determined. Because the predominant host hemolymph proteins, a putative apolipophorin I and the putative hexamerins, are readily digested by the serine proteinases present in the midguts of this parasitoid larva, these or similar proteins would provide an easily digested source of dietary amino acids that could be used for development of artificial diets for this beneficial insect. PMID:10899460

  17. Effects of Meat Cooking, and of Ingested Amount, on Protein Digestion Speed and Entry of Residual Proteins into the Colon: A Study in Minipigs

    OpenAIRE

    Bax, Marie-laure; Buffie?re, Caroline; Hafnaoui, Noureddine; Gaudichon, Claire; Savary-auzeloux, Isabelle; Dardevet, Dominique; Sante?-lhoutellier, Ve?ronique; Re?mond, Didier

    2013-01-01

    The speed of protein digestion impacts on postprandial protein anabolism. After exercise or in the elderly, fast proteins stimulate protein synthesis more efficiently than slow proteins. It has been shown that meat might be a source of fast proteins. However, cooking temperature, acting on the macrostructure and microstructure of the meat could affect both the speed, and efficiency, of protein digestion. This study aims to evaluate, in vivo, the effect of meat cooking on digestion parameters,...

  18. Ileal protein digestibility measurement in 15N-labelled rats

    International Nuclear Information System (INIS)

    After 15N -labelling over 7 days male albino rats received either a wheat or whole-egg diet (10 animals each) for 4 days. On the following day of the experiment 5 animals each continued to receive their diets as their morning meal (group 1 whole-egg, group 3 wheat) and 5 animals each after the previous feeding of a wheat diet received a 2.9 g whole-egg diet (group 2) and after the previous feeding of a whole-egg diet a 2.85 g wheat diet (group 4), resp. The morning meal was supplemented with chromium(III)oxide. The rats consumed their meals within 20 minutes. The animals were killed 3.5 hours after the beginning of feed intake. At that time the following relative amounts (in % of the intake) could be detected in the stomachs of groups 1 to 4:Cr2O3 = 22.5; 26.5; 57.5 and 64.2; dry matter = 25.4; 22.1; 43.2 and 38.5. The better coincidence between the whole-egg diet and Cr2O3 can be explained with hydrophobic qualities of Cr2O3 and low tendenc of Cr2O3 to separation from the whole-egg diet. In the first third of the small intestines less than 1% of Cr2O3 and a maximum of 3.5% of the dry matter could be detected. Between 20 and 36% of the Cr2O3 and 15 and 20% of the dry matter, resp. were ascertained in the small intestines as a whole; in the large intestines the values were 12 - 20% for Cr2O3 and 16 - 23% for2O3 and 16 - 23% for dry matter. Endogenous 15N -secretion could be ascertained in all parts of the digestive tract. The following ileal digestibility values were calculated for crude protein: whole-egg = 95.6%; whole-egg (wheat previously) = 95.5%; wheat = 94.1%; wheat (whole-egg previously) = 85.1%. It is a precondition for the application of this method that at the time of killing representative quotas of the diet sample to be tested can be detected both in the stomach and the large intestine so that the decrease of 15N -labelling in the ileum is actually caused by the test protein. (author)

  19. In Vitro Digestibilities of Six Rumen Protected Fat-Protein Supplement Formulas

    Directory of Open Access Journals (Sweden)

    L Hartati

    2012-01-01

    Full Text Available The aim of the research was to evaluate the efficacy of protection method of rumen protected fat-protein supplements. In vitro digestibility test was carried out to examine nutrients digestibility of different supplement formula based on the sources of protein and oil. The research used two sources of fat namely crude palm oil (CPO and fish oil (FO and three sources of protein namely milk skim, soy flour, and soybean meal. Thus there were 6 combinations that subjected in the in vitro digestibility test. The observed variables were the digestibility of dry matter (DM, organic matter (OM, crude fat (CF, and crude protein (CP. Results indicated that the method for protecting protein and fat was effective. This was showed by low nutrients digestibility in the rumen and high nutrients digestibility in the post rumen. In conclusion the combination between skim milk and CPO gave the best results among the other supplement formula.Keywords: rumen protected nutrient, fat-protein supplement, rumen digestibility, in vitro

  20. Integrative effects of feeding Aspergillus awamori and fructooligosaccharide on growth performance and digestibility in broilers: promotion muscle protein metabolism.

    Science.gov (United States)

    Saleh, Ahmed A; Amber, Khairy; El-Magd, Mohammed A; Atta, Mostafa S; Mohammed, Ahmed A; Ragab, Mohamed M; Abd El-Kader, Hanaa

    2014-01-01

    This study was conducted to show the effect of Aspergillus awamori (AA), fructooligosaccharide (FOS), and combined Aspergillus awamori and fructooligosaccharide (AA + FOS) on growth, digestibility, blood parameters, and expression of some growth-related genes. A total of 60 broiler chicks at the age of 15 d were divided into a control group (n = 15) and 3 treatment groups. The control group was fed a basal diet, and the treatment groups were fed basal diets supplemented with 0.05% AA, 0.05% FOS, and combined of 0.05% AA and 0.05% FOS. Results from measurement of growth performance and digestibility revealed a significant increase in the body weight gain with improved feed conversion rate in the experimental groups. Interestingly, dry matter digestibility (DMD) and crude protein utilization (CPU) were improved. In addition, plasma total cholesterol and low density lipoprotein-cholesterol (LDL-C) were decreased, while plasma high density lipoprotein-cholesterol (HDL-C) was increased by feeding AA, FOS, and AA + FOS. Expressions of growth hormone secretagogue receptor (GHSR), insulin-like growth factor 1 (IGF-1), and insulin-like growth factor 1 receptor (IGF1R) were increased in experimental groups. In conclusion, the supplementation of either Aspergillus awamori or fructooligosaccharide or both improves digestibility and growth performance probably by promoting skeletal muscle protein metabolism. PMID:24895630

  1. Influence of Non-fibrous Carbohydrate and Degradable Intake protein and Ruminal Fermentation ,Nutrien Digestion and performance of Local Sheep

    Directory of Open Access Journals (Sweden)

    Efka AR

    2001-05-01

    Full Text Available The objective of current study was to evaluate the impact dietary non-fibrous carbohydrate ( NFC and ruminally degradable intake protein (DIP concentration have on ruminal fermentation , nutrient digestion and performance of local sheep. The animal had a mean of liveweight 19.80 ±1.55 kg. four diets ,arranged in a 2x2 factorial ,were formulated to contain either 40 or 50 % NFC and 50 or 60 % of dietary crude protein as DIP .dietary DM contained 25 % Indonesian field grass and 75 % concentrate. Solvent –extracted or formaldehyd 2 % -treated soybean meal were used to alter DIP and corn or soybean hulls to alter NFC level. Percentage of energy and NDF digestion was similar ( p<0,01 as DIP level decreased in the diets. The soybean hulls was fermentable and total VFA concentration in the rumen increased ( p<0.01, but N-NH3 concentration was decreased ( p<0.01 as DIP level decreased in the diets. Daily live weight gain ( 146.29±25.84 g and body composition ( fat, water , protein and mineral was similar ( p<0.05 among diets. The preponderance ruminal fermentation ,nutrient digestion and performance of local sheeps did not be improved by sincronization of energy and nitrogen release but may more likely be limited by either energy or nitrogen alone. (Animal Production 3(2: 53-61 (2001Key Word : Carbohydrate, protein, rumen fermentation, nutrients digestion and performance

  2. Update: The Digestion and Absorption of Carbohydrate and Protein: Role of the Small Intestine.

    Science.gov (United States)

    Leese, H. J.

    1984-01-01

    Discusses the role of the small intestine in the digestion and absorption of carbohydrates and proteins. Indicates as outdated the view that these materials must be broken down to monomeric units before absorption and that the gut secretes a mixture of digestive juices which brings about absorption. (JN)

  3. FRACTIONATION OF DIGESTIVE PROTEINASES FROM TENEBRIO MOLITOR (COLEOPTERA: TENEBRIONIDAE) LARVAE AND ROLE IN PROTEIN DIGESTION

    Science.gov (United States)

    Tenebrio molitor larval digestive proteinases were purified and characterized by gel filtration chromatography combined with activity electrophoresis. Cysteine proteinases, consisting of at least six distinct activities, were found in three chromatographic peaks in anterior and posterior midgut chro...

  4. Rapid Online Non-Enzymatic Protein Digestion Combining Microwave Heating Acid Hydrolysis and Electrochemical Oxidation

    Science.gov (United States)

    Basile, Franco; Hauser, Nicolas

    2010-01-01

    We report an online non-enzymatic method for site-specific digestion of proteins to yield peptides that are well suited for collision induced dissociation (CID) tandem mass spectrometry (MS/MS). The method combines online microwave heating acid hydrolysis at aspartic acid and online electrochemical oxidation at tryptophan and tyrosine. The combined microwave/electrochemical (microwave/echem) digestion is reproducible and produces peptides with an average sequence length of 10 amino acids. This peptide length is similar to the average peptide length of 9 amino acids obtained by digestion of proteins with the enzyme trypsin. As a result, the peptides produced by this novel non-enzymatic digestion method, when analyzed by ESI-MS, produce protonated molecules with mostly +1 and +2 charge states. The combination of these two non-enzymatic methods overcomes shortcomings with each individual method in that: i) peptides generated by the microwave-hydrolysis method have an average amino acid length of 16 amino acids, and ii) the inability of the electrochemical-cleavage method to reproducibly digest proteins with molecular masses above 4 kDa. Preliminary results are presented on the application and utility of this rapid online digestion (total of 6 min digestion time) on a series of standard peptides and proteins as well as an E. coli protein extract. PMID:21138252

  5. Improving methodology in open vessel digestion with a graphite heating block (T7)

    International Nuclear Information System (INIS)

    Full text: Open block digestion systems have been very popular in environmental analysis over the past decades, but have consistently suffered from the major drawback of their sensitivity against corrosion and the subsequent risk of contamination. Therefore block digestion systems have not been considered state-of-the-art technology in trace and ultra trace sample preparation. Graphite block digestion systems are well established in North America and are recently becoming more frequently considered in Europe. These systems overcome the deficiencies of the traditional systems, made from stainless steel or aluminum, because the block is manufactured from graphite and typically coated with a fluoro-polymer to present the possibility metallic contamination from the surface of the system during the handling of the samples. Graphite block systems present an alternative to the current mainstream technology of open and closed vessel microwave assisted digestion systems, as they allow large numbers of samples to be digested simultaneously, thus overcoming one of the major weaknesses of closed vessel systems. More recently a number of improvements in the technology has been developed for graphite block digestion systems and studies have been performed to evaluate the effects of such improvements. The paper presented will deal with the technological improvements: monitoring and control of sample temperature vs. monitoring of block temperature, elimination of cross contamination effects during open vessel block digestion, evaporation of samples for pre-concentration or multiple digestion steps, addressing the needs of various labs and applications for block digesters. The effects of those developments will be discussed; application examples and finally an outlook into possible future trends for graphite block digestion systems will be given. (author)

  6. Influence of protein type on oxidation and digestibility of fish oil-in-water emulsions: gliadin, caseinate, and whey protein.

    Science.gov (United States)

    Qiu, Chaoying; Zhao, Mouming; Decker, Eric Andrew; McClements, David Julian

    2015-05-15

    The influence of three surface-active proteins on the oxidative stability and lipase digestibility of emulsified ?-3 oils was examined: deamidated wheat gliadin (gliadin); sodium caseinate (CN); whey protein isolate (WPI). Gliadin and WPI were more effective at inhibiting lipid oxidation (hydroperoxides and TBARS) of fish oil-in-water emulsions than CN. Protein oxidation during storage was determined by measuring the loss of tryptophan fluorescence. The CN-emulsions exhibited the highest loss of tryptophan fluorescence during aging, as well as the highest amount of lipid oxidation. Potential reasons for the differences in oxidative stability of the emulsions with different proteins include differences in interfacial film thickness, protein chelating ability, and antioxidant amino acids profiles. During in vitro digestion, gliadin-stabilized emulsions had the lowest digestion rate of the three proteins. These results have important implications for using proteins to fabricate emulsion-based delivery systems for ?-3 oils. PMID:25577077

  7. Immunohistochemical localization of translationally controlled tumor protein in the mouse digestive system.

    Science.gov (United States)

    Sheverdin, Vadim; Jung, Jiwon; Lee, Kyunglim

    2013-09-01

    Translationally controlled tumor protein (TCTP) is a housekeeping protein, highly conserved among various species. It plays a major role in cell differentiation, growth, proliferation, apoptosis and carcinogenesis. Studies reported so far on TCTP expression in different digestive organs have not led to any understanding of the role of TCTP in digestion, so we localized TCTP in organs of the mouse digestive system employing immunohistochemical techniques. Translationally controlled tumor protein was found expressed in all organs studied: tongue, salivary glands, esophagus, stomach, small and large intestines, liver and pancreas. The expression of TCTP was found to be predominant in epithelia and neurons of myenteric nerve ganglia; high in serous glands (parotid, submandibular, gastric, intestinal crypts, pancreatic acini) and in neurons of myenteric nerve ganglia, and moderate to low in epithelia. In epithelia, expression of TCTP varied depending on its type and location. In enteric neurons, TCTP was predominantly expressed in the processes. Translationally controlled tumor protein expression in the liver followed porto-central gradient with higher expression in pericentral hepatocytes. In the pancreas, TCTP was expressed in both acini and islet cells. Our finding of nearly universal localization and expression of TCTP in mouse digestive organs points to the hitherto unrecognized functional importance of TCTP in the digestive system and suggests the need for further studies of the possible role of TCTP in the proliferation, secretion, absorption and neural regulation of the digestive process and its importance in the physiology and pathology of digestive process. PMID:23834399

  8. Bayesian hierarchical reconstruction of protein profiles including a digestion model

    OpenAIRE

    Grangeat, Pierre; Szacherski, Pascal; Gerfault, Laurent; Giovannelli, Jean-franc?ois

    2012-01-01

    Introduction : Mass spectrometry approaches are very attractive to detect protein panels in a sensitive and high speed way. MS can be coupled to many proteomic separation techniques. However, controlling technological variability on these analytical chains is a critical point. Adequate information processing is mandatory for data analysis to take into account the complexity of the analysed mixture, to improve the measurement reliability and to make the technology user friend...

  9. Prebiotics affect nutrient digestibility but not faecal ammonia in dogs fed increased dietary protein levels.

    Science.gov (United States)

    Hesta, M; Roosen, W; Janssens, G P J; Millet, S; De Wilde, R

    2003-12-01

    An increased protein content and less digestible protein sources in the diet can induce bad faecal odour. The present study investigated the effect of adding prebiotics to dog diets enriched with animal-derived protein sources on apparent digestibilities and faecal ammonia concentration. In three subsequent periods eight healthy beagle dogs were fed a commercial dog diet that was gradually supplemented by up to 50 % with meat and bone meal (MBM), greaves meal (GM) or poultry meal (PM) respectively. Afterwards, 3 % fructo-oligosaccharides or 3 % isomalto-oligosaccharides were substituted for 3 % of the total diet. Supplementation with animal-derived protein sources did not decrease the apparent N digestibility significantly but oligosaccharides did. On the other hand the bacterial N content (% DM) in the faeces was highest in the oligosaccharide groups followed by the protein-supplemented groups and lowest in the control groups. When the apparent N digestibility was corrected for bacterial N no significant differences were noted anymore except for the GM group where the corrected N digestibility was still lower after oligosaccharide supplementation. The amount of faecal ammonia was significantly increased by supplementing with protein or oligosaccharides in the MBM and GM groups but not in the PM group. When apparent N digestibility is interpreted, a correction for bacterial N should be taken into account, especially when prebiotics are added to the diet. Oligosaccharides did not reduce the faecal ammonia concentrations as expected. PMID:14641959

  10. Influence of heat and shear induced protein aggregation on the in vitro digestion rate of whey proteins.

    Science.gov (United States)

    Singh, Tanoj K; Øiseth, Sofia K; Lundin, Leif; Day, Li

    2014-11-01

    Protein intake is essential for growth and repair of body cells, the normal functioning of muscles, and health related immune functions. Most food proteins are consumed after undergoing various degrees of processing. Changes in protein structure and assembly as a result of processing impact the digestibility of proteins. Research in understanding to what extent the protein structure impacts the rate of proteolysis under human physiological conditions has gained considerable interest. In this work, four whey protein gels were prepared using heat processing at two different pH values, 6.8 and 4.6, with and without applied shear. The gels showed different protein network microstructures due to heat induced unfolding (at pH 6.8) or lack of unfolding, thus resulting in fine stranded protein networks. When shear was applied during heating, particulate protein networks were formed. The differences in the gel microstructures resulted in considerable differences in their rheological properties. An in vitro gastric and intestinal model was used to investigate the resulting effects of these different gel structures on whey protein digestion. In addition, the rate of digestion was monitored by taking samples at various time points throughout the in vitro digestion process. The peptides in the digesta were profiled using SDS-polyacrylamide gel electrophoresis, reversed-phase-HPLC and LC-MS. Under simulated gastric conditions, whey proteins in structured gels were hydrolysed faster than native proteins in solution. The rate of peptides released during in vitro digestion differed depending on the structure of the gels and extent of protein aggregation. The outcomes of this work highlighted that changes in the network structure of the protein can influence the rate and pattern of its proteolysis under gastrointestinal conditions. Such knowledge could assist the food industry in designing novel food formulations to control the digestion kinetics and the release of biologically active peptides for desired health outcome. PMID:25205335

  11. An In-solution Ultrasonication-assisted Digestion Method for Improved Extracellular Matrix Proteome Coverage*

    OpenAIRE

    Hansen, Kirk C.; Kiemele, Lauren; Maller, Ori; O Brien, Jenean; Shankar, Aarthi; Fornetti, Jaime; Schedin, Pepper

    2009-01-01

    Epithelial cell behavior is coordinated by the composition of the surrounding extracellular matrix (ECM); thus ECM protein identification is critical for understanding normal biology and disease states. Proteomic analyses of ECM proteins have been hindered by the insoluble and digestion-resistant nature of ECM. Here we explore the utility of combining rapid ultrasonication- and s...

  12. Tube-Gel Digestion: A NOVEL PROTEOMIC APPROACH FOR HIGH THROUGHPUT ANALYSIS OF MEMBRANE PROTEINS*S

    OpenAIRE

    Lu, Xiaoning; Zhu, Haining

    2005-01-01

    This study describes a new protein digestion protocol in which a variety of detergents can be used to solubilize membrane proteins and facilitate trypsin digestion with higher efficiency. In this protocol, proteins are dissolved in solutions containing various detergents and directly incorporated into a polyacrylamide gel matrix without electrophoresis. Detergents are subsequently eliminated from the gel matrix while proteins are still immobilized in the gel matrix. After in-gel digestion of ...

  13. Digestion and microbial protein synthesis in sheep as affected by exogenous fibrolytic enzymes

    Scientific Electronic Library Online (English)

    W.F.J., van de Vyver; B.A., Useni.

    Full Text Available SciELO South Africa | Language: English Abstract in english The aim of the study was to determine the effect of an exogenous fibrolytic enzyme (EFE) on the ruminal disappearance of dry matter (DM), neutral detergent fibre (NDF), crude protein (CP) and on the microbial protein synthesis (MPS) in sheep of a milled substrate consisting of a 50 : 50 mixture of l [...] ucerne hay (LH) and wheat straw (WS). Four cannulated Döhne-Merino sheep were randomly assigned to two groups in a 2 x 2 cross-over experiment. Additional to the in situ study, the effects of an exogenous fibrolytic enzyme (EFE) on the in vitro gas production (GP) and ANKOM digestion systems on the mixture of milled LH and WS were determined. The substrate was pre-treated with distilled water (control) or EFE (treatment) 12 hours prior to incubation to allow interaction between the substrate and enzyme. Adding EFE significantly improved the in vitro cumulative gas production (GP), but it did not affect the in vitro MPS measured as purine derivates of the GP residues. The enzyme significantly increased the in vitro DM and NDF disappearances after 36 hours. However, the in situ disappearances of DM, NDF and CP of the roughage mixture were not affected by the EFE. Further, the in situ MPS was increased significantly with the EFE treatment of the mixed substrate. It is concluded that the improved ruminal digestion of DM, NDF and CP is not only due to direct enzyme hydrolysis, but also due to the improvement of MPS.

  14. Protein digestion: an overview of the available techniques and recent developments.

    Science.gov (United States)

    Switzar, Linda; Giera, Martin; Niessen, Wilfried M A

    2013-03-01

    Several proteomics approaches are available that are defined by the level (protein or peptide) at which analysis takes place. The most widely applied method still is bottom-up proteomics where the protein is digested into peptides that can be efficiently analyzed with a wide range of LC-MS or MALDI-TOF-MS instruments. Sample preparation for bottom-up proteomics experiments requires several treatment steps in order to get from the protein to the peptide level and can be very laborious. The most crucial step in such approaches is the protein digestion, which is often the bottleneck in terms of time consumption. Therefore, a significant gain in throughput may be obtained by speeding up the digestion process. Current techniques allow for reduction of the digestion time from overnight (~15 h) to minutes or even seconds. This advancement also makes integration into online systems feasible, thereby reducing the number of tedious sample handling steps and the risk of sample loss. In this review, an overview is given of the currently available digestion strategies and recent developments in the acceleration of the digestion process. Additionally, tailored approaches for classes of proteins that pose specific challenges are discussed. PMID:23368288

  15. Influence of whey protein-beet pectin conjugate on the properties and digestibility of ?-carotene emulsion during in vitro digestion.

    Science.gov (United States)

    Xu, Duoxia; Yuan, Fang; Gao, Yanxiang; Panya, Atikorn; McClements, David Julian; Decker, Eric Andrew

    2014-08-01

    The impact of a whey protein isolate (WPI)-beet pectin (BP) conjugate (formed by dry-heating) on the physical properties and digestibilities of ?-carotene and carrier oil in oil-in-water emulsions was studied when they passed through a model gastrointestinal system. ?-Carotene emulsions were stabilized by WPI, unconjugated and conjugated WPI-BP, separately. The emulsions were then passed through an in vitro digestion model and the mean droplet size, droplet distribution, zeta-potential, free fatty acids and ?-carotene released were measured. The stability to droplet flocculation and coalescence during digestion was increased for the WPI-BP conjugate stabilized emulsion. Addition of BP onto the WPI stabilized emulsions could inhibit the releases of carrier oil (MCT) and ?-carotene. The releases of free fatty acids and ?-carotene did not differ greatly between the unconjugated and conjugated WPI-BP stabilized emulsions. These results have important implications for protein-polysaccharide stabilized emulsions and conjugates used for the protection and delivery of bioactive compounds. PMID:24629983

  16. Surfactants in anaerobic digestion of cheese whey, poultry waste, and cattle dung for improved biomethanation

    Energy Technology Data Exchange (ETDEWEB)

    Desai, M.; Madamwar, D. [Sardar Patel Univ., Vallabh Vidyanagar (India)

    1994-05-01

    To obtain enriched methane content and improve the anaerobic digestion of a mixture of cattle dung, poultry waste and cheese whey, with enriched methane content, the effect of various surfactants was studied. Among the surfactants tested, Tween 80 and sodium lauryl sulphate showed the maximum enhancement in gas production as well as methane content, indicating better process performance. The Tween 80 dosed digester (300 {mu}L/L) produced about 3.5 L gas/L of digester/d with 70% methane. Results also indicated increased percent COD reduction in the presence of Tween 80. 13 refs., 2 figs.

  17. Dietary protein digestion and absorption are impaired during acute postexercise recovery in young men.

    Science.gov (United States)

    van Wijck, Kim; Pennings, Bart; van Bijnen, Annemarie A; Senden, Joan M G; Buurman, Wim A; Dejong, Cornelis H C; van Loon, Luc J C; Lenaerts, Kaatje

    2013-03-01

    Previously, we demonstrated that exercise can cause small intestinal injury, leading to loss of gut barrier function. The functional consequences of such exercise-induced intestinal injury on subsequent food digestion and absorption are unclear. The present study determined the impact of resistance-type exercise on small intestinal integrity and in vivo dietary protein digestion and absorption kinetics. Twenty-four young males ingested 20 g specifically produced intrinsically l-[1-(13)C]phenylalanine-labeled protein at rest or after performing a single bout of resistance-type exercise. Continuous intravenous infusions with l-[ring-(2)H5]phenylalanine were employed, and blood samples were collected regularly to assess in vivo protein digestion and absorption kinetics and to quantify plasma levels of intestinal fatty-acid binding protein (I-FABP) as a measure of small intestinal injury. Plasma I-FABP levels were increased after exercise by 35%, reaching peak values of 344 ± 53 pg/ml compared with baseline 254 ± 31 pg/ml (P < 0.05). In resting conditions, I-FABP levels remained unchanged. Dietary protein digestion and absorption rates were reduced during postexercise recovery when compared with resting conditions (P < 0.001), with average peak exogenous phenylalanine appearance rates of 0.18 ± 0.04 vs. 0.23 ± 0.03 mmol phenylalanine·kg lean body mass(-1)·min(-1), respectively. Plasma I-FABP levels correlated with in vivo rates of dietary protein digestion and absorption (rS = -0.57, P < 0.01). Resistance-type exercise induces small intestinal injury in healthy, young men, causing impairments in dietary protein digestion and absorption kinetics during the acute postexercise recovery phase. To the best of our knowledge, this is first evidence that shows that exercise attenuates dietary protein digestion and absorption kinetics during acute postexercise recovery. PMID:23283940

  18. Digestion of crude protein and organic matter of leaves by rumen microbes in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Ciszuk, A.; Murphy, M.

    1982-01-01

    22 leaf specimens, of which 6 were from an energy-woods project, were studied by incubation in vitro with rumen microbes or pepsin-hydrochloric acid. Several were also examined in situ using the nylon-bag technique. Many leaves, despite their low fiber and high crude protein content, gave low values for organic matter digestibility. The crude protein degradation by rumen microbes or pepsin-hydrochloric acid was low, on average, compared with hay. There was a wide variation among leaf specimens. Variation was also found as regards ammonia production in short-term (4 hours) incubation. No close correlation was found between crude protein content and crude protein degradation, or between the estimates of ruminal degradation and of pepsin-hydrochloric acid digestibility. This suggest that there are leaves that gives ruminants substantial amounts of digestible protein yet escape ruminal fermentation. (Refs. 12).

  19. Milk production response to varying protein supply is independent of forage digestibility in dairy cows.

    Science.gov (United States)

    Alstrup, L; Weisbjerg, M R; Hymøller, L; Larsen, M K; Lund, P; Nielsen, M O

    2014-07-01

    The aim of this experiment was to examine whether the positive response in milk production to increased crude protein (CP) supply in dairy cows was dependent on the digestibility of the forage. Forty-eight lactating Danish Holstein cows were used in a 4 × 4 Latin square design experiment with 4 rations: (1) high digestibility and high CP concentration (HdHp), (2) high digestibility and low CP concentration (HdLp), (3) low digestibility and high CP concentration (LdHp), and (4) low digestibility and low CP concentration (LdLp). All rations contained 30% corn silage, 25% grass-clover silage, and 45% concentrate on a dry matter (DM) basis. Different digestibilities were obtained by replacing a high-digestible grass-clover silage combined with a high-digestible corn silage with a low-digestible grass-clover silage combined with a low-digestible corn silage. Organic matter digestibilities were 79.8 and 74.7% in the high- and low-digestibility rations, respectively. Dietary CP concentration in the ration was increased by substituting barley and sugar beet pulp with rapeseed meal and soybean meal, whereby CP increased from 13.9 to 14.0% (Lp) to 15.7 to 16.0% (Hp). All cows were offered 3 kg of the same concentrate per day in the automatic milking system in addition to the mixed ration. Every feeding period lasted 3 wk, and DM intake and milk yield were measured in the last week in each period, and milk samples for determining milk composition, including fatty acid content, and blood samples were taken during the last 3d of each period. Dry matter intake increased by 2.2 kg/d on Hd compared with Ld and by 0.7 kg/d on Hp compared with Lp. The positive effect on DM intake was reflected in the energy-corrected milk (ECM) yield, as a higher ration digestibility increased the ECM yield by 1.7 kg/d and a higher CP concentration increased it by 1.2 kg/d. We detected no interaction between forage digestibility and CP concentration on milk production. Reduced digestibility was accompanied by an increase in the plasma level of glucose, suggesting that other nutrients were limiting to milk production. In conclusion, milk production responses to dietary CP supply appeared independent of forage digestibility. PMID:24835962

  20. Co-digestion of source segregated domestic food waste to improve process stability

    OpenAIRE

    Zhang, Yue; Banks, Charles J.; Heaven, Sonia

    2012-01-01

    Cattle slurry and card packaging were used to improve the operational stability of food waste digestion, with the aim of reducing digestate total ammoniacal nitrogen concentrations compared to food waste only. Use of cattle slurry could have major environmental benefits through reducing greenhouse gas emissions associated with current management practices; whilst card packaging is closely linked to food waste and could be co-collected as a source segregated material. Both options increase the...

  1. Tracking the fate of pasta (T. Durum semolina) immunogenic proteins by in vitro simulated digestion.

    Science.gov (United States)

    Mamone, Gianfranco; Nitride, Chiara; Picariello, Gianluca; Addeo, Francesco; Ferranti, Pasquale; Mackie, Alan

    2015-03-18

    The aim of the present study was to identify and characterize the celiacogenic/immunogenic proteins and peptides released during digestion of pasta (Triticum durum semolina). Cooked pasta was digested using a harmonized in vitro static model of oral-gastro-duodenal digestion. The course of pasta protein digestion was monitored by SDS-PAGE, and gluten proteins were specifically analyzed by Western blot using sera of celiac patients. Among the allergens, nonspecific lipid-transfer protein was highly resistant to gastro-duodenal hydrolysis, while other digestion-stable allergens such as ?-amylase/trypsin inhibitors were not detected being totally released in the pasta cooking water. To simulate the final stage of intestinal degradation, the gastro-duodenal digesta were incubated with porcine jejunal brush-border membrane hydrolases. Sixty-one peptides surviving the brush-border membrane peptidases were identified by liquid chromatography-mass spectrometry, including several gluten-derived sequences encrypting different motifs responsible for the induction of celiac disease. These results provide new insights into the persistence of wheat-derived peptides during digestion of cooked pasta samples. PMID:25682706

  2. Protein digestibility evaluations of meat and fish substrates using laboratory, avian, and ileally cannulated dog assays.

    Science.gov (United States)

    Faber, T A; Bechtel, P J; Hernot, D C; Parsons, C M; Swanson, K S; Smiley, S; Fahey, G C

    2010-04-01

    Meat and fish serve as important protein sources in the companion animal diet; however, limited protein digestibility data are available for assessing protein digestibility differences among good-quality protein sources. Beef loin, pork loin, chicken breast, pollock fillet, and salmon fillet were evaluated for composition, protein digestibility, and AA bioavailability using the immobilized digestive enzyme assay, cecectomized rooster assay, and ileally cannulated dog assay. Pollock contained the greatest amount of CP, total essential AA (TEAA), and total nonessential AA (TNEAA; DM basis; 96.9, 38.6, and 50.3%, respectively). Salmon contained the next greatest amounts (92.8, 36.4, and 44.6%), followed by chicken (90.3, 36.1, 43.2%). Beef had the least CP content (82.7%), but had slightly greater TEAA and TNEAA concentrations (33.9, 42.0%) compared with pork (86.2, 33.6, 41.3%). Immobilized digestive enzyme assay values were greatest for pollock fillet (0.71) and least for chicken breast (0.52). Beef loin, pork loin, and salmon fillet were similar (0.63, 0.62, and 0.64, respectively). Standardized TEAA and TNEAA digestibility coefficients, evaluated using the cecectomized rooster assay, were greatest (P 0.05) were found in ileal digestibility of protein. Values ranged from 88.9% for chicken to 90.5% for pork loin and pollock fillet. Ileal TEAA and TNEAA coefficients were not different among test substrates, with values between 91.7 and 92.7%, and 88.8 and 90.4%, respectively. Total tract CP apparent digestibility values ranged from 94.4 to 94.8%, with no differences noted among treatments. Despite marked differences in composition and predicted and standardized digestibility values, when the protein sources were added to diets at a concentration of approximately 30% (25% of total energy intake), no differences in test protein substrates were noted in either ileal or total tract nutrient digestibility. PMID:20023140

  3. Nanospray FAIMS fractionation provides significant increases in proteome coverage of unfractionated complex protein digests.

    Science.gov (United States)

    Swearingen, Kristian E; Hoopmann, Michael R; Johnson, Richard S; Saleem, Ramsey A; Aitchison, John D; Moritz, Robert L

    2012-04-01

    High-field asymmetric waveform ion mobility spectrometry (FAIMS) is an atmospheric pressure ion mobility technique that can be used to reduce sample complexity and increase dynamic range in tandem mass spectrometry experiments. FAIMS fractionates ions in the gas-phase according to characteristic differences in mobilities in electric fields of different strengths. Undesired ion species such as solvated clusters and singly charged chemical background ions can be prevented from reaching the mass analyzer, thus decreasing chemical noise. To date, there has been limited success using the commercially available Thermo Fisher FAIMS device with both standard ESI and nanoLC-MS. We have modified a Thermo Fisher electrospray source to accommodate a fused silica pulled tip capillary column for nanospray ionization, which will enable standard laboratories access to FAIMS technology. Our modified source allows easily obtainable stable spray at flow rates of 300 nL/min when coupled with FAIMS. The modified electrospray source allows the use of sheath gas, which provides a fivefold increase in signal obtained when nanoLC is coupled to FAIMS. In this work, nanoLC-FAIMS-MS and nanoLC-MS were compared by analyzing a tryptic digest of a 1:1 mixture of SILAC-labeled haploid and diploid yeast to demonstrate the performance of nanoLC-FAIMS-MS, at different compensation voltages, for post-column fractionation of complex protein digests. The effective dynamic range more than doubled when FAIMS was used. In total, 10,377 unique stripped peptides and 1649 unique proteins with SILAC ratios were identified from the combined nanoLC-FAIMS-MS experiments, compared with 6908 unique stripped peptides and 1003 unique proteins with SILAC ratios identified from the combined nanoLC-MS experiments. This work demonstrates how a commercially available FAIMS device can be combined with nanoLC to improve proteome coverage in shotgun and targeted type proteomics experiments. PMID:22186714

  4. Rapid Online Non-Enzymatic Protein Digestion Combining Microwave Heating Acid Hydrolysis and Electrochemical Oxidation

    OpenAIRE

    Basile, Franco; Hauser, Nicolas

    2010-01-01

    We report an online non-enzymatic method for site-specific digestion of proteins to yield peptides that are well suited for collision induced dissociation (CID) tandem mass spectrometry (MS/MS). The method combines online microwave heating acid hydrolysis at aspartic acid and online electrochemical oxidation at tryptophan and tyrosine. The combined microwave/electrochemical (microwave/echem) digestion is reproducible and produces peptides with an average sequence length of 10 amino acids. Thi...

  5. Facile Trypsin Immobilization in Polymeric Membranes for Rapid, Efficient Protein Digestion

    OpenAIRE

    Xu, Fei; Wang, Wei-han; Tan, Yu-jing; Bruening, Merlin L.

    2010-01-01

    Sequential adsorption of poly(styrene sulfonate) and trypsin in nylon membranes provides a simple, inexpensive method to create stable, microporous reactors for fast protein digestion. The high local trypsin concentration and short radial diffusion distances in membrane pores facilitate proteolysis in residence times of a few seconds, and the minimal pressure drop across the thin membranes allows their use in syringe filters. Membrane digestion and subsequent MS analysis of bovine serum album...

  6. Centrifugal Methods and Devices for Rapid In-Gel Digestion of Proteins

    OpenAIRE

    Lazarev, Alexander V.; Rejtar, Tomas; Dai, Shujia; Karger, Barry L.

    2009-01-01

    Modern proteomic research frequently relies upon separation of proteins in a polyacrylamide gel matrix followed by in-gel enzymatic digestion and extraction of peptides for subsequent analysis by mass spectrometry. In this work, we propose a novel semi-automated method of mechanical processing of gel bands by passing these bands through a specially designed centrifugal device termed a Gel Shredder prior to digestion and extraction of peptides. Such a device allows integrated washing, destaini...

  7. Digestive System

    Science.gov (United States)

    ... but these organs are still important for healthy digestion. The pancreas produces enzymes that help digest proteins, fats, and carbohydrates. It also makes a substance that neutralizes stomach acid. The liver produces bile , which helps the ...

  8. Kernel Composition, Starch Structure, and Enzyme Digestibility of Opaque-2 Maize and Quality Protein Maize

    Science.gov (United States)

    Objectives of this study were to understand how opaque-2 (o2) mutation and quality protein maize (QPM) affect maize kernel composition and starch structure, property, and enzyme digestibility. Kernels of o2 maize contained less protein (9.6−12.5%) than those of the wild-type (WT) counterparts (12...

  9. Protein digestion in red aak borer larvae, Enaphalodes rufulus

    Science.gov (United States)

    In the Ozark Mountains of Arkansas, Oklahoma, and Missouri, a recent outbreak of red oak borer, Enaphalodes rufulus (Haldeman), contributed to the death of tens of thousands of red oaks. To better understand nutrient digestion in E. rufulus larvae, biochemical analyses were used to characterize dige...

  10. Enrichment and desalting of tryptic protein digests and the protein depletion using boron nitride

    International Nuclear Information System (INIS)

    Highlights: • Protein tryptic digests were desalted and enriched utilizing hexagonal boron nitride. • Phosphopeptides were desalted with high recovery rates. • Boron nitride exhibits high wettability allowing fast sample preparation. • Boron nitride shows protein depletion capability applied for peptide purification. - Abstract: Sample preparation still remains a great challenge in modern bioanalysis and the interest in new efficient solid phase extraction (SPE) materials still remains high. In this work, hexagonal boron nitride (h-BN) is introduced as a new SPE material for the isolation and enrichment of peptides. The h-BN is isoelectronic and structurally similar to graphite. It has remarkable properties including good thermal conductivity, excellent thermal and chemical stability and a better oxidation resistance than graphite. BN attracts increasing interest because of its wide range of applicability. In the present work, the great potential of h-BN, as a new SPE-material, on the enrichment, preconcentration and desalting of tryptic digest of model proteins is demonstrated. A special attention was dedicated to the efficient enrichment of hydrophilic phosphopeptides. Two elution protocols were developed for the enrichment of peptides compatible for subsequent MALDI-MS and ESI-MS analysis. In addition, the recoveries of 5 peptides and 3 phosphopeptides with wide range of pI values utilizing h-BN materials with different surface areas were investigated. 84 106% recovery rate could be achieved using h-BN materials. The results were compared with those obtained using graphite and silica C18 under the same elution conditions, and lower recoveries were obtained. In addition, h-BN was found to have a capability of protein depletion, which is requisite for the peptide profiling

  11. Enrichment and desalting of tryptic protein digests and the protein depletion using boron nitride

    Energy Technology Data Exchange (ETDEWEB)

    Fischnaller, Martin; Köck, Rainer; Bakry, Rania, E-mail: rania.bakry@uibk.ac.at; Bonn, Günther K.

    2014-05-01

    Highlights: • Protein tryptic digests were desalted and enriched utilizing hexagonal boron nitride. • Phosphopeptides were desalted with high recovery rates. • Boron nitride exhibits high wettability allowing fast sample preparation. • Boron nitride shows protein depletion capability applied for peptide purification. - Abstract: Sample preparation still remains a great challenge in modern bioanalysis and the interest in new efficient solid phase extraction (SPE) materials still remains high. In this work, hexagonal boron nitride (h-BN) is introduced as a new SPE material for the isolation and enrichment of peptides. The h-BN is isoelectronic and structurally similar to graphite. It has remarkable properties including good thermal conductivity, excellent thermal and chemical stability and a better oxidation resistance than graphite. BN attracts increasing interest because of its wide range of applicability. In the present work, the great potential of h-BN, as a new SPE-material, on the enrichment, preconcentration and desalting of tryptic digest of model proteins is demonstrated. A special attention was dedicated to the efficient enrichment of hydrophilic phosphopeptides. Two elution protocols were developed for the enrichment of peptides compatible for subsequent MALDI-MS and ESI-MS analysis. In addition, the recoveries of 5 peptides and 3 phosphopeptides with wide range of pI values utilizing h-BN materials with different surface areas were investigated. 84–106% recovery rate could be achieved using h-BN materials. The results were compared with those obtained using graphite and silica C18 under the same elution conditions, and lower recoveries were obtained. In addition, h-BN was found to have a capability of protein depletion, which is requisite for the peptide profiling.

  12. Modelling of pepsin digestibility of myofibrillar proteins and of variations due to heating.

    Science.gov (United States)

    Kondjoyan, Alain; Daudin, Jean-Dominique; Santé-Lhoutellier, Véronique

    2015-04-01

    Digestibility of myofibrillar proteins by pepsin was determined by in vitro trials and mathematical modelling. A primary model was developed to predict in vitro digestion kinetics, and a secondary model based on the mechanisms of protein denaturation was then added to take into account the effect of meat heating. Model predictions agreed with measurements in the pH and pepsin concentration ranges 1.8-3.8 and 6-50 U mg(-1) respectively. The utility of the model is illustrated by a simple example where meat is assumed to be heated homogeneously, and myofibrillar proteins to be directly in contact with pepsin. The combined effects of heating time, temperature, enzyme concentration and pH modified the digestibility value, which also depends on residence time in the stomach. PMID:25442553

  13. Ostrich (Strutio camelus) meat protein quality and digestibility

    OpenAIRE

    Ls, Reis; Tc, Oliveira

    2008-01-01

    The purpose of the study was to evaluate ostrich meat protein quality, as its consumption has significantly increased in the last few years in Brazil. Male Wistar rats were distributed in groupe of six elements. The standard group received a casein-based diet, the control group received a protein-free diet, and the experimental group received ostrich meat diet as protein source. The evaluated biological parameters were protein efficiency ratio (PER), net protein ratio (NPR), net protein utili...

  14. Simulated gastrointestinal digestion reduces the allergic reactivity of shrimp extract proteins and tropomyosin.

    Science.gov (United States)

    Gámez, Cristina; Zafra, Ma Paz; Sanz, Verónica; Mazzeo, Carla; Ibáñez, Ma Dolores; Sastre, Joaquín; del Pozo, Victoria

    2015-04-15

    Shrimp are highly allergenic foods. Current management are limited to the avoidance of foods. Therefore, there is an unmet need for a safe and effective therapy using modified allergens. This study focuses on assessing the potential for modification of the allergenicity of shrimp proteins following heat treatment or simulated gastric digestion. Shrimp proteins do not reduce their IgE reactivity after heat treatment but it is reduced by simulated gastric digestion in a time- and dose-dependent manner. Tropomyosin in shrimp extract is worse digested than purified tropomyosin. After 60 min of 10 U/?g pepsin digestion, a strong inhibition was produced in the in vivo skin reactivity of shrimp extracts and in activation of basophils from allergic patients. Immunisation experiments performed in rabbits demonstrated that digested boiled shrimp extract is able to induce IgG antibodies that block the IgE binding to the untreated boiled shrimp extract in shrimp-allergic patients. Building on our observations, digestion treatment could be an effective method for reducing shrimp allergenicity while maintaining the immunogenicity. PMID:25466048

  15. Improved design of anaerobic digesters for household biogas production in indonesia: one cow, one digester, and one hour of cooking per day.

    Science.gov (United States)

    Usack, Joseph G; Wiratni, Wiratni; Angenent, Largus T

    2014-01-01

    A government-sponsored initiative in Indonesia to design and implement low-cost anaerobic digestion systems resulted in 21 full-scale systems with the aim to satisfy the cooking fuel demands of rural households owning at least one cow. The full-scale design consisted of a 0.3 m diameter PVC pipe, which was operated as a conventional plug-flow system. The system generated enough methane to power a cooking stove for ? 1 h. However, eventual clogging from solids accumulation inside the bioreactor proved to be a major drawback. Here, we improved the digester configuration to remedy clogging while maintaining system performance. Controlled experiments were performed using four 9-L laboratory-scale digesters operated at a temperature of 27 ± 1 °C, a volatile solids loading rate of 2.0 g VS · L(-1) · day(-1), and a 21-day hydraulic retention time. Two of the digesters were replicates of the original design (control digesters), while the other two digesters included internal mixing or effluent recycle (experimental digesters). The performance of each digester was compared based on methane yields, VS removal efficiencies, and steady-state solids concentrations during an operating period of 311 days. Statistical analyses revealed that internal mixing and effluent recycling resulted in reduced solids accumulation compared to the controls without diminishing methane yields or solids removal efficiencies. PMID:24715809

  16. Comparison of Digestion Protocols for Microgram Quantities of Enriched Protein Samples

    Energy Technology Data Exchange (ETDEWEB)

    Hervey, IV, William Judson [ORNL; Strader, Michael B [ORNL; Strader, Michael Brad [National Institutes of Health; Hurst, Gregory {Greg} B [ORNL

    2007-01-01

    Standard biochemical techniques that are used for protein enrichments, such as affinity isolation and density gradient centrifugation, frequently yield high nanogram to low microgram quantities at a significant expenditure of resources and time. The characterization of selected protein enrichments by the "shotgun" mass spectrometry approach is often compromised by the lack of effective and efficient in-solution proteolysis protocols specifically tailored for these small quantities of proteins. This study compares the results of five different digestion protocols that were applied to 2.5 g portions of protein isolates from two disparate sources: Rhodopseudomonas palustris 70S ribosomal proteins, and Bos taurus microtubule-associated proteins (MAPs). Proteolytic peptides produced according to each protocol in each type of protein isolate were analyzed by one-dimensional liquid chromatography-tandem mass spectrometry (LC-MS/MS). The effectiveness of each digestion protocol was assessed on the basis of three parameters: number of peptide identifications, number of protein identifications, and sequence coverage. The two protocols using a solvent containing 80% acetonitrile (CH3CN) for trypsin digestions performed as well as, and in some instances better than, protocols employing other solvents and chaotropes in both types of protein isolates. A primary advantage of the 80% CH3CN protocol is that it requires fewer sample manipulation steps.

  17. Comparison of digestion protocols for microgram quantities of enriched protein samples.

    Science.gov (United States)

    Hervey, W Judson; Strader, Michael Brad; Hurst, Gregory B

    2007-08-01

    Standard biochemical techniques that are used for protein enrichments, such as affinity isolation and density gradient centrifugation, frequently yield high-nanogram to low-microgram quantities at a significant expenditure of resources and time. The characterization of selected protein enrichments by the "shotgun" mass spectrometry approach is often compromised by the lack of effective and efficient in-solution proteolysis protocols specifically tailored for these small quantities of proteins. This study compares the results of five different digestion protocols that were applied to 2.5 mug portions of protein isolates from two disparate sources: Rhodopseudomonas palustris 70S ribosomal proteins, and Bos taurus microtubule-associated proteins (MAPs). Proteolytic peptides produced according to each protocol in each type of protein isolate were analyzed by one-dimensional liquid chromatography-tandem mass spectrometry (LC-MS/MS). The effectiveness of each digestion protocol was assessed on the basis of three parameters: number of peptide identifications, number of protein identifications, and sequence coverage. The two protocols using a solvent containing 80% acetonitrile (CH3CN) for trypsin digestions performed as well as, and in some instances better than, protocols employing other solvents and chaotropes in both types of protein isolates. A primary advantage of the 80% CH3CN protocol is that it requires fewer sample manipulation steps. PMID:17616116

  18. Investigation of protein digestibility and amino acid absorption in different segments of porcine digestive tract. 5

    International Nuclear Information System (INIS)

    In 4 pigs (live weight 63 kg) with re-entrance cannulae in the duodenum as well as in the ileocecal region intake, excretion and flow rate of protein, methionine and 35S were measured after supplementary oral application of 35S-methionine to a wheat-wheat gluten-lysine diet and a protein-free diet, respectively, over a period of 7 days. For both the N and the methionine metabolism patterns were outlined according to the measured values and with the aid of a compartment model. The endogenous and exogenous levels of proteins and methionine in the duodenum, ileum and feces were estimated

  19. Levels of digestible protein to surubim (Pseudoplatystoma sp. reared in net cages

    Directory of Open Access Journals (Sweden)

    Claucia Aparecida Honorato

    2014-10-01

    Full Text Available The Pseudoplatystoma sp. is species of carnivorous fish that require special attention in the diet offered. This work had the objective to determine the digestible protein requirement of juvenile the Pseudoplatystoma sp. reared in net cages. The test consisted of four isoenergetic diets (2606.69 ± 39.16 kcal kg-1 of digestible energy containing increasing levels of digestible protein (23, 24, 26 and 28%PD provided to juveniles of surubim (157.35±11.23g for five months. The parameters of growth, fillet composition, metabolic liver enzymes and morphometry of the intestine and liver were analyzed in completely randomized design with four treatments and four replicates. An increase of protein in the diet provided better weight gain. The metabolic liver enzymes increased in fish fed 24PD. The histopathological changes were not observed in the liver of the fish. The bowel histology showed adaptation to increased protein in the diet until the 26 level PD. juveniles of Pseudoplatystoma sp. Were demanding in digestible protein, showing the best results of production performance and nutrient use efficiency with the diet containing 28%PD.

  20. Effects of postruminal protein on fatty acid digestibility in dairy cows.

    Science.gov (United States)

    Goodling, L E; Grummer, R R

    1998-06-01

    Eight ruminally cannulated Holstein cows (four multiparous and four primiparous) were used in a replicated 4 x 4 Latin square with an extra period to determine the effects of postruminal protein on fatty acid digestibility. Samples were collected during the last 4 d of each 14-d period. Total mixed rations were composed of 41% alfalfa haylage, 42% corn silage, 12% concentrate based on corn, and 5% tallow. Cows were fed at 90% of ad libitum intake. Treatments were abomasal infusion of guar gum or guar gum plus urea, corn gluten meal, or blood meal in 12 L of water. The basal ration contained 12% crude protein (CP), and infusion of N sources increased CP to approximately 14%. Dry matter intake was similar for cows on all treatments. Milk, 3.5% fat-corrected milk, milk protein, and milk fat yields increased for cows receiving postruminal CP. Postruminal CP did not affect milk fat and protein percentages. Dry matter, organic matter, and CP digestibilities were greater in cows receiving postruminal CP. Total fatty acid and total C18 fatty acid digestibilities were not affected by treatment. Total C16 fatty acid and C18:0 fatty acid digestibilities were greater for cows receiving nonprotein N than for those receiving true protein. PMID:9684169

  1. The efficient approaches for enzymatic digestion of plant proteins.

    Czech Academy of Sciences Publication Activity Database

    Dy?ka, Filip; Bobá?, P.; Bobálová, Janette

    Brno : Ústav analytické chemie AV ?R, v. v. i, 2012 - (Foret, F.; K?enková, J.; Guttman, A.; Klepárník, K.; Bo?ek, P.), s. 267-270 ISBN 978-80-904959-1-3. [CECE 2012. International Interdisciplinary Meeting on Bioanalysis /9./. Brno (CZ), 01.11.2012-02.11.2012] R&D Projects: GA MŠk 1M0570; GA MŠk 1M06030; GA MŠk(CZ) EE2.3.20.0182 Institutional support: RVO:68081715 Keywords : ultrasonic * infrared * digestion Subject RIV: CB - Analytical Chemistry, Separation http://hdl.handle.net/11104/0215222

  2. Post-digestion ¹?O exchange/labeling for quantitative shotgun proteomics of membrane proteins.

    Science.gov (United States)

    Ye, Xiaoying; Luke, Brian T; Johann, Donald J; Chan, King C; Prieto, Darue A; Ono, Akira; Veenstra, Timothy D; Blonder, Josip

    2012-01-01

    The role of membrane proteins is critical for regulation of physiologic and pathologic cellular processes. Hence it is not surpassing that membrane proteins make ?70% of contemporary drug targets. Quantitative profiling of membrane proteins using mass spectrometry (MS)-based proteomics is critical in a quest for disease biomarkers and novel cancer drugs. Post-digestion (18)O exchange is a simple and efficient method for differential (18)O/(16)O stable isotope labeling of two biologically distinct specimens, allowing relative quantitation of proteins in complex mixtures when coupled with shotgun MS-based proteomics. Due to minimal sample consumption and unrestricted peptide tagging, (18)O/(16)O stable isotope labeling is particularly suitable for amount-limited protein specimens typically encountered in membrane and clinical proteomics. This chapter describes a protocol that relies on shotgun proteomics for quantitative profiling of the detergent-insoluble membrane proteins isolated from HeLa cells, differentially transfected with plasmids expressing HIV Gag protein and its myristylation-defective N-terminal mutant. Whilst this protocol depicts solubilization of detergent-insoluble membrane proteins coupled with post-digestion (18)O labeling, it is amenable to any complex membrane protein mixture. Described approach relies on solubilization and tryptic digestion of membrane proteins in a buffer containing 60% (v/v) methanol followed by differential (18)O/(16)O labeling of protein digests in 20% (v/v) methanol buffer. After mixing, the differentially labeled peptides are fractionated using off-line strong cation exchange (SCX) followed by on-line reversed phase nanoflow reversed-phase liquid chromatography (nanoRPLC)-MS identification/quantiation of peptides/proteins. The use of methanol-based buffers in the context of the post-digestion (18)O exchange/labeling eliminates the need for detergents or chaotropes that interfere with LC separations and peptide ionization. Sample losses are minimized because solubilization, digestion, and stable isotope labeling are carried out in a single tube, avoiding any sample transfer or buffer exchange between these steps. PMID:22665304

  3. Collaborative Peer Review. The Role of Faculty in Improving College Teaching. ERIC Digest.

    Science.gov (United States)

    Keig, Larry; Waggoner, Michael D.

    This digest, based on a full report of the same title, provides an examination of the improvement of college teaching and argues for the central involvement of faculty themselves in collaborative peer review and looks at some central issues and challenges to implementation. Since summative evaluation involved in tenure and promotion decisions…

  4. A simple MALDI plate functionalization by Vmh2 hydrophobin for serial multi-enzymatic protein digestions.

    Science.gov (United States)

    Longobardi, Sara; Gravagnuolo, Alfredo Maria; Funari, Riccardo; Della Ventura, Bartolomeo; Pane, Francesca; Galano, Eugenio; Amoresano, Angela; Marino, Gennaro; Giardina, Paola

    2015-01-01

    The development of efficient and rapid methods for the identification with high sequence coverage of proteins is one of the most important goals of proteomic strategies today. The on-plate digestion of proteins is a very attractive approach, due to the possibility of coupling immobilized-enzymatic digestion with direct matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF)-mass spectrometry (MS) analysis. The crucial step in the development of on-plate immobilization is however the functionalization of the solid surface. Fungal self-assembling proteins, the hydrophobins, are able to efficiently functionalize surfaces. We have recently shown that such modified plates are able to absorb either peptides or proteins and are amenable to MALDI-TOF-MS analysis. In this paper, the hydrophobin-coated MALDI sample plates were exploited as a lab-on-plate for noncovalent immobilization of enzymes commonly used in protein identification/characterization, such as trypsin, V8 protease, PNGaseF, and alkaline phosphatase. Rapid and efficient on-plate reactions were performed to achieve high sequence coverage of model proteins, particularly when performing multiple enzyme digestions. The possibility of exploiting this direct on-plate MALDI-TOF/TOF analysis has been investigated on model proteins and, as proof of concept, on entire whey milk proteome. PMID:25395204

  5. Ileal digestibility of amino acids of unheated and autoclaved pea protein concentrate in broilers.

    Science.gov (United States)

    Frikha, M; Valencia, D G; de Coca-Sinova, A; Lázaro, R; Mateos, G G

    2013-07-01

    The effects of autoclaving 2 varieties of micronized (fine grinding) pea protein concentrate (PPC) on the ileal digestibility (ID) of CP and amino acids (AA) were studied in broilers. There was a control diet based on fermented soybean meal (FSBM) and 4 extra diets in which the FSBM was substituted on a CP basis by PPC from 2 different pea cultivars (PPC-1 and PPC-2), either unheated or autoclaved. Chicks were fed a common diet from 1 to 17 d of age and, then, their respective experimental diets from 18 to 21 d of age. Each treatment was replicated 6 times. Autoclaving reduced trypsin inhibitor activity (TIA) but had little effect on the saponin content of the PPC. The apparent ID (AID) of CP was similar for the FSBM and the unheated PPC and lower for both than for the autoclaved PPC. Autoclaving improved (P standardized ID (SID) of most indispensable AA was similar for the FSBM and the PPC-2 and higher for both than for the PPC-1 (P < 0.05). For Lys, the lowest SID value was observed for the FSBM and the highest for the PPC-2 either unheated or autoclaved. It is concluded that the ID of the AA of the PPC improved with heating and was in general higher for the autoclaved PPC than for the FSBM. Consequently, heat processed PPC is a good alternative to FSBM and unheated PPC in starter diets for broilers. PMID:23776273

  6. Staphylococcus aureus Typing by Digestion of Protein A Coding Gene Using Bsp143I

    OpenAIRE

    Fatemeh Shakeri; Ezzat Allah Ghaemi; Maya Babai Kochkaksaraei

    2014-01-01

    Background: Protein A is the virulence factors of Staphylococcus aureus rolling in its pathogenesis, and its gene is used for typing. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with different enzymes has been used for this action. Objectives: In this study, we used Bsp143I enzyme for digestion of the gene, coding protein A (spa gene) in S. aureus. The bacteria were isolated from patients and healthy carriers in Gorgan, north of Iran. Patients and Met...

  7. Relationship between in vitro assays and standardized ileal amino acid digestibility of animal protein meals in broilers.

    Science.gov (United States)

    Rochell, S J; Kuhlers, D L; Dozier, W A

    2013-01-01

    Two identical trials were conducted to determine the relationship of a novel digestive enzyme assay, Poultry Complete IDEA (PC IDEA), and the pepsin digestibility assay with standardized ileal amino acid digestibility (SIAAD) of 20 animal protein meals (APM) fed to broilers from 25 to 30 d of age. Animal protein meals included 10 meat and bone meals (MBM) consisting of bovine, porcine, or mixed bovine and porcine raw materials (BP), and 10 animal protein blends containing animal proteins from various species. Treatments consisted of 20 semi-purified diets containing 1 APM as the sole source of dietary amino acids (AA), and 1 N-free diet to determine endogenous ileal AA flow. With the exception of the N-free diet, diets were formulated to contain 20% CP. In each trial, 756 Ross × Ross 708 male broilers were housed in battery cages and randomly assigned to 21 dietary treatments on d 25 (12 birds per cage; 3 replicate cages), and ileal digesta were collected on d 30 for determination of SIAAD. Pepsin digestibility and PC IDEA were determined for APM samples from each experimental diet (3 replicates per trial; 6 total replicates). Pepsin digestibility and PC IDEA were both correlated (P < 0.001) with SIAAD for each AA. Multiple linear regression of PC IDEA and pepsin digestibility on SIAAD resulted in the following equations: % Lys SIAAD = [-9.65 + (0.38 × % PC IDEA predicted Lys digestibility) + (0.69 × % pepsin digestibility)], % Met SIAAD = [-35.95 + (0.62 × % PC IDEA predicted Met digestibility) + (0.75 × % pepsin digestibility)], % Thr SIAAD = [-77.5 + (0.39 × % PC IDEA predicted Thr digestibility) + (1.37 × % pepsin digestibility)]. Values of R(2) were 0.46, 0.47, and 0.55 for Lys, Met, and Thr, respectively. The relatively low R(2) values may have been due to the limited range in SIAAD observed for the 20 APM, and additional data on APM varying in SIAAD are needed. PMID:23243243

  8. Comparison of Chemical Composition and Protein Digestibility, Carotenoids, Tanins and Alkaloids Content of Wild Lupinus Varieties Flour

    OpenAIRE

    Jorge Yanez-Fernandez; Hernandez-chavez, Juan F.; Jorge Martinez-Herrera; Norma Guemes-Vera; Alfonso Totosaus

    2012-01-01

    Proximate composition, carotenoids, tannins, quinolizidine alkaloids and in vitro protein digestibility were determined in flours of two wild lupines seeds recollected at central region of Mexico. Varieties identified as Lupinus barkeri and Lupinus montanus were compared with a domesticated cultivated Lupinus albus crop. Although total protein content resulted significantly (p0.05) difference were found in in vitro protein digestibility. Ash and crude fiber contents were significantly (p

  9. Oxidation of sarcoplasmic proteins during processing of Cantonese sausage in relation to their aggregation behaviour and in vitro digestibility.

    Science.gov (United States)

    Sun, Weizheng; Zhao, Mouming; Yang, Bao; Zhao, Haifeng; Cui, Chun

    2011-07-01

    The physicochemical changes of sarcoplasmic proteins, especially oxidation behaviour, were measured to determine their mechanism of action on in vitro protein digestibility during Cantonese sausage processing. The results indicated that carbonyl level increased (pprotein aggregation were highly (pcorrelated with carbonyl level and protein surface hydrophobicity (H(0)), indicating that protein oxidation and thermal denaturation could induce protein aggregation, leading to secondary structural changes. The analysis of in vitro digestibility showed no correlation between pepsin activity and protein oxidation, due to the biphasic response of sarcoplasmic proteins toward proteolysis. However, a highly significant (pcorrelation was observed with trypsin and ?-chymotrypsin activity, indicating that protein oxidation induced the changes in H(0), protein aggregation and secondary structure, which further influenced in vitro digestibility. PMID:21353394

  10. Effect of carnosic acid, quercetin and ?-tocopherol on lipid and protein oxidation in an in vitro simulated gastric digestion model.

    Science.gov (United States)

    Raes, Katleen; Doolaege, Evelyne H A; Deman, Steven; Vossen, Els; De Smet, Stefaan

    2015-03-01

    Carnosic acid, quercetin and ?-tocopherol are well-known antioxidants in many biological systems. However, their antioxidative effect during food digestion against lipid and protein oxidation is not well known. Therefore, in this study, an in vitro simulated gastric digestion model was used to investigate their stability during gastrointestinal conditions and their antioxidative properties during low pH digestion. In general, the stability of the antioxidants in the different steps of digestion was in the order of ?-tocopherol?>?quercetin?>?carnosic acid. Salivary components, as well as the acidity of the gastric juice, were responsible for the reduction in antioxidants. Both ?-tocopherol and quercetin were able to lower lipid oxidation during digestion, while the effect on protein oxidation was not clear. In contrast, carnosic acid did not have any effect on lipid oxidation and tended to stimulate protein oxidation. This study clearly demonstrated that the environmental conditions are of major importance to the properties of antioxidant compounds. PMID:25578758

  11. Enzymatic digestion and chromatographic analysis of arsenic species released from proteins.

    Science.gov (United States)

    Lu, Meiling; Wang, Hailin; Li, Xing-Fang; Lu, Xiufen; Le, X Chris

    2009-05-01

    A method combining gel filtration chromatography (GFC), protease digestion, and ion pair chromatography with inductively coupled plasma mass spectrometry detection was developed for the determination of arsenic species bound to proteins. The method was first established by examining the interactions of two model proteins, metallothionein (MT) and hemoglobin, with three reactive trivalent arsenic species. It was then successfully applied to the speciation of arsenic in red blood cells of rats. Inorganic arsenite (iAs(III)), monomethylarsonous acid (MMA(III)), and dimethylarsinous acid (DMA(III)) were efficiently released from the proteins by protease digestion at pH 8.0, with the recovery ranging from 93% to 106%. There was no oxidation of iAs(III) or MMA(III) during the protease digestion process. Up to 61% DMA(III) (the least stable arsenic species) was unchanged, and the rest was oxidized to the pentavalent dimethylarsinic acid (DMA(V)). The arsenic species in the red blood cells of control rats was present as DMA(III) complex with hemoglobin. The method enabling the determination of the specific arsenic species that bind to cellular proteins is potentially useful for studying arsenic distribution, metabolism, and toxicity. PMID:19327778

  12. Studies on voltammetric determination of cadmium in samples containing native and digested proteins

    Energy Technology Data Exchange (ETDEWEB)

    Drozd, Marcin; Pietrzak, Mariusz, E-mail: mariusz@ch.pw.edu.pl; Malinowska, El?bieta

    2014-03-01

    Highlights: • Proteins exhibit diverse impact on the DPASV cadmium signals. • Proteins subjected to HNO{sub 3} introduce less interference, than the native ones. • Optimal amount of SDS depends on the kind of protein. • Presence of thiolated coating agents of QDs do not influence the analysis. - Abstract: This work focuses on determination of cadmium ions using anodic stripping voltammetry (ASV) on thin film mercury electrode in conditions corresponding to those obtained after digestion of cadmium-based quantum dots and their conjugates. It presents the impact of selected proteins, including potential receptors and surface blocking agents on the voltammetric determination of cadmium. Experiments regarding elimination of interferences related to proteins presence using sodium dodecyl sulfate (SDS) are also shown. Effect of SDS on selected analytical parameters and simplicity of analyses carried out was investigated in the framework of current studies. The significant differences of influence among tested proteins on ASV cadmium determination, as well as the variability in SDS effectiveness as the antifouling agent were observed and explained. This work is especially important for those, who design new bioassays and biosensors with a use of quantum dots as electrochemical labels, as it shows what problems may arise from presence of native and digested proteins in tested samples.

  13. Studies on voltammetric determination of cadmium in samples containing native and digested proteins

    International Nuclear Information System (INIS)

    Highlights: • Proteins exhibit diverse impact on the DPASV cadmium signals. • Proteins subjected to HNO3 introduce less interference, than the native ones. • Optimal amount of SDS depends on the kind of protein. • Presence of thiolated coating agents of QDs do not influence the analysis. - Abstract: This work focuses on determination of cadmium ions using anodic stripping voltammetry (ASV) on thin film mercury electrode in conditions corresponding to those obtained after digestion of cadmium-based quantum dots and their conjugates. It presents the impact of selected proteins, including potential receptors and surface blocking agents on the voltammetric determination of cadmium. Experiments regarding elimination of interferences related to proteins presence using sodium dodecyl sulfate (SDS) are also shown. Effect of SDS on selected analytical parameters and simplicity of analyses carried out was investigated in the framework of current studies. The significant differences of influence among tested proteins on ASV cadmium determination, as well as the variability in SDS effectiveness as the antifouling agent were observed and explained. This work is especially important for those, who design new bioassays and biosensors with a use of quantum dots as electrochemical labels, as it shows what problems may arise from presence of native and digested proteins in tested samples

  14. Anaerobic digestion of carbohydrate and protein-based wastewaters in fluidized bed bioreactors

    Scientific Electronic Library Online (English)

    M., Fuentes; N. J., Scenna; P. A., Aguirre; M. C., Mussati.

    2007-12-01

    Full Text Available SciELO Argentina | Language: English Abstract in english The present work is focused on evaluating the anaerobic digestion of complex substrates (proteins and carbohydrates) in anaerobic fluidized bed reactors (AFBRs). In a previous paper, an AFBR model was adjusted using only the experimental data obtained during the acclimatization stage (glucose and ac [...] etate-based feeding) of two mesophilic bioreactors. Here, the sensitivity of that model to represent the digestion processes of proteins such as gelatin and albumin, and sucrose as a carbohydrate, is analyzed. Some kinetic expressions and parameters for the enzymatic hydrolysis processes need to be modified to represent the real behavior of bioreactors. Under the analyzed operating conditions, a hydraulic retention time higher than 1 day is suggested to ensure good system efficiency.

  15. Anaerobic digestion of cheese whey, poultry waste and cattle dung -a study of the use of mixture of adsorbents and/or surfactants to improve digester performance

    Energy Technology Data Exchange (ETDEWEB)

    Desai, Manik; Madamwar, Datta (Sardar Patel Univ., Gujarat (India). Post Graduate Dept. of Biosciences)

    1994-01-01

    This paper describes the improvement of anaerobic digestion process due to the addition of various surfactants and adsorbents in different combinations. Among the different combinations tested mixture of silica gel (2000 mg litre[sup -1]) and sodium lauryl sulphate (300 mg litre[sup -1]) showed maximum enhancement in gas production as well as methane content indicating better process performance. (author)

  16. Phaseolin type and heat treatment influence the biochemistry of protein digestion in the rat intestine

    OpenAIRE

    Montoya, Carlos A.; Leterme, Pascal; Beebe, Stephen; Souffrant, Wolfgang B.; Molle, Daniel; Lalles, Jean-paul

    2008-01-01

    The study aimed to investigate the in vivo digestion of Phaseolus vulgaris phaseolin types differing in their subunit pattern composition. Diets contained either casein as the sole source of protein or a mixture (1:1) of casein and pure Sanilac (S), Tendergreen (T) or Inca (I) phaseolin either unheated or heated. Rats were fed for 11 d with the experimental diets. Their ileal content and mucosa were collected and prepared for electrophoresis, Western blotting, densitometry and MS. Differences...

  17. Development of an open-tubular trypsin reactor for on-line digestion of proteins

    OpenAIRE

    Stigter, E. C. A.; Jong, G. J.; Bennekom, W. P.

    2007-01-01

    A study was initiated to construct a micro-reactor for protein digestion based on trypsin-coated fused-silica capillaries. Initially, surface plasmon resonance was used both for optimization of the surface chemistry applied in the preparation and for monitoring the amount of enzyme that was immobilized. The highest amount of trypsin was immobilized on dextran-coated SPR surfaces which allowed the covalent coupling of 11 ng mm?2 trypsin. Fused-silica capillaries were modified in a similar m...

  18. Digestibility of extruded proteins and metabolic transit of N ? -carboxymethyllysine in rats.

    Science.gov (United States)

    Alamir, Issam; Niquet-Leridon, Céline; Jacolot, Philippe; Rodriguez, Camille; Orosco, Martine; Anton, Pauline M; Tessier, Frédéric J

    2013-06-01

    Milk proteins are frequently used as supplements in fortified foods. However, processing produces chemical changes which likely affect the nutritional advantage. This study was intended to explore the possible difference in digestibility between extruded and non-extruded caseins and how the dietary N (?) -carboxymethyllysine (CML) is metabolised. Normal rats were randomized into either an extruded protein diet (EP) or the same with unextruded proteins (UEP), for two periods of 2 weeks at 7 to 9 and 11 to 13 weeks of age. However, no difference in protein digestibility was detected between the two diets, either in young or in adult animals, despite a 9.4-fold higher level of CML and an 8.5-fold higher level of lysinoalanine in the EP than in the UEP. No diet-related changes were observed in plasma CML, either protein bound or free. Amounts of 38 and 48 % of the orally absorbed CML were excreted in urine and faeces, respectively, in UEP-fed rats. Lower rates of excretion were found in the EP-fed rats (23 and 37 %, respectively). A second animal study using a single oral dose of free CML (400 ?g/rat) was set up to measure the systemic concentration of CML every hour from 0 to 4 h. It revealed that protein-bound CML was not affected by the oral dose of CML, and the highest free CML level found in the circulation was 600 ng/mL. Extruded proteins, therefore, appear to be well digested, and CML rapidly eliminated. Since its elimination is, however, incomplete, the question of its biodistribution and metabolism remains open. PMID:23160731

  19. NUTRALYS® pea protein: characterization of in vitro gastric digestion and in vivo gastrointestinal peptide responses relevant to satiety

    Directory of Open Access Journals (Sweden)

    Joost Overduin

    2015-04-01

    Design: Under in vitro simulated gastric conditions, the digestion of NUTRALYS® pea protein was compared to that of two dairy proteins, slow-digestible casein and fast-digestible whey. In vivo, blood glucose and gastrointestinal hormonal (insulin, ghrelin, cholecystokinin [CCK], glucagon-like peptide 1 [GLP-1], and peptide YY [PYY] responses were monitored in nine male Wistar rats following isocaloric (11 kcal meals containing 35 energy% of either NUTRALYS® pea protein, whey protein, or carbohydrate (non-protein. Results: In vitro, pea protein transiently aggregated into particles, whereas casein formed a more enduring protein network and whey protein remained dissolved. Pea-protein particle size ranged from 50 to 500 µm, well below the 2 mm threshold for gastric retention in humans. In vivo, pea-protein and whey-protein meals induced comparable responses for CCK, GLP-1, and PYY, that is, the anorexigenic hormones. Pea protein induced weaker initial, but equal 3-h integrated ghrelin and insulin responses than whey protein, possibly due to the slower gastric breakdown of pea protein observed in vitro. Two hours after meals, CCK levels were more elevated in the case of protein meals compared to that of non-protein meals. Conclusions: These results indicate that 1 pea protein transiently aggregates in the stomach and has an intermediately fast intestinal bioavailability in between that of whey and casein; 2 pea-protein- and dairy-protein-containing meals were comparably efficacious in triggering gastrointestinal satiety signals.

  20. Minimizing asynchronism to improve the performances of anaerobic co-digestion of food waste and corn stover.

    Science.gov (United States)

    Zhou, Qi; Shen, Fei; Yuan, Hairong; Zou, Dexun; Liu, Yanping; Zhu, Baoning; Jaffu, Muhanmad; Chufo, Akiber; Li, Xiujin

    2014-08-01

    To investigate the existence of the asynchronism during the anaerobic co-digestion of different substrates, two typical substrates of food waste and corn stover were anaerobically digested with altering organic loadings (OL). The results indicated that the biodegradability of food waste and corn stover was calculated to be 81.5% and 55.1%, respectively, which was main reason causing the asynchronism in the co-digestion. The asynchronism was minimized by NaOH-pretreatment for corn stover, which could improve the biodegradability by 36.6%. The co-digestion with pretreatment could increase the biomethane yield by 12.2%, 3.2% and 0.6% comparing with the co-digestion without pretreatment at C/N ratios of 20, 25 and 30 at OL of 35 g-VS/L, respectively. The results indicated that the digestibility synchronism of food waste and corn stover was improved through enhancing the accessibility and digestibility of corn stover. The biomethane production could be increased by minimizing the asynchronism of two substrates in co-digestion. PMID:24880810

  1. An H/D Exchange- and Protease Digestion-Based Screening Assay for Protein-Ligand Binding Detection

    OpenAIRE

    Hopper, Erin D.; Pittman, Adrianne M. C.; Tucker, Chandra L.; Campa, Michael J.; Patz, Edward F.; Fitzgerald, Michael C.

    2009-01-01

    A protease digestion strategy was incorporated into single-point SUPREX (stability of unpurified proteins from rates of H/D exchange), which is an H/D exchange- and mass spectrometry-based assay for the detection of protein-ligand binding. Single-point SUPREX is an abbreviated form of SUPREX in which protein-ligand binding interactions are detected by measuring the increase in a protein’s thermodynamic stability upon ligand binding. The new protease digestion protocol provides a noteworthy ...

  2. Efficient Digestion and Mass Spectral Analysis of Vesicular Glutamate Transporter 1: A Recombinant Membrane Protein Expressed in Yeast

    OpenAIRE

    Cox, Holly D.; Chao, Chih-kai; Patel, Sarjubhai A.; Thompson, Charles M.

    2008-01-01

    Attempts to characterize recombinant integral membrane proteins (IMPs) by mass spectrometry are frequently hindered by several factors including the detergents required for extraction and purification that interferes with analysis, poor solubility, incomplete digestion, and limited identification of the transmembrane domain-spanning peptides. The goal of this study was to examine and develop methods for purification of an IMP that are amenable to downstream digestion of the protein and peptid...

  3. Histochemical changes of carbohydrate and protein contents in the digestive gland cells of the land snail Monacha cartusiana following starvation

    OpenAIRE

    Sharaf, Hesham M.

    2009-01-01

    The present study was designed to investigate histochemically the detection of carbohydrate and protein in the normally feeding snails and after 15 and 30 days of starvation. Generally, abundant carbohydrate and protein materials were detected in the component cells of the digestive gland of normally feeding snails. The results of this investigation revealed a pronounced decline of carbohydrates in the digestive gland cells of Monacha cartusiana snails after starvation. Severe decline in car...

  4. Ileal and faecal protein digestibility measurement in humans and other non-ruminants - a comparative species view

    OpenAIRE

    Hendriks, W. H.; Baal, J.; Bosch, G.

    2012-01-01

    A comparative non-ruminant species view of the contribution of the large intestinal metabolism to inaccuracies in nitrogen and amino acid absorption measurements is provided to assess potential implications for the determination of crude protein/amino acid digestibility in adult humans consuming lower digestible protein sources. Most of the amino acids in the hindgut are constituents of the microorganisms and significant microbial metabolism of dietary and endogenous amino acids occu...

  5. The effect of physiological conditions on the surface structure of proteins: Setting the scene for human digestion of emulsions

    Science.gov (United States)

    Maldonado-Valderrama, J.; Gunning, A. P.; Ridout, M. J.; Wilde, P. J.; Morris, V. J.

    2009-10-01

    Understanding and manipulating the interfacial mechanisms that control human digestion of food emulsions is a crucial step towards improved control of dietary intake. This article reports initial studies on the effects of the physiological conditions within the stomach on the properties of the film formed by the milk protein ( ? -lactoglobulin) at the air-water interface. Atomic force microscopy (AFM), surface tension and surface rheology techniques were used to visualize and examine the effect of gastric conditions on the network structure. The effects of changes in temperature, pH and ionic strength on a pre-formed interfacial structure were characterized in order to simulate the actual digestion process. Changes in ionic strength had little effect on the surface properties. In isolation, acidification reduced both the dilatational and the surface shear modulus, mainly due to strong repulsive electrostatic interactions within the surface layer and raising the temperature to body temperature accelerated the rearrangements within the surface layer, resulting in a decrease of the dilatational response and an increase of surface pressure. Together pH and temperature display an unexpected synergism, independent of the ionic strength. Thus, exposure of a pre-formed interfacial ? -lactoglobulin film to simulated gastric conditions reduced the surface dilatational modulus and surface shear moduli. This is attributed to a weakening of the surface network in which the surface rearrangements of the protein prior to exposure to gastric conditions might play a crucial role.

  6. 75 FR 82428 - VASRD Improvement Forum-Updating Disability Criteria for the Genitourinary System, Digestive...

    Science.gov (United States)

    2010-12-30

    ...Updating Disability Criteria for the Genitourinary System, Digestive System, Dental Conditions, and Infectious Diseases...Updating Disability Criteria for the Genitourinary System, Digestive System, Dental Conditions, and Infectious...

  7. Identification of a Low Digestibility ?-Conglutin in Yellow Lupin (Lupinus luteus L.) Seed Meal for Atlantic Salmon (Salmo salar L.) by Coupling 2D-PAGE and Mass Spectrometry

    OpenAIRE

    Ogura, Takahiro; Herna?ndez, Adria?n; Aizawa, Tomoko; Ogihara, Jun; Sunairi, Michio; Alcaino, Javier; Salvo-garrido, Haroldo; Maureira-butler, Iva?n J.

    2013-01-01

    The need of quality protein in the aquaculture sector has forced the incorporation of alternative plant proteins into feeding diets. However, most plant proteins show lower digestibility levels than fish meal proteins, especially in carnivorous fishes. Manipulation of protein content by plant breeding can improve the digestibility rate of plant proteins in fish, but the identification of low digestibility proteins is essential. A reduction of low digestibility proteins will not only increase ...

  8. Induced mutants for cereal grain protein improvement

    International Nuclear Information System (INIS)

    Out of 17 papers and one summary presented, six dealing with the genetic improvement of seed protein using ionizing radiations fall within the INIS subject scope. Other topics discussed were non-radiation induced mutants used for cereal grain protein improvement

  9. Protein improvement in crop plants

    International Nuclear Information System (INIS)

    There are compelling reasons for attempting to increase the quality and quantity of protein available in crop plants through plant breeding, despite the fact that some critics have argued that no worldwide protein shortage exists. What used to be thought of as a 'protein gap' has now come to be considered in terms of protein-calorie malnutrition. This is only right since protein and calorie nutrition are inextricable. t the moment there are still unanswered questions as to the precise protein requirements of humans as a function of age, health and ambient conditions. There are, in addition, some indications that the incidence of Kwashiorkor (protein deficiency disease) is increasing in different parts of the world. At a recent meeting of the Protein Advisory Group of the United Nations System, Dr. Jean Mayer, an eminent human nutritionist of Harvard University, U.S.A., indicated the reasons for concern for the current food situation generally, and the protein food supply in particular. These factors include: - Immoderate continuing human population increases, most pronounced in some poor developing countries. - The highly accelerated consumption of animal foods associated with increasing affluence in the richer countries of the world. The production of such foods as meat demands great expenditures of grain, which is an inefficient mode of obtaining the required calories and protein for human consumption. - The over-exploitation of many of the world's fishery resourcesn of many of the world's fishery resources resulting in reduced yields, perhaps irreversibly, of some fishes. - Recent price increases in petroleum and fertilizer products which have imposed a major obstacle to increasing crop production. - The apparent alteration of climates in places like Africa, Asia and other parts of the Northern hemisphere which may put significant restrictions on crop production. hey are cogent reasons to be seriously concerned about these matters. (author)

  10. Kicking in the Guts: Schistosoma mansoni Digestive Tract Proteins are Potential Candidates for Vaccine Development

    Science.gov (United States)

    Figueiredo, Barbara Castro-Pimentel; Ricci, Natasha Delaqua; de Assis, Natan Raimundo Gonçalves; de Morais, Suellen Batistoni; Fonseca, Cristina Toscano; Oliveira, Sergio Costa

    2015-01-01

    Schistosomiasis is a debilitating disease that represents a major health problem in at least 74 tropical and subtropical countries. Current disease control strategies consist mainly of chemotherapy, which cannot prevent recurrent re-infection of people living in endemic area. In the last decades, many researchers made a remarkable effort in the search for an effective vaccine to provide long-term protection. Parasitic platyhelminthes of Schistosoma genus, which cause the disease, live in the blood vessels of definitive hosts where they are bathed in host blood for many years. Among the most promising molecules as vaccine candidates are the proteins present in the host–parasite interface, so numerous tegument antigens have been assessed and the achieved protection never got even close to 100%. Besides the tegument, the digestive tract is the other major site of host–parasite interface. Since parasites feed on blood, they need to swallow a considerable amount of blood for nutrient acquisition. Host blood ingested by schistosomes passes through the esophagus and reaches the gut where many peptidases catalyze the proteolysis of blood cells. Recent studies show the emergence of antigens related to the parasite blood feeding, such as esophageal gland proteins, proteases, and other proteins related to nutrient uptake. Herein, we review what is known about Schistosoma mansoni digestive tract proteins, emphasizing the ones described as potential vaccine candidates. PMID:25674091

  11. Behaviour of whey protein emulsion gel during oral and gastric digestion: effect of droplet size.

    Science.gov (United States)

    Guo, Qing; Ye, Aiqian; Lad, Mita; Dalgleish, Douglas; Singh, Harjinder

    2014-06-21

    A set of whey protein stabilized-emulsion gels with different droplet size distributions (D4,3 = ?1, 6 and 12 ?m) was produced, and the mechanical properties of the gels in the linear viscoelastic region and at large deformation were measured, along with the physicochemical and structural changes of the gels during oral mastication and gastric digestion. The gels containing 1 ?m oil droplets had an aggregated particle structure with proteins coating at oil droplets whereas the gels containing 12 ?m oil droplets had a particle-filled structure with spatially continuous matrix. During oral processing, the release of oil droplets from the gels increased as the droplet size increased, with coalescence being seen in gels containing oil droplets of 6 and 12 ?m diameter. Under gastric digestion, high degrees of coalescence and phase separation of oil droplets occurred in the gels containing 6 and 12 ?m oil droplets because of oil droplet release from the gel matrix; this led to slow gastric emptying. The gels were finally broken down into peptide aggregates and oil droplets (or free oil). The gels, containing 1 ?m oil droplets disintegrated into various particles of several to several tens of microns with a low degree of oil droplet release and coalescence. Protein breakdown was slower in these gels, suggesting that the protein structures of the gel matrices were affected by the sizes of the incorporated oil droplets. PMID:24763731

  12. Non-Starch Polysaccharides (NSP Enzyme Improves the Nutrient Digestibility of Weaned Piglet Fed a Cottonseed Meal-Based Diets

    Directory of Open Access Journals (Sweden)

    Xiaosong Wu

    2012-01-01

    Full Text Available NSP is the major factor introducing a low digestibility of cottonseed meal in piglets. The present study was conducted to determine the effect of NSP enzyme on the nutrient digestibility of weaned piglets fed a cottonseed meal-based diets. About 15 weaned piglets (14±1.4 kg were selected and ileal fistula surgery was done to determine the DE, CP, DM and NSP’s apparent digestibility. The results indicated that NSP enzyme improved piglet’s apparent digestibility (p<0.05 in varying degrees; pectinase resulted in obvious degradation of the cell wall and increased piglets’ DE (26.69% and CP of ileal digesta (20.33% and compound enzyme was better in improving cottonseed meal based diets than single enzyme.

  13. In situ identification of the synthrophic protein fermentative Coprothermobacter spp. involved in the thermophilic anaerobic digestion process.

    Science.gov (United States)

    Gagliano, Maria Cristina; Braguglia, Camilla Maria; Rossetti, Simona

    2014-09-01

    Thermophilic bacteria have recently attracted great attention because of their potential application in improving different biochemical processes such as anaerobic digestion of various substrates, wastewater treatment or hydrogen production. In this study we report on the design of a specific 16S rRNA-targeted oligonucleotide probe for detecting members of Coprothermobacter genus characterized by a strong protease activity to degrade proteins and peptides. The newly designed CTH485 probe and helper probes hCTH429 and hCTH439 were optimized for use in fluorescence in situ hybridization (FISH) on thermophilic anaerobic sludge samples. In situ probing revealed that thermo-adaptive mechanisms shaping the 16S rRNA gene may affect the identification of thermophilic microorganisms. The novel developed FISH probe extends the possibility to study the widespread thermophilic syntrophic interaction of Coprothermobacter spp. with hydrogenotrophic methanogenic archaea, whose establishment is a great benefit for the whole anaerobic system. PMID:25041640

  14. Tegoprens in anaerobic digestion of a mixture of cheese whey, poultry waste, and cattle dung for improved biomethanation

    Energy Technology Data Exchange (ETDEWEB)

    Patel, C.; Sastry, V.; Madamwar, D. [Sardar Patel Univ., Gujarat (India)

    1996-01-01

    To obtain enriched methane content and improve the anaerobic digestion of a mixture of cattle dung, poultry waste, and cheese whey, the effect of various doses of Tegoprens: T-3012, T-3099, T-5842, T-5843, T-5851, T-5852 has been studied, in bench-scale digesters. Among them, Tegoprens 3022 showed more than a 45% increase in gas production with higher methane content. 18 refs., 1 fig.

  15. The influence of protein fractions from bovine colostrum digested in vivo and in vitro on human intestinal epithelial cell proliferation.

    Science.gov (United States)

    Morgan, Alison J; Riley, Lisa G; Sheehy, Paul A; Wynn, Peter C

    2014-02-01

    Colostrum consists of a number of biologically active proteins and peptides that influence physiological function and development of a neonate. The present study investigated the biological activity of peptides released from first day bovine colostrum through in vitro and in vivo enzymatic digestion. This was assessed for proliferative activity using a human intestinal epithelial cell line, T84. Digestion of the protein fraction of bovine colostrum in vitro was conducted with the enzymes pepsin, chymosin and trypsin. Pepsin and chymosin digests yielded protein fractions with proliferative activity similar to that observed with undigested colostrum and the positive control foetal calf serum (FCS). In contrast trypsin digestion significantly (P<0·05) decreased colostral proliferative activity when co-cultured with cells when compared with undigested colostrum. The proliferative activity of undigested colostrum protein and abomasal whey protein digesta significantly increased (P<0·05) epithelial cell proliferation in comparison to a synthetic peptide mix. Bovine colostrum protein digested in vivo was collected from different regions of the gastrointestinal tract (GIT) in newborn calves fed either once (n=3 calves) or three times at 12-h intervals (n=3 calves). Digesta collected from the distal duodenum, jejunum and colon of calves fed once, significantly (P<0·05) stimulated cell proliferation in comparison with comparable samples collected from calves fed multiple times. These peptide enriched fractions are likely to yield candidate peptides with potential application for gastrointestinal repair in mammalian species. PMID:24433585

  16. Digestibility and digestive organ development in indigenous and improved chickens and ducks fed diets with increasing inclusion levels of cassava leaf meal.

    Science.gov (United States)

    Borin, K; Lindberg, J E; Ogle, R B

    2006-06-01

    Growing indigenous Cambodian chickens and ducks, and broiler chickens and White Pekin ducks were fed diets containing 0%, 7%, 14% and 20% of cassava leaf meal (CLM) to study the effects of CLM level on diet digestibility and gastrointestinal tract (GIT) and organ development. The coefficient of total tract apparent digestibility (CTTAD) of dry matter (DM) and intake of digestible DM decreased with increased dietary CLM. DM and digestible DM intake was higher for local breeds than for the corresponding exotic breeds, and higher for ducks than for chickens (p 0.05). Weight of small intestine, caeca, gizzard and pancreas, expressed as per kg body weight, increased with increased CLM in the diet (p < 0.001). There was no consistent diet effect on liver weight. Length of small intestine and caeca, expressed on a mass-specific basis, increased with dietary CLM content (p < 0.001). When expressed as per kg body weight small intestine, proventriculus, gizzard, pancreas and liver weights, and small intestine length, were higher in ducks than in chickens (p < 0.001), and were higher in the indigenous than in the improved breeds (p < 0.01), except for small intestine weights, which were similar. However chickens had higher weight of caeca (p < 0.001) and colon (p < 0.01) in absolute units and per kg body weight. PMID:16684144

  17. Highly Stable Trypsin-Aggregate Coatings on Polymer Nanofibers for Repeated Protein Digestion

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Byoung Chan; Lopez-Ferrer, Daniel; Lee, Sang-mok; Ahn, Hye-kyung; Nair, Sujith; Kim, Seong H.; Kim, Beom S.; Petritis, Konstantinos; Camp, David G.; Grate, Jay W.; Smith, Richard D.; Koo, Yoon-mo; Gu, Man Bock; Kim, Jungbae

    2009-04-01

    A stable and robust trypsin-based biocatalytic system was developed and demonstrated for proteomic applications. The system utilizes polymer nanofibers coated with trypsin aggregates for immobilized protease digestions. After covalently attaching an initial layer of trypsin to the polymer nanofibers, highly concentrated trypsin molecules are crosslinked to the layered trypsin by way of a glutaraldehyde treatment. This new process produced a 300-fold increase in trypsin activity compared with a conventional method for covalent trypsin immobilization and proved to be robust in that it still maintained a high level of activity after a year of repeated recycling. This highly stable form of immobilized trypsin was also resistant to autolysis, enabling repeated digestions of bovine serum albumin over 40 days and successful peptide identification by LC-MS/MS. Finally, the immobilized trypsin was resistant to proteolysis when exposed to other enzymes (i.e. chymotrypsin), which makes it suitable for use in “real-world” proteomic applications. Overall, the biocatalytic nanofibers with enzyme aggregate coatings proved to be an effective approach for repeated and automated protein digestion in proteomic analyses.

  18. Electrochemical detection of protein kinase activity based on carboxypeptidase Y digestion triggered signal amplification.

    Science.gov (United States)

    Yin, Huanshun; Wang, Xinxu; Guo, Yunlong; Zhou, Yunlei; Ai, Shiyun

    2015-04-15

    An effective assay method for monitoring protein kinase activity and screening inhibitors is greatly beneficial to kinase-related drug discovery, early diagnosis of diseases, and therapeutic effect evaluation. Herein, we develop a simple electrochemical method for detecting the activity of casein kinase II (CK2) based on phosphorylation against carboxypeptidase Y (CPY) digestion triggered signal amplification, where CK2 catalyzed phosphorylation event protects the substrate peptide from the digestion of CPY, maintains the repulsive force of the substrate peptide towards the redox probe, and results in a weak electrochemical signal. Whereas, without phosphorylation, the substrate peptide is digested by CPY and a strong electrochemical signal is obtained. The detection feasibility is demonstrated for the assay of CK2 activity with low detection limit of 0.047unit/mL. Moreover, the biosensor was used for the analysis of kinase inhibition. Based on the electrochemical signal dependent inhibitor concentration, the IC50 value of ellagic acid was estimated to be 39.77nM. The proposed method is also successfully applied to analyze CK2 activity in cell lysates, proving the applicability in complex biological samples. PMID:25460885

  19. Apparent Digestibility Coefficients of Protein in Selected Feedstuffs for Juvenile Nile Tilapia (Oreochromis niloticus Linnaeus, 1758

    Directory of Open Access Journals (Sweden)

    Kenan Koprucu

    2004-01-01

    Full Text Available Apparent Digestibility Coefficients (ADC of protein in anchovy (Engraulis encrasicolus, corn (Zea mays gluten, soybean (Glycine hispida, gammarid (Gammarus kischineffensis and crayfish (Astacus leptodactylus leptodactylus exoskeleton meals was determined for juvenile Nile tilapia, Oreochromis niloticus. In each of five trials, a reference diet was mixed with test ingredients in a 70:30 ratio to produce a series of test diets. Chromic oxide was added as a marker to the reference diet. The ADC for protein were; anchovy (90.2%, corn gluten (88.5%, soybean (87.6%, gammarid (76.3% and crayfish exoskeleton (70.9% meals (p<0.05 for each case, except for anchovy, corn gluten and soybean meals. The results of the study indicated that O. niloticus may efficiently utilise gammarid and crayfish exoskeleton meals as dietary protein source, like anchovy, corn gluten and soybean meals.

  20. Improving recombinant protein purification yield

    Science.gov (United States)

    Production of adequate amounts of recombinant proteins is essential for antibody production, biochemical activity study, and structural determination during the post-genomic era. It’s technologically challenging and a limiting factor for tung oil research because analytical reagents such as high qua...

  1. Feed transit and apparent protein, phosphorus and energy digestibility of practical feed ingredients by Senegalese sole (Solea senegalensis)

    OpenAIRE

    Dias, Jorge; Yu?fera, Manuel; Valente, Luisa M. P.; Rema, Paulo

    2010-01-01

    A study was conducted with Senegalese sole (Solea senegalensis) to determine the apparent digestibility coefficients (ADCs) of protein, phosphorus and energy in practical feed ingredients. The digestible energy (DE) content was also evaluated. Test ingredients were anchovy fishmeal, solvent-extracted soybean meal, corn gluten and wheat meal. Due to their low cohesion, sole faeces were collected directly in posterior intestine by dissection. The feed transit time along the gastrointestinal tra...

  2. Protein--RNA interaction in the rat liver 5S rRNA-protein L5 complex studied by digestion with ribonucleases.

    OpenAIRE

    Gross, B.; Welfle, H.; Bielka, H.

    1985-01-01

    Protein-RNA interactions in the 5S rRNA-protein L5 complex from rat liver ribosomes were studied by limited digestion of free and protein bound 5S rRNA with ribonuclease A and T1. In the complex with protein L5 the digestion of 5S rRNA by ribonuclease T1 is decreased at G37 and G89, whereas U38 and C39, and to a lower extent also C10 and U12 become accessible for ribonuclease A.

  3. The impact of dietary protein levels on nutrient digestibility and water and nitrogen balances in eventing horses.

    Science.gov (United States)

    Oliveira, C A A; Azevedo, J F; Martins, J A; Barreto, M P; Silva, V P; Julliand, V; Almeida, F Q

    2015-01-01

    This study was performed to evaluate the impact of dietary protein levels on nutrient digestibility and water and nitrogen balances in conditioning eventing horses. Twenty-four Brazilian Sport Horses, male and female (8.0 to 15.0 yr; 488 ± 32 kg BW), were used in a randomized design with 4 levels of CP diets: 7.5%, 9.0%, 11.0%, and 13.0%. A digestion assay was performed with partial feces collection over 4 d, followed by 1 d of total urine collection. Data were submitted to regression analysis and adjusted to linear and quadratic models (P linear increase as a function of increasing protein level in diets. A quadratic response (P linear effect on ADF digestibility coefficients, digestible DM and protein intake, and CP/DE ratio according to dietary protein levels. There was no impact of dietary protein levels on daily water intake, total water intake, or fecal water excretion. Urinary excretion values showed a linear increase in response to increased dietary protein levels, but no impact was observed on water balance, with an average of 8.4 L/d. Nitrogen intake (NI), N absorption (NA), and urinary N increased linearly as a function of increasing dietary protein levels. There was no impact of dietary protein levels on N retention (NR), with an average of 7.5 g N/d. Nitrogen retention as a percentage of NI or NA showed no significant changes in the function of dietary protein levels. There was an impact of dietary protein levels on the digestibility coefficient of CP, NDF, ADF, and digestible protein intake on conditioning eventing horses. The 11.6% CP level in the diet provided an intake of 2.25 g CP/kg BW and 0.37 g N/kg BW, and this intake was the most appropriate for the conditioning of intensely exercised horses, considering the responses related to NI, NA, and the estimated NR to NA ratio. The NDF and ADF responses indicated that dietary fiber was more digested with an increased amount of N in the digestive tract. PMID:25568371

  4. Improved coverage of cDNA-AFLP by sequential digestion of immobilized cDNA

    Science.gov (United States)

    Weiberg, Arne; Pöhler, Dirk; Morgenstern, Burkhard; Karlovsky, Petr

    2008-01-01

    Background cDNA-AFLP is a transcriptomics technique which does not require prior sequence information and can therefore be used as a gene discovery tool. The method is based on selective amplification of cDNA fragments generated by restriction endonucleases, electrophoretic separation of the products and comparison of the band patterns between treated samples and controls. Unequal distribution of restriction sites used to generate cDNA fragments negatively affects the performance of cDNA-AFLP. Some transcripts are represented by more than one fragment while other escape detection, causing redundancy and reducing the coverage of the analysis, respectively. Results With the goal of improving the coverage of cDNA-AFLP without increasing its redundancy, we designed a modified cDNA-AFLP protocol. Immobilized cDNA is sequentially digested with several restriction endonucleases and the released DNA fragments are collected in mutually exclusive pools. To investigate the performance of the protocol, software tool MECS (Multiple Enzyme cDNA-AFLP Simulation) was written in Perl. cDNA-AFLP protocols described in the literatur and the new sequential digestion protocol were simulated on sets of cDNA sequences from mouse, human and Arabidopsis thaliana. The redundancy and coverage, the total number of PCR reactions, and the average fragment length were calculated for each protocol and cDNA set. Conclusion Simulation revealed that sequential digestion of immobilized cDNA followed by the partitioning of released fragments into mutually exclusive pools outperformed other cDNA-AFLP protocols in terms of coverage, redundancy, fragment length, and the total number of PCRs. Primers generating 30 to 70 amplicons per PCR provided the highest fraction of electrophoretically distinguishable fragments suitable for normalization. For A. thaliana, human and mice transcriptome, the use of two marking enzymes and three sequentially applied releasing enzymes for each of the marking enzymes is recommended. PMID:18851732

  5. Changes in nitrogenous and other chemical constituents, protein fractions and in vitro protein digestibility of germinating fluted pumpkin (Telfairia occidentalis Hook) seed.

    Science.gov (United States)

    Giami, S Y; Chibor, B S; Edebiri, K E; Achinewhu, S C

    1999-01-01

    The effect of 7 days of germination on levels of nitrogenous and other nutrition related parameters, protein fractions and in vitro protein digestibility of fluted pumpkin (Telfairia occidentalis) seed was studied. The non-protein nitrogen gradually increased and the protein nitrogen content decreased during germination. Albumin and globulin fractions were found to be the major seed proteins of fluted pumpkin seeds, constituting about 58.6% of the total protein of the ungerminated (raw) seeds. The protein fractions, albumin and glutelin, were observed to increase by 61.5% and 57.0%, respectively, while a 54.6% decrease was noted in the prolamine fraction. The globulin fraction increased at the beginning of germination but decreased at the end. Germination significantly (p< or =0.05) increased the crude protein, nitrogen solubility and in vitro protein digestibility but decreased the fat, phytic acid and polyphenol contents of the seeds. PMID:10540985

  6. Comparison of standardized ileal amino acid digestibilities in protein supplements and cereal grains for weaned pigs.

    Science.gov (United States)

    Sauer, N; Eklund, M; Hoerner, S; Rademacher, M; Mosenthin, R

    2012-12-01

    Standardized ileal digestibility (SID) of AA in protein ingredients and grains was determined in weaned piglets (5 kg initial BW) using the difference method. Animals were fitted with a simple ileal T-cannula on day 24 or 25 of age. A synthetic diet based on corn (Zea mays) starch and casein was either supplemented with an extruded soybean (Glycine max) meal (ESM), rice (Oryza sativa) protein concentrate (RPC), full fat heat-treated soybeans (SB), corn, barley (Hordeum vulgare), or wheat (Triticum aestivum). The contribution of CP and AA from casein and from the assay feed ingredients to the assay diet averaged 50% each. Each diet was fed to 6 animals according to a row-column design with 3 periods. Apparent ileal digestibility was corrected for basal ileal endogenous losses of AA to obtain SID values. Between RPC and soy products, differences (P 0.05) between barley and wheat, except for Met (P < 0.05). Furthermore, SID of Arg, His, Ile, Phe, and Trp was lower (P < 0.05) in corn than wheat. In conclusion, SID of most AA in the present study with piglets was lower than tabulated SID values determined in studies with grower-finisher pigs. Furthermore, the use of RPC, despite its high CP content, in diets for weaned piglets is limited due to its low SID of AA. PMID:23365298

  7. Improved biogas production from whole stillage by co-digestion with cattle manure.

    Science.gov (United States)

    Westerholm, Maria; Hansson, Mikael; Schnürer, Anna

    2012-06-01

    Whole stillage, as sole substrate or co-digested with cattle manure, was evaluated as substrate for biogas production in five mesophilic laboratory-scale biogas reactors, operating semi-continuously for 640 days. The process performance was monitored by chemical parameters and by quantitative analysis of the methanogenic and acetogenic population. With whole stillage as sole substrate the process showed clear signs of instability after 120 days of operation. However, co-digestion with manure clearly improved biogas productivity and process stability and indicated increased methane yield compared with theoretical values. The methane yield at an organic loading rate (OLR) at 2.8 g VS/(L×day) and a hydraulic retention time (HRT) of 45 days with a substrate mixture 85% whole stillage and 15% manure (based on volatile solids [VS]) was 0.31 N L CH(4)/gVS. Surprisingly, the abundance of the methanogenic and acetogenic populations remained relatively stable throughout the whole operation and was not influenced by process performance. PMID:22464422

  8. Histochemical studies on enzyme-digested protein plugs of patients with chronic pancreatitis: a preliminary report.

    Directory of Open Access Journals (Sweden)

    Harada,Hideo

    1983-06-01

    Full Text Available Sulfated acidic mucopolysaccharides have been found to be significant components of "protein plugs" in patients with chronic pancreatitis. The precise identification of the mucopolysaccharides and their distribution within the protein plugs may clarify the pathogenesis of the plugs. Pure pancreatic juice from five patients with chronic pancreatitis was obtained by endoscopic retrograde catheterization of the papilla of Vater. Enzymes for digestion of the plugs included hyaluronidase of the bovine testes and streptomyces hyalurolyticus, chondroitinase ABC and AC, and sialidase (neuraminidase. Our study indicated that: I Sialic acid is distributed throughout the plugs and may be a major component, followed by a lesser amount of chondroitin sulfate B. 2 Chondroitin sulfate A, C, D and E and chondroitin may be minor components. 3 Hyaluronic acid is negligible in the plugs.

  9. Uric Acid Determination in Poultry Excreta and its Application to Correcting Protein Digestibility Values: Spectrophotometry with Ultraviolet Detection vs HPLIC

    Directory of Open Access Journals (Sweden)

    R. R. Marquardt

    2002-10-01

    Full Text Available Evaluation of uric acid contents of poultry excreta was examined for use in the estimation of apparent protein digestibility. Uric acid was determined either by high-performance liquid chromatography or by spectrophotometric method. A high-pressure liquid chromatographic assay was used with absorbance measured at 285 nm. The method used a reverse-phase system with a C18-bonded column (250*4.6 mm, i.d.. The mobile phase was distilled deionized water. Injection volume was 20 l with a flow rate of 1 ml/min. Correlation (r between the two methods for uric acid content of excreta from birds fed diets with different methionine levels was 0.976. The uric acid values were then used to determine apparent protein digestibility. The apparent protein digestibility values, when corrected for uric acid nitrogen, were comparable with those calculated from amino acid analysis.

  10. The Effects of Different Energy and Protein Ratio to Sheepâ??s Nutrient Intake and Digestibility

    Directory of Open Access Journals (Sweden)

    Agung Purnomoadi

    2013-06-01

    Full Text Available Normal 0 false false false IN X-NONE X-NONE MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;} The objective of this research was to study the effects of different energy and protein ratio towards sheep�s nutrient intake and digestibility. Twenty four male sheep�s, 6 � 7 months old with initial average live weight 13+1.56 kg, coefficient variant11.78% were used in this research. The complete feed ration which consisted of King Grass (Pennisetum purpureum, soybean powder, rice bran, dried cassava and molasses was used in this research. Protein content on each component was 10, 12 and 14% and total digestible nutrients (TDN 60 and 65%, respectively. Dry matter (DM and organic matter (OM intake, DM and OM digestibility were studied in this research. Analysis of variance (ANOVA was employed to analyze the data. Test of Small Difference (P Dry matter and OM ration intake showed significant different among treatments (P0.05 among crude protein and TDN treatments. Different energy and protein ration treatments caused different DM and OM intake but were not cause different in DM and OM digestibility. Based on the research results, a study on the effects of different ration�s energy and protein ratio towards N efficiency should be conducted in order to increase cattle productivity. Normal 0 false false false IN X-NONE X-NONE MicrosoftInternetExplorer4 Doi: http://dx.doi.org/10.12777/ijse.4.2.75-79 [How to cite this article: Mawati, S., Soedarsono, S., Sunarso, S. & Purnomoadi, A. (2013. The Effects of Different Energy and Ratio to Sheep�s Nutrient Intake and Digestibility. International Journal of Science and Engineering, 4(2,76-79. Doi: http://dx.doi.org/10.12777/ijse.4.2.75-79] /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;}

  11. Amaranth-protein interaction in food system and its impact on tryptic digestibility

    International Nuclear Information System (INIS)

    Amaranth, a food color, is used in variety of food products to attract consumers, especially children. The purpose of the present study was to identify the component present in the food system which acts as a carrier of color and its distribution. The protein is the most possible candidate for color-conjugates and this was first explored by staining the resolved food proteins on PAGE simultaneously and separately with Amaranth as well as by Coomassie brilliant blue R250. It is the most widely used dye for protein assay. The color intensity of the Amaranth-protein complexes was slightly less than those of Coomassie brilliant blue R250, although the bands stained by Amaranth were very sharp, clearly separated and distinct. The staining procedure followed for Amaranth was quick. The impact of tryptic digestibility on amaranth-protein complexes has illustrated that dye may safely be used without any adverse effect. The possible moode of conjugation between amino acid and azo-bond is also discussed. (author)

  12. Effects of synchronizing starch and protein degradation in rumen on fermentation, nutrient utilization and total tract digestibility in sheep

    OpenAIRE

    Birgul Bozan; Gulay Deniz; Bulent Haluk Gulmez; Hakan Biricik; Ibrahim Ismet Turkmen; Hidir Gencoglu

    2010-01-01

    The objective of this experiment was to investigate the effects of synchronizing the dietary starch and crude protein (CP) degradation in the rumen on nutrient utilization, fermentation, and total tract digestibility in sheep. The four diets were formulated with different rates of starch and CP release in the rumen but with similar metabolic energy, starch, and CP. The diets were slowly degradable starch, slowly degradable protein; slowly degradable starch, rapidly degradable protein; rapidly...

  13. Identification of a low digestibility ?-Conglutin in yellow lupin (Lupinus luteus L.) seed meal for atlantic salmon (Salmo salar L.) by coupling 2D-PAGE and mass spectrometry.

    Science.gov (United States)

    Ogura, Takahiro; Hernández, Adrián; Aizawa, Tomoko; Ogihara, Jun; Sunairi, Michio; Alcaino, Javier; Salvo-Garrido, Haroldo; Maureira-Butler, Iván J

    2013-01-01

    The need of quality protein in the aquaculture sector has forced the incorporation of alternative plant proteins into feeding diets. However, most plant proteins show lower digestibility levels than fish meal proteins, especially in carnivorous fishes. Manipulation of protein content by plant breeding can improve the digestibility rate of plant proteins in fish, but the identification of low digestibility proteins is essential. A reduction of low digestibility proteins will not only increase feed efficiency, but also reduce water pollution. Little is known about specific digestible protein profiles and/or molecular identification of more bioavailable plant proteins in fish diets. In this study, we identified low digestibility L. luteus seed proteins using Atlantic salmon (Salmo salar) crude digestive enzymes in an in vitro assay. Low digestibility proteins were identified by comparing SDS-PAGE banding profiles of digested and non-digested lupin seed proteins. Gel image analysis detected a major 12 kDa protein band in both lupin meal and protein isolate digested products. The 12 kDa was confirmed by 2D-PAGE gels and the extracted protein was analyzed with an ion trap mass spectrometer in tandem mass mode. The MS/MS data showed that the 12 kDa low digestibility protein was a large chain ?conglutin, a common seed storage protein of yellow lupin. Comparison of the protein band profiles between lupin meal and protein isolates showed that the isolatation process did not affect the low digestibility of the 12 kDa protein. PMID:24278278

  14. A Comparative Study of In-Gel Digestions Using Microwave and Pressure-Accelerated Technologies

    OpenAIRE

    Alvarado, Rudy; Tran, Diana; Ching, Bonnie; Phinney, Brett S.

    2010-01-01

    One of the most popular methods to prepare tryptic peptides for bottom-up proteomic analysis is in-gel digestion. To date, there have been few studies comparing various digestion methods. In this study, we compare the efficiency of several popular in-gel digestion methods, along with new technologies that may improve digestion efficiency, using a human epidermoid carcinoma cell lysate protein standard. The efficiency of each protocol was based on the average number of proteins identified and ...

  15. On-plate digestion using a commercial micro-fraction collector for nano-HPLC MALDI-TOF/TOF protein analysis

    OpenAIRE

    Vitorino, Rui; Guedes, Sofia; Tomer, Kenneth; Domingues, Pedro; Duarte, Jose?; Amado, Francisco

    2008-01-01

    A new method for on-plate protein digestion and MALDI MS analysis is proposed involving an automated one-step sample separation using nanoflow-HPLC followed by nanoliter fraction collection and on-plate digestion with trypsin. This procedure uses a commercial automatic nanoliter fraction collection system for on-line spotting of the eluent onto a MALDI target. After protein digestion, the reaction is stopped by the addition of acidified matrix using the same automated system. Collected spots ...

  16. Dietary protein hydrolysate and trypsin inhibitor effects on digestive capacities and performances during early-stages of spotted wolffish: Suggested mechanisms

    OpenAIRE

    Savoie, A.; Le Francois, N. R.; Lamarre, S. G.; Blier, P. U.; Beaulieu, L.; Cahu, Chantal

    2011-01-01

    Growth rate is dependent upon adequate provision of amino acids especially in newly-hatched fish which experience very high growth rate. The replacement of a fraction of protein content by partially hydrolyzed (pre-digested) proteins was carried out and the digestive capacities and performances of larval/juvenile spotted wolffish (Anarhichas minor) were measured. The goal of this study was to verify whether the scope for growth is principally dictated by the proteolytic capacity of the digest...

  17. Comparison of electron beam and gamma ray irradiations effects on ruminal crude protein and amino acid degradation kinetics, and in vitro digestibility of cottonseed meal

    Science.gov (United States)

    Ghanbari, F.; Ghoorchi, T.; Shawrang, P.; Mansouri, H.; Torbati-Nejad, N. M.

    2012-06-01

    This study was conducted to compare effects of electron beam (EB) and gamma ray (GR) treatments at doses of 25, 50 and 75 kGy on ruminal degradation kinetics of crude protein (CP), amino acid (AA), and in vitro digestibility of cottonseed meal (CSM). Ionizing radiations of EB and GR had significant effects (Pdegradability characteristics of CSM. Effective ruminal degradability (ERD) of CP was lower in EB and GR irradiated CSM (P0.05). Irradiation processing caused decrement in AA degradation after 16 h of ruminal incubation (Pdegradability of AA (Pvitro digestibility of CSM numerically. This study showed that EB could cause CP and AA bypass rumen as well as GR. Therefore, ionizing irradiation processing can be used as an efficient method in improving nutritional value of CSM.

  18. Digestibilidade intestinal verdadeira da proteína de alimentos para ruminantes True small intestinal protein digestibility of ruminant feeds

    Directory of Open Access Journals (Sweden)

    Antonio Ferriani Branco

    2006-08-01

    Full Text Available A digestibilidade intestinal verdadeira de diferentes classes de alimentos usados em dietas para ruminantes foi avaliada por meio das técnicas in situ e in vitro. Foram utilizados dois bovinos machos castrados (450 kg PV com cânulas implantadas no rúmen para incubação in situ de concentrados protéicos de origens animal e vegetal e energéticos, resíduos da agroindústria e alimentos volumosos. Avaliou-se a digestibilidade intestinal verdadeira dos alimentos submetidos à digestão apenas com pepsina ou com pepsina + pancreatina, precedida ou não da incubação ruminal. A incubação ruminal diminuiu a digestibilidade intestinal verdadeira da proteína de 24 dos 30 alimentos testados, com exceção da farinha de penas, da aveia preta, do grão de milho triturado a 2,5 mm e dos fenos de aveia e tifton, para os quais ocorreu aumento, e do farelo de girassol, para o qual não houve efeito da incubação ruminal. A digestibilidade intestinal da proteína não-degradada no rúmen (PNDR, na maioria dos alimentos utilizados em dietas para ruminantes, é menor que a da proteína original do alimento. Entre os alimentos avaliados, 29 apresentaram maior digestibilidade intestinal verdadeira quando incubados com pepsina + pancreatina, evidenciando a importância da etapa de digestão abomasal sobre as proteínas dos alimentos (com exceção à aveia preta. A digestibilidade intestinal dos alimentos é variável e, portanto, deve ser considerada na formulação de dietas para atendimento das exigências de proteína metabolizável.The true protein digestibility in the small intestine of different ruminant feeds were measured using in situ and in vitro techniques. Two steers with average body weight of 450 kg and fitted with ruminal cannulas were used for in situ incubation of different feeds. The following feedstuffs were evaluated: animal and vegetable protein concentrate, energy concentrate, fiber by-products, and forage. Protein truly digested in the small intestine was estimated by pepsin or pepsin/pancreatin incubation with or without previous ruminal incubation. Out of 30 evaluated feeds, ruminal incubation decreased the true protein digestibility in the small intestine of 24 feeds, increased that of feather meal, black oat, 2.5-mm particle size corn grain, and oat and Tifton hays and had no effect on sunflower meal. These results showed that the small intestinal digestibility of rumen-undegradable protein from most analyzed feeds was lower than that of the original feed protein. With the exception of black oat, all remaining feeds (29 had greater true small intestinal protein digestibility after incubation with pepsin plus pancreatin showing the contribution of abomasal digestion on feed protein. Because true small intestinal protein digestibility differed among feeds, is important to take it into account on diet formulation in order to accurately determine the requirements of metabolizable protein of ruminants.

  19. Phosphopeptide enrichment by covalent chromatography after derivatization of protein digests immobilized on reversed-phase supports.

    Science.gov (United States)

    Nika, Heinz; Nieves, Edward; Hawke, David H; Angeletti, Ruth Hogue

    2013-09-01

    A rugged sample-preparation method for comprehensive affinity enrichment of phosphopeptides from protein digests has been developed. The method uses a series of chemical reactions to incorporate efficiently and specifically a thiol-functionalized affinity tag into the analyte by barium hydroxide catalyzed ?-elimination with Michael addition using 2-aminoethanethiol as nucleophile and subsequent thiolation of the resulting amino group with sulfosuccinimidyl-2-(biotinamido) ethyl-1,3-dithiopropionate. Gentle oxidation of cysteine residues, followed by acetylation of ?- and ?-amino groups before these reactions, ensured selectivity of reversible capture of the modified phosphopeptides by covalent chromatography on activated thiol sepharose. The use of C18 reversed-phase supports as a miniaturized reaction bed facilitated optimization of the individual modification steps for throughput and completeness of derivatization. Reagents were exchanged directly on the supports, eliminating sample transfer between the reaction steps and thus, allowing the immobilized analyte to be carried through the multistep reaction scheme with minimal sample loss. The use of this sample-preparation method for phosphopeptide enrichment was demonstrated with low-level amounts of in-gel-digested protein. As applied to tryptic digests of ?-S1- and ?-casein, the method enabled the enrichment and detection of the phosphorylated peptides contained in the mixture, including the tetraphosphorylated species of ?-casein, which has escaped chemical procedures reported previously. The isolates proved highly suitable for mapping the sites of phosphorylation by collisionally induced dissociation. ?-Elimination, with consecutive Michael addition, expanded the use of the solid-phase-based enrichment strategy to phosphothreonyl peptides and to phosphoseryl/phosphothreonyl peptides derived from proline-directed kinase substrates and to their O-sulfono- and O-linked ?-N-acetylglucosamine (O-GlcNAc)-modified counterparts. Solid-phase enzymatic dephosphorylation proved to be a viable tool to condition O-GlcNAcylated peptide in mixtures with phosphopeptides for selective affinity purification. Acetylation, as an integral step of the sample-preparation method, precluded reduction in recovery of the thiolation substrate caused by intrapeptide lysine-dehydroalanine cross-link formation. The solid-phase analytical platform provides robustness and simplicity of operation using equipment readily available in most biological laboratories and is expected to accommodate additional chemistries to expand the scope of solid-phase serial derivatization for protein structural characterization. PMID:23997662

  20. Allergenicity of potato proteins and of their conjugates with galactose, galactooligosaccharides, and galactan in native, heated, and digested forms.

    Science.gov (United States)

    Seo, Sooyoun; L'Hocine, Lamia; Karboune, Salwa

    2014-04-23

    The effect of glycation of potato proteins on their immunoreactivity was studied by using a pool of human sera with specific IgE to potato proteins. Patatin conjugates were more immunoreactive than protease inhibitors ones. To better understand this behavior, the changes in patatin structure upon glycation and heat treatment were investigated. Patatin demonstrated an increase in total immunoreactivity when glycated with galactose and galactooligosaccharides. However, galactan conjugation to patatin resulted in a decrease in immunoreactivity by restricting IgE's access to the epitopes. Although the heat treatment resulted in a decrease in patatin's immunoreactivity through aggregation, it was less effective when patatin conjugates were used due to the decrease in aggregation and the secondary structural changes. Upon digestion, native patatin exhibited the largest decrease in immunoreactivity resulting from the disruption of both conformational and sequential epitopes. Patatin conjugates were less digested and had higher IgE-immunoreactivity as compared to the digested patatin. PMID:24661320

  1. Effect of palm kernel cake as protein source in a concentrate diet on intake, digestibility and live weight gain of goats fed Napier grass.

    Science.gov (United States)

    Rahman, Mohammad Mijanur; Abdullah, Ramli Bin; Wan Embong, Wan Khadijah; Nakagawa, Toshinori; Akashi, Ryo

    2013-03-01

    The effects of palm kernel cake (PKC) as a protein source in a concentrate diet (comprising 35 % crushed maize, 30 % rice bran, 32 % PKC, 2 % vitamin mineral premix and 1 % salt) were examined on intake, live weight (LW) gain and digestibility in female goats (average LW of 12.4?±?2.6 kg). Four goats were randomly allocated to each of the four treatment diets: (a) Napier grass (Pennisetum purpureum) offered ad libitum (T1), (b) T1 + concentrate at 0.5 % of LW (T2), (c) T1 + concentrate at 1.0 % of LW (T3) and (d) T1 + concentrate at 2.0 % of LW (T4). A 7-day digestibility trial and an 82-day growth experiment were conducted. No differences were observed among diets for intakes of roughage dry matter (DM), total DM, organic matter (OM) and neutral detergent fibre (NDF). The crude protein (CP) intake increased (P??0.05) among treatments. The digestibility of dietary NDF decreased (P??0.05) difference between T2 and T3 diets. Supplementing a basal diet of Napier grass with PKC-based concentrate improved CP intake and LW gain. The PKC-based concentrate diet can therefore be exploited for the use of local feed resources for goat production; however, further research is required to achieve the best growth response. PMID:23096766

  2. Improving bioorthogonal protein ubiquitylation by click reaction.

    Science.gov (United States)

    Schneider, Daniel; Schneider, Tatjana; Rösner, Daniel; Scheffner, Martin; Marx, Andreas

    2013-06-15

    Posttranslational modification of proteins with ubiquitin (ubiquitylation) regulates numerous cellular processes. Besides functioning as a signal for proteasomal degradation, ubiquitylation has also non-proteolytic functions by altering the biochemical properties of the modified protein. To investigate the effect(s) of ubiquitylation on the properties of a protein, sufficient amounts of homogenously and well-defined ubiquitylated proteins are required. Here, we report on the elaboration of a method for the generation of high amounts of site-specifically mono-ubiquitylated proteins. Firstly, a one-step affinity purification scheme was developed for ubiquitin containing the unnatural amino acid azidohomoalanine at the C-terminal position. This ubiquitin was conjugated in a click reaction to recombinant DNA polymerase ?, equipped with an alkyne function at a distinct position. Secondly, addition of defined amounts of SDS to the reaction significantly improved product formation. With these two technical improvements, we have developed a straight forward procedure for the efficient generation of site-specifically ubiquitylated proteins that can be used to study the effect(s) of ubiquitylation on the activities/properties of a protein. PMID:23611767

  3. In vivo assessment of the ability of condensed tannins to interfere with the digestibility of plant protein in sheep

    Energy Technology Data Exchange (ETDEWEB)

    Andrabi, S.M.; Ritchie, M.M.; Stimson, C.; Horadagoda, A.; Hyde, M.; McNeill, D.M. [MC Franklin Laboratory, Faculty of Veterinary Science, University of Sydney, Camden, NSW (Australia)]. E-mail: karalue@bigpond.net.au

    2005-08-19

    To assess the effect of condensed tannin (CT) astringency on the digestibility of protein post-ruminally, CT were purified from four types (accessions) of Mulga (Acacia aneura), and a Leucaena pallida and dosed into sheep as complexes with {sup 15}N-plant protein (tannin-protein complexes, TPC), together with an indigestible marker, chromium-EDTA (Cr-EDTA). Each CT treatment dose comprised 12 mg CT, 10 mg {sup 15}N-protein, and 2.77 mg of Cr. A protein-only control (same as the other TPC solutions but without any CT) treatment was also included to make a total of six treatments. Treatments were applied in two 6 x 6 Latin Square designs with 72 h between each infusion for each sheep. In the first, the solutions were infused post-ruminally via an abomasal cannula. In the second they were dosed directly into the mouth following an intra-nasal dose of an analogue of vasopressin in an attempt to stimulate the oesophageal groove reflex to direct the solutions more efficiently toward the small intestine. Results showed no detectable effect of CT type on the in vivo digestibility of the {sup 15}N-protein. Protein digestibilities were uniformly high, indicative of complete dissociation of the TPC. There was no correlation between protein digestibility, mouth to faeces, and protein digestibility, abomasum to faeces (P > 0.05). In vivo digestibility was also uncorrelated with CT astringencies defined in vitro (P > 0.05). Astringency in vitro was defined as the mg of CT required to achieve half-maximal precipitation of 0.5 mg of protein (bovine serum albumin). In vitro, the most astringent CT (A. aneura 883558), had at least 1.6 times the astringency of the weakest CT (A. aneura 842394). The A. aneura with the weakest CT also contained less than 1/3 the total amount of CT/g leaf dry matter than that with the strongest, highlighting the scope for selection of more nutritionally useful types of A. aneura. Limitations of the in vivo protocol used are discussed and it is concluded that the effect of CT astringency on the availability of protein post-ruminally is minimal. (author)

  4. In vivo assessment of the ability of condensed tannins to interfere with the digestibility of plant protein in sheep

    International Nuclear Information System (INIS)

    To assess the effect of condensed tannin (CT) astringency on the digestibility of protein post-ruminally, CT were purified from four types (accessions) of Mulga (Acacia aneura), and a Leucaena pallida and dosed into sheep as complexes with 15N-plant protein (tannin-protein complexes, TPC), together with an indigestible marker, chromium-EDTA (Cr-EDTA). Each CT treatment dose comprised 12 mg CT, 10 mg 15N-protein, and 2.77 mg of Cr. A protein-only control (same as the other TPC solutions but without any CT) treatment was also included to make a total of six treatments. Treatments were applied in two 6 x 6 Latin Square designs with 72 h between each infusion for each sheep. In the first, the solutions were infused post-ruminally via an abomasal cannula. In the second they were dosed directly into the mouth following an intra-nasal dose of an analogue of vasopressin in an attempt to stimulate the oesophageal groove reflex to direct the solutions more efficiently toward the small intestine. Results showed no detectable effect of CT type on the in vivo digestibility of the 15N-protein. Protein digestibilities were uniformly high, indicative of complete dissociation of the TPC. There was no correlation between protein digestibility, mouth to faeces, and protein digestibility, abomasum to faeces (P > 0.05). In vivo digestibility was also uncorrelated with CT astringencies defined in vitro (P > 0.05). Astringency in vitro was defined as the mg stringency in vitro was defined as the mg of CT required to achieve half-maximal precipitation of 0.5 mg of protein (bovine serum albumin). In vitro, the most astringent CT (A. aneura 883558), had at least 1.6 times the astringency of the weakest CT (A. aneura 842394). The A. aneura with the weakest CT also contained less than 1/3 the total amount of CT/g leaf dry matter than that with the strongest, highlighting the scope for selection of more nutritionally useful types of A. aneura. Limitations of the in vivo protocol used are discussed and it is concluded that the effect of CT astringency on the availability of protein post-ruminally is minimal. (author)

  5. In vitro study on digestion of pumpkin oil cake protein hydrolysate: evaluation of impact on bioactive properties.

    Science.gov (United States)

    Vaštag, Zužana; Popovi?, Ljiljana; Popovi?, Senka; Peri?in-Star?evi?, Ivana; Krimer-Maleševi?, Vera

    2013-06-01

    In this work, a simulated gastrointestinal digestion of pumpkin oil cake protein hydrolysate prepared by alcalase (AH) was studied to evaluate the impact of the main gastrointestinal proteases on its antiradical and angiotensin I-converting enzyme (ACE) inhibitory activity. The in vitro digestion was performed in a model system under optimized reaction conditions, first by pepsin and then with ?-chymotrypsin and trypsin, simultaneously. The treatment with the gastrointestinal proteases led to a significant increase of the degree of hydrolysis, up to 55.95 ± 3.1% in the final digest. After the digestion, the 2,2-azinobis3-ethylbenzo-thiazoline-6-sulphonic acid radical cation activity of AH was increased from 7.59 ± 0.1 to 10.25 ± 0.3 mM trolox equivalent antioxidant coefficient/mg (p 0.05) in the final digest. These results showed an advantage of AH to increase the antiradical and resist ACE inhibitory activity during digestion by main gastrointestinal proteases, appearing as promising bioactive food ingredient. PMID:23216435

  6. The Protein Composition of the Digestive Fluid from the Venus Flytrap Sheds Light on Prey Digestion Mechanisms*

    OpenAIRE

    Schulze, Waltraud X.; Sanggaard, Kristian W.; Kreuzer, Ines; Knudsen, Anders D.; Bemm, Felix; Thøgersen, Ida B.; Bra?utigam, Andrea; Thomsen, Line R.; Schliesky, Simon; Dyrlund, Thomas F.; Escalante-perez, Maria; Becker, Dirk; Schultz, Jo?rg; Karring, Henrik; Weber, Andreas

    2012-01-01

    The Venus flytrap (Dionaea muscipula) is one of the most well-known carnivorous plants because of its unique ability to capture small animals, usually insects or spiders, through a unique snap-trapping mechanism. The animals are subsequently killed and digested so that the plants can assimilate nutrients, as they grow in mineral-deficient soils. We deep sequenced the cDNA from Dionaea traps to obtain transcript libraries, which were used in the mass spectrometry-based identification of the pr...

  7. Simulated gastrointestinal digestion, intestinal permeation and plasma protein interaction of white, green, and black tea polyphenols.

    Science.gov (United States)

    Tenore, Gian Carlo; Campiglia, Pietro; Giannetti, Daniela; Novellino, Ettore

    2015-02-15

    The gastrointestinal digestion, intestinal permeation, and plasma protein interaction of polyphenols from a single tea cultivar at different stages of processing (white, green, and black teas) were simulated. The salivary phase contained 74.8-99.5% of native polyphenols, suggesting potential bioavailability of significant amounts of antioxidants through the oral mucosal epithelium that might be gastric sensitive and/or poorly absorbed in the intestine. White tea had the highest content and provided the best intestinal bioaccessibility and bioavailability for catechins. Since most of native catechins were not absorbed, they were expected to accumulate in the intestinal lumen where a potential inhibition capacity of cellular glucose and cholesterol uptake was assumed. The permeated catechins (approximately, 2-15% of intestinal levels) significantly bound (about 37%) to plasma HDLs, suggesting a major role in cholesterol metabolism. White tea and its potential nutraceuticals could be effective in the regulation of plasma glucose and cholesterol levels. PMID:25236233

  8. Immobilized trypsin on hydrophobic cellulose decorated nanoparticles shows good stability and reusability for protein digestion.

    Science.gov (United States)

    Sun, Xiuxia; Cai, Xiangdong; Wang, Ren-Qi; Xiao, Jianxi

    2015-05-15

    The preparation of biocatalysts based on immobilized trypsin is of great importance for both proteomic research and industrial applications. Here, we have developed a facile method to immobilize trypsin on hydrophobic cellulose-coated silica nanoparticles by surface adsorption. The immobilization conditions for the trypsin enzyme were optimized. The as-prepared biocatalyst was characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, and elemental analysis. In comparison with free enzyme, the immobilized trypsin exhibited greater resistances against thermal inactivation and denaturants. In addition, the immobilized trypsin showed good durability for multiple recycling. The general applicability of the immobilized trypsin for proteomic studies was confirmed by enzymatic digestion of two widely used protein substrates: bovine serum albumin (BSA) and cytochrome c. The surface adsorption protocols for trypsin immobilization may provide a promising strategy for enzyme immobilization in general, with great potential for a range of applications in proteomic studies. PMID:25700866

  9. EnzymePredictor: a tool for predicting and visualizing enzymatic cleavages of digested proteins.

    Science.gov (United States)

    Vijayakumar, Vaishnavi; Guerrero, Andrés N; Davey, Norman; Lebrilla, Carlito B; Shields, Denis C; Khaldi, Nora

    2012-12-01

    Mass spectrometric analysis of peptides contained in enzymatically digested hydrolysates of proteins is increasingly being used to characterize potentially bioactive or otherwise interesting hydrolysates. However, when preparations containing mixtures of enzymes are used, from either biological or experimental sources, it is unclear which of these enzymes have been most important in hydrolyzing the sample. We have developed a tool to rapidly evaluate the evidence for which enzymes are most likely to have cleaved the sample. EnzymePredictor, a web-based software, has been developed to (i) identify the protein sources of fragments found in the hydrolysates and map them back on it, (ii) identify enzymes that could yield such cleavages, and (iii) generate a colored visualization of the hydrolysate, the source proteins, the fragments, and the predicted enzymes. It tabulates the enzymes ranked according to their cleavage counts. The provision of odds ratio and standard error in the table permits users to evaluate how distinctively particular enzymes may be favored over other enzymes as the most likely cleavers of the samples. Finally, the method displays the cleavage not only according to peptides, but also according to proteins, permitting evaluation of whether the cleavage pattern is general across all proteins, or specific to a subset. We illustrate the application of this method using milk hydrolysates, and show how it can rapidly identify the enzymes or enzyme combinations used in generating the peptides. The approach developed here will accelerate the identification of enzymes most likely to have been used in hydrolyzing a set of mass spectrometrically identified peptides derived from proteins. This has utility not only in understanding the results of mass spectrometry experiments, but also in choosing enzymes likely to yield similar cleavage patterns. EnzymePredictor can be found at http://bioware.ucd.ie/?enzpred/Enzpred.php. PMID:23098558

  10. Using guanidine-hydrochloride for fast and efficient protein digestion and single-step affinity-purification mass spectrometry

    DEFF Research Database (Denmark)

    Poulsen, Jon Wriedt; Madsen, Christian Toft

    2013-01-01

    Protein digestion is an integral part of the "shotgun" proteomics approach and commonly requires overnight incubation prior to mass spectrometry analysis. Quadruplicate "shotgun" proteomic analysis of whole yeast lysate demonstrated that Guanidine-Hydrochloride (Gnd-HCl) protein digestion can be optimally completed within 30 min with endoprotease Lys-C. No chemical artifacts were introduced when samples were incubated in Gnd-HCl at 95 °C, making Gnd-HCl an appropriate digestion buffer for shotgun proteomics. Current methodologies for investigating protein-protein interactions (PPIs) often require several preparation steps, which prolongs any parallel operation and high-throughput interaction analysis. Gnd-HCl allow the efficient elution and subsequent fast digestion of PPIs to provide a convenient high-throughput methodology for affinity-purification mass spectrometry (AP-MS) experiments. To validate the Gnd-HCl approach, label-free PPI analysis of several GFP-tagged yeast deubiquitinating enzymes was performed. The identification of known interaction partners demonstrates the utility of the optimized Gnd-HCl protocol that is also scalable to the 96 well-plate format.

  11. Thermal wet oxidation improves anaerobic biodegradability of raw and digested biowaste

    DEFF Research Database (Denmark)

    Lissens, G.; Thomsen, Anne Belinda

    2004-01-01

    Anaerobic digestion of solid biowaste generally results in relatively low methane yields of 50-60% of the theoretical maximum. Increased methane recovery from organic waste would lead to reduced handling of digested solilds, lower methane emissions to the environment, and higher green energy profits. The objective of this research was to enhance the anaerobic biodegradability and methane yields from different biowastes (food waste, yard waste, and digested biowaste already treated in a full-scale biogas plant (DRANCO, Belgium)) by assessing thermal wet oxidation. The biodegradability of the waste was evaluated by using biochemical methane potential assays and continuous 3-L methane reactors. Wet oxidation temperature and oxygen pressure (T, 185-220 degreesC; O-2 pressure, 0-12 bar; t, 15 min) were varied for their effect on total methane yield and digestion kinetics of digested biowaste. Measured methane yields for raw yard waste, wet oxidized yard waste, raw food waste, and wet oxidized food waste were 345, 685, 536, and 571 mL of CH4/g of volatile suspended solids, respectively. Higher oxygen pressure during wet oxidation of digested biowaste considerably increased the total methane yield and digestion kinetics and permitted lignin utilization during a subsequent second digestion. The increase of the specific methane yield for the full-scale biogas plant by applying thermal wet oxidation was 35-40%, showing that there is still a considerable amount of methane that can be harvested from anaerobic digested biowaste.

  12. Environmental sustainability of an energy self-sufficient sewage treatment plant: Improvements through DEMON and co-digestion.

    Science.gov (United States)

    Schaubroeck, Thomas; De Clippeleir, Haydée; Weissenbacher, Norbert; Dewulf, Jo; Boeckx, Pascal; Vlaeminck, Siegfried E; Wett, Bernhard

    2015-05-01

    It is still not proven that treatment of sewage in a wastewater treatment plant (WWTP) is (in every case) environmentally friendly. To address this matter, we have applied a state-of-the-art life cycle assessment (LCA) to an energy self-sufficient WWTP in Strass (Austria), its supply chain and the valorization of its 'products': produced electricity out of biogas from sludge digestion and the associated stabilized digestate, applied as agricultural fertilizer. Prominent aspects of our study are: a holistic environmental impact assessment, measurement of greenhouse gas emissions (including N2O), and accounting for infrastructure, replacement of conventional fertilizers and toxicity of metals present in the stabilized digestate. Additionally, the environmental sustainability improvement by implementing one-stage partial nitritation/anammox (e.g. DEMON(®)) and co-digestion was also assessed. DEMON on the digesters reject water leads to a considerable saving of natural resources compared to nitritiation/denitritation (about 33% of the life cycle resource input), this through the lowering of sludge consumption for N-removal, and thus increasing electricity production via a higher sludge excess. However, its N2O emission could be restrained through further optimization as it represents a large share (30-66%) of the plants' damaging effect on human health, this through climate change. The co-substrate addition to the digester resulted in no significant improvement of the digestion process but induced net electricity generation. If respective amounts of conventional fertilizers are replaced, the land application of the stabilized digestate is environmentally friendly through prevention of natural resource consumption and diversity loss, but possibly not regarding human health impact due the presence of toxic heavy metals, mainly Zn, in the digestate. The outcomes show that the complete life cycle results in a prevention of resource extraction from nature and a potential mitigation of diversity loss (though for some impact categories no quantification of associated diversity loss is possible) but it also leads to a damaging effect on human health, mainly via climate change and heavy metal toxicity. Since it is for now impossible to aggregate the impact to these different aspects in a sound manner, it is not yet possible to consider in this case the studied system as environmentally friendly. Generally, the field of LCA needs further development to present a better and single outcome. PMID:25727156

  13. Comparative evaluation of standardized ileal amino acid digestibility in protein supplements for piglets.

    Science.gov (United States)

    Eklund, M; Sauer, N; Hörner, S; Rademacher, M; Mosenthin, R

    2012-12-01

    Standardized ileal digestibility (SID) of AA was determined in 6 protein ingredients for piglets. A basal diet based on corn (Zea mays) starch and casein was supplemented with fluid-bed-dried porcine intestinal mucosa hydrolysate, spray-dried porcine intestinal mucosa hydrolysate, soy (Glycine max) protein concentrate, 2 batches of soy protein, or full-fat soybeans. The SID of AA did not differ between the 4 soybean products (P > 0.05). Compared to most SID values in the 4 soybean products, SID of AA were lower in the 2 porcine intestinal mucosa hydrolysates (P ? 0.05). In conclusion, although the initial trypsin inhibitor contents in the raw soybeans have not been determined, high SID values in the 4 soybean products indicate that the different processing procedures used to manufacture these products were efficient to inactivate trypsin inhibitors. For most AA in the 2 porcine intestinal mucosa hydrolysates, drying procedure did not affect SID of AA, but SID values were generally lower compared to the 4 soybean products. PMID:23365314

  14. Assessment of digestibility improving enzymes potential to reduce greenhouse gas emissions in broiler production

    DEFF Research Database (Denmark)

    Bundgaard, Anja Marie; Dalgaard, Randi

    2012-01-01

    The objective of this study was to examine the potential of digestibility improving enzymes to reduce greenhouse gas (GHG) emissions in broiler production. The product examined was a new enzyme called Axtra XAP, developed by DuPont, Danisco Animal Nutrition. Two scenarios were compared: one where Axtra XAP was not included in the diet and one where Axtra XAP was included in the diet. Axtra XAP facilitated higher inclusion rates of cheaper (and possibly more environmentally friendly) feed ingredients that have a lower nutritional value in the diet. Axtra XAP’s environmental improvement potential was documented through a Life Cycle Assessment (LCA) by applying a consequential approach including indirect land use changes (ILUC). The findings showed that Axtra XAP could reduce GHG emissions from broiler production by 5%. A sensitivity analysis was conducted to assess the robustness of the results and it showed that the result varied substantially. The most important parameters were the inclusion or exclusion of ILUC and changes in the feed formulation.

  15. Vegetable processing wastes addition to improve swine manure anaerobic digestion: Evaluation in terms of methane yield and SEM characterization

    International Nuclear Information System (INIS)

    Highlights: ? Vegetable waste as co-substrate for swine manure anaerobic digestion. ? Two hydraulic retention times of 25 and 15 d, respectively. ? SEM characterization of anaerobic sludges to observe microbial composition. ? Vegetable waste as co-substrate increases methane yields up to three times. ? Microbial composition changes after 120 d of digestion. -- Abstract: The effect of adding vegetable waste as a co-substrate in the anaerobic digestion of swine manure was investigated. The study was carried out at laboratory scale using semi-continuous stirred tank reactors working at 37 °C. Organic loading rates (OLRs) of 0.4 and 0.6 g VS L?1 d?1 were evaluated, corresponding to hydraulic retention times (HRTs) of 25 and 15 d, respectively. The addition of vegetable wastes (50% dw/dw) resulted in an improvement of 3 and 1.4-fold in methane yields at HRTs of 25 and 15 d, respectively. Changes on microbial morphotypes were studied by Scanning Electron Microscopy (SEM). Samples analyzed were sludge used as inoculum and digestate obtained from swine manure anaerobic reactors. SEM pictures demonstrated that lignocellulosic material was not completely degraded. Additionally, microbial composition was found to change to cocci and rods morphotypes after 120 d of anaerobic digestion.

  16. Effect of Undegraded Protein and Energy Level on Intake and Digestibility of Nutrient and Blood Metabolite in Dairy Cows

    OpenAIRE

    Bp, Widyobroto; Sps, Budhi; Agus, A.

    2008-01-01

    An experiment was conducted to determine the effect of undegraded protein and energy level on intake and digestibility of nutrient and glucose and urea blood content in dairy cows. The benefit of the research was to inform about the utilization of undegraded protein and energy level to optimize nutrient utilization in dairy cattle. The experiment was conducted in 4 month in the Department of Animal Nutrition and Feed Science, Faculty of Animal Science Gadjah Mada University. The experiment us...

  17. Chemical Composition, Starch Digestibility and Antioxidant Capacity of Tortilla Made with a Blend of Quality Protein Maize and Black Bean

    OpenAIRE

    Bello-pe?rez, Luis A.; Guzma?n-maldonado, Salvador H.; Deisy Hervert-Hernández; Isabel Goñi; Grajales-garci?a, Eva M.; Perla Osorio-Díaz

    2011-01-01

    Tortilla and beans are the basic components in the diet of people in the urban and rural areas of Mexico. Quality protein maize is suggested for tortilla preparation because it presents an increase in lysine and tryptophan levels. Beans contain important amounts of dietary fiber. The objective of this study was to prepare tortilla with bean and assesses the chemical composition, starch digestibility and antioxidant capacity using a quality protein maize variety. Tortilla with bean had higher ...

  18. EVALUATION OF APPARENT DIGESTIBILITY COEFFICIENTS OF DIFFERENT DIETARY PROTEIN LEVELS WITH AND WITHOUT FISH MEAL FOR LABEO ROHITA

    Directory of Open Access Journals (Sweden)

    Y. GUL, M. SALIM AND B. RABBANI

    2007-07-01

    Full Text Available The apparent digestibility coefficients of three protein levels of reference diets (without fish meal and test diets (with fish meal were estimated for Labeo rohita. Three test diets (test diet-I, test diet-II and test diet-III and three reference diets (reference diet-I, reference diet-II and reference diet-III having 28, 30 and 32% protein levels were prepared. Chromic oxide was used as an internal marker in the experimental diets for the evaluation of digestibility of protein levels. The differences in apparent digestibility for dry matter, crude protein, crude fat and gross energy of reference and test diets were highly significant (P<0.01. The apparent digestibility coefficients (ADCs of dry matter were 32.10 ± 0.30 and 35.30 ± 0.30% for reference and test diets -III. The ADCs of crude protein and crude fat were 77.92 ± 0.10, 69.23 ± 0.06 and 75.77 ± 0.05, 70.40 ± 0.22% for reference and test diets-III, respectively. The values of ADCs of gross energy were maximum for reference diet-III (52.28 ± 0.25% and test diet-III (48.65 ± 0.78%. The ADCs of dry matter were 19.80 ± 1.40 and 18.15 ± 0.05% for reference and test diet-I. The ADCs of crude protein and crude fat were 73.93 ± 0.05, 68.43 ± 1.58 and 59.49 ± 1.13, 57.02 ± 2.42% for reference and test diet-I respectively. The ADCs values of gross energy were for reference diet-I (42.43 ± 0.36% and test diet-I (39.09 ± 1.35%. The better ADCs for dry matter, crude protein, crude fat and gross energy were at 32% protein level.

  19. Formation of a phorbol ester-binding fragment from protein kinase C by proteolytic digestion

    International Nuclear Information System (INIS)

    When washed human platelets were disrupted by sonication in the presence of ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, both the catalytic and [3H]phorbol-12,13-dibutyrate (PDBu)-binding activities of protein kinase C were recovered in the soluble fraction and were not separable from each other upon several column chromatographies. Platelet protein kinase C required diacylglycerol, Ca2+, and phospholipid for its activation and showed a molecular weight of about 87,000 as estimated by gel filtration analysis. However, when platelets were first incubated with 2 microM Ca2+-ionophore A23187 for 5 min at 37 degrees C in the medium containing 3 mM CaCl2 and then disrupted under the same conditions, the catalytic and [3H]phorbol-12,13-dibutyrate-binding activities were separately recovered in the soluble and particulate fractions, respectively; moreover, the catalytic activity recovered in the soluble fraction became independent of diacylglycerol, Ca2+, and phospholipid, and showed a molecular weight of about 50,000 as estimated by gel filtration analysis. The kinetic properties of this Mr 50,000 enzyme were similar to those of the catalytic fragment of rat brain protein kinase C described previously. In a cell-free system, digestion with trypsin of protein kinase C highly purified from rat brain caused the generation of a fragment which had no catalytic activity but showed full [3H]phorbol-12,13-dibutyr full [3H]phorbol-12,13-dibutyrate-binding activity. The molecular weight of this fragment was estimated to be about 35,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results indicate that protein kinase C consists of at least two functionally different domains, a hydrophobic phorbol ester- or diacylglycerol-binding and hydrophilic catalytic domains

  20. Effect of radiation processing on antinutrients, in-vitro protein digestibility and protein efficiency ratio bioassay of legume seeds

    International Nuclear Information System (INIS)

    The effects of irradiation (dose levels of 5, 7.5 and 10 kGy) on nutritive characteristics of peas (Pisum satinum L), cowpeas (Vigna unguiculata L.Walp), lentils (Lens culinaris Med), kidneybeans (Phaseolus vulgaris L), and chickpeas (Cicer arietinum L) were examined. Analyses included proximate composition, levels of anti-nutrients (phytic acid, tannins), available lysine (AL), in vitro protein digestibility (IVPD), and protein efficiency ratio (PER) in the growing rat. The results showed that moisture, crude protein, crude fat, crude fiber, and ash were unchanged by the irradiation. Radiation processing significantly (p<0.05) reduced the levels of phytic acid (PA), tannins (TN), and AL. IVPD and PER were significantly enhanced in a dose-dependent manner, relative to unirradiated control samples, for all legumes. The data sets for each legume exhibited high correlation coefficients between radiation dose and PA, TN, AL, IVPD, and PER. These results demonstrate the benefits of irradiation on the nutritional properties of these legumes

  1. Characterization, in vitro Trypsin Digestibility and Antioxidant Activity of Fermented Soybean Protein Meal with Lactobacillus plantarum Lp6

    Directory of Open Access Journals (Sweden)

    O.S. Gbadamosi

    2009-01-01

    Full Text Available In this study, soybean protein meal was subjected to solid state fermentation with Lactobacillus plantarum Lp6 either in the presence or absence of a protease. The extracts were investigated for changes in mineral composition, amino acid composition, in vitro trypsin digestibility, DPPH radical scavenging activities and electrophoretic pattern. The amino acid and mineral element compositions showed significant (pin vitro trypsin digestibility and showed a single polypeptide with estimated molecular weight of 14.4 kDa in the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE assay.

  2. Ruminal, Intestinal, and Total Digestibilities of Nutrients in Cows Fed Diets High in Fat and Undegradable Protein

    DEFF Research Database (Denmark)

    Palmquist, D.L.; Weisbjerg, Martin Riis

    1993-01-01

    To study relationships of high undegradable intake protein and dietary fat on intestinal AA supply, the ruminal, intestinal, and total digestibilities of diets with or without added fat (5% of DM) and animal protein (blood meal: hydrolyzed feather meal, 1:1; 8% of DM) were examined with four cows in a 2×2 factorial design in a 4×4 Latin square experiment. Ruminal degradabilities were 14.9 and 18.6%, and intestinal digestibilities were 98.9 and 68.3%, respectively, for CP in blood meal and feather meal. Treatment effects on ruminal digestibilities were small. Protein supplementation increased total N intake by 29%, duodenal AA N flow by 39%, and AA N absorbed by 37%; absorption of Leu and Lys increased 60 and 33%, and absorption of ne and Met increased 11 and 7%, respectively. Measured duodenal AA N flow (Cr2O3 marker) was 33% higher in cows cannulated adjacent to the pylorus compared with cows cannulated 100-cm distal to the pylorus, but only when cows were fed protein-supplemented diets; the estimates from those diets caused calculated microbial protein efficiency to exceed theoretical values. We postulated that blood meal and feather meal segregated near the pylorus, yielding high estimates of duodenal AA N flow. Removal of data for protein-supplemented diets obtained from cows cannulated at the pylorus yielded estimates of microbial protein synthetic efficiency consistent with literature values. Microbial synthesis of AA N was related linearly to ruminal digestion of carbohydrate. Location of intestinal cannulas may influence accuracy of nutrient flow estimates.

  3. Ruminal, intestinal, and total digestibilities of nutrients in cows fed diets high in fat and undegradable protein.

    Science.gov (United States)

    Palmquist, D L; Weisbjerg, M R; Hvelplund, T

    1993-05-01

    To study relationships of high undegradable intake protein and dietary fat on intestinal AA supply, the ruminal, intestinal, and total digestibilities of diets with or without added fat (5% of DM) and animal protein (blood meal: hydrolyzed feather meal, 1:1; 8% of DM) were examined with four cows in a 2 x 2 factorial design in a 4 x 4 Latin square experiment. Ruminal degradabilities were 14.9 and 18.6%, and intestinal digestibilities were 98.9 and 68.3%, respectively, for CP in blood meal and feather meal. Treatment effects on ruminal digestibilities were small. Protein supplementation increased total N intake by 29%, duodenal AA N flow by 39%, and AA N absorbed by 37%; absorption of Leu and Lys increased 60 and 33%, and absorption of Ile and Met increased 11 and 7%, respectively. Measured duodenal AA N flow (Cr2O3 marker) was 33% higher in cows cannulated adjacent to the pylorus compared with cows cannulated 100-cm distal to the pylorus, but only when cows were fed protein-supplemented diets; the estimates from those diets caused calculated microbial protein efficiency to exceed theoretical values. We postulated that blood meal and feather meal segregated near the pylorus, yielding high estimates of duodenal AA N flow. Removal of data for protein-supplemented diets obtained from cows cannulated at the pylorus yielded estimates of microbial protein synthetic efficiency consistent with literature values. Microbial synthesis of AA N was related linearly to ruminal digestion of carbohydrate. Location of intestinal cannulas may influence accuracy of nutrient flow estimates. PMID:8505426

  4. Staphylococcus aureus Typing by Digestion of Protein A Coding Gene Using Bsp143I

    Directory of Open Access Journals (Sweden)

    Fatemeh Shakeri

    2014-06-01

    Full Text Available Background: Protein A is the virulence factors of Staphylococcus aureus rolling in its pathogenesis, and its gene is used for typing. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP with different enzymes has been used for this action. Objectives: In this study, we used Bsp143I enzyme for digestion of the gene, coding protein A (spa gene in S. aureus. The bacteria were isolated from patients and healthy carriers in Gorgan, north of Iran. Patients and Methods: DNAs of 128 S. aureus subjects (53 from healthy carriers and 75 from patients were extracted and amplified using specific primers of the spa gene. The product was digested by Bsp143I enzyme and its pattern was assessed by gel electrophoresis. Results: There were seven spa types among the tested S. aureus samples, among which six types differed in the repeated X region of the spa gene, but the seventh type had a deletion on one of BSP143I restriction sites. The frequency of spa types among isolated S. aureus samples as well as healthy carriers was six and five, respectively. S. aureus isolated from wounds showed the most diverse spa types (five among clinical samples. Types 1, 2 and 4 were observed in all clinical samples, while only one case of type 3 was identified among patients, whereas this type constituted over 32% of the isolates among carriers. We found seven and four spa types among methicillin-resistant S. aureus (MRSA and methicillin-sensitive S. aureus (MSSA isolates, respectively. Conclusions: Our results showed that typing the spa gene using PCR-RFLP using Bsp143I was an acceptable method for typing S. aureus. Furthermore, this survey showed that the types in healthy carriers and MSSA were more variable than patient and MRSA isolates, respectively. We used the Bsp143I enzyme, which was not used in any previous studies on the spa gene. The results of this study suggested that we can use PCR-RFLP of spa gene by Bsp143I for molecular typing and sequencing of S. aureus, instead of relatively expensive methods. This method is relatively rapid and inexpensive, and can be accomplished in centers with conventional molecular facilities. Keywords: Staphylococcal Protein A; Staphylococcus aureus; Methicillin-Resistant Staphylococcus aureus

  5. Effects of dietary energy density and digestible protein:energy ratio on de novo lipid synthesis from dietary protein in gilthead sea bream (Sparus aurata) quantified with stable isotopes

    DEFF Research Database (Denmark)

    Ekmann, Kim SchØn; Dalsgaard, Anne Johanne Tang

    2013-01-01

    The effects of varying dietary digestible protein (DP) and digestible energy (DE) content on performance, nutrient retention efficiency and the de novo lipogenesis of DP origin were examined in triplicate groups of gilthead sea bream (Sparus aurata), fed nine extruded experimental diets. In order to trace the metabolic fate of dietary protein, 1·8% fishmeal was replaced with isotope-labelled whole protein (.98% 13C). The experiment was divided into a growth period lasting 89 d, growing fish from approximately 140 to 350 g, followed by a 3 d period feeding isotope-enriched diets. Isotope ratio MS was applied to quantify the 13C enrichment of whole-body lipid from dietary DP. Between 18·6 and 22·4% of the carbon derived from protein was recovered in the lipid fraction of the fish, and between 21·6 and 30·3% of the total lipid deposited could be attributed to dietary protein. DP retention was significantly improved by reductions in dietary DP:DE ratio, while the opposite was true for apparent digestible lipid retention. Both overall DE retention and whole-body proximate composition of whole fish were largely unaffected by dietary treatments, while feed conversion ratios were significantly improved with increasing dietary energy density. The present study suggests that gilthead sea bream efficiently utilises dietary nutrients over a wide range of DP:DE ratios and energy densities. In addition, they appear to endeavour a certain body energy status rather than maximising growth, which in the present trial was apparent from inherently high de novo lipogenesis originating from DP

  6. Anaerobic digestate from biogas production as a resource for improving soil fertility: effects on crop yield and soil properties

    Science.gov (United States)

    Pastorelli, Roberta; Lagomarsino, Alessandra; Vignozzi, Nadia; Valboa, Giuseppe; Papini, Rossella; Fabiani, Arturo; Simoncini, Stefania; Mocali, Stefano; Piccolo, Raimondo

    2013-04-01

    Soil fertility is fundamental in determining crops productivity in all farming systems. Production of biogas through anaerobic digestion of energy crops generates residues that can represent a valuable resource to sustain and improve soil fertility and to increase soil organic matter content. Residues from anaerobic digestion contain organic fractions and available nutrients, that can thus be returned to the cultivation soil as fertilizer and soil conditioner. However, some unknown aspects of digested residues utilization remain to explore: i) the nutrient supply and the real potential for mineral fertilization substitution, ii) the impact on the structure and functioning of soil microbial communities, iii) the direct and indirect effects on soil structure, organic matter and C mineralization. The aim of the present research was to gain a better understanding of these aspects, evaluating the effects of anaerobic digestate application on soil properties and maize yield. With the main focus of comparing mineral fertilization (250 Kg N ha-1) with digested residues addition (at the dose of 25 % and 50 % of mineral fertilizer), a triplicate sets of plots were designed in a field experiment on a silty-clay loam soil in the southern Po Valley (Italy). The amount of applied residues was calculated according to its N content in order to fertilizer each plots with the same amount of total nitrogen. Residues from digestion showed a N content of 0.4 % (60 % as N-NH4) and a C/N ratio of 3. Changes in soil quality after residues application were studied with a holistic approach, involving microbiological, physical and chemical aspects of soil fertility. In particular, we determined: the abundance and diversity of bacterial and fungal soil communities; the soil organic matter content, its distribution within soil aggregates and the C mineralization potential; cation exchange capacity; the main macro and micro nutrients; bulk density; aggregate stability. No significant differences among treatments were registered in the above ground maize biomass. Molecular analysis conducted on microbial soil communities suggested that the application of digested residues to soil contributes to substantial modifications of both bacterial and fungal community structure. Soil organic C and total N increased in soils treated with digested residues addition, with no significant differences between the two doses of digestate. Cation exchange capacity did not show significant differences among treatments, remaining stable during the maize vegetative cycle. Differently, some variations occurred in the exchangeable cation pool. In particular, K content increased under digestate treatments, while Na and Mg contents increased with time irrespective of the fertilization treatment. No significant variations were observed in soil microelement levels, except for an increase in Zn content at the highest digestate dose. Moreover, digested residue addition had a positive impact on aggregates stability. From the first results, the absence of negative effects in plant productivity and soil fertility after residues application, at both doses, is a promising indication for the potential use of anaerobic digestate as substitute of mineral fertilizers.

  7. Effects of physical form and urea treatment of rice straw on rumen fermentation, microbial protein synthesis and nutrient digestibility in dairy steers.

    Science.gov (United States)

    Gunun, P; Wanapat, M; Anantasook, N

    2013-12-01

    This study was designed to determine the effect of physical form and urea treatment of rice straw on rumen fermentation, microbial protein synthesis and nutrient digestibility. Four rumen-fistulated dairy steers were randomly assigned according to a 2 (2 factorial arrangement in a 4 (4 Latin square design to receive four dietary treatments. Factor A was roughage source: untreated rice straw (RS) and urea-treated (3%) rice straw (UTRS), and factor B was type of physical form of rice straw: long form rice straw (LFR) and chopped (4 cm) rice straw (CHR). The steers were offered the concentrate at 0.5% body weight (BW) /d and rice straw was fed ad libitum. DM intake and nutrient digestibility were increased (pnitrogen (NH3-N) and blood urea nitrogen (BUN) were increased (p0.05), except propionic acid (C3) was increased (pNitrogen (N) balance was affected by urea treatment (price straw improved feed intake, digestibility, rumen fermentation and efficiency of microbial N synthesis in crossbred dairy steers. PMID:25049759

  8. Improving the prediction of yeast protein function using weighted protein-protein interactions

    OpenAIRE

    Saloma Nahed H; Ahmed Khaled S; Kadah Yasser M

    2011-01-01

    Abstract Background Bioinformatics can be used to predict protein function, leading to an understanding of cellular activities, and equally-weighted protein-protein interactions (PPI) are normally used to predict such protein functions. The present study provides a weighting strategy for PPI to improve the prediction of protein functions. The weights are dependent on the local and global network topologies and the number of experimental verification methods. The proposed methods were applied ...

  9. Effects of electron beam irradiation on chemical composition, antinutritional factors, ruminal degradation and in vitro protein digestibility of canola meal

    International Nuclear Information System (INIS)

    The aim of the present study was to determine the impact of electron beam (EB) irradiation at doses of 15, 30 and 45 kGy on the nutritional value of canola meal. The phytic acid and total glucosinolate content of EB-irradiated canola meal decreased as irradiation doses increased (P<0.01). From in situ results, irradiation of canola meal at doses of 45 kGy decreased (P<0.05) the effective degradibility of crude protein (CP) by 14%, compared with an untreated sample. In vitro CP digestibility of EB-irradiated canola meal at doses of 15 and 30 kGy was improved (P<0.05). Electrophoresis results showed that napin and cruciferin sub-units of 30 and 45 kGy EB-irradiated canola meal were more resistant to degradation, compared with an untreated sample. Electron beam irradiation was effective in protecting CP from ruminal degradation and reducing antinutritional factors of irradiated canola meal.

  10. Effect of Undegraded Protein and Energy Level on Intake and Digestibility of Nutrient and Blood Metabolite in Dairy Cows

    Directory of Open Access Journals (Sweden)

    BP Widyobroto

    2008-05-01

    Full Text Available An experiment was conducted to determine the effect of undegraded protein and energy level on intake and digestibility of nutrient and glucose and urea blood content in dairy cows. The benefit of the research was to inform about the utilization of undegraded protein and energy level to optimize nutrient utilization in dairy cattle. The experiment was conducted in 4 month in the Department of Animal Nutrition and Feed Science, Faculty of Animal Science Gadjah Mada University. The experiment used 4 female rumen fistulated dry cows (Friesian Holstein Crossbreed of 3.0-3.5 years old and 350–400 kg body weight. The treatments of this experiment were T1: 20% of undegraded protein (UDP and 70% energy from requirement; T2: 20% undegraded protein and 120% energy from requirement; T3: 30% undegraded protein and 70% energy from requirement; and T4: 30% undegraded protein and 120% energy from requirement with Latin square design. Variables observed were intake and digestibility of dry matter (DM, organic matter (OM, crude protein (CP, neutral detergent fiber (NDF, Acid detergent fiber (ADF, and glucose and urea blood content. Collected data were analyzed by analysis of variances, and further differences were tested by orthogonal contrast. Results of the research showed that the level of UDP in the rations did not affect DM, OM, CP intake, rumen degradable protein (RDP, NDF and ADF; however level of energy had significant affect on DM, OM, CP, RDP and ADF intake. The level of UDP resulted in non significant difference in DM; but significantly different on CP digestibility. Digestibility of DM and OM were significantly influenced by the level of energy, but it did not influence NDF and ADF digestibility. Cattle treated with the high energy rations had higher blood glucose concentration than cattle treated with the low energy rations. Blood urea in cattle fed low UDP rations was higher than cattle fed high UDP rations. (Animal Production 10(2: 96-101 (2008 Key Words: Undegraded protein, nitrogen, energy, blood metabolite

  11. Effects of synchronizing starch and protein degradation in rumen on fermentation, nutrient utilization and total tract digestibility in sheep

    Directory of Open Access Journals (Sweden)

    Birgul Bozan

    2010-01-01

    Full Text Available The objective of this experiment was to investigate the effects of synchronizing the dietary starch and crude protein (CP degradation in the rumen on nutrient utilization, fermentation, and total tract digestibility in sheep. The four diets were formulated with different rates of starch and CP release in the rumen but with similar metabolic energy, starch, and CP. The diets were slowly degradable starch, slowly degradable protein; slowly degradable starch, rapidly degradable protein; rapidly degradable starch, rapidly degradable protein; and rapidly degradable starch, slowly degradable protein. The diets were fed to four cannulated sheep ad libi- tumin two equal portions, using a 4x4 Latin square design. Dry matter intake (DM was not influenced by either the rate of starch or protein degradation. There was no significant effect of dietary treatment on the digestibility of DM, organic matter, starch, CP, neutral detergent fiber or acid detergent fiber in the rumen and total tract. Ruminal pH was greater for sheep fed slowly degrad- able starch diets than rapidly degradable starch (P< 0.05. Ruminal total volatile fatty acid concentrations were not affected by treatments but the molar proportions of propionic acid were greater for sheep fed rapid- ly degradable starch diets than slowly degradable starch diets (P< 0.05. The ratios of acetic acid (A to pro- pionic acid (P were higher for sheep fed slowly degradable starch diets than rapidly degradable starch diets (P< 0.05. Ruminal ammonia-N concentrations were not affected from the degradability characteristics of protein. Rumen pH and A:P were higher in diets containing slowly degradable starch than in diets rapidly degradable starch. Propionic acid was higher in diets containing rapidly degradable starch than in diets con- taining slowly degradable starch. Rumen fermentation and utilization of nutrients in the rumen affected starch degradability more than protein degradability. Synchronizing starch and protein degradation in rumen had no effect on the intake, digestibility of nutrients in sheep.

  12. Evaluation of protein fractionation and ruminal and intestinal digestibility of corn milling co-products.

    Science.gov (United States)

    Kelzer, J M; Kononoff, P J; Tedeschi, L O; Jenkins, T C; Karges, K; Gibson, M L

    2010-06-01

    Novel corn milling co-products developed from technological advancements in ethanol production vary widely in chemical composition and nutrient availability. The objectives of this study were to characterize feed protein fractions and evaluate differences in rumen-undegradable protein (RUP) and its digestible fraction (dRUP), amino acid concentration, and in vitro gas production of 7 corn milling co-products. The crude protein (CP; % of dry matter) of co-products was 12.7 for germ, 26.9 for dried distillers grains plus solubles that had no heat exposure before fermentation (DDGS1), 45.4 for high-protein dried distillers grains (HPDDG), 12.7 for bran, 30.2 for wet distillers grains plus solubles (WDGS), 23.1 for wet corn gluten feed (WCGF), and 26.0 for dried distillers grains plus solubles that had heat exposure before fermentation (DDGS2). Two ruminally and duodenally fistulated Holstein steers weighing 663+/-24 kg were used to determine RUP and dRUP with the in situ and mobile bag techniques. Samples of each feed were ruminally incubated for 16 h, and mobile bags were exposed to simulated abomasal digestion before insertion into the duodenum and subsequent collection in the feces. Protein fractions A, B(1), B(2), B(3), and C were characterized as follows (% CP): germ=30.0, 15.0, 38.1, 13.5, 3.4; DDGS1=17.0, 7.0, 67.0, 4.8, 4.2; HPDDG=7.4, 0.6, 82.4, 8.8, 0.8; bran=33.5, 4.0, 54.3, 6.0, 2.2; WDGS=18.6, 2.4, 53.1, 11.0, 14.9; WCGF=36.6, 15.9, 33.2, 10.1, 4.1; and DDGS2=17.9, 2.1, 41.1, 11.1, 27.9. The proportions of RUP and dRUP were different and are reported as follows (% CP): DDGS2=56.3, 91.9; HPDDG=55.2, 97.7; WDGS=44.7, 93.1; DDGS1=33.2, 92.1; bran=20.7, 65.8; germ=16.5, 66.8; and WCGF=11.5, 51.1. The concentrations of Lys and Met in the RUP were different and are listed as follows (% CP): germ=2.9, 2.0; DDGS1=1.9, 2.0; HPDDG=2.0, 3.2; bran=3.2, 1.5; WDGS=1.9, 2.3; WCGF=3.5, 1.6; and DDGS2=1.9, 2.4. In vitro gas production (mL/48h) was highest for germ (52.1) followed by bran (50.1), WDGS (40.7), DDGS2 (40.1), WCGF (39.0), DDGS1 (38.6), and HPDDG (37.5). Comparison of co-products defined differences in chemical composition, protein fractionation, ruminal availability, and microbial fermentation. PMID:20494190

  13. Effects of Heat-oxidized Soy Protein Isolate on Growth Performance and Digestive Function of Broiler Chickens at Early Age.

    Science.gov (United States)

    Chen, X; Chen, Y P; Wu, D W; Wen, C; Zhou, Y M

    2015-04-01

    This study was conducted to investigate effects of heat-oxidized soy protein isolate (HSPI) on growth performance, serum biochemical indices, apparent nutrient digestibility and digestive function of broiler chickens. A total of 320 1-day-old Arbor Acres chicks were randomly divided into 4 groups with 8 replicates of 10 birds, receiving diets containing soy protein isolate (SPI, control group) or the same amount of SPI heated in an oven at 100°C for 1, 4, or 8 h, for 21 days, respectively. The results indicated that compared with the control group, body weight gain and feed intake of birds fed diet containing SPI heated for 8 h were significantly lower (pheated for 4 or 8 h at d 21 (pheated for 8 h (pheated for 4 or 8 h had a decreased lipase activity in anterior intestinal content at d 14 and 21 (pheated for 1 h at d 21 (pheated for 8 h at d 21 (pheated for 8 h (pheated for 4 h also exhibited lower CP apparent digestibility from d 15 to 17 (p<0.05). It was concluded that HSPI inclusion can exert a negative influence on the growth performance of broilers, which was likely to result from the simultaneously compromised digestive function. PMID:25656201

  14. An aptamer-based trypsin reactor for on-line protein digestion with electrospray ionization tandem mass spectrometry.

    Science.gov (United States)

    Xiao, Peng; Lv, Xuefei; Wang, Shanshan; Iqbal, Javed; Qing, Hong; Li, Qin; Deng, Yulin

    2013-10-15

    An immobilized trypsin reactor that was based on aptamers has been developed and applied for the first time for proteomic digestion. Briefly, 25 single-stranded DNA aptamers that were specific for trypsin were obtained after SELEX (systematic evolution of ligands by exponential enrichment) selection. TApt.23 (no. 23 trypsin aptamer), which had the lowest dissociation constant (Kd) value (0.0123 ?M), was amino-modified and subsequently grafted to an amino-modified silica surface with glutaraldehyde. The results indicated that 14.65±0.35 ?g of trypsin could be immobilized on 10mg of TApt.23-silica when an optimized borate buffer was used. Subsequently, a trypsin reactor was fabricated by using a PEEKsil micro column. Compared with in-solution digestion, the aptamer-based trypsin reactor exhibited similar results for protein identification but used a much shorter digestion time (?30 min). An on-line analysis platform, which included a trypsin reactor coupled to a high-performance liquid chromatography tandem mass spectrometry device, was built through a 6-port valve and achieved efficient protein digestion compared with in-solution and off-line methods. Compared with irreversible covalent enzyme immobilization, the aptamer-based carrier enables more rapid and convenient immobilized trypsin elution as well as re-immobilization of the enzyme. This superior reactor demonstrated that an aptamer could become a more widely used method for enzyme immobilization and other applications. PMID:23831476

  15. Low temperature anaerobic digestion of mixtures of llama, cow and sheep manure for improved methane production

    Energy Technology Data Exchange (ETDEWEB)

    Alvarez, Rene [IIDEPROQ, UMSA, Plaza del Obelisco 1175, La Paz (Bolivia)]|[Department of Chemical Engineering, Lund University, P.O. Box 124, 221 00 Lund (Sweden); Liden, Gunnar [Department of Chemical Engineering, Lund University, P.O. Box 124, 221 00 Lund (Sweden)

    2009-03-15

    Biogas production in anaerobic digestion in farm-scale units is typically performed under mesophilic conditions when used for producing domestic fuel and stabilizing animal waste for the use of digested manure as a fertilizer. Previous studies on the digestion of llama and cow manure have shown the feasibility of producing biogas under altiplano conditions (low pressure and low temperature) and of llama manure as a promising feedstock. The present study concerns the utilization of various mixtures of feedstocks from the Bolivian altiplano under low temperature conditions (18-25 C). Laboratory scale experiments were performed on the digestion of mixtures of llama, sheep and cow manure in a semi-continuous process using ten 2-L stainless steel digesters to determine the effects of organic loading rate (OLR) and the feed composition. The semi-continuous operation of mixture of llama-cow-sheep manure proved to be a reliable system, which could be operated with good stability. The results suggest that in a system digesting a mixture of llama-cow-sheep manure at low temperature (18-25 C) the maximum OLR value is between 4 and 6 kg VS m{sup 3} d{sup -1}. The methane yields obtained in the mixture experiments were in the range 0.07-0.14 m{sup 3} kg{sup -1} VS added, with a methane concentration in the gas of between 47 and 55%. (author)

  16. Effect of Protein Level, Main Protein and non Forage Fiber Source on Digestibility, N Balance and Energy Value of Sheep Rations

    OpenAIRE

    Ch. Milis; Liamadis, D.

    2007-01-01

    in vivo digestion trials were conducted, by using a latin square 4x4 experimental design with castrated rams, to evaluate the effects of diet?s CP level, N degradability and Non-Forage Fiber Source (NFFS) on nutrient digestibility, N balance and energy value of sheep rations. In each trial, rams were fed at maintenance level four isocaloric-isonitrogenous and isofibrous rations, differing in main protein and/or NFFS source. At the first trial mean CP/ME ratio of the diets was 17 g MJ-1 ME and...

  17. Using context to improve protein domain identification

    Directory of Open Access Journals (Sweden)

    Llinás Manuel

    2011-03-01

    Full Text Available Abstract Background Identifying domains in protein sequences is an important step in protein structural and functional annotation. Existing domain recognition methods typically evaluate each domain prediction independently of the rest. However, the majority of proteins are multidomain, and pairwise domain co-occurrences are highly specific and non-transitive. Results Here, we demonstrate how to exploit domain co-occurrence to boost weak domain predictions that appear in previously observed combinations, while penalizing higher confidence domains if such combinations have never been observed. Our framework, Domain Prediction Using Context (dPUC, incorporates pairwise "context" scores between domains, along with traditional domain scores and thresholds, and improves domain prediction across a variety of organisms from bacteria to protozoa and metazoa. Among the genomes we tested, dPUC is most successful at improving predictions for the poorly-annotated malaria parasite Plasmodium falciparum, for which over 38% of the genome is currently unannotated. Our approach enables high-confidence annotations in this organism and the identification of orthologs to many core machinery proteins conserved in all eukaryotes, including those involved in ribosomal assembly and other RNA processing events, which surprisingly had not been previously known. Conclusions Overall, our results demonstrate that this new context-based approach will provide significant improvements in domain and function prediction, especially for poorly understood genomes for which the need for additional annotations is greatest. Source code for the algorithm is available under a GPL open source license at http://compbio.cs.princeton.edu/dpuc/. Pre-computed results for our test organisms and a web server are also available at that location.

  18. Maize silage supplementation to lower quality grass silage improves the intake, apparent digestibility and N retention in wether sheep

    Directory of Open Access Journals (Sweden)

    Ivana Mati?

    2009-12-01

    Full Text Available The objective of this experiment was to study the effects of interactions between lower quality grass silage (GS dominated by orchardgrass (Dactylis glomerata L. and maize silage (MS (Zea mays L. on ad libitum intake, digestibility and N retention in wether sheep. The study consisted of four feeding treatments involving GS and MS alone, and GS and MS mixtures in ratios of 67:33 or 33:67 (DM basis fed twice daily. The GS was harvested at the beginning of flowering of orchardgrass (about 35 % of plants were in flower while the MS used was of lower DM and starch concentration (264 g kg-1 fresh weight and 211 g kg-1 DM respectively. Mean DM content of GS was 408 g kg-1 fresh weight. MS was lower in crude protein (CP (62 g kg-1 DM than the GS (98 g kg-1 DM (P<0.001. GS contained larger quantities of neutral detergent fibre (NDF and acid detergent fiber (ADF (P<0.001 than MS. Inclusion of MS in the diet (33 % vs. 67 % had positive linear effects on fresh matter (FM voluntary intake (P<0.05, digestibility of DM, organic matter (OM, digestible OM in the DM (D-value (P<0.05, starch digestibility (P<0.05, N intake (P<0.01, N output in faeces (P<0.05 and absorbed N (P<0.05. A positive associative effect of GS and MS was observed for all intake parameters measured (FM, DM, OM, NDF (quadratic, P<0.05 to P<0.01, digestibility of DM, ADF (quadratic, P<0.05, CP (quadratic, P<0.01, N intake and absorbed N (quadratic, P<0.01. It was concluded that positive interactions of GS and MS were recorded for a limited number of parameters because of the lower quality MS than required for the full benefit of two forages fed together.

  19. Efecto ahorrativo de la proteína usando niveles altos de energía y obtención de la relación optima energía digestible/proteína digestible en dietas para el crecimiento de Oreochromis niloticus (L) / Protein-sparing effect with high energy levels and obtaining the optimum digestible energy/digestible protein ratio in growth diets to Oreochromis niloticus (L.)

    Scientific Electronic Library Online (English)

    Felix Walter, Gutierrez; Máximo, Quispe; Luz, Valenzuela.

    2013-12-01

    Full Text Available Se evaluó el efecto ahorrativo de la proteína usando dietas altas en energía. Se utilizó un diseño factorial para medir la interacción de dos niveles de proteína (30% y 35%) y dos niveles de energía digestible (3.3 y 3.7 kcal/g de alimento) sobre la ganancia (GP), conversión alimenticia (CA), proteí [...] na retenida (PR), energía retenida (ER) y la relación de eficiencia proteica (REP) en la tilapia del Nilo todos machos (Oreochromis niloticus). En la composición de las dietas se emplearon harina de anchoveta y harina de torta de soya como fuentes de proteína y maíz amarillo duro y subproducto de trigo como fuentes de energía. El aceite de pescado fue añadido para ajustar los niveles de energía requeridos en las dietas experimentales. Se encontraron interacciones altamente significativas (P Abstract in english The sparing effect of the dietary protein using diets with high energy levels was evaluated. A factorial design was used to examine the interaction of two levels of protein (30 and 35%) and two levels of digestible energy (3.3 and 3.7 kcal/g) on the weight gain (WG), food conversion FC), retained pr [...] otein (PR), retained energy (ER) and protein efficiency ratio (PER) in “sex reversal Nile tilapia” ( Oreochromis niloticus). Diets contained anchovy and soybean meal as protein sources and yellow corn and wheat bran as energy sources. The fish oil was used to adjust the energy levels. There were highly significant interactions (P

  20. Optimization of Immobilized Gallium (III) Ion Affinity Chromatography for Selective Binding and Recovery of Phosphopeptides from Protein Digests

    OpenAIRE

    Aryal, Uma K.; Olson, Douglas J. H.; Ross, Andrew R. S.

    2008-01-01

    Although widely used in proteomics research for the selective enrichment of phosphopeptides from protein digests, immobilized metal-ion affinity chromatography (IMAC) often suffers from low specificity and differential recovery of peptides carrying different numbers of phosphate groups. By systematically evaluating and optimizing different loading, washing, and elution conditions, we have developed an efficient and highly selective procedure for the enrichment of phosphopeptides using a comme...

  1. Kinetic properties of mouse pancreatic lipase-related protein-2 suggest the mouse may not model human fat digestion

    OpenAIRE

    Xiao, Xunjun; Ross, Leah E.; Miller, Rita A.; Lowe, Mark E.

    2011-01-01

    Genetically engineered mice have been employed to understand the role of lipases in dietary fat digestion with the expectation that the results can be extrapolated to humans. However, little is known about the properties of mouse pancreatic triglyceride lipase (mPTL) and pancreatic lipase-related protein-2 (mPLRP2). In this study, both lipases were expressed in Pichia Pastoris GS115, purified to near homogeneity, and their properties were characterized. Mouse PTL displayed the kinetics typica...

  2. Effect of plants containing secondary compounds with palm oil on feed intake, digestibility, microbial protein synthesis and microbial population in dairy cows.

    Science.gov (United States)

    Anantasook, N; Wanapat, M; Cherdthong, A; Gunun, P

    2013-06-01

    The objective of this study was to determine the effect of rain tree pod meal with palm oil supplementation on feed intake, digestibility, microbial protein synthesis and microbial populations in dairy cows. Four, multiparous early-lactation Holstein-Friesian crossbred (75%) lactating dairy cows with an initial body weight (BW) of 405±40 kg and 36±8 DIM were randomly assigned to receive dietary treatments according to a 4×4 Latin square design. The four dietary treatments were un-supplementation (control), supplementation with rain tree pod meal (RPM) at 60 g/kg, supplementation with palm oil (PO) at 20 g/kg, and supplementation with RPM at 60 g/kg and PO at 20 g/kg (RPO), of total dry matter intake. The cows were offered concentrates, at a ratio of concentrate to milk production of 1:2, and chopped 30 g/kg of urea treated rice straw was fed ad libitum. The RPM contained condensed tannins and crude saponins at 88 and 141 g/kg of DM, respectively. It was found that supplementation with RPM and/or PO to dairy cows diets did not show negative effects on feed intake and ruminal pH and BUN at any times of sampling (p>0.05). However, RPM supplementation resulted in lower crude protein digestibility, NH3-N concentration and number of proteolytic bacteria. It resulted in greater allantoin absorption and microbial crude protein (p<0.05). In addition, dairy cows showed a higher efficiency of microbial N supply (EMNS) in both RPM and RPO treatments. Moreover, NDF digestibility and cellulolytic bacteria numbers were highest in RPO supplementation (p<0.05) while, supplementation with RPM and/or PO decreased the protozoa population in dairy cows. Based on this study, supplementation with RPM and/or PO in diets could improve fiber digestibility, microbial protein synthesis in terms of quantity and efficiency and microbial populations in dairy cows. PMID:25049855

  3. Digestibilidade intestinal verdadeira da proteína de alimentos para ruminantes / True small intestinal protein digestibility of ruminant feeds

    Scientific Electronic Library Online (English)

    Antonio Ferriani, Branco; Sabrina Marcantonio, Coneglian; Fábio José, Maia; Kátia Cylene, Guimarães.

    1788-17-01

    Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese A digestibilidade intestinal verdadeira de diferentes classes de alimentos usados em dietas para ruminantes foi avaliada por meio das técnicas in situ e in vitro. Foram utilizados dois bovinos machos castrados (450 kg PV) com cânulas implantadas no rúmen para incubação in situ de concentrados protéi [...] cos de origens animal e vegetal e energéticos, resíduos da agroindústria e alimentos volumosos. Avaliou-se a digestibilidade intestinal verdadeira dos alimentos submetidos à digestão apenas com pepsina ou com pepsina + pancreatina, precedida ou não da incubação ruminal. A incubação ruminal diminuiu a digestibilidade intestinal verdadeira da proteína de 24 dos 30 alimentos testados, com exceção da farinha de penas, da aveia preta, do grão de milho triturado a 2,5 mm e dos fenos de aveia e tifton, para os quais ocorreu aumento, e do farelo de girassol, para o qual não houve efeito da incubação ruminal. A digestibilidade intestinal da proteína não-degradada no rúmen (PNDR), na maioria dos alimentos utilizados em dietas para ruminantes, é menor que a da proteína original do alimento. Entre os alimentos avaliados, 29 apresentaram maior digestibilidade intestinal verdadeira quando incubados com pepsina + pancreatina, evidenciando a importância da etapa de digestão abomasal sobre as proteínas dos alimentos (com exceção à aveia preta). A digestibilidade intestinal dos alimentos é variável e, portanto, deve ser considerada na formulação de dietas para atendimento das exigências de proteína metabolizável. Abstract in english The true protein digestibility in the small intestine of different ruminant feeds were measured using in situ and in vitro techniques. Two steers with average body weight of 450 kg and fitted with ruminal cannulas were used for in situ incubation of different feeds. The following feedstuffs were eva [...] luated: animal and vegetable protein concentrate, energy concentrate, fiber by-products, and forage. Protein truly digested in the small intestine was estimated by pepsin or pepsin/pancreatin incubation with or without previous ruminal incubation. Out of 30 evaluated feeds, ruminal incubation decreased the true protein digestibility in the small intestine of 24 feeds, increased that of feather meal, black oat, 2.5-mm particle size corn grain, and oat and Tifton hays and had no effect on sunflower meal. These results showed that the small intestinal digestibility of rumen-undegradable protein from most analyzed feeds was lower than that of the original feed protein. With the exception of black oat, all remaining feeds (29) had greater true small intestinal protein digestibility after incubation with pepsin plus pancreatin showing the contribution of abomasal digestion on feed protein. Because true small intestinal protein digestibility differed among feeds, is important to take it into account on diet formulation in order to accurately determine the requirements of metabolizable protein of ruminants.

  4. [Ruminal digestion and intestinal absorption of lupine proteins extruded in the lactating cow].

    Science.gov (United States)

    Benchaar, C; Bayourthe, C; Moncoulon, R; Vernay, M

    1991-01-01

    Four lactating cows fitted with permanent ruminal, duodenal and ileal cannulae were used to study the effect of extrusion of whole lupin seeds at 195 degrees C (Lupinus albus cv Lublanc) on organic matter (OM) and nitrogen (N) degradation in the rumen and their flow to and absorption from the small intestine. Raw whole lupin seeds (RWLS) and extruded whole lupin seeds (EWLS) were fed in diets containing 15.5% crude protein and composed of 22.6% whole lupin seeds, 56.5% corn silage, 10.2% corn grain and 10.7% Italian ray-grass on a DM basis, supplemented with vitamins and minerals. Chromium ethylenediaminotetraacetic (Cr-EDTA) and ytterbium chloride (YbCl3) were used as liquid and particulate markers respectively, while purines and 15N ammonium sulfate were utilized as bacterial markers. Cows fed EWLS had a similar ruminal ammonia N and volatile fatty acid concentrations and efficiency of bacterial protein synthesis compared to those fed the RWLS diet. Total tract OM and N digestion were not affected by inclusion of EWLS instead of RWLS; the corresponding mean values were 70 and 71%. Apparent degradation of OM and N in the rumen were 44 and 64% for diets containing RWLS, and 40 and 39% for EWLS diets. Feeding diets including EWLS both increased non ammonia N and dietary N flow to the duodenum compared with diets containing RWLS (472 vs 357 g/d) and (263 vs 153 g/d) respectively. Absorption from the small intestine (g/d and % entering) of dietary N was higher for EWLS diets (146 vs 62 g/d; 34 vs 15%). The PDIA, PDIE and PDIN contents (g/kg of DM) of RWLS were 18, 94 and 245 respectively; the corresponding values after extrusion were 145, 220 and 220. PMID:1777057

  5. Determination of protein and amino acid digestibility in foods including implications of gut microbial amino acid synthesis.

    Science.gov (United States)

    Fuller, Malcolm

    2012-08-01

    To meet the protein and amino acid requirements of individuals and of populations requires information not only about their requirements but also about the capacity of available foods to meet those requirements. Most of our current knowledge of the digestibility of food proteins and the methods to estimate it has been derived from work with animals. Because the microbiota of the large intestine alter the amino acid composition of the digesta, and because only trivial quantities of amino acids are absorbed intact from the large intestine, the current method of choice for assessing amino acid digestibility is ileal digestibility corrected for basal endogenous losses, that is, standardized ileal digestibility. For protein as a whole, however, because nitrogen absorbed in forms other than as amino acids can contribute to the nitrogen economy, the absorption of nitrogen over the whole digestive tract is the more appropriate measure. Most of the methods developed for estimating ileal amino acid outflow in animals are not directly applicable to man: the exception is the use of volunteers with an ileostomy. The flow and composition of ileal digesta in human subjects can also be measured by the infusion of a marker and withdrawal of samples through a naso-intestinal tube. However, this method is too demanding for routine use and is likely to be restricted to validating the application to humans of digestibility data obtained either from animals, of which the pig seems most suitable, or in vitro methods. Microbial activity in the gastrointestinal (GI) tract is not confined to the large intestine: the numbers and metabolic activity of the upper GI microbiota lead to substantial amounts of microbial protein leaving the ileum. It appears however that a large proportion of the amino acids used by the upper GI microbiota are preformed - from the diet or from endogenous materials - rather than from de novo synthesis. Although there are still uncertainties about the impact of microbial activity in the upper GI tract, the amino acid composition of ileal digesta provides the best available basis for estimating the proportion of dietary amino acids available for metabolism. PMID:23107534

  6. Paradata for 'Effects of traditional cooking methods on some antinutrients and in vitro protein digestibility of dry bean varieties (Phaseolus vulgaris L.) grown in Turkey'

    Science.gov (United States)

    This record contains paradata for the resource 'Effects of traditional cooking methods on some antinutrients and in vitro protein digestibility of dry bean varieties (Phaseolus vulgaris L.) grown in Turkey'

  7. Thermostable trypsin conjugates immobilized to biogenic magnetite show a high operational stability and remarkable reusability for protein digestion

    Science.gov (United States)

    Pe?ová, M.; Šebela, M.; Marková, Z.; Poláková, K.; ?uda, J.; Šafá?ová, K.; Zbo?il, R.

    2013-03-01

    In this work, magnetosomes produced by microorganisms were chosen as a suitable magnetic carrier for covalent immobilization of thermostable trypsin conjugates with an expected applicability for efficient and rapid digestion of proteins at elevated temperatures. First, a biogenic magnetite was isolated from Magnetospirillum gryphiswaldense and its free surface was coated with the natural polysaccharide chitosan containing free amino and hydroxy groups. Prior to covalent immobilization, bovine trypsin was modified by conjugating with ?-, ?- and ?-cyclodextrin. Modified trypsin was bound to the magnetic carriers via amino groups using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysulfosuccinimide as coupling reagents. The magnetic biomaterial was characterized by magnetometric analysis and electron microscopy. With regard to their biochemical properties, the immobilized trypsin conjugates showed an increased resistance to elevated temperatures, eliminated autolysis, had an unchanged pH optimum and a significant storage stability and reusability. Considering these parameters, the presented enzymatic system exhibits properties that are superior to those of trypsin forms obtained by other frequently used approaches. The proteolytic performance was demonstrated during in-solution digestion of model proteins (horseradish peroxidase, bovine serum albumin and hen egg white lysozyme) followed by mass spectrometry. It is shown that both magnetic immobilization and chemical modification enhance the characteristics of trypsin making it a promising tool for protein digestion.

  8. Chemical composition, starch digestibility and antioxidant capacity of tortilla made with a blend of quality protein maize and black bean.

    Science.gov (United States)

    Grajales-García, Eva M; Osorio-Díaz, Perla; Goñi, Isabel; Hervert-Hernández, Deisy; Guzmán-Maldonado, Salvador H; Bello-Pérez, Luis A

    2012-01-01

    Tortilla and beans are the basic components in the diet of people in the urban and rural areas of Mexico. Quality protein maize is suggested for tortilla preparation because it presents an increase in lysine and tryptophan levels. Beans contain important amounts of dietary fiber. The objective of this study was to prepare tortilla with bean and assesses the chemical composition, starch digestibility and antioxidant capacity using a quality protein maize variety. Tortilla with bean had higher protein, ash, dietary fiber and resistant starch content, and lower digestible starch than control tortilla. The hydrolysis rate (60 to 50%) and the predicted glycemic index (88 to 80) of tortilla decreased with the addition of bean in the blend. Extractable polyphenols and proanthocyanidins were higher in the tortilla with bean than control tortilla. This pattern produced higher antioxidant capacity of tortilla with bean (17.6 ?mol Trolox eq/g) than control tortilla (7.8 ?mol Trolox eq/g). The addition of bean to tortilla modified the starch digestibility and antioxidant characteristics of tortilla, obtaining a product with nutraceutical characteristics. PMID:22312252

  9. Chemical Composition, Starch Digestibility and Antioxidant Capacity of Tortilla Made with a Blend of Quality Protein Maize and Black Bean

    Directory of Open Access Journals (Sweden)

    Luis A. Bello-Pérez

    2011-12-01

    Full Text Available Tortilla and beans are the basic components in the diet of people in the urban and rural areas of Mexico. Quality protein maize is suggested for tortilla preparation because it presents an increase in lysine and tryptophan levels. Beans contain important amounts of dietary fiber. The objective of this study was to prepare tortilla with bean and assesses the chemical composition, starch digestibility and antioxidant capacity using a quality protein maize variety. Tortilla with bean had higher protein, ash, dietary fiber and resistant starch content, and lower digestible starch than control tortilla. The hydrolysis rate (60 to 50% and the predicted glycemic index (88 to 80 of tortilla decreased with the addition of bean in the blend. Extractable polyphenols and proanthocyanidins were higher in the tortilla with bean than control tortilla. This pattern produced higher antioxidant capacity of tortilla with bean (17.6 ?mol Trolox eq/g than control tortilla (7.8 ?mol Trolox eq/g. The addition of bean to tortilla modified the starch digestibility and antioxidant characteristics of tortilla, obtaining a product with nutraceutical characteristics.

  10. Improving anaerobic sewage sludge digestion by implementation of a hyper-thermophilic prehydrolysis step

    DEFF Research Database (Denmark)

    Lu, Jingquan; Gavala, Hariklia N.

    2008-01-01

    The present study focuses on a two-step process for treatment and stabilisation of primary sludge. The process consists of a hyperthermophilic hydrolysis step operated at 70 degrees C and a hydraulic retention time (HRT) of 2 clays followed by a thermophilic (55 degrees C) anaerobic digestion step at a HRT of 13 days. A one-step anaerobic digester operated at 55 degrees C and 15 days HRT Was used as a reference process. The two-step process was characterized by a 12% higher organic suspended solids removal efficiency and better pathogen reduction effect than the conventional one-step digestion. The microbial community of the digester fed with pre-treated sludge was characterised by it higher activity compared to that of the digester treating raw sludge. Moreover, the pre-treatment of the primary sludge resulted up to 48% increase of the methane potential (20.09 and 13.56 mmol CH4 g(-)VS(-1) with and Without pre-treatment respectively) and up to 115% increase of the methane production rate. Finally it was shown that the extra energy requirements for the operation of a pre-treatment step would be covered by the energy Produced from the extra methane production and in addition there would be it significant energy Surplus of 2.17 kJ d(-1) For the system tested.

  11. Eco-efficient post treatment of digestate from farm and collective biogas plants to improve nutrients (N&P) recycling

    DEFF Research Database (Denmark)

    Trémier, Anne; Béline, Fabrice

    Anaerobic digestion (AD) of organic waste from agriculture and others sectors is a widely used technology which shows increasing implementation due to its capacity to produce renewable energy and also to reduce greenhouse gas emissions from waste management. The development of AD is also an opportunity to improve nutrient recycling from organic waste through the development of an eco-effi cient post-treatment system. In this context, LCA was applied to evaluate the sustainability of diff erent raw digestate post-treatment technologies regarding recycling of nutrients from agricultural and organic waste to agricultural soils for decreased resource depletion and climate mitigation. Substitution of the use of N and P mineral fertilizers with recycled soil health improver or organic fertilizers products as function of fi ve diff erent post-treatment technologies and raw digestate characteristics was evaluated. A particular attention was carried to (1) the gaseous emissions (NH3 and N2O) from process (post-treatment) but also after land spreading and (2) the carbon cycle considering the CO2 carbon costs of fertiliser production and the soil carbon sequestration benefit.

  12. Research on crude protein and digestibility of Arnica montana L. using conventional NIR spectrometry and hyperspectral imaging NIR

    OpenAIRE

    Dale, Laura; Fernandez, Juan Antonio; Vermeulen, Philippe; Lecler, Bernard; Bogdan, Anca Dorina; Pacurar, Florin; Rotar, Ioan; Thewis, Andre?; Baeten, Vincent

    2012-01-01

    Arnica montana L. (AM) is considered a medicinal plant, used as hay in feed ration. The aim of this study is to assess the prediction of protein content and in vitro organic matter digestibility value in grass mixtures containing Arnica montana L., and in a second step to check if these values have a positive or negative influence in the mixtures. Crude protein has been selected because it is one of the most important quality parameters of forages as nutritional element used in animal feeding...

  13. Influence of Non-fibrous Carbohydrate and Degradable Intake protein and Ruminal Fermentation ,Nutrien Digestion and performance of Local Sheep

    OpenAIRE

    Ar, Efka; Sno, Suwandyastuti; Ning Iriyanti

    2001-01-01

    The objective of current study was to evaluate the impact dietary non-fibrous carbohydrate ( NFC) and ruminally degradable intake protein (DIP) concentration have on ruminal fermentation , nutrient digestion and performance of local sheep. The animal had a mean of liveweight 19.80 ±1.55 kg. four diets ,arranged in a 2x2 factorial ,were formulated to contain either 40 or 50 % NFC and 50 or 60 % of dietary crude protein as DIP .dietary DM contained 25 % Indonesian field grass and 75 % concent...

  14. New technologies in proteomics: automated excision and digestion of fluorescently stained protein gel spots and identification of proteins by MALDI-QTOF mass spectrometry

    International Nuclear Information System (INIS)

    This presentation is an overview of the techniques and systems we use for large-scale proteomics in plants. This includes a variety of 2D gel electrophoresis methods, and automation of spot-cutting, in-gel digestion, nanoLC-MS/MS tandem mass spectrometry and SEQUEST based protein identification. Examples and applications of several new technologies will also be shown, including the use of a robotic spot cutter in conjunction with high sensitivity fluorescent protein staining and the identification of proteins by redundant peptide identification in a high-throughput fashion using a hybrid MALDI-Qtof mass spectrometer

  15. Comparison of electron beam and gamma ray irradiations effects on ruminal crude protein and amino acid degradation kinetics, and in vitro digestibility of cottonseed meal

    International Nuclear Information System (INIS)

    This study was conducted to compare effects of electron beam (EB) and gamma ray (GR) treatments at doses of 25, 50 and 75 kGy on ruminal degradation kinetics of crude protein (CP), amino acid (AA), and in vitro digestibility of cottonseed meal (CSM). Ionizing radiations of EB and GR had significant effects (P0.05). Irradiation processing caused decrement in AA degradation after 16 h of ruminal incubation (P<0.05). EB irradiation was more effective than GR irradiation in lessening the ruminal degradability of AA (P<0.05). EB and GR treatments at a dose of 75 kGy increased in vitro digestibility of CSM numerically. This study showed that EB could cause CP and AA bypass rumen as well as GR. Therefore, ionizing irradiation processing can be used as an efficient method in improving nutritional value of CSM. - Highlights: ? Irradiation was effective on reducing ruminal degradability of cottonseed meal. ? Ionizing radiations, especially electron beam, lessened ruminal degradability of amino acid substantially. ? Irradiation processing could be used as a safe and efficient method in improving nutritional value of cottonseed meal.

  16. Improved method for trypsin digestion of Paraplast sections before immunofluorescence staining.

    OpenAIRE

    Rowse-eagle, D.; Watson, H. D.; Tignor, G. H.

    1981-01-01

    Formalin-fixed, Paraplast-embedded tissue sections mounted on subbed slides were digested with crude trypsin in modified Sorensen phosphate buffer containing sodium fluoride before immunofluorescence staining. This method prevented detachment of sections from slides during processing, thereby permitting immunofluorescence and histological examinations of adjacent serial sections.

  17. Inaccuracy of AOAC method 2009.01 with amyloglucosidase for measuring non-digestible oligosaccharides and proposal for an improvement of the method.

    Science.gov (United States)

    Tanabe, Kenichi; Nakamura, Sadako; Oku, Tsuneyuki

    2014-05-15

    We wished to clarify the inaccuracy of AOAC method 2009.01 for the measurement of non-digestible oligosaccharides and to propose an improved method using porcine intestinal enzymes. Amyloglucosidase used in AOAC method 2009.01 scarcely hydrolyses sucrose, palatinose and panose (which are readily digested by intestinal enzymes). Hence, oligosaccharides could not be measured accurately by AOAC method 2009.01. To confirm the inaccuracy of the method, we used porcine intestinal enzymes instead of amyloglucosidase. Using the improved method, fructooligosaccharide and galactooligosaccharide were measured accurately as non-digestible oligosaccharides, but sucrose, palatinose, panose and isomaltooligosaccharide were not. The improved method hydrolysed digestible oligosaccharides into monosaccharides. These results demonstrate that the inaccuracy of AOAC method 2009.01 for oligosaccharide measurement is due to incomplete hydrolysis by amyloglucosidase. We propose that amyloglucosidase should be replaced with porcine intestinal enzymes for such measurements. PMID:24423568

  18. Effects of ethanol and protein deficiency on pancreatic digestive and lysosomal enzymes.

    OpenAIRE

    Apte, M. V.; Wilson, J. S.; Korsten, M. A.; Mccaughan, G. W.; Haber, P. S.; Pirola, R. C.

    1995-01-01

    The pathogenesis of alcoholic pancreatitis is not fully understood. An increase in pancreatic digestive and lysosomal enzyme synthesis because of ethanol consumption could contribute to the development of pancreatic injury in alcoholics. This study aimed, firstly, to determine the effect of ethanol on the content and messenger RNA levels of pancreatic digestive enzymes and on the messenger RNA level of the lysosomal enzyme cathepsin B, and secondly, to examine the influence of concomitant pro...

  19. The heat treatment and the gelation are strong determinants of the kinetics of milk proteins digestion and of the peripheral availability of amino acids.

    Science.gov (United States)

    Barbé, Florence; Ménard, Olivia; Le Gouar, Yann; Buffière, Caroline; Famelart, Marie-Hélène; Laroche, Béatrice; Le Feunteun, Steven; Dupont, Didier; Rémond, Didier

    2013-02-15

    This study aimed to determine the kinetics of milk protein digestion and amino acid absorption after ingestion of four dairy matrices by six minipigs: unheated or heated skim milk and corresponding rennet gels. Digestive contents and plasma samples were collected over a 7 h-period after meal ingestion. Gelation of milk slowed down the outflow of the meal from the stomach and the subsequent absorption of amino acids, and decreased their bioavailability in peripheral blood. The gelled rennet matrices also led to low levels of milk proteins at the duodenum. Caseins and ?-lactoglobulin, respectively, were sensitive and resistant to hydrolysis in the stomach with the unheated matrices, but showed similar digestion with the heated matrices, with a heat-induced susceptibility to hydrolysis for ?-lactoglobulin. These results suggest a significant influence of the meal microstructure (resulting from heat treatment) and macrostructure (resulting from gelation process) on the different steps of milk proteins digestion. PMID:23194515

  20. High-moisture air-tight storage of barley and wheat improves nutrient digestibility

    DEFF Research Database (Denmark)

    Poulsen, H D; Blaabjerg, K

    2012-01-01

    12Barley (Hordeum vulgare) and wheat (Triticum aestivum) are often stored dry with 14% or less moisture, which during rainy periods may require that grains are dried after harvest. The hypothesis is that air-tight storage of high-moisture barley and wheat will increase nutrient digestibility due to chemical conversions prior to feeding. The objective was to evaluate the effect of high moisture compared to dry storage of barley and wheat on digestibility of P and CP. The crops were grown on 1 field keeping other factors constant. Half of the grains was harvested in the morning after a rainy day and stored in air-tight silos (DM, %: barley, 85.2; wheat, 82.8) and the other half was harvested later the same day (windy and sunny) and stored dry (DM, %: barley, 89.8; wheat, 88.3). After 6 mo of storage, 1 low- and 1 high-moisture diet were prepared with a barley:wheat ratio of 1:1 mixed with soybean (Glycine max) meal and rapeseed cake to produce a compound diet without inorganic P and microbial phytase. Sixteen 45-kg pigs housed in metabolism crates were fed either the low- or the high-moisture diet for 5 d for adaptation and 7 d for total collection of feces. Digestibility of P was 12% higher (P <0.01) and of CP was 4% higher (P = 0.08) in the high-moisture diet. Phytase activity of dry-stored grain was lower (P <0.01) and phytate P was 4% higher in the high-moisture stored grain vs. the grains stored dry. Overall, high-moisture storage increased digestibility of P and CP when the grain was fed to finishing pigs. Therefore, high-moisture air-tight storage saved energy (without drying) and at the same time enhanced P digestibility and increased the nutritional value of grain probably through enzymatic activity during storage.

  1. Thermophilic co-digestion of pig manure and crude glycerol: process performance and digestate stability.

    Science.gov (United States)

    Astals, S; Nolla-Ardèvol, V; Mata-Alvarez, J

    2013-07-10

    Anaerobic co-digestion has been widely used to enhance biogas production of digesters and, therefore, to improve the anaerobic plants economic feasibility. In the present study, glycerol, a by-product of the biodiesel industry, was used as a co-substrate for pig manure. The results showed that the thermophilic anaerobic co-digestion of pig manure supplemented with 3% of glycerol, on weight basis, was satisfactory. The specific biogas production of the co-digester was 180% higher than the one obtained by the reference digester, which was only fed with pig manure. The improvement was related to the double of the organic loading rate, the high biodegradability of the crude glycerol, the slight reduction of the free ammonia concentration and the optimisation of the carbon-to-nitrogen ratio. Moreover, the analysis of the organic matter (protein, lipids, carbohydrates and fibers) of the influent and the effluent of both digesters together with their biogas flow rates indicated that the microorganisms in the co-digester obtained large amounts of nutrients from the glycerol, whereas the microorganisms of the reference digester mainly produced biogas from the particulate matter. However, the digestate obtained from the co-digester cannot be directly applied as soil fertiliser or conditioner due to the presence of high levels of biodegradable matter, which may exert negative impacts on the plant-soil system. Thus, a longer hydraulic retention time, a reduction of the glycerol concentration and/or a post-treatment is required if the digestate is to be used as soil fertiliser or conditioner. In contrast, pig manure digestate can be directly applied on land. PMID:23685137

  2. Strategies in protein sequencing and characterization: multi-enzyme digestion coupled with alternate CID/ETD tandem mass spectrometry.

    Science.gov (United States)

    Nardiello, Donatella; Palermo, Carmen; Natale, Anna; Quinto, Maurizio; Centonze, Diego

    2015-01-01

    A strategy based on a simultaneous multi-enzyme digestion coupled with electron transfer dissociation (ETD) and collision-induced dissociation (CID) was developed for protein sequencing and characterization, as a valid alternative platform in ion-trap based proteomics. The effect of different proteolytic procedures using chymotrypsin, trypsin, a combination of both, and Lys-C, was carefully evaluated in terms of number of identified peptides, protein coverage, and score distribution. A systematic comparison between CID and ETD is shown for the analysis of peptides originating from the in-solution digestion of standard caseins. The best results were achieved with a trypsin/chymotrypsin mix combined with CID and ETD operating in alternating mode. A post-database search validation of MS/MS dataset was performed, then, the matched peptides were cross checked by the evaluation of ion scores, rank, number of experimental product ions, and their relative abundances in the MS/MS spectrum. By integrated CID/ETD experiments, high quality-spectra have been obtained, thus allowing a confirmation of spectral information and an increase of accuracy in peptide sequence assignments. Overlapping peptides, produced throughout the proteins, reduce the ambiguity in mapping modifications between natural variants and animal species, and allow the characterization of post translational modifications. The advantages of using the enzymatic mix trypsin/chymotrypsin were confirmed by the nanoLC and CID/ETD tandem mass spectrometry of goat milk proteins, previously separated by two-dimensional gel electrophoresis. PMID:25479873

  3. / Effects of different sources of protein on digestive characteristics, microbial efficiency, and nutrient flow in dairy goats

    Scientific Electronic Library Online (English)

    Nivea Regina de Oliveira, Felisberto; Marcelo Teixeira, Rodrigues; Marco Aurélio Delmondes, Bomfim; Rogério da Silva, Matos; Ana Gabriela Pombo Celles, Cordeiro; Márcia Maria Cândido da, Silva.

    2228-22-01

    Full Text Available [...] Abstract in english Diets formulated with protein sources presenting different resistance to ruminal degradation were compared by evaluating ruminal parameters, production and microbial efficiency and nutrients flow to the omasum in goats. Eight rumen cannulated non-lactating, non-pregnant goats were distributed in a 4 [...] × 4 Latin square design with two replicates. Treatments consisted of four diets where different sources of plant protein accounted for the major protein source named soybean meal, source of higher ruminal degradability, and three other sources of higher resistance of degradation: roasted soybean, corn gluten meal, and cottonseed cake. Amounts of rumen protein were similar among rations; however, flows of dry matter, protein and non-fiber carbohydrate to omasum were higher for diets with protein source with reduced rumen degradation rate. Higher values of rumen ammonia were obtained by using ration with soybean meal as major source of protein. Higher values of pH were obtained for rations with roasted soybean e cottonseed cake. Regarding kinetic of transit, similar values were found among rations. Diets with protein sources presenting reduced ruminal degradation increase nutrients flow to the omasum in goats and alter digestive parameters such as pH and ammonia without compromising bacteria growth and efficiency, which grants their use for dairy goats with similar efficiency to rations using more degradable sources of protein.

  4. The influence of irradiation treatment on gossypol and protein digestibility of decorticated cottonseed meal for Broiler chicks

    International Nuclear Information System (INIS)

    The Effect of gamma irradiation (5-20 kGy) on the chemical composition, soluble protein, available lysine, gossypol, in-vitro and in vivo digestibility of decorticated cottonseed meal (CSM) and the performance of broilers fed processed CSM were investigated. Radiation treatment of CSM at dose levels of 5, 10, 15 and 20 kGy did not affect moisture, crude protein, ether extract, crude fiber, ash and NFW content. However, the treatment reduced soluble protein by 4.18, 7.82, 10.91, 13.46%, respectively. Fluorodinitrobenzene-available lysine concentration reduced by 4.48, 8.12, 17.65 and 21.29% respectively. The concentrations of total gossypol decreased by 7.97, 15.78, 19.92 and 25.39% and free gossypol by 3.74, 7.85, 18.43 and 28.01%, respectively. The digestibility of CSM protein was increased by 5.3, 9.0, 16.6 and 17.2%, for in-vitro digestibility and by 4.8, 7.3, 9.4 and 12.9% for in-vivo due to treatment with 5,10, 15 and 20 kGy, respectively. From 7 to 42 days, arbor acres broilers were freely given iso energetic and iso nitrogenous : corn-soy control diet or diets containing 300 g kg-1 raw or irradiated CSM at 4, 15 or 20 kGy. Intake of raw CSM affected growth, feed intake and feed conversion as compared with birds receiving the control diet

  5. Characterization of condensed tannins purified from legume forages: chromophore production, protein precipitation, and inhibitory effects on cellulose digestion.

    Science.gov (United States)

    McAllister, Tim A; Martinez, Tomas; Bae, Hee Dong; Muir, Alister D; Yanke, L Jay; Jones, Graham A

    2005-09-01

    To identify simple screening tools for selecting condensed tannin (CT)-containing forages as candidate sources for further study, CT were isolated from nine legumes, and their molecular weights (MW), chromophore production, capacity to precipitate bovine serum albumin (BSA) and Fraction 1 protein (Rubisco) isolated from alfalfa, and inhibition of filter paper digestion were compared. Sources were as follows: leaves of sericea lespedeza (Lespedeza cuneata Dum.-Cours.), crown vetch (Coronilla varia L.), and sainfoin (Onobrychis viciifolia Scop.); stems of hedysarum (Hedysarum alpinum L.); seeds of alfalfa (Medicago sativa L.); and whole plants of birdsfoot trefoil (Lotus corniculatus var. corniculatus L.) and three varieties of big trefoil (Lotus pedunculatus Cav.), viz., Lotus uliginosus Schkuhr, L. uliginosus var. glabriusculus, and L. uliginosus var. villosus. Molecular weights and sizes (degrees of polymerization) of the CT varied considerably within and among plant species. Average MW ranged from 3036 Da (crown vetch) to 7143 Da (lespedeza). All CT exhibited greater capacity (w/w basis) to bind alfalfa Rubisco than BSA. Relative astringencies (microg CT required to precipitate 1 mg protein) against BSA ranged from 262.5 for CT from lespedeza to 435.5 for CT from L. corniculatus, and against Rubisco, from 49.6 (sainfoin) to 108.2 (alfalfa seed). Including CT at 300 microg/ml in cultures of Fibrobacter succinogenes reduced digestion of cellulose filter paper by 19.8% (sainfoin) to 92.4% (crown vetch) and increased the specific activity of cell-associated endoglucanase. There were no correlations between inhibitory effects of CT on filter paper digestion and (1) chromophore formation during CT assay by butanol-HCl, vanillin-HCl, or H2SO4; (2) precipitation of BSA or alfalfa Rubisco; and (3) MW of CT. The most inhibitory CT for cellulose digestion included those with broad and with narrow MW distributions. Sainfoin was the most desirable source of CT, as it had the highest capacity to bind alfalfa protein and was least inhibitory to cellulose digestion by F. succinogenes. This study suggests that these properties are not easily defined via chemical means, and that biological assays using rumen bacteria may help identify those CT with properties of nutritional interest. PMID:16132212

  6. Effects of Heat-oxidized Soy Protein Isolate on Growth Performance and Digestive Function of Broiler Chickens at Early Age

    Science.gov (United States)

    Chen, X.; Chen, Y. P.; Wu, D. W.; Wen, C.; Zhou, Y. M.

    2015-01-01

    This study was conducted to investigate effects of heat-oxidized soy protein isolate (HSPI) on growth performance, serum biochemical indices, apparent nutrient digestibility and digestive function of broiler chickens. A total of 320 1-day-old Arbor Acres chicks were randomly divided into 4 groups with 8 replicates of 10 birds, receiving diets containing soy protein isolate (SPI, control group) or the same amount of SPI heated in an oven at 100°C for 1, 4, or 8 h, for 21 days, respectively. The results indicated that compared with the control group, body weight gain and feed intake of birds fed diet containing SPI heated for 8 h were significantly lower (p<0.05). Serum urea nitrogen concentration was higher in the broilers fed diet containing SPI heated for 4 or 8 h at d 21 (p<0.05). In contrast, serum glucose content was decreased by HSPI substitution at d 21 (p<0.05). The relative pancreas weight in HSPI groups was higher than that in the control group at d 21 (p<0.05). Meanwhile, the opposite effect was observed for relative weight of anterior intestine and ileum in broilers fed a diet containing SPI heated for 8 h (p<0.05). Birds fed diets containing SPI heated for 4 or 8 h had a decreased lipase activity in anterior intestinal content at d 14 and 21 (p<0.05), respectively. In addition, the same effect was also noted in broilers given diets containing SPI heated for 1 h at d 21 (p<0.05). Similarly, amylase, protease and trypsin activity in anterior intestinal content were lower in broilers fed diets containing SPI heated for 8 h at d 21 (p<0.05). The apparent digestibility of dry matter (DM) from d 8 to 10 and DM, crude protein (CP), and ether extract from d 15 to 17 were lower in broilers fed diets containing SPI heated for 8 h (p<0.05). Besides, birds given diets containing SPI heated for 4 h also exhibited lower CP apparent digestibility from d 15 to 17 (p<0.05). It was concluded that HSPI inclusion can exert a negative influence on the growth performance of broilers, which was likely to result from the simultaneously compromised digestive function. PMID:25656201

  7. Low-cost additive improved silage quality and anaerobic digestion performance of napiergrass.

    Science.gov (United States)

    Lianhua, Li; Feng, Zhen; Yongming, Sun; Zhenhong, Yuan; Xiaoying, Kong; Xianyou, Zhou; Hongzhi, Niu

    2014-12-01

    Effects of molasses-alcoholic wastewater on the ensiling quality of napiergrass were investigated at ambient temperature, and its anaerobic digestion performance was assessed at mesophilic temperature. Results showed that the molasses-alcoholic wastewater had positive effect on silage quality and anaerobic digestion performance. Lower pH values of 5.20-5.28, lower NH3-N contents of 32.65-36.60 g/kg and higher lactic acid contents of 56-61 mg/kg FM were obtained for the silage samples with molasses-alcoholic wastewater addition. Higher specific biogas yield of 273 mL/g VS was obtained for the sample with 11% molasses-alcoholic wastewater added. Therefore 11% molasses-alcoholic wastewater addition was recommended. PMID:25443806

  8. Dietary actinidin from kiwifruit (Actinidia deliciosa cv. Hayward) increases gastric digestion and the gastric emptying rate of several dietary proteins in growing rats.

    Science.gov (United States)

    Montoya, Carlos A; Hindmarsh, Jason P; Gonzalez, Lucrecia; Boland, Mike J; Moughan, Paul J; Rutherfurd, Shane M

    2014-04-01

    Dietary actinidin influences the extent to which some dietary proteins are digested in the stomach, and it is hypothesized that the latter modulation will in turn affect their gastric emptying rate (GE). In this study, the effect of dietary actinidin on GE and gastric digestion of 6 dietary protein sources was determined in growing rats. Each dietary protein source [beef muscle, gelatin, gluten, soy protein isolate (SPI), whey protein isolate, and zein] was included in 2 semisynthetic diets as the sole nitrogen source. For each protein source, 1 of the 2 diets contained actinidin [76.5 U/g dry matter (DM)] in the form of ground freeze-dried green kiwifruit (Actinidia deliciosa cv. Hayward), whereas the other diet contained freeze-dried gold kiwifruit (Actinidia chinensis cv. Hort16A), which is devoid of actinidin (3.4 U/g DM). For both diets, dietary kiwifruit represented 20% of the diet on a DM basis. The real-time GE was determined in rats gavaged with a single dose of the diets using magnetic resonance spectroscopy over 150 min (n = 8 per diet). Gastric protein digestion was determined based on the free amino groups in the stomach chyme collected from rats fed the diets (n = 8 per diet) that were later killed. GE differed across the protein sources [e.g., the half gastric emptying time (T(½)) ranged from 157 min for gluten to 266 min for zein] (P digestion of beef muscle (0.6-fold), gluten (3.2-fold), and SPI (0.6-fold) and increased the GE of the diets containing beef muscle (43% T(½)) and zein (23% T(½); P protein digestion and DM retained in the stomach (r = -0.67; P protein digestion and accelerated the GE for several dietary protein sources. GE may be influenced by gastric protein digestion, and dietary actinidin can be used to modulate GE and protein digestion in the stomach of some dietary protein sources but not others. PMID:24431326

  9. Effects of milk proteins on release properties and particle morphology of ?-carotene emulsions during in vitro digestion.

    Science.gov (United States)

    Liu, Yuwei; Lei, Fei; Yuan, Fang; Gao, Yanxiang

    2014-11-01

    In the present study, ?-lactoglobulin, sodium caseinate, lactalbumin and lactoferrin were used to prepare ?-carotene emulsions. The milk protein-stabilized emulsions were explored using an in vitro release model to elucidate the effects of different milk proteins on ?-carotene release properties in the stomach, duodenum and small intestine, respectively. Notable changes in the droplet size and size distribution were observed among these four oil-in-water (O/W) milk protein emulsions. In the gastric environment, the highest ?-carotene release rate (2.9%) was achieved in ?-lactoglobulin emulsion with a remarkable change in the particle size. In the simulated intestine, the best ?-carotene micellarization potency (92%) was observed in ?-lactoglobulin emulsion and its droplet diameter moderately increased from 215 nm to 471 nm. Moreover, substantial release of ?-carotene was found in the small intestine for the four types of emulsions. It was concluded that ?-carotene release in different digestive stages was characterized by the emulsion interfacial composition. PMID:25215854

  10. Enzymatic protein digests do not assist in E. coli discrimination at the strain level using mass spectrometry

    Directory of Open Access Journals (Sweden)

    Ricardo J. Carreira

    2013-01-01

    Full Text Available Different procedures for matrix assisted laser desorption ionization time of flight mass spectrometry-based E. coli classification at the strain level using the enzymatic digestion of proteins from the cell lysate have been studied. The effects of ultrasonic energy as well as the effects of protein reduction and protein alkylation in the sample treatment and in the subsequent classification were assessed. The final optimal method for classification was then compared with an intact cell-based approach in a different set of samples. Our results show that E. coli classification at the strain level is possible as 12 different strains were correctly classified using intact cell analysis. Overall, the confidence level in classification was higher when the analysis was performed with the intact cell approach.

  11. Energy, phosphorus, and amino acid digestibility of high-protein distillers dried grains and corn germ fed to growing pigs.

    Science.gov (United States)

    Widmer, M R; McGinnis, L M; Stein, H H

    2007-11-01

    Three experiments were conducted to measure energy, P, and AA digestibility in 2 novel co-products from the ethanol industry [i.e., high-protein distillers dried grains (HP DDG) and corn germ]. These products are produced by dehulling and degerming corn before it enters the fermentation process. Experiment 1 was an energy balance experiment conducted to measure DE and ME in HP DDG, corn germ, and corn. Six growing pigs (initial BW, 48.9 +/- 1.99 kg) were placed in metabolism cages and fed diets based on corn, corn and HP DDG, or corn and corn germ. Pigs were allotted to a replicated, 3 x 3 Latin square design. The DE and ME in corn (4,056 and 3,972 kcal/kg of DM, respectively) did not differ from the DE and ME in corn germ (3,979 and 3,866 kcal/kg of DM, respectively). However, HP DDG contained more (P < 0.05) energy (4,763 kcal of DE/kg of DM and 4,476 kcal of ME/kg of DM) than corn or corn germ. Experiment 2 was conducted to measure apparent total tract digestibility (ATTD) and true total tract digestibility of P in HP DDG and corn germ. Thirty growing pigs (initial BW, 33.2 +/- 7.18 kg) were placed in metabolism cages and fed a diet based on HP DDG or corn germ. A P-free diet was used to measure endogenous P losses. Pigs were assigned to treatments in a randomized complete block design, with 10 replications per treatment. The ATTD and the retention of P were calculated for the diets containing HP DDG and corn germ, and the endogenous loss of P was estimated from pigs fed the P-free diet. The ATTD was lower (P < 0.05) in corn germ (28.6%) than in the HP DDG (59.6%). The retention of P was also lower (P < 0.05) in pigs fed corn germ (26.7%) than in pigs fed HP DDG (58.9%). The endogenous loss of P was estimated to be 211 +/- 39 mg per kg of DMI. The true total tract digestibility of P for HP DDG and corn germ was calculated to be 69.3 and 33.7%, respectively. In Exp. 3, apparent ileal digestibility and standardized ileal digestibility values of CP and AA in HP DDG and corn germ were measured using 6 growing pigs (initial BW, 78.2 +/- 11.4 kg) allotted to a replicated, 3 x 3 Latin square design. The apparent ileal digestibility for CP and all AA except Arg and Pro, and the standardized ileal digestibility for CP and all AA except Arg, Lys, Gly, and Pro were greater (P < 0.05) in HP DDG than in corn germ. It was concluded that HP DDG has a greater digestibility of energy, P, and most AA than corn germ. PMID:17644788

  12. Competition between bound and free peptides in an ELISA-based procedure that assays peptides derived from protein digests

    Directory of Open Access Journals (Sweden)

    Pace Umberto

    2006-05-01

    Full Text Available Abstract Background We describe an ELISA-based method that can be used to identify and quantitate proteins in biological samples. In this method, peptides in solution, derived from proteolytic digests of the sample, compete with substrate-attached synthetic peptides for antibodies, also in solution, generated against the chosen peptides. The peptides used for the ELISA are chosen on the basis of their being (i products of the proteolytic (e.g. tryptic digestion of the protein to be identified and (ii unique to the target protein, as far as one can know from the published sequences. Results In this paper we describe the competition assay and we define the optimal conditions for the most effective assay. We have performed an analysis of the kinetics of interaction between the four components of the assay: the plastic substratum to which the peptide is bound, the bound peptide itself, the competing added peptide, and the antibody that is specific for the peptide and we compare the results of theoretical simulations to the actual data in some model systems. Conclusion The data suggest that the peptides bind to the plastic substratum in more than one conformation and that, once bound, the peptide displays different affinities for the antibody, depending on how it has bound to the plate

  13. A peptidomic analysis of human milk digestion in the infant stomach reveals protein-specific degradation patterns.

    Science.gov (United States)

    Dallas, David C; Guerrero, Andrés; Khaldi, Nora; Borghese, Robyn; Bhandari, Aashish; Underwood, Mark A; Lebrilla, Carlito B; German, J Bruce; Barile, Daniela

    2014-06-01

    In vitro digestion of isolated milk proteins results in milk peptides with a variety of actions. However, it remains unclear to what degree protein degradation occurs in vivo in the infant stomach and whether peptides previously annotated for bioactivity are released. This study combined nanospray LC separation with time-of-flight mass spectrometry, comprehensive structural libraries, and informatics to analyze milk from 3 human mothers and the gastric aspirates from their 4- to 12-d-old postpartum infants. Milk from the mothers contained almost 200 distinct peptides, demonstrating enzymatic degradation of milk proteins beginning either during lactation or between milk collection and feeding. In the gastric samples, 649 milk peptides were identified, demonstrating that digestion continues in the infant stomach. Most peptides in both the intact milk and gastric samples were derived from ?-casein. The numbers of peptides from ?-casein, lactoferrin, ?-lactalbumin, lactadherin, ?-casein, serum albumin, bile salt-associated lipase, and xanthine dehydrogenase/oxidase were significantly higher in the gastric samples than in the milk samples (P < 0.05). A total of 603 peptides differed significantly in abundance between milk and gastric samples (P < 0.05). Most of the identified peptides have previously identified biologic activity. Gastric proteolysis occurs in the term infant in the first 2 wk of life, releasing biologically active milk peptides with immunomodulatory and antibacterial properties of clinical relevance to the proximal intestinal tract. Data are available via ProteomeXchange (identifier PXD000688). PMID:24699806

  14. Development of an efficient on-chip digestion system for protein analysis using MALDI-TOF MS.

    Science.gov (United States)

    Lee, Jeonghoon; Soper, Steven A; Murray, Kermit K

    2009-12-01

    A solid-phase trypsin microreactor was constructed and operated with electrokinetically-driven flow for the digestion of proteins and coupled off-line with MALDI-TOF MS. The bioreactor was fabricated from poly(methyl methacrylate), PMMA, by hot embossing using a mold master prepared by micro-milling. The solid-phase bioreactor consisted of a 4 cm long, 200 microm wide, and 50 microm deep microfluidic channel that was populated with an array of 50 microm diameter micropost structures with a 50 microm inter-post spacing. The bioreactor was prepared by covalently attaching the proteolytic enzyme, trypsin, to the UV-modified surface of the PMMA microstructures using the appropriate coupling reagents. The performance of the system was evaluated using a set of proteins. The bioreactor provided efficient digestion of cytochrome c at a field strength of 375 V/cm, producing a reaction time of approximately 20 s to produce 97% sequence coverage for protein identification. Bovine serum albumin (BSA), phosphorylase b, and beta-casein were also assessed and the sequence coverages were 46, 63, and 79%, respectively, using the same reactor residence time. Furthermore, Escherichia coli was used as a model to demonstrate the feasibility of fingerprint analysis for intact cells using this solid-phase bioreactor. PMID:19918612

  15. Impact of dehulling on the physico-chemical properties and in vitro protein digestion of common beans (Phaseolus vulgaris L.).

    Science.gov (United States)

    Romano, A; Giosafatto, C V L; Masi, P; Mariniello, L

    2015-04-01

    The objective of this study was to understand the effect of dehulling on the microstructural, physico-chemical characteristics, and in vitro protein digestion of common bean flours with particular regard to differences between adults and infants. The microstructure of flour samples from undehulled (WB) and manually dehulled (SB) beans, observed through scanning electron microscopy, showed that WB starch granules appeared to be surrounded by an integral matrix, while the SB starch granule structure was still visible although covered by protein clusters. The starch granules were oval and spherical, with heterogeneous sizes ranging from 19 to 30 ?m in diameter. Particle size analysis determined with a laser diffraction particle size analyzer showed similar bimodal particle size distributions of small (1-25 ?m) and large (>100 ?m) granules, though the particle size of WB was obviously higher than SB. Color and other physico-chemical analyses showed that dehulling had significant (P flour protein digestion which could be utilized in various food applications. PMID:25778680

  16. Rumen Degradability and Post-ruminal Digestion of Dry Matter, Nitrogen and Amino Acids of Three Protein Supplements.

    Science.gov (United States)

    Gao, Wei; Chen, Aodong; Zhang, Bowen; Kong, Ping; Liu, Chenli; Zhao, Jie

    2015-04-01

    This study evaluated the in situ ruminal degradability, and subsequent small intestinal digestibility (SID) of dry matter, crude protein (CP), and amino acids (AA) of cottonseed meal (CSM), sunflower seed meal (SFSM) and distillers dried grains with solubles (DDGS) by using the modified three-step in vitro procedure. The ruminal degradability and subsequent SID of AA in rumen-undegradable protein (RUP-AA) varied among three protein supplements. The result show that the effective degradability of DM for SFSM, CSM, and DDGS was 60.8%, 56.4%, and 41.0% and their ruminal fermentable organic matter was 60.0%, 55.9%, and 39.9%, respectively. The ruminal degradable protein (RDP) content in CP for SFSM, CSM, and DDGS was 68.3%, 39.0%, and 32.9%, respectively, at the ruminal solid passage rate of 1.84%/h. The SFSM is a good source of RDP for rumen micro-organisms; however, the SID of RUP of SFSM was lower. The DDGS and CSM are good sources of RUP for lambs to digest in the small intestine to complement ruminal microbial AA of growing lambs. Individual RUP-AA from each protein source was selectively removed by the rumen micro-organisms, especially for Trp, Arg, His, and Lys (p<0.01). The SID of individual RUP-AA was different within specific RUP origin (p<0.01). Limiting amino acid was Leu for RUP of CSM and Lys for both RUP of SFSM and DDGS, respectively. Therefore, different protein supplements with specific limitations should be selected and combined carefully in growing lambs ration to optimize AA balance. PMID:25656208

  17. Effect of different concentrations of protein on the digestive system of juvenile silver catfish Efeito de diferentes concentrações de proteína sobre o sistema digestivo de juvenis de jundiá

    OpenAIRE

    Melo, J. F. B.; Lundstedt, L. M.; Moraes, G.; Inoue, L. A. K. A.

    2012-01-01

    Aspects of digestion in juvenile silver catfish Rhamdia quelen fed with four levels of protein (20, 27, 34 and 41%) were studied. The studied traits were digestive enzymes (nonspecific proteases, trypsin, chymotrypsin, amylase and lipase) and weight gain. The experiment lasted 60 days; 160 juvenile catfish with initial body weight of 32 grams were transferred to the experimental system of 500L tanks with re-circulated and thermostatized water. The average weight of silver catfish increased wi...

  18. A Quantitative Study of the Effects of Chaotropic Agents, Surfactants, and Solvents on the Digestion Efficiency of Human Plasma Proteins by Trypsin

    OpenAIRE

    Proc, Jennifer L.; Kuzyk, Michael A.; Hardie, Darryl B.; Yang, Juncong; Smith, Derek S.; Jackson, Angela M.; Parker, Carol E.; Borchers, Christoph H.

    2010-01-01

    Plasma biomarkers studies are based on the differential expression of proteins between different treatment groups or between diseased and control populations. Most mass spectrometry-based methods of protein quantitation, however, are based on the detection and quantitation of peptides, not intact proteins. For peptide-based protein quantitation to be accurate, the digestion protocols used in proteomic analyses must be both efficient and reproducible. There have been very few studies, however,...

  19. Internal amino acid sequence analysis of proteins separated by one- or two-dimensional gel electrophoresis after in situ protease digestion on nitrocellulose.

    OpenAIRE

    Aebersold, R. H.; Leavitt, J.; Saavedra, R. A.; Hood, L. E.; Kent, S. B.

    1987-01-01

    We have developed a general two-step method for obtaining peptide fragments for sequence analysis from picomole quantities of proteins separated by gel electrophoresis. After separation by one- or two-dimensional polyacrylamide gel electrophoresis, proteins are electrophoretically transferred (electroblotted) onto nitrocellulose, the protein-containing regions are detected by reversible staining and are cut out, and each protein is digested in situ by proteolytic enzymes such as trypsin or st...

  20. Improving nutrient fixation and dry matter content of an ammonium-rich anaerobic digestion effluent by struvite formation and clay adsorption.

    Science.gov (United States)

    Estevez, Maria M; Linjordet, Roar; Horn, Svein J; Morken, John

    2014-01-01

    The anaerobic digestion (AD) of organic wastes that contain nitrogen leads to its mineralization, yielding a digestate rich in ammonium (NH(4)(+)), an important fertilizing nutrient. The applicability of AD digestate as fertilizer can be improved by fixating the nutrients and increasing its dry matter content. Methods for the fixation and recovery of the digestate's NH(4)(+) and possible also PO(4)(3-) include struvite precipitation and adsorption in clay materials such as bentonite. These techniques were tested in batch experiments employing the liquid fraction of a digestate originating from the AD of a substrate mix containing lignocellulose, cattle manure and fish industrial waste. The concentration of NH(4)(+)-N in this digestate was 2,300 mg L?¹. Struvite precipitation conditions at a molar ratio of 1.2:1:1 (Mg²?:NH(4)(+):PO(4)(3-)) and pH 9.5 were best in terms of simultaneous removal of NH(4)(+)-N (88%), PO(4)(3-) (60%) and soluble chemical oxygen demand (44%). Bentonite adsorption gave comparably high removal levels for NH(4)(+)-N (82%) and PO(4)(3-) (52%). Analysis of the precipitates' morphology and elemental composition confirmed their struvite and bentonite nature. Dry matter content was increased from 5.8% in the AD digestate to 27% and 22% in the struvite and bentonite sludges, respectively. PMID:25051482

  1. Investigation on the effects of dietary protein reduction with constant ratio of digestible sulfur amino acids and threonine to lysine on performance, egg quality and protein retention in two strains of laying hens

    Directory of Open Access Journals (Sweden)

    Farhad Foroudi

    2013-01-01

    Full Text Available An experiment was conducted to determine the possibility of using various levels of crude protein (CP by providing laying hens with constant levels of digestible sulfur amino acid, threonine and lysine to improve performance and egg quality. The experiment was conducted as a completely randomized block design in a factorial arrangement (4 × 2 with 8 replicates of 10 hens in each. Factors included 4 levels of CP (18.5%, 17.5%, 16.5% and 15.5% and 2 strains (LSL and Hy-Line W-36 of laying hens. Hens were fed experimental diets from 25 to 33 weeks of age. Production performance was measured for eight weeks and egg quality characteristics were determined at 29 and 33 weeks of age. Protein reduction decreased egg weight, egg mass and hen body weight linearly (P?0.01. Egg production was not affected by protein reduction but feed efficiency, and average daily feed intake increased significantly (P?0.01. Lohmann Selected Leghorn laying hens showed significantly higher egg production, egg weight, egg mass, weight gain, feed efficiency and feed intake compared to the W-36 laying hens (P?0.01. Shell thickness increased linearly as protein levels decreased (P?0.05. There were significant differences between two strains on the egg quality characteristics (P?0.01. Significant (P?0.05 CP × strain interactions were observed for hen weight, albumen height, Haugh units, yolk and shell percentage. Based on the results of this experiment, a reduction in dietary protein level (from 18.5% to 15.5%, without any alteration in digestible TSAA and Thr: Lys ratio, led to inferior egg mass and feed conversion ratio during the peak production period.

  2. Dewaterability of thermophilically digested biosolids: effects of temperature and cellular polymeric substances

    International Nuclear Information System (INIS)

    Thermophilic processes digest sludge at high temperatures to produce Class A biosolids.Recent research work revealed that digestion temperature is the predominant factor affecting dewaterability of thermophilic biosolids. This paper presents findings of a laboratory study that investigated how various digestion temperatures affect dewaterability of digested biosolids, studied the phase partition of the substances affecting dewaterability in digested biosolids, and tested the role of cellular polymeric substances in affecting dewaterability.Secondary sludges were digested at 40-70oC or 22oC for up to 12 days. Centrate from thermophilically digested biosolids were treated with protease and boiling. This study found that, during the first few hours of digestion, higher temperatures resulted in more rapid and more significant deterioration in dewaterability than lower digestion temperatures. Continued digestion resulted in either improved (60oC or 70oC), or unchanged (40oC or 50oC), or gradually deteriorated dewaterability (22oC). The substances affecting dewaterability were primarily located in the liquid phase of thermophilically digested biosolids. Boiling treatment did not result in significant changes in dewaterability. Protease treatment of the liquid phase of thermophilic biosolids improved dewaterability by 13-19%. Such an improvement confirmed the role of proteins in affecting dewaterabie role of proteins in affecting dewaterability. (author)

  3. Improving bioorthogonal protein ubiquitylation by click reaction

    OpenAIRE

    Schneider, Daniel; Schneider, Tatjana; Ro?sner, Daniel; Scheffner, Martin; Marx, Andreas

    2013-01-01

    Posttranslational modification of proteins with ubiquitin (ubiquitylation) regulates numerous cellular processes. Besides functioning as a signal for proteasomal degradation, ubiquitylation has also non-proteolytic functions by altering the biochemical properties of the modified protein. To investigate the effect(s) of ubiquitylation on the properties of a protein, sufficient amounts of homogenously and well-defined ubiquitylated proteins are required. Here, we report on the elaboration of a ...

  4. Experimental and clinical studies on simultaneous fat and protein tolerance digestion-absorption test using 131I-triolein and 125I-RISA

    International Nuclear Information System (INIS)

    Simultaneous fat and protein tolerance digestion-absorption test using 131I-triolein and 125I-RISA was investigated experimentally in rats and clinically in 148 inpatients with various diseases. The results were as follows: Animal experiments. Fat absorption was markedly impaired in the exclusion of bile from the intestine. Each function of fat and protein absorption was independently detected. These animal experiments gave the clinical evaluation of this test for fat and protein digestion-absorption function. Clinical investigations. The test meal was prepared without using commercial Tween 80 as an emulgent. Either fat or protein absorption was markedly impaired in some patients with various diseases. Inpatients with various diseases investigated were divided into 3 groups by correlation coefficient between fecal excretion ratios of fat and protein. Each function of fat and protein digestion-absorption was able to be detected independently by this test simultaneously. This test can further be used as a labolatory aid in evaluating fat and protein digestion-absorption function in patients with malabsorption syndrome. (author)

  5. Acid and rennet gels exhibit strong differences in the kinetics of milk protein digestion and amino acid bioavailability.

    Science.gov (United States)

    Barbé, Florence; Ménard, Olivia; Le Gouar, Yann; Buffière, Caroline; Famelart, Marie-Hélène; Laroche, Béatrice; Feunteun, Steven Le; Rémond, Didier; Dupont, Didier

    2014-01-15

    This study aimed at determining the kinetics of milk protein digestion and amino acid absorption after ingestion by six multi-canulated mini-pigs of two gelled dairy matrices having the same composition, similar rheological and structural properties, but differing by their mode of coagulation (acidification/renneting). Duodenal, mid-jejunal effluents and plasma samples were collected at different times during 7h after meal ingestion. Ingestion of the acid gel induced a peak of caseins and ?-lactoglobulin in duodenal effluents after 20min of digestion and a peak of amino acids in the plasma after 60min. The rennet gel induced lower levels of both proteins in the duodenum (with no defined peak) as well as much lower levels of amino acids in the plasma than the acid gel. Plasma ghrelin concentrations suggested a potentially more satiating effect of the rennet gel compared to the acid gel. This study clearly evidences that the gelation process can significantly impact on the nutritive value of dairy products. PMID:24054204

  6. A novel protein mixture containing vegetable proteins renders enteral nutrition products non-coagulating after in vitro gastric digestion (online first)

    OpenAIRE

    Braak, C. C. M.; Klebach, M.; Abrahamse, E.; Minor, M.; Knol, J.; Hofman, Z.; Ludwig, T.

    2012-01-01

    Background & aims: Non-coagulation of protein from enteral nutrition (EN) in the stomach is considered to improve gastric emptying and may result in reduced upper gastrointestinal complications such as reflux and aspiration pneumonia. For the development of a new EN protein mixture with reduced gastric coagulation, the coagulating properties of individual proteins, a novel blend of four proteins (P4 protein blend) and commercial EN products were investigated. Methods: A semi-dynamic, computer...

  7. Determination of denaturated proteins and biotoxins by on-line size-exclusion chromatography-digestion-liquid chromatography-electrospray mass spectrometry.

    Science.gov (United States)

    Carol, Jeroen; Gorseling, Maarten C J K; de Jong, Camiel F; Lingeman, Henk; Kientz, Charles E; van Baar, Ben L M; Irth, Hubertus

    2005-11-01

    A multidimensional analytical method for the rapid determination and identification of proteins has been developed. The method is based on the size-exclusion fractionation of protein-containing samples, subsequent on-line trypsin digestion and desalination, and reversed-phase high-performance liquid chromatography-electrospray mass spectrometry detection. The present system reduces digestion times to 20 min and the total analysis time to less than 100 min. Using bovine serum albumin and myoglobin as model proteins, optimization of key parameters such as digestion times and interfacing conditions between the different pretreatment steps was performed. The automated system was tested for the identification of infectious disease agents such as cholera toxin and staphylococcal enterotoxin B. This resulted typically in a positive identification by a total sequence coverage of approximately 40%. PMID:16185643

  8. Gut health immunomodulatory and anti-inflammatory functions of gut enzyme digested high protein micro-nutrient dietary supplement-Enprocal

    Directory of Open Access Journals (Sweden)

    Kanwar Rupinder K

    2009-01-01

    Full Text Available Abstract Background Enprocal is a high-protein micro-nutrient rich formulated supplementary food designed to meet the nutritional needs of the frail elderly and be delivered to them in every day foods. We studied the potential of Enprocal to improve gut and immune health using simple and robust bioassays for gut cell proliferation, intestinal integrity/permeability, immunomodulatory, anti-inflammatory and anti-oxidative activities. Effects of Enprocal were compared with whey protein concentrate 80 (WPC, heat treated skim milk powder, and other commercially available milk derived products. Results Enprocal (undigested and digested (Enprocal D selectively enhanced cell proliferation in normal human intestinal epithelial cells (FHs74-Int and showed no cytotoxicity. In a dose dependent manner Enprocal induced cell death in Caco-2 cells (human colon adencarcinoma epithelial cells. Digested Enprocal (Enprocal D: gut enzyme cocktail treated maintained the intestinal integrity in transepithelial resistance (TEER assay, increased the permeability of horseradish peroxidase (HRP and did not induce oxidative stress to the gut epithelial cells. Enprocal D upregulated the surface expression of co-stimulatory (CD40, CD86, CD80, MHC I and MHC II molecules on PMA differentiated THP-1 macrophages in coculture transwell model, and inhibited the monocyte/lymphocyte (THP-1/Jurkat E6-1 cells-epithelial cell adhesion. In cytokine secretion analyses, Enprocal D down-regulated the secretion of proinflammatory cytokines (IL-1? and TNF-? and up-regulated IFN-?, IL-2 and IL-10. Conclusion Our results indicate that Enprocal creates neither oxidative injury nor cytotoxicity, stimulates normal gut cell proliferation, up regulates immune cell activation markers and may aid in the production of antibodies. Furthermore, through downregulation of proinflammatory cytokines, Enprocal appears to be beneficial in reducing the effects of chronic gut inflammatory diseases such as inflammatory bowel disease (IBD. Stimulation of normal human fetal intestinal cell proliferation without cell cytotoxicity indicates it may also be given as infant food particularly for premature babies.

  9. Studies on the digestibility of microbial cell protein in buffalo calves

    International Nuclear Information System (INIS)

    The digestibility of cell wall, cell contents and mixed whole 35S-rumen bacteria and 14C-protozoal cells was determined in buffalo calves. The intact cells as well as their fractions were injected in a single dose into the rumen after treating with formaldehyde to protect degradation in the rumen and radioactivity excreted in the faeces was measured for six consecutive days. The indigestibility of mixed whole rumen bacteria, cell wall and cell contents was observed to be 87.13 +- 0.61, 92.77 +- 0.83 and 83.62 +- 1.10 percent, respectively. The digestibility coefficients of mixed whole rumen protozoa, cell wall, cell content and Holotrics were 90.95 +- 0.39, 94.39 +- 1.08, 89.01 +- 0.97 and 93.16 +- 0.69 percent, respectively. (auth.)

  10. Rapid and Efficient Protein Digestion using Trypsin Coated Magnetic Nanoparticles under Pressure Cycles

    OpenAIRE

    Lee, Byoungsoo; Lopez-ferrer, Daniel; Kim, Byoung Chan; Na, Hyon Bin; Park, Yong Il; Weitz, Karl K.; Warner, Marvin G.; Hyeon, Taeghwan; Lee, Sang-won; Smith, Richard D.; Kim, Jungbae

    2010-01-01

    Trypsin-coated magnetic nanoparticles (EC-TR/NPs), prepared via a simple multilayer random crosslinking of the trypsin molecules onto magnetic nanoparticles, were highly stable and could be easily captured using a magnet after the digestion was complete. EC-TR/NPs showed a negligible loss of trypsin activity after multiple uses and continuous shaking, while the conventional immobilization of covalently-attached trypsin on NPs resulted in a rapid inactivation under the same conditions due to t...

  11. Highly stable trypsin-aggregate coatings on polymer nanofibers for repeated protein digestion

    OpenAIRE

    Kim, Byoung Chan; Lopez-ferrer, Daniel; Lee, Sang-mok; Ahn, Hye-kyung; Nair, Sujith; Kim, Seong H.; Kim, Beom Soo; Petritis, Konstantinos; Camp, David G.; Grate, Jay W.; Smith, Richard D.; Koo, Yoon-mo; Gu, Man Bock; Kim, Jungbae

    2009-01-01

    A stable and robust trypsin-based biocatalytic system was developed and demonstrated for proteomic applications. The system utilizes polymer nanofibers coated with trypsin aggregates for immobilized protease digestions. After covalently attaching an initial layer of trypsin to the polymer nanofibers, highly concentrated trypsin molecules are crosslinked to the layered trypsin by way of a glutaraldehyde treatment. This process produced a 300-fold increase in trypsin activity compared with a co...

  12. High-sensitivity LC-MS/MS quantification of peptides and proteins in complex biological samples: the impact of enzymatic digestion and internal standard selection on method performance.

    Science.gov (United States)

    Bronsema, Kees J; Bischoff, Rainer; van de Merbel, Nico C

    2013-10-15

    Two important aspects of peptide and protein quantification by LC-MS/MS, the enzymatic digestion step and the internal standardization approach, were systematically investigated with a small protein, salmon calcitonin, which could be analyzed both without and with digestion. Quantification of undigested salmon calcitonin, after solid-phase extraction from plasma, resulted in a lower limit of quantification of 10 pg/mL, while introduction of a tryptic digestion step, followed by quantification of a signature peptide, increased this to 50 pg/mL. The sensitivity was reduced by interferences in the selected reaction monitoring (SRM) transition of the signature peptide due to the increase in sample complexity caused by the digestion and a less selective SRM transition of the signature peptide as compared to undigested salmon calcitonin. Eight internal standardization approaches were compared with respect to accuracy and precision in workflows with and without digestion. Analogue and stable-isotope-labeled (SIL) internal standards were evaluated including an in-house created (18)O-labeled peptide, a cleavable SIL peptide, and an internal standard created by differential derivatization of the signature peptide. We conclude that the best internal standard for the workflows both with and without digestion was the SIL form of the analyte, although the use of several SIL signature peptides and a differentially derivatized signature peptide also resulted in methods with performances which meet the FDA guidelines. PMID:24010948

  13. Improvement of protein content in winter wheat with physicochemical mutagens

    International Nuclear Information System (INIS)

    Induced mutation is one of the approaches to improve the protein content of wheat. In present experiments the wheat mutation contained high protein, gluten and lysine were obtained by means of treating dry seeds with different physicochemical mutation. The procedure of breeding for improving the qiality of wheat was found

  14. Cryoconservation of peptide extracts from trypsin digestion of proteins for proteomic analysis in a hospital biobank facility.

    Science.gov (United States)

    Mateos, Jesús; Pintor-Iglesias, Alejandra; Fernández-Puente, Patricia; García-Camba, Marta; Ruiz-Romero, Cristina; Doménech, Nieves; Blanco, Francisco J

    2014-04-01

    We tested a semiautomated protocol for the proper storage and conservation in a hospital biobank of tryptic peptide extracts coming from samples with low and high protein complexity for subsequent mass spectrometry analysis. Low-complexity samples (serum albumin, serotransferrin. and alpha-S1-casein) were loaded in replicates in SDS-PAGE and subjected to standard in-gel trypsin digestion. For LC-MALDI-TOF/TOF analysis, purified ?-galactosidase and human serum samples were in-solution digested following standard procedures and desalted with C18 stage-tips. In both cases, peptides extracts were aliquoted in individually 2D coded tubes, vacuum-dried, barcode-read, and stored in an automated -20 °C freezer in the Biobank facility. Samples were kept dried at -20 °C until the corresponding time-point of analysis, then reconstituted in the proper buffer and analyzed by either MALDI-TOF/TOF (peptide fingerprinting and MS/MS) or LC-MALDI-TOF/TOF following a highly reproducible pattern to ensure the reproducibility of the results. Protein identification was done with either Mascot or Protein Pilot as search engines using constant parameters. Over a period of 1 year we checked six different time points at days 0, 7, 30, 90, 180, and 365. We compared MS and MS/MS protein score, number of identified peptides, and coverage of the identified proteins. In the low complexity samples, the number of peptides detected gradually decreased over time, especially affecting the MS score. However, two of the three proteins - serum albumin and serotransferrin - were identified by both PMF and MS/MS at day 90. By day 180, only MS/MS identification in some replicates was possible. By LC-MS/MS, ?-galactosidase and the most abundant serum proteins were identified with good scores at all time points even by day 365, with no detectable peptide loss or decrease in the fragmentation efficiency, although a progressive decrease in peptide intensity indicates that detection of low abundant proteins could not be optimal after very long periods of time. Our results encourage us to use the biobank facility in the future for long-term storage - up to 3 months - of dried peptide extracts. PMID:24521361

  15. Nutritive value of digested cattle slurry from a biogas plant in sheep

    Energy Technology Data Exchange (ETDEWEB)

    Saxena, K.K.; Ranjhan, S.K.

    1983-01-01

    When animal waste is processed through an anaerobic digester, the quality and quantity of protein improves during fermentation because of the increase in the microbial population in the digested slurry, making the slurry a potential animal feed. Tests indicate that oven-dried as well as sun-dried slurry was palatable when mixed with wheat bran and could be fed to sheep in a maintenance ration of up to 30% slurry without any adverse effects. The digestible crude protein and total digestible nutrients of the slurry were 7 and 40%, respectively. Furthermore, the slurry showed no evidence of pathogenic organisms.

  16. Degradation of Cry1Ab protein from genetically modified maize (MON810) in relation to total dietary feed proteins in dairy cow digestion.

    Science.gov (United States)

    Paul, Vijay; Guertler, Patrick; Wiedemann, Steffi; Meyer, Heinrich H D

    2010-08-01

    To investigate the relative degradation and fragmentation pattern of the recombinant Cry1Ab protein from genetically modified (GM) maize MON810 throughout the gastrointestinal tract (GIT) of dairy cows, a 25 months GM maize feeding study was conducted on 36 lactating Bavarian Fleckvieh cows allocated into two groups (18 cows per group) fed diets containing either GM maize MON810 or nearly isogenic non-GM maize as the respective diet components. All cows were fed a partial total mixed ration (pTMR). During the feeding trial, 8 feed (4 transgenic (T) and 4 non-transgenic (NT) pTMR) and 42 feces (26 T and 18 NT) samples from the subset of cows fed T and NT diets, and at the end of the feeding trial, digesta contents of rumen, abomasum, small intestine, large intestine and cecum were collected after the slaughter of six cows of each feeding group. Samples were analyzed for Cry1Ab protein and total protein using Cry1Ab specific ELISA and bicinchoninic acid assay, respectively. Immunoblot analyses were performed to evaluate the integrity of Cry1Ab protein in feed, digesta and feces samples. A decrease to 44% in Cry1Ab protein concentration from T pTMR to the voided feces (9.40 versus 4.18 mug/g of total proteins) was recorded. Concentrations of Cry1Ab protein in GIT digesta of cows fed T diets varied between the lowest 0.38 mug/g of total proteins in abomasum to the highest 3.84 mug/g of total proteins in rumen. Immunoblot analysis revealed the extensive degradation of recombinant Cry1Ab protein into a smaller fragment of around 34 kDa in GIT. The results of the present study indicate that the recombinant Cry1Ab protein from MON810 is increasingly degraded into a small fragment during dairy cow digestion. PMID:19888668

  17. The influence of lupin seed germination on the chemical composition and standardized ileal digestibility of protein and amino acids in pigs.

    Science.gov (United States)

    Chilomer, K; Kasprowicz-Potocka, M; Gulewicz, P; Frankiewicz, A

    2013-08-01

    The germination process can modify the chemical composition of nutrients in seeds, which can influence the digestibility and utilization of sprouts in animal diets compared to raw seeds. The aims of research were to provide controlled germination process of lupin seeds, monitor the changes in seed composition and determine the influence of the germination on the coefficients of standardized ileal digestibility (SID) of crude protein and amino acids in growing pigs, compared to raw lupin seeds. The seeds of two lupin species were used: yellow (RYL) (Lupinus luteus, cv. Lord) and blue (RBL) (Lupinus angustifolius, cv. Graf). Germination was provided in the dark at 24°C for 4 days. Nutritional and antinutritional compositions of raw and germinated seeds (GYL and GBL, respectively) were analysed. Digestibility study was performed on pigs with an average body weight of 25 kg, and the pigs were surgically fitted with a T-cannula in the distal ileum, with chromic oxide as an indicator. Seed germination increased the crude protein and fibre concentrations, but reduced the levels of the ether extract, nitrogen-free extracts and all amino acids in protein. The content of alkaloids and raffinose family oligosaccharides decreased in both lupin species. Germination had no positive impact (p>0.05) on the SID of crude protein and amino acids. Germination of lupin seeds negatively influenced the SID of lysine and methionine (plupins compared to raw seeds; however, no positive effect was observed on the coefficients of the standardized ileal apparent digestibility of protein and amino acids. PMID:22540870

  18. Dietary protein hydrolysate and trypsin inhibitor effects on digestive capacities and performances during early-stages of spotted wolffish: suggested mechanisms.

    Science.gov (United States)

    Savoie, A; Le François, N R; Lamarre, S G; Blier, P U; Beaulieu, L; Cahu, C

    2011-04-01

    Growth rate is dependent upon adequate provision of amino acids especially in newly-hatched fish which experience very high growth rate. The replacement of a fraction of protein content by partially hydrolyzed (pre-digested) proteins was carried out and the digestive capacities and performances of larval/juvenile spotted wolffish (Anarhichas minor) were measured. The goal of this study was to verify whether the scope for growth is principally dictated by the proteolytic capacity of the digestive system by examining the effect of protein hydrolysates (PH) and trypsin inhibitor dietary inclusion on protein digestion/assimilation capacities, growth and survival. Four experimental diets were examined: C (control) I (supplemented with 750 mg/kg soybean trypsin inhibitor (SBTI)) H (supplemented with 20% PH) and HI (supplemented with 20% PH and 750 mg/kg SBTI). Protein hydrolysate supplementation gave significantly higher body mass than control at day 15 post-hatching. Unexpectedly, at day 30 and 60, fish administered diet HI (containing trypsin inhibitor) were heavier than the other groups. Suggested mechanisms are presented and discussed. The main conclusions of this study are that wolffish larval stage lasts roughly 15 days and that juvenile growth is linked to proteolytic capacity, but also very likely to absorption capacity of peptides and amino acids. PMID:21220041

  19. Digestibility and nitrogen balance of diets containing non conventional vegetable proteins fed to pigs of genetic strains suitable for outdoor systems

    Directory of Open Access Journals (Sweden)

    L. Sardi

    2010-04-01

    Full Text Available The study was carried out to evaluate on growing pigs kept in metabolism crates, either belonging to a “traditional” cross-breed (Duroc x Large White – DLW or a local breed (Cinta Senese – CS, the digestibility and the nitrogen balance of non-conventional vegetable protein sources (field beans, pea and sunflower cake when compared to soybean meal. The four diets were formulated so as to supply the same crude protein amount. CS pigs showed lower apparent digestibility coefficients (ADCs for crude protein (81,38% vs 82,65% and crude fibre (35,97% vs 40,65%. Nitrogen balance was more favourable in DLW pigs (32,12% vs 19,67% which showed a higher nitrogen retention (+69%.With respect to the protein source, no differences were observed for nitrogen balance. Sunflower diet showed the higher ADC for ether extract (+20% compared with soybean diet. Crude fibre digestibility was lowered in sunflower (-33% and field beans (-13% diets. Sunflower and field beans diets showed the lowest gross energy digestibility coefficients. Field pea diet had the highest ADC for crude fibre (+ 12% leading to ADCs for organic matter and gross energy comparable to those of soybean meal. It is concluded that field bean, sunflower cake and field pea could be considered as valuable non-conventional protein sources in swine nutrition.

  20. Matrix-assisted laser desorption ionization mass spectrometry imaging for peptide and protein analyses: a critical review of on-tissue digestion.

    Science.gov (United States)

    Cillero-Pastor, Berta; Heeren, Ron M A

    2014-02-01

    Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) has established itself among the plethora of mass spectrometry applications. In the biomedical field, MALDI-MSI is being more frequently recognized as a new method for the discovery of biomarkers and targets of treatment, classification of diseased and healthy tissues, or prediction of the outcome of a pathology. The technology has been used to study the localization of proteolytic peptides directly on tissue sections. A direct correlation between the detected peptides and the distribution and identity of the original precursor protein is the ultimate goal of any MALDI-MSI experiment. Enzymatic digestion protocols are commonly used to reveal the protein signature of these complex tissues. Considerations that pertain to methods of sample preparation, on-tissue digestion, data analysis, and visualization will be addressed. This review will also discuss selected applications of on-tissue digestion combined with the MALDI-MSI technology in biomedicine. PMID:24087847

  1. Effect of simulated processing on the antioxidant capacity and in vitro protein digestion of fruit juice-milk beverage model systems.

    Science.gov (United States)

    He, Zhiyong; Yuan, Bo; Zeng, Maomao; Tao, Guanjun; Chen, Jie

    2015-05-15

    The effects of simulated processing (pH adjustment and thermal treatment) on the antioxidant capacity and in vitro protein digestion of fruit juice-milk beverage (FJMB) models consisting of whey protein (WP), and chlorogenic acid (CHA) or catechin (CAT) were investigated. Results indicated that CAT was more susceptible to processing than CHA, and showed a significant (p antioxidant activity of FJMB. Pasteurization (63°C/30 min) and pH adjustment (pH 3.7 or pH 6.8) had either non-significant or slight effects on FJMB's antioxidant capacity, while sterilization significantly (p0.05) by pasteurization, whereas sterilization initially accelerated WP digestion but did not change its overall digestibility. PMID:25577106

  2. Phytase improves apparent total tract digestibility of phosphorus and calcium in piglets fed diets with adequate or reduced phosphorus content.

    Science.gov (United States)

    Kühn, I; Partanen, K

    2012-12-01

    The effect of a thermotolerant 6-phytase produced by Trichoderma reesei on performance and apparent total tract digestibility (ATTD) of P and Ca was evaluated in 192 weaned piglets (randomized block design; 16 replicates; 2 piglets each). Diets based on wheat (Triticum aestivum), barley (Hordeum vulgare), oat (Avena sativa), soybean (Glycine max) meal, and whey protein with adequate [positive control (PC)] or reduced [negative control (NC)] Ca and P levels were fed for 46 d after weaning. The PC and NC diets contained 8.0 and 6.4 g/kg Ca and 2.9 and 1.9 g/kg digestible P, respectively. Pelleted diets contained 0, 500, or 1000 phytase units (FTU)/kg. Growth performance and G:F were measured during starter (25 d) and weaner pig (21 d) periods. The ATTD of Ca and P was determined by spot sampling at the end of the weaner pig period (8 pens per treatment over 5 consecutive d). Data were analyzed using a mixed model with random block effect and fixed effect of dietary P and phytase level and their interaction. Dietary P level did not affect ADG or G:F of piglets over the entire feeding period (P > 0.10) whereas phytase increased G:F (P 0.10). A P × phytase interaction (P < 0.05) was observed for ATTD of P, more so for NC (48, 61, and 68%, respectively) than PC diets (52, 62, and 61%). The ATTD of Ca was higher (P < 0.05) for PC than NC diets (68 vs. 58%) and increased quadratically by phytase (61, 65, and 63%). In conclusion, the phytase tested enhanced piglet performance during the postweaning period and increased ATTD of P and Ca. PMID:23365327

  3. Different Dietary Levels of Protein to Lipid Ratio Affected Digestive Efficiency, Skeletal Growth, and Muscle Protein in Rainbow Trout Families

    OpenAIRE

    Torrissen, Krisna Rungruangsak; Stien, Lars Helge; Daae, Britt S.; Va?gseth, Tone; Thorsheim, Grethe B.; Tobin, Declan; Ritola, Ossi

    2009-01-01

    Normal Protein (NP) and High Protein (HP) diets were provided to rainbow trout families from juvenile to maturity. Muscle protein concentration increased during growth and the protein to lipid (P/L) ratio was doubled at late stage with respect to the dietary P/L ratio. The HP-diet fish showed higher protein deposition in body and white muscle, and had lower condition factor due to protein deposition associated more with body length than body weight. Fish growth rates were decreased at maturat...

  4. Influence of Processing on Dietary Fiber, Tannin and in Vitro Protein Digestibility of Pearl Millet

    Directory of Open Access Journals (Sweden)

    Florence Suma Pushparaj

    2011-10-01

    Full Text Available From the nutritional point of view, data on dietary fiber content, tannin and in vitro protein digestibility of processed millet is of importance, because millets are never eaten raw. Effects of commonly used traditional methods on dietary fiber, tannin content and %IVPD of two locally available pearl millet varieties (Kalukombu and Maharashtra Rabi Bajra were investigated. The millet was subjected to various processing methods like milling (whole flour, semi refined flour and bran rich fraction roasting, boiling, pressure cooking & germination respectively. Processing had little effect on the total dietary fiber (TDF content in both varieties; however the bran rich fraction showed highest TDF content of around 29%. Tannins effectively lowered upon boiling and pressure cooking respectively, but significantly increased (P ? 0.05 upon germination. Although the % IVPD of the millet (45.5 – 49.3 g/100g was low, it significantly increased upon milling (bran rich fraction, roasting and germination respectively.

  5. Short Term (14 Days Consumption of Insoluble Wheat Bran Fibre-Containing Breakfast Cereals Improves Subjective Digestive Feelings, General Wellbeing and Bowel Function in a Dose Dependent Manner

    Directory of Open Access Journals (Sweden)

    Louise Dye

    2013-04-01

    Full Text Available This study investigated whether increasing insoluble (predominantly wheat bran fibre over 14 days improves subjective digestive feelings, general wellbeing and bowel function. A single centre, multi-site, open, within subjects design with a 14 day non-intervention (baseline monitoring period followed by a 14 day fibre consumption (intervention period was performed. 153 low fibre consumers (<15 g/day AOAC 985.29 completed a daily symptom diary for 14 days after which they consumed one bowl of ready-to-eat breakfast cereal containing at least 5.4 g fibre (3.5 g from wheat bran for 14 days and completed a daily symptom diary. Significant improvements were demonstrated in subjective perception of bowel function (e.g., ease of defecation and digestive feelings (bloating, constipation, feeling sluggish and digestive discomfort. Significant improvements were also found in subjective perception of general wellbeing (feeling less fat, more mentally alert, slim, happy and energetic whilst experiencing less stress, mental and physical tiredness, difficulty concentrating and fewer headaches. In general, improvements in study outcomes increased with increasing cereal/fibre consumption. However, consuming an additional minimum 5.4 g of fibre (3.5 g wheat bran per day was shown to deliver measurable and significant benefits for digestive health, comfort and wellbeing. Encouraging consumption of relatively small amounts of wheat bran could also provide an effective method of increasing overall fibre consumption.

  6. SIMULATING PROTEIN DIGESTION ON TROUT A RAPID AND INEXPENSIVE METHOD FOR DOCUMENTING FISH MEAL QUALITY AND SCREENING NOVEL PROTEIN SOURCES FOR USE IN AQUAFEEDS

    OpenAIRE

    Bassompierre, M.; Kjar, A.; Ewen McLean

    1997-01-01

    A novel in vitro digestion system, which simulated rainbow trout gastric and intestinal digestion was developed. The method was employed to evaluate the impact of the gastric phase of digestion upon degradation of three fish meals od differing quality. Results illustrated that two-phase gastric-intestinal digestion increased the discriminatory powers of the system when compared to one-step intestinal digestion. A comparison of the system with pH-STAT methods demonstrated that the in vitro tec...

  7. Compositions and methods for improved protein production

    Science.gov (United States)

    Bodie, Elizabeth A. (San Carlos, CA); Kim, Steve (San Francisco, CA)

    2012-07-10

    The present invention relates to the identification of novel nucleic acid sequences, designated herein as 7p, 8k, 7E, 9G, 8Q and 203, in a host cell which effect protein production. The present invention also provides host cells having a mutation or deletion of part or all of the gene encoding 7p, 8k, 7E, 9G, 8Q and 203, which are presented in FIG. 1, and are SEQ ID NOS.: 1-6, respectively. The present invention also provides host cells further comprising a nucleic acid encoding a desired heterologous protein such as an enzyme.

  8. Compositions and methods for improved protein production

    Energy Technology Data Exchange (ETDEWEB)

    Bodie, Elizabeth A.; Kim, Steve Sungjin

    2014-06-03

    The present invention relates to the identification of novel nucleic acid sequences, designated herein as 7p, 8k, 7E, 9G, 8Q and 203, in a host cell which effect protein production. The present invention also provides host cells having a mutation or deletion of part or all of the gene encoding 7p, 8k, 7E, 9G, 8Q and 203, which are presented in FIG. 1, and are SEQ ID NOS.: 1-6, respectively. The present invention also provides host cells further comprising a nucleic acid encoding a desired heterologous protein such as an enzyme.

  9. Mass Spectrometry Characterization of the Thermal Decomposition/Digestion (TDD) at Cysteine in Peptides and Proteins in the Condensed Phase

    Science.gov (United States)

    Basile, Franco; Zhang, Shaofeng; Kandar, Sujit Kumar; Lu, Liang

    2011-11-01

    We report on the characterization by mass spectrometry (MS) of a rapid, reagentless and site-specific cleavage at the N-terminus of the amino acid cysteine (C) in peptides and proteins induced by the thermal decomposition at 220-250 °C for 10 s in solid samples. This thermally induced cleavage at C occurs under the same conditions and simultaneously to our previously reported thermally induced site-specific cleavage at the C-terminus of aspartic acid (D) (Zhang, S.; Basile, F. J. Proteome Res. 2007, 6, (5), 1700-1704). The C cleavage proceeds through cleavage of the nitrogen and ?-carbon bond (N-terminus) of cysteine and produces modifications at the cleavage site with an amidation (-1 Da) of the N-terminal thermal decomposition product and a -32 Da mass change of the C-terminal thermal decomposition product, the latter yielding either an alanine or ?-alanine residue at the N-terminus site. These modifications were confirmed by off-line thermal decomposition electrospray ionization (ESI)-MS, tandem MS (MS/MS) analyses and accurate mass measurements of standard peptides. Molecular oxygen was found to be required for the thermal decomposition and cleavage at C as it induced an initial cysteine thiol side chain oxidation to sulfinic acid. Similar to the thermally induced D cleavage, missed cleavages at C were also observed. The combined thermally induced digestion process at D and C, termed thermal decomposition/digestion (TDD), was observed on several model proteins tested under ambient conditions and the site-specificity of the method confirmed by MS/MS.

  10. Effects of Temperature during Moist Heat Treatment on Ruminal Degradability and Intestinal Digestibility of Protein and Amino Acids in Hempseed Cake.

    Science.gov (United States)

    Karlsson, L; Ruiz-Moreno, M; Stern, M D; Martinsson, K

    2012-11-01

    The objective of this study was to evaluate ruminal degradability and intestinal digestibility of crude protein (CP) and amino acids (AA) in hempseed cake (HC) that were moist heat treated at different temperatures. Samples of cold-pressed HC were autoclaved for 30 min at 110, 120 or 130°C, and a sample of untreated HC was used as the control. Ruminal degradability of CP was estimated, using the in situ Dacron bag technique; intestinal CP digestibility was estimated for the 16 h in situ residue using a three-step in vitro procedure. AA content was determined for the HC samples (heat treated and untreated) of the intact feed, the 16 h in situ residue and the residue after the three-step procedure. There was a linear increase in RUP (p = 0.001) and intestinal digestibility of RUP (p = 0.003) with increasing temperature during heat treatment. The 130°C treatment increased RUP from 259 to 629 g/kg CP, while intestinal digestibility increased from 176 to 730 g/kg RUP, compared to the control. Hence, the intestinal available dietary CP increased more than eight times. Increasing temperatures during heat treatment resulted in linear decreases in ruminal degradability of total AA (p = 0.006) and individual AA (pdegradability of total AA from 837 to 471 g/kg, while intestinal digestibility increased from 267 to 813 g/kg of rumen undegradable AA, compared with the control. There were differences between ruminal AA degradability and between intestinal AA digestibility within all individual HC treatments (p<0.001). It is concluded that moist heat treatment at 130°C did not overprotect the CP of HC and could be used to shift the site of CP and AA digestion from the rumen to the small intestine. This may increase the value of HC as a protein supplement for ruminants. PMID:25049517

  11. Evanescent wave-initiated photopolymerisation as a new way to create monolithic open-tubular capillary columns: use as enzymatic microreactor for on-line protein digestion.

    Czech Academy of Sciences Publication Activity Database

    Abele, S.; Smejkal, Petr; Yavorska, O.; Foret, František; Macka, M.

    2010-01-01

    Ro?. 135, ?. 3 (2010), s. 477-481. ISSN 0003-2654 R&D Projects: GA MŠk LC06023; GA AV ?R KAN400310651 Institutional research plan: CEZ:AV0Z40310501 Keywords : photopolymerisation * light- emitting -diode * on-line protein digestion Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 3.913, year: 2010

  12. In vitro digestion of short-dough biscuits enriched in proteins and/or fibres, using a multi-compartmental and dynamic system (1): Viscosity measurement and prediction.

    Science.gov (United States)

    Villemejane, C; Wahl, R; Aymard, P; Denis, S; Michon, C

    2015-09-01

    The effects of biscuit composition on the viscosity generated during digestion were investigated. A control biscuit, one with proteins, one with fibres, and one with both proteins and fibres were digested under the same conditions, using the TNO intestinal model (TIM-1). The TIM-1 is a multi-compartmental and dynamic in vitro system, simulating digestion in the upper tract (stomach and small intestine) of healthy adult humans. Digesta were collected at different times, in the different compartments of the TIM-1 (stomach, duodenum, jejunum and ileum) and viscosity was measured with a dynamic rheometer. Results showed a marked effect of biscuit composition on chyme viscosity. Highest viscosity was obtained with biscuits containing viscous soluble fibres, followed by those enriched in both proteins and fibres, then by protein-enriched and control biscuits. The viscosity was maintained throughout the gut up to the ileal compartment. A prediction of the evolution of the chyme viscosity in each compartment of the TIM-1 was built, based on model curves describing the evolution of the viscosity as a function of biscuit concentration, and on dilution factors measured by spectrophotometry on a blank digestion. PMID:25842308

  13. The effects of inulin supplementation of diets with or without hydrolysed protein sources on digestibility, faecal characteristics, haematology and immunoglobulins in dogs.

    Science.gov (United States)

    Verlinden, A; Hesta, M; Hermans, J M; Janssens, G P J

    2006-11-01

    Dogs with food allergy are often treated by giving a diet with hydrolysed protein sources. Prebiotics might also be successful in prevention and treatment of allergic disease through their effect on the colonic microflora, analogous to studies on probiotics in allergic children. The present study was set up to investigate the effect of supplementing inulin (IN) to commercial hypoallergenic dog diets on apparent nutrient digestibility, faecal characteristics, haematology and Ig in dogs. Supplementation of 3 % IN did not affect faecal pH, food and water intake and urine production. Compared with the intact protein diet with a limited number of ingredients (L), the diet with a hydrolysed protein source (H) resulted in an increased water intake (Pdogs fed the L+IN diet. Total faecal IgA concentrations were lower in the H group (P<0.05) because of lower antigenic stimulation of hydrolysed protein, which implies that hydrolysed protein is really hypoallergenic. The present study indicates that the use of hydrolysed protein diets for canine food allergy treatment can affect digestibility and that combination with IN affected apparent protein digestibility but not IgA response. PMID:17092385

  14. Development of a bioreactor based on trypsin immobilized on monolithic support for the on-line digestion and identification of proteins.

    Science.gov (United States)

    Calleri, E; Temporini, C; Perani, E; Stella, C; Rudaz, S; Lubda, D; Mellerio, G; Veuthey, J L; Caccialanza, G; Massolini, G

    2004-08-01

    The preparation and characterization of a new trypsin-based bioreactor is here described for on-line protein digestion and peptide analysis. Trypsin was immobilized on an epoxy-modified silica monolithic support with a single reaction step and the amount of immobilized enzyme was found to be 66.07 mg (+/-11.75 S.D.)/column (n = 6). The bioreactor was coupled through a switching valve to an analytical column for the on-line digestion, peptide separation and identification of test proteins by ESI-MS-MS. The influence of various parameters (flow rate, temperature, buffer pH and molarity, etc.) on enzymatic activity was investigated by an experimental design and the mostly significant factor was found to be the flow rate. The efficacy of the reported on-line bioreactor for tryptic mapping is reported for somatostatin and myoglobin, selected as model compounds. Tryptic peptide maps obtained by on-line digestion of myoglobin were compared to those obtained by traditional off-line digestion. Sequence coverage obtained with the on-line protocol (21 peptides, 75.16% coverage of myoglobin sequence) was found to be comparable to the one obtained with the off-line protocol (18 peptides, 76.47% coverage). Sensitivity for myoglobin digestion and identification was 0.1 mg/ml. The reproducibily of the peptide maps in terms of retention time was from 1.53 to 4.31%, R.S.D. PMID:15378884

  15. Digestibilidade de alimentos protéicos e energéticos para fêmeas de beta Digestibility of protein feedstuffs and energetic feedstuffs for female beta

    Directory of Open Access Journals (Sweden)

    Jener Alexandre Sampaio Zuanon

    2007-08-01

    Full Text Available Foram avaliados os coeficientes de digestibilidade aparente de MS, PB, energia bruta (EB e EE de alimentos protéicos (farelo de soja e farinha de peixe e energéticos (fubá de milho e farelo de trigo para beta (Betta splendens. Fêmeas adultas foram alojadas em gaiolas e mantidas em dois aquários de fibra de vidro (30 peixes/aquário para alimentação e dois para coleta de fezes, ambos de formato cônico e com capacidade para 30 L. Os resultados dos coeficientes de digestibilidade aparente de MS, PB, EB e EE foram, respectivamente, de 69,43; 72,52; 67,91 e 55,50% para farelo de soja; 60,67; 51,15; 75,55 e 58,26% para farinha de peixe; 63,88; 87,16; 77,61 e 50,40% para fubá de milho; e 61,06; 93,37; 58,17 e 65,51% para farelo de trigo. Os resultados obtidos neste estudo permitem otimizar a formulação de dietas práticas balanceadas, economicamente viáveis para a espécie.Apparent digestibility coefficients of dry matter, crude protein, gross energy and ether extract from protein feeds: soybean meal and fish meal and from energy feeds: corn and wheat middlings were evaluated for Siamese fighting fish (Betta splendens. Adult female fish were stocked in cages and held in two fiberglass aquaria (30 fish/aquarium for feeding and two aquaria for collect fecal samples, both with conic shape with 30L. The results of apparent digestibility coefficients for DM, CP, GE and EE were respectively 69.43, 72.52, 67.91 and 55.50% for soybean meal; 60.67, 51.15, 75.55 and 58.26% for fish meal; 63.88, 87.16, 77.61 and 50.40% for corn and 61.06, 93.37, 58.17 and 65.51% for wheat middlings. The determined digestibility values are essential to economically viable practical diet formulation for Siamese fish fighting.

  16. Effect on digestion and performance of dietary protein content and of increased substitution of lucerne hay with soya-bean protein concentrate in starter diets for young rabbits.

    Science.gov (United States)

    Chamorro, S; Gómez-Conde, M S; Pérez de Rozas, A M; Badiola, I; Carabaño, R; De Blas, J C

    2007-06-01

    The aim of this work was to study the effect of protein source / availability on the intestinal microbiota, digestive traits and nutritional performance of early-weaned rabbits. The effects of supplemental antibiotics in the drinking water were also evaluated. Four isoenergetic and isofibrous diets were formulated: a control diet with a high protein (207 g/kg dry matter (DM)) and lucerne hay content (HPHL), a diet with low crude protein (CP) (179 g/kg DM) and high lucerne hay content (LPHL) and low protein diets in which the lucerne hay in diet LPHL was replaced partially (LPML) or totally (LPLL) with soya-bean protein concentrate. Rabbits, weaned at 25 days (52 per diet), were fed the experimental diets for a 2-week period and thereafter received a commercial diet until 56 days of age. The incidence of mortality was investigated using 70 animals per diet without supplemental medication. The profile of the ileal microbiota was studied at 35 days of age in rabbits treated (18 per diet) or not (12 per diet) with antibiotic. As expected, supplementation with antibiotics effectively reduced fattening mortality rate and microbial biodiversity. However, lowering of also the dietary CP content led to a reduction in the mortality rate ( P rabbits and limit the numbers of potentially harmful bacteria in the lower gut. Modulation of dietary CP should be contemplated as a strategy to increase the intestinal health in rabbits. PMID:22444465

  17. Digestibilidade aparente de dietas práticas com diferentes relações energia: proteína em juvenis de pirarucu Apparent digestibility coefficient of practical diets with different energy: protein ratios for pirarucu juveniles

    Directory of Open Access Journals (Sweden)

    Eduardo Akifumi Ono

    2008-02-01

    Full Text Available O objetivo deste trabalho foi determinar a digestibilidade aparente de nutrientes e energia de dietas para juvenis de pirarucu, Arapaima gigas. Foram testadas oito dietas, contendo quatro relações energia:proteína (11, 10,1, 9, 8 kcal energia digestível por grama de proteína bruta e duas fontes de energia não-protéica (óleo de soja e gordura de aves, em esquema fatorial 4x2, com três repetições. Foram estocados 240 juvenis de pirarucu com peso de 96,8±2,3 g, distribuídos em 24 tanques cilíndricos com fundo cônico, adaptados para a coleta de fezes (sistema Guelph modificado. Os peixes foram alimentados duas vezes ao dia até a saciedade aparente com as dietas experimentais contendo 0,5% de óxido de cromo, como marcador inerte para determinação dos coeficientes de digestibilidade aparente. As dietas com a relação energia:proteína de 9 kcal energia digestível por grama de proteína bruta apresentaram os menores coeficientes de digestibilidade aparente da matéria seca, proteína bruta e extrativo não nitrogenado. O maior coeficiente de digestibilidade aparente da gordura foi obtido com o uso do óleo de soja. A relação energia:proteína na dieta influencia os coeficientes de digestibilidade aparente dos macronutrientes e energia no pirarucu.The objective of this work was to determine the apparent digestibility of nutrients and energy of diets for pirarucu (Arapaima gigas juveniles. Eight experimental diets containing four energy:protein ratios (11, 10.1, 9, 8 kcal digestible energy per gram of crude protein and two non-protein energy sources (soybean oil and poultry fat were tested in a 4x2 factorial scheme, in triplicates. Two hundred and forty pirarucu juveniles weighting 96.8±2.3 g were distributed in 24 cylindrical-conical fiberglass tanks, adapted for feces collection (modified Guelph system. Fish were fed twice a day to apparent satiation, with experimental diets containing 0.5% of chromium oxide as inert marker in order to determine the apparent digestibility coefficients. Diets containing an energy:protein ratio of 9 kcal digestible energy per gram of crude protein resulted in significantly lower apparent digestibility coefficient for dry matter, crude protein and non-nitrogenous extract. The highest apparent digestibility coefficients for crude fat was obtained with soybean oil. The dietary energy:protein ratio influences the nutrient and energy apparent digestibility coefficients to pirarucu juveniles.

  18. Digestibilidade aparente de dietas práticas com diferentes relações energia: proteína em juvenis de pirarucu / Apparent digestibility coefficient of practical diets with different energy: protein ratios for pirarucu juveniles

    Scientific Electronic Library Online (English)

    Eduardo Akifumi, Ono; Érica da Silva Santiago, Nunes; Jimmy Carlos Campos, Cedano; Manoel, Pereira Filho; Rodrigo, Roubach.

    2008-02-01

    Full Text Available O objetivo deste trabalho foi determinar a digestibilidade aparente de nutrientes e energia de dietas para juvenis de pirarucu, Arapaima gigas. Foram testadas oito dietas, contendo quatro relações energia:proteína (11, 10,1, 9, 8 kcal energia digestível por grama de proteína bruta) e duas fontes de [...] energia não-protéica (óleo de soja e gordura de aves), em esquema fatorial 4x2, com três repetições. Foram estocados 240 juvenis de pirarucu com peso de 96,8±2,3 g, distribuídos em 24 tanques cilíndricos com fundo cônico, adaptados para a coleta de fezes (sistema Guelph modificado). Os peixes foram alimentados duas vezes ao dia até a saciedade aparente com as dietas experimentais contendo 0,5% de óxido de cromo, como marcador inerte para determinação dos coeficientes de digestibilidade aparente. As dietas com a relação energia:proteína de 9 kcal energia digestível por grama de proteína bruta apresentaram os menores coeficientes de digestibilidade aparente da matéria seca, proteína bruta e extrativo não nitrogenado. O maior coeficiente de digestibilidade aparente da gordura foi obtido com o uso do óleo de soja. A relação energia:proteína na dieta influencia os coeficientes de digestibilidade aparente dos macronutrientes e energia no pirarucu. Abstract in english The objective of this work was to determine the apparent digestibility of nutrients and energy of diets for pirarucu (Arapaima gigas) juveniles. Eight experimental diets containing four energy:protein ratios (11, 10.1, 9, 8 kcal digestible energy per gram of crude protein) and two non-protein energy [...] sources (soybean oil and poultry fat) were tested in a 4x2 factorial scheme, in triplicates. Two hundred and forty pirarucu juveniles weighting 96.8±2.3 g were distributed in 24 cylindrical-conical fiberglass tanks, adapted for feces collection (modified Guelph system). Fish were fed twice a day to apparent satiation, with experimental diets containing 0.5% of chromium oxide as inert marker in order to determine the apparent digestibility coefficients. Diets containing an energy:protein ratio of 9 kcal digestible energy per gram of crude protein resulted in significantly lower apparent digestibility coefficient for dry matter, crude protein and non-nitrogenous extract. The highest apparent digestibility coefficients for crude fat was obtained with soybean oil. The dietary energy:protein ratio influences the nutrient and energy apparent digestibility coefficients to pirarucu juveniles.

  19. Digestive physiology of the pig symposium: G protein-coupled receptors in nutrient chemosensation and gastrointestinal hormone secretion.

    Science.gov (United States)

    Liou, A P

    2013-05-01

    The gastrointestinal tract is a highly effective and efficient organ system that digests and absorbs nutrients, contributes to the regulation of glucose homeostasis, and signals postprandial satiety. A network of enteroendocrine cells orchestrates these events through the release of neuropeptide hormones secreted in response to the specific nutrient components within the intraluminal milieu. Nutrient chemosensing by these cells is mediated by cell membrane proteins that have been localized to hormone-producing cells. However, functional studies of the nutrient detection abilities of the endocrine cell population have been limited due to its rare and singly distributed cell type. Recent technological advances have enabled investigations with primary endocrine cells that promise to enhance our current understanding of enteroendocrine cell biology. This review focuses on a particular subset of chemosensing receptors, the G protein-coupled receptors (GPCR), that have been identified as putative nutrient sensors of the major macronutrients, lipids, proteins, and carbohydrates by enteroendocrine cells. The contributions of these receptors in directly activating and stimulating hormone secretion in several subsets of enteroendocrine cells will be discussed, based on evidence gathered by functional studies in animal models, in vitro studies in endocrine cell lines, and newly described findings in primary endocrine cells. Key insights in chemosensory detection and hormone secretion from enteroendocrine cells may help further the studies in larger animal models and guide the formulation of feed or supplements to influence the gastrointestinal signals regulating optimal food intake, absorptive capacity, and growth. PMID:23230119

  20. Ambient DESI and LESA-MS analysis of proteins adsorbed to a biomaterial surface using in-situ surface tryptic digestion

    OpenAIRE

    Rao, Wei; Celiz, Adam D.; Scurr, David J.; Alexander, Morgan R.; Barrett, David A.

    2013-01-01

    The detection and identification of proteins adsorbed onto biomaterial surfaces under ambient conditions has significant experimental advantages but has proven to be difficult to achieve with conventional measuring technologies. In this study, we present an adaptation of desorption electrospray ionization (DESI) and liquid extraction surface analysis (LESA) mass spectrometry (MS) coupled with in-situ surface tryptic digestion to identify protein species from a biomaterial surface. Cytochrome ...

  1. Proteomic strategy for identifying mollusc shell proteins using mild chemical degradation and trypsin digestion of insoluble organic shell matrix: a pilot study on Haliotis tuberculata.

    Science.gov (United States)

    Bédouet, Laurent; Marie, Arul; Berland, Sophie; Marie, Benjamin; Auzoux-Bordenave, Stéphanie; Marin, Frédéric; Milet, Christian

    2012-08-01

    A successful strategy for the identification of shell proteins is based on proteomic analyses where soluble and insoluble fractions isolated from organic shell matrix are digested with trypsin with the aim of generating peptides, which are used to identify novel shell proteins contained in databases. However, using trypsin as a sole degradative agent is limited by the enzyme's cleavage specificity and is dependent upon the occurrence of lysine and arginine in the shell protein sequence. To bypass this limitation, we investigated the ability of trifluoroacetic acid (TFA), a low-specificity chemical degradative agent, to generate clusters of analyzable peptides from organic shell matrix, suitable for database annotation. Acetic acid-insoluble fractions from Haliotis tuberculata shell were processed by trypsin followed by TFA digestion. The hydrolysates were used to annotate an expressed sequence tag library constructed from the mantle tissue of Haliotis asinina, a tropical abalone species. The characterization of sequences with repeat motifs featured in some of the shell matrix proteins benefited from TFA-induced serial cutting, which can result in peptide ladder series. Using the degradative specificities of TFA and trypsin, we were able to identify five novel shell proteins. This pilot study indicates that a mild chemical digestion of organic shell matrix combined with trypsin generates peptides suitable for proteomic analysis for better characterization of mollusc shell matrix proteins. PMID:22160345

  2. Ambient DESI and LESA-MS Analysis of Proteins Adsorbed to a Biomaterial Surface Using In-Situ Surface Tryptic Digestion

    Science.gov (United States)

    Rao, Wei; Celiz, Adam D.; Scurr, David J.; Alexander, Morgan R.; Barrett, David A.

    2013-12-01

    The detection and identification of proteins adsorbed onto biomaterial surfaces under ambient conditions has significant experimental advantages but has proven to be difficult to achieve with conventional measuring technologies. In this study, we present an adaptation of desorption electrospray ionization (DESI) and liquid extraction surface analysis (LESA) mass spectrometry (MS) coupled with in-situ surface tryptic digestion to identify protein species from a biomaterial surface. Cytochrome c, myoglobin, and BSA in a combination of single and mixture spots were printed in an array format onto Permanox slides, followed by in-situ surface digestion and detection via MS. Automated tandem MS performed on surface peptides was able to identify the proteins via MASCOT. Limits of detection were determined for DESI-MS and a comparison of DESI and LESA-MS peptide spectra characteristics and sensitivity was made. DESI-MS images of the arrays were produced and analyzed with imaging multivariate analysis to automatically separate peptide peaks for each of the proteins within a mixture into distinct components. This is the first time that DESI and LESA-MS have been used for the in-situ detection of surface digested proteins on biomaterial surfaces and presents a promising proof of concept for the use of ambient MS in the rapid and automated analysis of surface proteins.

  3. Improved tagging strategy for protein identification in mammalian cells

    Directory of Open Access Journals (Sweden)

    Reiser Jakob

    2005-09-01

    Full Text Available Abstract Background The tagging strategy enables full-length endogenous proteins in mammalian cells to be expressed as green fluorescent fusion proteins from their authentic promoters. Results We describe improved genetic tools to facilitate protein tagging in mammalian cells based on a mobile genetic element that harbors an artificial exon encoding a protein tag. Insertion of the artificial exon within introns of cellular genes results in expression of hybrid proteins consisting of the tag sequence fused in-frame to sequences of a cellular protein. We have used lentiviral vectors to stably introduce enhanced green fluorescent protein (EGFP tags into expressed genes in target cells. The data obtained indicate that this strategy leads to bona fide tripartite fusion proteins and that the EGFP tag did not affect the subcellular localization of such proteins. Conclusion The tools presented here have the potential for protein discovery, and subsequent investigation of their subcellular distribution and role(s under defined physiological conditions, as well as for protein purification and protein-protein interaction studies.

  4. Improved Design of Anaerobic Digesters for Household Biogas Production in Indonesia: One Cow, One Digester, and One Hour of Cooking per Day

    OpenAIRE

    Usack, Joseph G.; Wiratni Wiratni; Angenent, Largus T.

    2014-01-01

    A government-sponsored initiative in Indonesia to design and implement low-cost anaerobic digestion systems resulted in 21 full-scale systems with the aim to satisfy the cooking fuel demands of rural households owning at least one cow. The full-scale design consisted of a 0.3?m diameter PVC pipe, which was operated as a conventional plug-flow system. The system generated enough methane to power a cooking stove for ?1?h. However, eventual clogging from solids accumulation inside the bior...

  5. Mucus digestion improves the detection of Chlamydia trachomatis and Neisseria gonorrhoeae on the cobas 4800.

    Science.gov (United States)

    Miller, Amanda; Bromhead, Collette; Jones, Mark; Tustin, Paul

    2012-09-01

    We compared the performance of cobas 4800 PCR for Neisseria gonorrhoeae with culture and found an association between pipetting errors and positive culture, caused by mucopurulent discharge. The problem may be resolved by pretreating samples with Sputasol, thus improving detection of both Chlamydia trachomatis and Neisseria gonorrhoeae. PMID:22902672

  6. Quantifying raft proteins in neonatal mouse brain by 'tube-gel' protein digestion label-free shotgun proteomics

    OpenAIRE

    Stephen, Tint G.; Halligan Brian; Li Man; Wakim Bassam; Yu Hongwei; Patel Shailendra B

    2007-01-01

    Abstract Background The low concentration and highly hydrophobic nature of proteins in lipid raft samples present significant challenges for the sensitive and accurate proteomic analyses of lipid raft proteins. Elimination of highly enriched lipids and interfering substances from raft samples is generally required before mass spectrometric analyses can be performed, but these procedures often lead to excessive protein loss and increased sample variability. For accurate analyses of the raft pr...

  7. Concentrados protéicos para bovinos: 2. Digestão pós-ruminal da matéria seca e da proteína / Protein concentrates for bovines: 2. Post-ruminal digestion of protein and dry matter

    Scientific Electronic Library Online (English)

    N.M., Rodriguez; J.F.C., Moreira; P.C.C., Fernandes; C.M., Veloso; E.O.S., Saliba; I., Borges; L.C., Gonçalves; A.L.C.C., Borges.

    2003-06-01

    Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese O objetivo deste trabalho foi avaliar a digestão pós-ruminal de fontes protéicas. Utilizou-se a técnica de sacos de náilon móveis introduzidos no duodeno e colhidos nas fezes. Os alimentos introduzidos no intestino foram anteriormente incubados em sacos de náilon no rúmen ou foram separados das fraç [...] ões solúveis em água. O farelo de soja foi a fonte protéica de maior degradabilidade ruminal (73%) e altíssima digestibilidade intestinal (98%). O glúten de milho foi a fonte protéica menos degradada no rúmen (16%) e sua parcela sobrepassante foi altamente digerida no intestino (95%). Os farelos de algodão e de mamona mostraram características semelhantes com digestibilidade total da proteína de 95% e em torno de 65% de degradabilidade ruminal. O farelo de palmiste foi menos digerido (87% digestibilidade total quando incubado). Farinhas de origem animal foram menos digeridas no intestino e também no rúmen (fora o glúten), com digestibilidades totais de 75% para farinha de carne e ossos, 85% para farinha de penas e vísceras e farinha de peixe I, 67% para farinha de peixe II e apenas 46% para farinha de sangue. Abstract in english This work aimed to evaluate post ruminal digestion of protein sources, by mobile nylon bags introduced in the duodenum of fistulated steers and collected from faeces. The feedstuffs were previously incubated in the rumen or separated the water soluble fraction before the intestinal digestion test. T [...] he soybean meal at 5% passage rate showed the highest ruminal protein degradability (73%) and highest intestinal digestibility (98%). Gluten meal was the least rumen degradable protein (16%) and showed a high digestible undegradable protein (95% if rumen incubated previously). Cottonseed meal and castorbean meal had similar patterns of digestion with total digestible protein of 95% and about 70% of rumen degradability. Rumen incubated palmist meal was less digestible (87% of total digestibility). Animal origin meals were less digestible either in the rumen or in the intestine, with total digestibility of 75% for meat and bone meal, 85% for feather and viscera meal and for fish meal I, 67 % for fish meal II and only 46% for blood meal.

  8. Improvement of protein quality in grain legumes. An overview on mutational improvement of protein quality in pigeon pea

    International Nuclear Information System (INIS)

    Grain legumes provide an essential part of the diet in developing countries in terms of protein. Besides increasing production also improving the grain quality would be an important objective. The paper discusses the methodology for protein improvement in seeds of pigeon pea. Variety Hy-2 was irradiated with 20, 25, 30 and 35 Kr of gamma radiation and the M2 population was screened for total sulfur content

  9. Ileal digestibility of broilers fed diets supplemented with exogenous enzymes

    Directory of Open Access Journals (Sweden)

    Nei André Arruda Barbosa

    2014-12-01

    Full Text Available The experiment was conducted to evaluate the effect of enzyme efficiency in diets with and without nutrient reduction on the ileal digestibility of broilers. A total of 1440 male broiler chicks (Cobb ® were distributed in a completely randomized design with 4 treatments in a factorial 2 x 2 (two control diets versus supplemented or not with enzyme with 8 replicates of 45 birds in each experimental unit. The control diets were: positive control (with nutritional levels recommended in the literature for each phase and a negative control (with reduced energy, calcium and phosphorus. Enzyme supplementation consisted of a combination of enzymes, phytase (100g/t and complex of amylase, protease and xylanase (500g/t. We evaluated the digestibility of dry matter, crude protein and energy of birds of age 22 and 43 days old. The reduction of nutrients in the diets caused a reduction in the negative control digestible energy and protein digestibility, however, the addition of enzyme mix in the negative control increased the digestible energy in two phases, and the digestibility of protein at 22 days old. The addition of enzymes improved the digestibility of nutrients in diets with nutrient reduction.

  10. 'Stealth' lipid-based formulations: poly(ethylene glycol)-mediated digestion inhibition improves oral bioavailability of a model poorly water soluble drug.

    Science.gov (United States)

    Feeney, Orlagh M; Williams, Hywel D; Pouton, Colin W; Porter, Christopher J H

    2014-10-28

    For over 20years, stealth drug delivery has been synonymous with nanoparticulate formulations and intravenous dosing. The putative determinants of stealth in these applications are the molecular weight and packing density of a hydrophilic polymer (commonly poly(ethylene glycol) (PEG)) that forms a steric barrier at the surface of the nanoparticle. The current study examined the potential translation of the concepts learned from stealth technology after intravenous administration to oral drug delivery and specifically, to enhance drug exposure after administration of oral lipid-based formulations (LBFs) containing medium-chain triglycerides (MCT). MCT LBFs are rapidly digested in the gastrointestinal tract, typically resulting in losses in solubilisation capacity, supersaturation and drug precipitation. Here, non-ionic surfactants containing stealth PEG headgroups were incorporated into MCT LBFs in an attempt to attenuate digestion, reduce precipitation risk and enhance drug exposure. Stealth capabilities were assessed by measuring the degree of digestion inhibition that resulted from steric hindrance of enzyme access to the oil-water interface. Drug-loaded LBFs were assessed for maintenance of solubilising capacity during in vitro digestion and evaluated in vivo in rats. The data suggest that the structural determinants of stealth LBFs mirror those of parenteral formulations, i.e., the key factors are the molecular weight of the PEG in the surfactant headgroup and the packing density of the PEG chains at the interface. Interestingly, the data also show that the presence of labile ester bonds within a PEGylated surfactant also impact on the stealth properties of LBFs, with digestible surfactants requiring a PEG Mw of ~1800g/mol and non-digestible ether-based surfactants ~800g/mol to shield the lipidic cargo. In vitro evaluation of drug solubilisation during digestion showed stealth LBFs maintained drug solubilisation at or above 80% of drug load and reduced supersaturation in comparison to digestible counterparts. This trend was also reflected in vivo, where the relative bioavailability of drug after administration in two stealth LBFs increased to 120% and 182% in comparison to analogous digestible (non-stealth) formulations. The results of the current study indicate that self-assembled "stealth" LBFs have potential as a novel means of improving LBF performance. PMID:25058571

  11. Digestibility of Nutrients on Broiler Given Various Protein Sources and Two Levels of Quebracho Tannin

    OpenAIRE

    Rusdi

    2006-01-01

    Suatu penelitian telah dilaksanakan yang bertujuan untuk melihat pengaruh sumber protein dan kandungan tannin dalam ransum terhadap kecernaan nutrien pada ayam broiler. Sebanyak enampuluh empat ayam telah digunakan dan ditempatkan secara acak dalam faktorial 2x4 sebanyak delapan ulangan. Protein yang digunakan meliputi gelatin, tepung ikan, tepung bulu dan campuran tepung daging dan kedelai. Hasil penelitian menunjukkan bahwa Quebracho Tanin (QT) menekan kecernaan nutrien pada broiler. Su...

  12. [Protein and energy value of spiruline blue algae supplemented by amino acids: digestive and metabolic utilization by the growing rat].

    Science.gov (United States)

    Vermorel, M; Toullec, G; Dumond, D; Pion, R

    1975-01-01

    Protein and energy value of 6 samples of "Spirulina" was studied on growing rats in 1972 and 1973. Sample RL 1(Spirulina platensis, originating from Tchad) was grown in artifical conditions in a laboratory. Others samples (Spirulina maxima) were grown in the solar evaporator near Mexico, washed and dried either on heated rollers (MR8, MR13) or by spraying (MA 7, MA10). Sample MA10 D corresponds to sample MA10, bleached by ethanol plus acetone (Baron, 1975). Each Spirulina sample was the only protein source of balanced, starch diets. The diets were supplemented in essential amino acids (E.A.A.) according to the requirements of growing rats (table 1). The ratios [(digestible nitrogen/metabolisable energy (EM] of the Spirulina diets were similar to that of the control diets containing herring meal. The diets were fed to groups of 15 to 17 growing rats. Energy and nitrogen balances were established by the comparative slaughter technique. Blood and muscle samples were taken at slaughter for the determination of free amino acids levels. PMID:824996

  13. The effect of dent versus flint maize genotype on site and the extent of starch and protein digestion, ruminal fermentation and microbial protein synthesis in the rumen of sheep

    OpenAIRE

    Verbic, Joz?e; Babnik, Drago; Z?nidars?ic-pongrac, Vida; Resnik, Mojca; Gregorcic, Ana; Kmecl, Veronika

    2005-01-01

    Ruminal and whole tract digestibility of protein and starch, microbial protein yield in the rumen and molar proportions of volatile fatty acids (VFA) in the rumen fluid and faeces were determined in sheep that were given either maize silage derived from the dent (DTS) or flint type hybrid (FTS). Degradabilities in the rumen were determined by means of the in sacco method using three sheep and microbial protein yield in the rumen by means of urinary purine derivative excretion using four sheep...

  14. A two-stage spin cartridge for integrated protein precipitation, digestion and SDS removal in a comparative bottom-up proteomics workflow.

    Science.gov (United States)

    Crowell, Andrew M J; MacLellan, Dawn L; Doucette, Alan A

    2014-10-12

    Protein precipitation with organic solvent is an effective means of depleting contaminants such as sodium dodecyl sulfate (SDS), while maintaining high analyte recovery. Here, we report the use of a disposable two-stage spin cartridge to facilitate isolation of the precipitated protein, with subsequent enzyme digestion and peptide cleanup in the cartridge. An upper filtration cartridge retains over 95% of the protein (10?g BSA), with 99.75% detergent depleted from a sample initially containing 2% SDS. Following precipitation, a plug attached to the base of the filtration cartridge retains the solution to enable tryptic digestion in the vial, while a solid phase extraction cartridge attached to the base of the filter facilitates peptide cleanup post-digestion. A GELFrEE fractionated Escherichia coli proteome extract processed with the spin cartridge yields similar protein identifications compared to controls (226 vs 216 for control), and with an increased number of unique peptides (1753 vs 1554 for control). The device is applied to proteome characterization of rat kidneys experiencing a surgically induced ureteral tract obstruction, revealing several statistically altered proteins, consistent with the morphology and expected pathophysiology of the disease. PMID:25316050

  15. P164-M Optimization of the Gel Shredder: A Centrifugal Device for Automated In-gel Digestion with Improved Peptide Recovery.

    OpenAIRE

    Lazarev, A.; Rejtar, T.; Karger, B. L.

    2007-01-01

    Proteomic workflow frequently involves gel separations. Poor peptide recovery from in-gel digestion procedures is at large influenced by the properties of polyacrylamide gel, which restricts access of proteolytic enzymes to the protein substrate as well as prevents complete extraction of certain peptides. We have designed an optimized centrifugal device which allows integrated washing, destaining and shredding of gel bands into uniform blocks of controlled size, roughly 100 ?m, prior to the ...

  16. Polyacrylamide gel miniaturization improves protein visualization and autoradiographic detection

    International Nuclear Information System (INIS)

    Polyacrylamide gels shrink to one-quarter of their original area when soaked in a 50% (w/v) solution of polyethylene glycol. Gel miniaturization improves the contrast of protein bands, with four valuable consequences. (i) A 5- to 10-fold increase in sensitivity for Coomassie blue is observed. (ii) Gels are more durable; i.e., they resist tearing when wet and they do not crack during drying under vacuum. (iii) Shrunken gels give sharper photographic images and provide better interlane protein band comparisons. (iv) Condensed protein bands lead to an increased sensitivity for detecting low-abundance, radioactively-labeled proteins by fluorography

  17. Improvement of sewage sludge quality: Combination of ozone and anaerobic digestion processes for the removal of Polycyclic Aromatic Hydrocarbons (PAH5) present in sludge

    Energy Technology Data Exchange (ETDEWEB)

    Bernal-Martinez, A.; Carrere, H.; Patureau, D.; Delgenes, J.P.

    2003-07-01

    Land application of sewage sludge is a choice disposal route for sustainable development. However, sludge quality has to be controlled in order to limit potential risks owing to the accumulation in agricultural soil of trace metals or trace organic pollutants. Amongst these organic compounds, Polycyclic Aromatic Hydrocarbons (PAHs) are particularly monitored because of their toxic and carcinogenic properties. Previous results have shown the partial removal of PAHs (50%) during anaerobic sludge stabilisation. The aim of this study was to optimize PAHs removal by combining anaerobic digestion with ozonation. Ozonation of anaerobically digested sludge improved the PAHs removal rate (61%). An additional enhancement of the PAHs removal rate was obtained by the addition of hydrogen peroxide during ozonation process. Similar performances (up to 81%) ware achieved by the use of surfactants which improved PAHs solubility in the aqueous phase. Finally, in order to evaluate the feasibility of the combination of the anaerobic digestion with ozonation, the biodegradability of the digested sludge ozonized in presence of surfactants and H{sub 2}O{sub 2} was measured. (author)

  18. Intestinal digestibility of enriched-protein fodders measured by mobile bag incubated with or without pepsin-HCl and three-step techniques

    Scientific Electronic Library Online (English)

    P., Paengkoum; S., Traiyakun; S., Paengkoum.

    Full Text Available Ruminal, intestinal and total tract digestibility of dry matter (DM) and crude protein (CP) of leucaena (Leucaena leucocephala), Madras thorn (Pithecellobium dulce) and moringa (Moringa oleifera) fodders were measured in this study, using nylon bag and mobile bag techniques. Three cattle were fitted [...] with permanent rumen and duodenal cannulae. Intestinal digestibility was measured using the mobile nylon bag (MNB) technique with or without incubation in a pepsin-HCl solution, and a three-step in vitro technique. The rate of ruminal disappearances of DM and CP, and the potential degradation of CP from nylon bags of both Madras thorn and moringa fodders were significantly higher than that for leucaena fodder. Potential degradation (A+B) values of CP were 45.6%, 54.2% and 52.8% for leucaena, Madras thorn and moringa fodders, respectively. Average DM and CP digestibility in the intestine and total tract for both Madras thorn and moringa fodders were significantly higher than for leucaena fodder. Average digestibility of DM and CP in the intestine and total tract measured using MNB without pepsin-HCl solution was significantly lower than with pepsin-HCl and with the three-step methods. These data suggest that the results of in vivo and in vitro methods for estimating intestinal digestibility are similar, though in all methods the incubation in a pepsin-HCl solution is necessary.

  19. Facilitating protein disulfide mapping by a combination of pepsin digestion, electron transfer higher energy dissociation (EThcD), and a dedicated search algorithm SlinkS.

    Science.gov (United States)

    Liu, Fan; van Breukelen, Bas; Heck, Albert J R

    2014-10-01

    Disulfide bond identification is important for a detailed understanding of protein structures, which directly affect their biological functions. Here we describe an integrated workflow for the fast and accurate identification of authentic protein disulfide bridges. This novel workflow incorporates acidic proteolytic digestion using pepsin to eliminate undesirable disulfide reshuffling during sample preparation and a novel search engine, SlinkS, to directly identify disulfide-bridged peptides isolated via electron transfer higher energy dissociation (EThcD). In EThcD fragmentation of disulfide-bridged peptides, electron transfer dissociation preferentially leads to the cleavage of the S-S bonds, generating two intense disulfide-cleaved peptides as primary fragment ions. Subsequently, higher energy collision dissociation primarily targets unreacted and charge-reduced precursor ions, inducing peptide backbone fragmentation. SlinkS is able to provide the accurate monoisotopic precursor masses of the two disulfide-cleaved peptides and the sequence of each linked peptide by matching the remaining EThcD product ions against a linear peptide database. The workflow was validated using a protein mixture containing six proteins rich in natural disulfide bridges. Using this pepsin-based workflow, we were able to efficiently and confidently identify a total of 31 unique Cys-Cys bonds (out of 43 disulfide bridges present), with no disulfide reshuffling products detected. Pepsin digestion not only outperformed trypsin digestion in terms of the number of detected authentic Cys-Cys bonds, but, more important, prevented the formation of artificially reshuffled disulfide bridges due to protein digestion under neutral pH. Our new workflow therefore provides a precise and generic approach for disulfide bridge mapping, which can be used to study protein folding, structure, and stability. PMID:24980484

  20. Improved microwave-assisted wet digestion procedures for accurate Se determination in fish and shellfish by flow injection-hydride generation-atomic absorption spectrometry

    International Nuclear Information System (INIS)

    Accurate determination of Se in biological samples, especially fish and shellfish, by hydride generation techniques has generally proven troublesome owing to the presence of organoselenium that cannot readily converted into inorganic selenium under usual oxidising conditions. Further improvements in the oxidation procedures are needed so as to obtain accurate concentration values when this type of samples is analyzed. Microwave-assisted wet digestion (MAWD) procedures of seafood based on HNO3 or the mixture HNO3/H2O2 and further thermal reduction of the Se(VI) formed to Se(IV) were evaluated. These procedures were as follows: (I) without H2O2 and without heating to dryness; (II) without H2O2 and with heating to dryness; (III) with H2O2 and without heating to dryness; (IV) with H2O2 and with heating to dryness. In general, low recoveries of selenium are obtained for several marine species (e.g., crustaceans and cephalopods), which may be ascribed to the presence of Se forms mainly associated with nonpolar proteins and lipids. Post-digestion UV irradiation proved very efficient since not only complete organoselenium decomposition was achieved but also the final step required for prereduction of Se(VI) into Se(IV) (i.e. heating at 90 deg. C for 30 min in 6 M HCl) could be avoided. With the MAWD/UV procedure, the use of strong oxidising agentocedure, the use of strong oxidising agents (persuphate, etc.) or acids (e.g. perchloric acid) which are typically applied prior to Se determination by hydride generation techniques is overcome, and as a result, sample pre-treatment is significantly simplified. The method was successfully validated against CRM DOLT-2 (dogfish liver), CRM DORM-2 (dogfish muscle) and CRM TORT-2 (lobster hepatopancreas). Automated ultrasonic slurry sampling with electrothermal atomic absorption spectrometry was also applied for comparison. Total Se contents in ten seafood samples were established. Se levels ranged from 0.7 to 2.9 ?g g-1

  1. Effects of corn-based diet starch content and corn particle size on lactation performance, digestibility, and bacterial protein flow in dairy cows.

    Science.gov (United States)

    Fredin, S M; Ferraretto, L F; Akins, M S; Bertics, S J; Shaver, R D

    2015-01-01

    An experiment was conducted to determine the effects of dietary starch content in corn-based diets and corn particle size on lactation performance, nutrient digestibility, and bacterial protein flow in dairy cows using the omasal and reticular sampling technique. Eight ruminally cannulated lactating multiparous Holstein cows were used in a replicated 4×4 Latin square design with a 2×2 factorial arrangement of treatments. Treatments were fine (FG; mean particle size=552µm) and coarse (CG; 1,270µm) ground dry shelled corn in normal- (NS) and reduced- (RS) starch diets fed as total mixed rations. The NS and RS rations contained 27 and 18% starch (dry matter basis), respectively, and were formulated by partially replacing corn with soy hull pellets. Mean dry matter intake was unaffected by treatment (23.2kg/d). Cows fed NS diets produced 1.9kg/d more milk and 0.06kg/d more milk protein compared with cows fed RS diets. Cows fed NSFG and RSCG diets produced more fat-corrected milk than did cows fed NSCG and RSFG diets. Milk urea concentration was decreased for cows fed NS diets (12.4mg/dL) compared with RS diets (13.5mg/dL). Ruminal digestibility of neutral detergent fiber (NDF; % of NDF intake) determined by the omasal sampling technique was increased in cows fed RS diets compared with NS diets (43.4 vs. 34.9%), and total-tract digestibility of NDF (% of NDF intake) was increased in cows fed RS diets compared with those fed NS diets (50.1 vs. 43.1%). Ruminal digestibility of starch (% of starch intake) determined by the omasal sampling technique was greater in cows fed NS diets compared with those fed RS diets (85.6 vs. 81.6%). Total-tract starch digestion was increased in cows fed RS diets compared with those fed NS diets (96.9 vs. 94.6%) and in cows fed FG diets compared with those fed CG diets (98.0 vs. 93.5%). Bacterial protein flow was unaffected by treatment. The omasal and reticular sampling techniques resulted in similar treatment effects for nutrient flow and digestibility, although nutrient flow was lower and nutrient digestibility was greater in cows when sampled by the omasal technique compared with the reticular technique. Cows fed FG diets had greater ruminal propionate, lower acetate:propionate ratio, and lower pH. Feeding NS diets increased milk and protein yields and feeding finely ground corn increased ruminal propionate concentration. PMID:25468693

  2. Relationships between leucine and the pancreatic exocrine function for improving starch digestibility in ruminants.

    Science.gov (United States)

    Liu, K; Liu, Y; Liu, S M; Xu, M; Yu, Z P; Wang, X; Cao, Y C; Yao, J H

    2015-04-01

    Four Holstein heifers (215±7kg; means ± SD), fitted with one pancreatic pouch, duodenal re-entrant cannulas, and duodenal infusion catheters, were used in this experiment. In phase 1, the 24-h profile of pancreatic fluid was determined. Pancreatic fluid flow peaked 1h after feeding, but peaks of similar magnitude also occurred before the morning feed, necessitating 24-h collection of pancreatic fluid to estimate daily excretion. In phase 2, the effects of duodenal infusions of 0, 10, 20, or 30g of leucine on pancreatic fluid flow were determined in a 4×4 Latin square design. The leucine was infused for 12h in 2,500mL of the infusate, and samples of pancreatic fluid and jugular blood were collected in 1-h intervals from the beginning of the infusion for 36h. The results showed that the secretion rate of pancreatic fluid (mL/h) was significantly higher in 10-g leucine group than the other groups (mL/h). Protein concentration (mg/mL) in pancreatic fluid was elevated proportional to the amount of leucine infused. Leucine infusions increased both the concentration (U/mL) and secretion rate (U/h) of ?-amylase. Infusion of 10g of leucine also increased the secretion rates (U/h) of trypsin, chymotrypsin, and lipase, but did not change their concentrations. No significant effects of leucine infusions on plasma glucose and insulin concentrations were found. The results indicate that leucine could act as a nutrient signal to stimulate ?-amylase production and pancreatic exocrine function in dairy heifers. PMID:25648818

  3. Impact of immune system stimulation on the ileal nutrient digestibility and utilisation of methionine plus cysteine intake for whole-body protein deposition in growing pigs.

    Science.gov (United States)

    Rakhshandeh, Anoosh; Htoo, John K; Karrow, Neil; Miller, Stephen P; de Lange, Cornelis F M

    2014-01-14

    The impact of immune system stimulation (ISS) on the ileal nutrient digestibility and utilisation of dietary methionine plus cysteine (SAA) intake for whole-body protein deposition (PD) was evaluated in growing pigs. For this purpose, sixty barrows were used in two experiments: thirty-six pigs in Expt I and twenty-four pigs in Expt II. Pigs were feed restricted and assigned to five levels of dietary SAA allowance (three and two levels in Expt I and II, respectively) from SAA-limiting diets. Following adaptation, pigs at each dietary SAA level were injected with either increasing amounts of Escherichia coli lipopolysaccharide (ISS+; eight and six pigs per dietary SAA level in Expt I and II, respectively) or saline (ISS - ; four and six pigs in Expt I and II, respectively) while measuring the whole-body nitrogen (N) balance. After N-balance observations, pigs were euthanised, organs were removed and ileal digesta were collected for determining nutrient digestibility. Ileal digestibility of gross energy, crude protein and amino acids was not affected by ISS (P>0·20). ISS reduced PD at all levels of dietary SAA intake (P0·20). Physiological and metabolic changes associated with systemic ISS had no effect on the ileal digestibility of nutrients per se, but altered SAA requirements for PD in growing pigs. PMID:23803219

  4. Actinidin from kiwifruit (Actinidia deliciosa cv. Hayward) increases the digestion and rate of gastric emptying of meat proteins in the growing pig.

    Science.gov (United States)

    Montoya, Carlos A; Rutherfurd, Shane M; Olson, Trent D; Purba, Ajitpal S; Drummond, Lynley N; Boland, Mike J; Moughan, Paul J

    2014-03-28

    The present study aimed to investigate the effect of dietary actinidin on the kinetics of gastric digestion of beef muscle proteins and on the rate of stomach emptying in growing pigs. For this purpose, 120 pigs (mean body weight 28 (sd 2·9) kg) were fed beef muscle protein-based diets containing either actinidin (fresh green kiwifruit pulp or gold kiwifruit pulp supplemented with purified actinidin) or no actinidin (fresh gold kiwifruit pulp or green kiwifruit pulp with inactivated actinidin). Additionally, fifteen pigs were fed with a protein-free diet to determine the endogenous protein flow. Pigs were euthanised at exactly 0·5, 1, 3, 5 and 7 h postprandially (n 6 per time point for each kiwifruit diet and n 3 for protein-free diet). Stomach chyme was collected for measuring gastric retention, actinidin activity, individual beef muscle protein digestion based on SDS-PAGE and the degree of hydrolysis based on the appearance of free amino groups. The stomach emptying of DM and N was faster when actinidin was present in the diet (Pdigestion of beef muscle protein (P34 kDa; Pdigestion of several beef muscle proteins. PMID:24252432

  5. Antioxidant and antimicrobial activity of lecithin free egg yolk protein preparation hydrolysates obtained with digestive enzymes

    OpenAIRE

    Aleksandra Zambrowicz; Marta Pokora; Ewelina Eckert; Marek Szo?tysik; Anna D?browska; Józefa Chrzanowska; Tadeusz Trziszka

    2012-01-01

    ABSTRACT:Several biological activities have now been associated with egg protein- derived peptides, including antihypertensive, antimicrobial, immunomodulatory, anticancer and antioxidantactivities, highlighting the importance of these biopeptides in human health, and disease prevention and treatment. Special attention has been given to peptides with antioxidant and antimicrobial activities as a new source of natural preservatives in food industry. In this study, the antioxidant properti...

  6. Latex proteins from the plant Calotropis procera are partially digested upon in vitro enzymatic action and are not immunologically detected in fecal material.

    Science.gov (United States)

    Ramos, Márcio V; Aguiar, Valéria C; da Silva Xavier, Ana A; Lima, Michael W; Bandeira, Glaís P; Etchells, J Peter; Nogueira, Nádia A P; Alencar, Nylane M N

    2006-06-01

    Soluble proteins from the latex of Calotropis procera (LP) were investigated in vitro and in vivo for digestibility as the latex has previously been shown to produce considerable toxic effects on animals. The latex is also an important biologically active compound that displays antiinflammatory and antidiarrhea properties. The proteins were digested by the action of trypsin, pepsin or chemotrypsin as revealed by gel filtration and SDS-PAGE analysis. Furthermore, the full LP digestion was easily achieved by protease treatment. Rabbit polyclonal antibodies raised against LP failed to detect cross-reactive molecules in fecal material of experimental rats following 35 consecutive days of LP consumption in water. Similar patterns of electrophoresis were observed for the negligible amounts of protein observed in the fecal extracts of control and test animals. No death or toxic effects were observed among animals. Taken together these results suggest that harmful and toxic effects on animals of the latex from C. procera are present in its rubber and low molecular weight fractions rather than its protein content. PMID:16581200

  7. Exigência de treonina, com base no conceito de proteína ideal, de alevinos de tilápia-do-nilo / Digestible threonine requirement of Nile tilapia fingerlings using ideal protein concept

    Scientific Electronic Library Online (English)

    Marcos Antonio Delmondes, Bomfim; Eduardo Arruda Teixeira, Lanna; Juarez Lopes, Donzele; Moisés, Quadros; Felipe Barbosa, Ribeiro; Wagner Azis Garcia de, Araújo.

    2077-20-01

    Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese Avaliou-se o efeito dos níveis de treonina digestível, com base no conceito de proteína ideal, em rações com 1,35% de lisina digestível sobre o desempenho de alevinos de tilápia-do-nilo (Oreochromis niloticus). Utilizaram-se 432 alevinos revertidos de tilápia, linhagem tailandesa, com peso inicial d [...] e 1,64 ± 0,03 g, em delineamento inteiramente ao acaso, com seis tratamentos, cada um com seis repetições, e doze peixes por unidade experimental. Os tratamentos consistiram de cinco rações com diversas relações treonina:lisina digestível (69,0; 74,0; 79,0; 84,0 e 89,0%) e relação metionina + cistina:lisina digestível de 70,0% e uma ração com relação treonina:lisina digestível de 84,0% e de metionina + cistina:lisina digestível de 75,0%, todas isoenergéticas e isoprotéicas. Os peixes foram mantidos em 36 aquários de 130 L, dotados de abastecimento de água, temperatura controlada e aeração individuais, e alimentados à vontade, seis vezes ao dia, durante 30 dias. Avaliaram-se o desempenho, a composição corporal, as deposições de proteína e gordura e a eficiência de retenção de nitrogênio dos peixes. O consumo de ração, o teor de proteína corporal e a deposição de proteína corporal aumentaram de forma linear de acordo com a relação treonina:lisina na ração, contudo, não foram influenciados pelo aumento da relação metionina + cistina:lisina da ração. Os níveis de treonina total ou digestível que proporcionaram os melhores resultados de desempenho em alevinos de tilápia-do-nilo foram de 1,11 e 0,99%, enquanto os níveis que promoveram melhor deposição de proteína corporal foram de 1,43 e 1,28%, que correspondem às relações treonina:lisina total de 71,0 e 69,0% e treonina:lisina digestível de 90,0 e 89,0%. Abstract in english The effects of digestible threonine levels, based on ideal protein concept in diets with 1.35% of digestible lysine on Nile tilapia (Oreochromis niloticus) fingerlings performance. Four hundred thirty two reverted Nile tilapia, Thailand strain, with initial weight of 1.64 ± 0.03 g, were allotted to [...] completely randomized design, with six diets and six replications with twelve fishes each. The five diets, isoenergetic isoproteic, consisted of various threonine:digestible lysine ratio (69.0, 74.0, 79.0, 84.0, and 89.0%) and methionine + cystine:digestible lysine ratio of 70.0% and one diet with threonine:digestible lysine of 84.0% and methionine + cystine:digestible lysine of 75.0%. The fish were kept in 36 aquariums of 130 L, equipped with water supply, controlled temperature and individual aeration, and ad libitum fed six times a day for 30 days. Performance, body composition, protein and fat depositions and nitrogen retention efficiency of fishes were evaluated. Consumption of feed, body protein content and body protein deposition increased in a linear fashion in accordance to threonine:digestible lysine in the diet, however, was not affected by the increase of dietary methionine + cystine:digestible lysine. Total or digestible threonine levels that provided the best results of performance of Nile tilapia fingerlings were 1.11 and 0.99%, while the levels that promoted better body protein deposition were 1.43 and 1.28%, which correspond to threonine:total lysine ratio of 71.0 and 69.0% and threonine:digestible lysine of 90.0 and 89.0%.

  8. Exigência de treonina, com base no conceito de proteína ideal, de alevinos de tilápia-do-nilo Digestible threonine requirement of Nile tilapia fingerlings using ideal protein concept

    Directory of Open Access Journals (Sweden)

    Marcos Antonio Delmondes Bomfim

    2008-12-01

    Full Text Available Avaliou-se o efeito dos níveis de treonina digestível, com base no conceito de proteína ideal, em rações com 1,35% de lisina digestível sobre o desempenho de alevinos de tilápia-do-nilo (Oreochromis niloticus. Utilizaram-se 432 alevinos revertidos de tilápia, linhagem tailandesa, com peso inicial de 1,64 ± 0,03 g, em delineamento inteiramente ao acaso, com seis tratamentos, cada um com seis repetições, e doze peixes por unidade experimental. Os tratamentos consistiram de cinco rações com diversas relações treonina:lisina digestível (69,0; 74,0; 79,0; 84,0 e 89,0% e relação metionina + cistina:lisina digestível de 70,0% e uma ração com relação treonina:lisina digestível de 84,0% e de metionina + cistina:lisina digestível de 75,0%, todas isoenergéticas e isoprotéicas. Os peixes foram mantidos em 36 aquários de 130 L, dotados de abastecimento de água, temperatura controlada e aeração individuais, e alimentados à vontade, seis vezes ao dia, durante 30 dias. Avaliaram-se o desempenho, a composição corporal, as deposições de proteína e gordura e a eficiência de retenção de nitrogênio dos peixes. O consumo de ração, o teor de proteína corporal e a deposição de proteína corporal aumentaram de forma linear de acordo com a relação treonina:lisina na ração, contudo, não foram influenciados pelo aumento da relação metionina + cistina:lisina da ração. Os níveis de treonina total ou digestível que proporcionaram os melhores resultados de desempenho em alevinos de tilápia-do-nilo foram de 1,11 e 0,99%, enquanto os níveis que promoveram melhor deposição de proteína corporal foram de 1,43 e 1,28%, que correspondem às relações treonina:lisina total de 71,0 e 69,0% e treonina:lisina digestível de 90,0 e 89,0%.The effects of digestible threonine levels, based on ideal protein concept in diets with 1.35% of digestible lysine on Nile tilapia (Oreochromis niloticus fingerlings performance. Four hundred thirty two reverted Nile tilapia, Thailand strain, with initial weight of 1.64 ± 0.03 g, were allotted to completely randomized design, with six diets and six replications with twelve fishes each. The five diets, isoenergetic isoproteic, consisted of various threonine:digestible lysine ratio (69.0, 74.0, 79.0, 84.0, and 89.0% and methionine + cystine:digestible lysine ratio of 70.0% and one diet with threonine:digestible lysine of 84.0% and methionine + cystine:digestible lysine of 75.0%. The fish were kept in 36 aquariums of 130 L, equipped with water supply, controlled temperature and individual aeration, and ad libitum fed six times a day for 30 days. Performance, body composition, protein and fat depositions and nitrogen retention efficiency of fishes were evaluated. Consumption of feed, body protein content and body protein deposition increased in a linear fashion in accordance to threonine:digestible lysine in the diet, however, was not affected by the increase of dietary methionine + cystine:digestible lysine. Total or digestible threonine levels that provided the best results of performance of Nile tilapia fingerlings were 1.11 and 0.99%, while the levels that promoted better body protein deposition were 1.43 and 1.28%, which correspond to threonine:total lysine ratio of 71.0 and 69.0% and threonine:digestible lysine of 90.0 and 89.0%.

  9. MSACompro: improving multiple protein sequence alignment by predicted structural features.

    Science.gov (United States)

    Deng, Xin; Cheng, Jianlin

    2014-01-01

    Multiple Sequence Alignment (MSA) is an essential tool in protein structure modeling, gene and protein function prediction, DNA motif recognition, phylogenetic analysis, and many other bioinformatics tasks. Therefore, improving the accuracy of multiple sequence alignment is an important long-term objective in bioinformatics. We designed and developed a new method MSACompro to incorporate predicted secondary structure, relative solvent accessibility, and residue-residue contact information into the currently most accurate posterior probability-based MSA methods to improve the accuracy of multiple sequence alignments. Different from the multiple sequence alignment methods that use the tertiary structure information of some sequences, our method uses the structural information purely predicted from sequences. In this chapter, we first introduce some background and related techniques in the field of multiple sequence alignment. Then, we describe the detailed algorithm of MSACompro. Finally, we show that integrating predicted protein structural information improved the multiple sequence alignment accuracy. PMID:24170409

  10. Legume proteins, their nutritional improvement and screening techniques

    International Nuclear Information System (INIS)

    In assessing the nutritional limitation of legume proteins it is essential to consider both sulphur amino acids, methionine and cysteine. The possibility of using total seed sulphur as a criteria for screening for improved protein quality is discussed. In some species when relatively large amounts of S-methyl-cysteine are present, total sulphur determinations would be invalid unless that amino acid were extracted with ethanol before the sulphur determination. Methods for sulphur determination are discussed and evaluated. (author)

  11. Antioxidant and antimicrobial activity of lecithin free egg yolk protein preparation hydrolysates obtained with digestive enzymes

    Directory of Open Access Journals (Sweden)

    Aleksandra Zambrowicz

    2012-12-01

    Full Text Available ABSTRACT:Several biological activities have now been associated with egg protein- derived peptides, including antihypertensive, antimicrobial, immunomodulatory, anticancer and antioxidantactivities, highlighting the importance of these biopeptides in human health, and disease prevention and treatment. Special attention has been given to peptides with antioxidant and antimicrobial activities as a new source of natural preservatives in food industry. In this study, the antioxidant properties of the egg-yolk protein by-product (YP hydrolysates were evaluated based on their radical scavenging capacity (DPPH, Fe2+chelating effect and ferric reducing power (FRAP. Furthermore, antimicrobial properties of obtained hydrolysates against Bacillus species were studied. The degrees (DHs of hydrolysis for 4h hydrolysates were: 19.1%, 13.5% and 13.0%, for pepsin, chymotrypsin and trypsin, respectively. Pepsin was the most effective in producing the free amino groups (1410.3 ?molGly/g. The RP-HPLC profiles of the protein hydrolysates showed differences in the hydrophobicity of the generated peptides.Trypsin hydrolysate obtained after 4h reaction demonstrated the strongest DPPH free radical scavenging activity (0.85 µmol Troloxeq/mg. Trypsin and chymotrypsin hydrolysates obtained after 4h reaction exhibited 4 times higher ferric reducing capacity than those treated bypepsin. The hydrolysis products obtained from YP exhibited significant chelating activity. The 4h trypsin hydrolysate exhibited weak antimicrobial activity against B. subtilis B3; B. cereus B512; B. cereus B 3p and B. laterosporum B6.

  12. Improvement of microwave-assisted digestion of milk powder with diluted nitric acid using oxygen as auxiliary reagent

    Science.gov (United States)

    Bizzi, Cezar A.; Barin, Juliano S.; Garcia, Edivaldo E.; Nóbrega, Joaquim A.; Dressler, Valderi L.; Flores, Erico M. M.

    2011-05-01

    The feasibility of using diluted HNO 3 solutions under oxygen pressure for decomposition of whole and non-fat milk powders and whey powder samples has been evaluated. Digestion efficiency was evaluated by determining the carbon content in solution (digests) and the determination of Ca, Cd, Cu, Fe, K, Mg, Mn, Mo, Na, Pb and Zn was performed by inductively coupled plasma optical emission spectrometry and Hg by chemical vapor generation coupled to inductively coupled plasma mass spectrometry. Samples (up to 500 mg) were digested using HNO 3 solutions (1 to 14 mol L - 1 ) and the effect of oxygen pressure was evaluated between 2.5 and 20 bar. It was possible to perform the digestion of 500 mg of milk powder using 2 mol L - 1 HNO 3 with oxygen pressure ranging from 7.5 to 20 bar with resultant carbon content in digests lower than 1700 mg L - 1 . Using optimized conditions, less than 0.86 mL of concentrated nitric acid (14 mol L - 1 ) was enough to digest 500 mg of sample. The accuracy was evaluated by determination of metal concentrations in certified reference materials, which presented an agreement better than 95% (Student's t test, P < 0.05) for all the analytes.

  13. AMINO ACID METABOLISM IN COWS DURING THE TRANSITION PERIOD IN BALANCING DIET ON THE EXCHANGE PROTEIN AND DIGESTIBLE AMINO ACIDS

    Directory of Open Access Journals (Sweden)

    Ryadchikov V. G.

    2014-02-01

    Full Text Available Application of a factorial method for determining the needs in metabolic protein and essential amino acids, helps to deepen knowledge on physiology of protein and amino acid supply and allow to improve the standards for dairy cows during the transition period; in insufficient of metabolic protein and essential amino acids increased coefficients of their transformation into net protein and absorptive amino acids as a result of mobilization of body of cows; with an optimal protein nutrition their transformation in net milk protein, lysine and methionine accordingly amounted to 0.67, 0,83 and 0,82. The most significant changes in the concentration of methionine, proline, glutamate, glutamine, glycine were observed in cows before calving and immediately after birth, stabilization of their level starts with a 24 lactation day, that is connected with the peculiarities of the feeding behavior of the cows and the gradual intensification of the processes of metabolism and milk production. To control the status of protein metabolism we have offered benchmarks compositions of free amino acids in cows’ blood plasma phases: 21-0 days before calving, 0-21 and 22-120 days after calving

  14. Improved hybrid optimization algorithm for 3D protein structure prediction.

    Science.gov (United States)

    Zhou, Changjun; Hou, Caixia; Wei, Xiaopeng; Zhang, Qiang

    2014-07-01

    A new improved hybrid optimization algorithm - PGATS algorithm, which is based on toy off-lattice model, is presented for dealing with three-dimensional protein structure prediction problems. The algorithm combines the particle swarm optimization (PSO), genetic algorithm (GA), and tabu search (TS) algorithms. Otherwise, we also take some different improved strategies. The factor of stochastic disturbance is joined in the particle swarm optimization to improve the search ability; the operations of crossover and mutation that are in the genetic algorithm are changed to a kind of random liner method; at last tabu search algorithm is improved by appending a mutation operator. Through the combination of a variety of strategies and algorithms, the protein structure prediction (PSP) in a 3D off-lattice model is achieved. The PSP problem is an NP-hard problem, but the problem can be attributed to a global optimization problem of multi-extremum and multi-parameters. This is the theoretical principle of the hybrid optimization algorithm that is proposed in this paper. The algorithm combines local search and global search, which overcomes the shortcoming of a single algorithm, giving full play to the advantage of each algorithm. In the current universal standard sequences, Fibonacci sequences and real protein sequences are certified. Experiments show that the proposed new method outperforms single algorithms on the accuracy of calculating the protein sequence energy value, which is proved to be an effective way to predict the structure of proteins. PMID:25069136

  15. Total and partial digestibility, rates of digestion obtained with rumen evacuation and microbial protein synthesis in bovines fed fresh or ensiled sugar cane and corn silage / Digestibilidade total e parcial, taxas de digestão obtidas com o esvaziamento ruminal e síntese de proteína microbiana em bovinos alimentados com cana-de-açúcar fresca ou ensilada e silagem de milho

    Scientific Electronic Library Online (English)

    Gustavo Chamon de Castro, Menezes; Sebastião de Campos, Valadares Filho; Felipe Antunes, Magalhães; Rilene Ferreira Diniz, Valadares; Lays Débora, Mariz; Edenio, Detmann; Odilon Gomes, Pereira; Maria Ignez, Leão.

    1104-11-01

    Full Text Available Avaliaram-se os consumos, as digestibilidades ruminal e intestinal e as taxas de passagem e de digestão de nutrientes em bovinos alimentados com dietas constituídas de silagem de milho, cana-de-açúcar triturada e fornecida in natura, cana triturada e ofertada após 72 horas de armazenamento, cana-de- [...] açúcar ensilada com 1 % de cal e sem tratamento e um mesmo concentrado fixado em 1% do peso corporal. Todos os volumosos foram corrigidos com ureia/sulfato de amônio para conterem 10% de proteína bruta. Utilizaram-se cinco bovinos fistulados no rúmen, com peso médio de 240 kg ± 15 kg, distribuídos em um quadrado latino 5 × 5. Foram realizadas coletas totais de fezes, abomasal e dois esvaziamentos pela manhã. Os animais alimentados com dietas à base de silagem de milho apresentaram maiores digestibilidades ruminal da proteína e intestinal do extrato etéreo, taxas de ingestão, passagem e de digestão da matéria seca, justificando os maiores consumos de matéria seca e fibra em detergente neutro corrigida para cinzas e proteína (FDNcp). As maiores taxas de passagem nos animais alimentados com dietas à base de cana-de-açúcar in natura justificam os maiores consumos de matéria seca e FDNcp em relação ao observado com o fornecimento de silagens de cana-de-açúcar. Animais consumindo dietas contendo silagem de milho apresentam maiores taxas de passagem da matéria seca e digestão da FDNcp. Dietas contendo cana-de-açúcar in natura, armazenada ou não, favorecem o consumo e a taxa de passagem da matéria seca, em relação a dietas com cana ensilada. O uso de cal na ensilagem não melhora a digestibilidade dos nutrientes nem a taxa de passagem da dieta. A cana-de-açúcar armazenada por 72 horas possui características digestíveis semelhantes às da cana-de-açúcar in natura. Abstract in english It was evaluated intake, rumen and intestinal digestibility and passage and digestion rates in bovines fed diets constituted of corn silage, crushed sugar cane and given fresh, crushed sugar cane and given after 72 hours of storage, ensiled sugar cane with 1% of calcium oxide and with no treatment a [...] nd a same concentrate fixed at 1% of body weight. All roughage was corrected to contain 10% of crude protein. It was used five rumen-fistulated bovine with average weight of 240 ± 15 kg, distributed in a 5 × 5 Latin square. Abomasum and total fecal collection and two rumen evacuations were carried out in the morning. Animals fed corn silage based diet presented greater rumen digestibility of the protein and intestinal digestibility of the ether extract, greater intake and passage of dry matter, justifying greater intakes of dry matter and neutral detergent fiber corrected for protein and ash (NDFap). The greatest passage rates in animals fed fresh sugar cane based diet justify greater intakes of dry matter and NDFap in relation to the one observed with sugar cane silage supply. Animal consuming corn silage diets present greater dry matter passage rate and NDFap digestion. Diets with fresh sugar cane, stored or not, favor dry matter passage rate and intake, in relation to ensiled sugar cane. The use of calcium oxide in the ensilage does not improve nutrient digestibility neither passage rate of the diet. Sugar cane stored for 72 hours has digestible traits similar to the ones of fresh sugar cane.

  16. Total and partial digestibility, rates of digestion obtained with rumen evacuation and microbial protein synthesis in bovines fed fresh or ensiled sugar cane and corn silage Digestibilidade total e parcial, taxas de digestão obtidas com o esvaziamento ruminal e síntese de proteína microbiana em bovinos alimentados com cana-de-açúcar fresca ou ensilada e silagem de milho

    Directory of Open Access Journals (Sweden)

    Gustavo Chamon de Castro Menezes

    2011-05-01

    Full Text Available It was evaluated intake, rumen and intestinal digestibility and passage and digestion rates in bovines fed diets constituted of corn silage, crushed sugar cane and given fresh, crushed sugar cane and given after 72 hours of storage, ensiled sugar cane with 1% of calcium oxide and with no treatment and a same concentrate fixed at 1% of body weight. All roughage was corrected to contain 10% of crude protein. It was used five rumen-fistulated bovine with average weight of 240 ± 15 kg, distributed in a 5 × 5 Latin square. Abomasum and total fecal collection and two rumen evacuations were carried out in the morning. Animals fed corn silage based diet presented greater rumen digestibility of the protein and intestinal digestibility of the ether extract, greater intake and passage of dry matter, justifying greater intakes of dry matter and neutral detergent fiber corrected for protein and ash (NDFap. The greatest passage rates in animals fed fresh sugar cane based diet justify greater intakes of dry matter and NDFap in relation to the one observed with sugar cane silage supply. Animal consuming corn silage diets present greater dry matter passage rate and NDFap digestion. Diets with fresh sugar cane, stored or not, favor dry matter passage rate and intake, in relation to ensiled sugar cane. The use of calcium oxide in the ensilage does not improve nutrient digestibility neither passage rate of the diet. Sugar cane stored for 72 hours has digestible traits similar to the ones of fresh sugar cane.Avaliaram-se os consumos, as digestibilidades ruminal e intestinal e as taxas de passagem e de digestão de nutrientes em bovinos alimentados com dietas constituídas de silagem de milho, cana-de-açúcar triturada e fornecida in natura, cana triturada e ofertada após 72 horas de armazenamento, cana-de-açúcar ensilada com 1 % de cal e sem tratamento e um mesmo concentrado fixado em 1% do peso corporal. Todos os volumosos foram corrigidos com ureia/sulfato de amônio para conterem 10% de proteína bruta. Utilizaram-se cinco bovinos fistulados no rúmen, com peso médio de 240 kg ± 15 kg, distribuídos em um quadrado latino 5 × 5. Foram realizadas coletas totais de fezes, abomasal e dois esvaziamentos pela manhã. Os animais alimentados com dietas à base de silagem de milho apresentaram maiores digestibilidades ruminal da proteína e intestinal do extrato etéreo, taxas de ingestão, passagem e de digestão da matéria seca, justificando os maiores consumos de matéria seca e fibra em detergente neutro corrigida para cinzas e proteína (FDNcp. As maiores taxas de passagem nos animais alimentados com dietas à base de cana-de-açúcar in natura justificam os maiores consumos de matéria seca e FDNcp em relação ao observado com o fornecimento de silagens de cana-de-açúcar. Animais consumindo dietas contendo silagem de milho apresentam maiores taxas de passagem da matéria seca e digestão da FDNcp. Dietas contendo cana-de-açúcar in natura, armazenada ou não, favorecem o consumo e a taxa de passagem da matéria seca, em relação a dietas com cana ensilada. O uso de cal na ensilagem não melhora a digestibilidade dos nutrientes nem a taxa de passagem da dieta. A cana-de-açúcar armazenada por 72 horas possui características digestíveis semelhantes às da cana-de-açúcar in natura.

  17. Isolation and identification of a bacterium from marine shrimp digestive tract: A new degrader of starch and protein

    Science.gov (United States)

    Li, Jiqiu; Tan, Beiping; Mai, Kangsen

    2011-09-01

    It is a practical approach to select candidate probiotic bacterial stains on the basis of their special traits. Production of digestive enzyme was used as a trait to select a candidate probiotic bacterial strain in this study. In order to select a bacterium with the ability to degrade both starch and protein, an ideal bacterial strain STE was isolated from marine shrimp ( Litopenaeus vannamei) intestines by using multiple selective media. The selected isolate STE was identified on the basis of its morphological, physiological, and biochemical characteristics as well as molecular analyses. Results of degradation experiments confirmed the ability of the selected isolate to degrade both starch and casein. The isolate STE was aerobic, Gram-negative, rod-shaped, motile and non-spore-forming, and had catalase and oxidase activities but no glucose fermentation activity. Among the tested carbon/nitrogen sources, only Tween40, alanyl-glycine, aspartyl-glycine, and glycyl-l-glutamic acid were utilized by the isolate STE. Results of homology comparison analyses of the 16S rDNA sequences showed that the isolate STE had a high similarity to several Pseudoalteromonas species and, in the phylogenetic tree, grouped with P. ruthenica with maximum bootstrap support (100%). In conclusion, the isolate STE was characterized as a novel strain belonging to the genus Pseudoalteromonas. This study provides a further example of a probiotic bacterial strain with specific characteristics isolated from the host gastrointestinal tract.

  18. Optimization of Immobilized Gallium (III) Ion Affinity Chromatography for Selective Binding and Recovery of Phosphopeptides from Protein Digests

    Science.gov (United States)

    Aryal, Uma K.; Olson, Douglas J.H.; Ross, Andrew R.S.

    2008-01-01

    Although widely used in proteomics research for the selective enrichment of phosphopeptides from protein digests, immobilized metal-ion affinity chromatography (IMAC) often suffers from low specificity and differential recovery of peptides carrying different numbers of phosphate groups. By systematically evaluating and optimizing different loading, washing, and elution conditions, we have developed an efficient and highly selective procedure for the enrichment of phosphopeptides using a commercially available gallium(III)-IMAC column (PhosphoProfile, Sigma). Phosphopeptide enrichment using the reagents supplied with the column is incomplete and biased toward the recovery and/or detection of smaller, singly phosphorylated peptides. In contrast, elution with base (0.4 M ammonium hydroxide) gives efficient and balanced recovery of both singly and multiply phosphorylated peptides, while loading peptides in a strong acidic solution (1% trifluoracetic acid) further increases selectivity toward phosphopeptides, with minimal carryover of nonphosphorylated peptides. 2,5-Dihydroxybenzoic acid, a matrix commonly used when analyzing phosphopeptides by matrix-assisted laser desorption/ionization mass spectrometry was also evaluated as an additive in loading and eluting solvents. Elution with 50% acetonitrile containing 20 mg/mL dihydroxybenzoic acid and 1% phosphoric acid gave results similar to those obtained using ammonium hydroxide as the eluent, although the latter showed the highest specificity for phosphorylated peptides. PMID:19183793

  19. Phytic acid, in vitro protein digestibility, dietary fiber, and minerals of pulses as influenced by processing methods.

    Science.gov (United States)

    Chitra, U; Singh, U; Rao, P V

    1996-06-01

    The objective of this project was to determine the effect of various types of processing on selected nutrition related parameters of commonly consumed Indian pulses and soybean. Germination reduced the phytic acid content of chickpea and pigeonpea seeds by over 60%, and that of mung bean, urd bean, and soybean by about 40%. Fermentation reduced phytic acid contents by 26-39% in all these legumes with the exception of pigeonpea in which it was reduced by more than 50%. Autoclaving and roasting were more effective in reducing phytic acid in chickpea and pigeonpea than in urd bean, mung bean, and soybean. Germination and fermentation greatly increased the in vitro protein digestibility (IVPD). IVPD was only slightly increased by roasting and autoclaving of all legumes. Germination and fermentation also remarkably decreased the total dietary fiber (TDF) in all legumes. Autoclaving and roasting resulted in slight increases in TDF values. All the processing treatments had little effect on calcium, magnesium and iron contents. PMID:8983057

  20. Robust enzyme design: Bioinformatic tools for improved protein stability.

    Science.gov (United States)

    Suplatov, Dmitry; Voevodin, Vladimir; Švedas, Vytas

    2015-03-01

    The ability of proteins and enzymes to maintain a functionally active conformation under adverse environmental conditions is an important feature of biocatalysts, vaccines, and biopharmaceutical proteins. From an evolutionary perspective, robust stability of proteins improves their biological fitness and allows for further optimization. Viewed from an industrial perspective, enzyme stability is crucial for the practical application of enzymes under the required reaction conditions. In this review, we analyze bioinformatic-driven strategies that are used to predict structural changes that can be applied to wild type proteins in order to produce more stable variants. The most commonly employed techniques can be classified into stochastic approaches, empirical or systematic rational design strategies, and design of chimeric proteins. We conclude that bioinformatic analysis can be efficiently used to study large protein superfamilies systematically as well as to predict particular structural changes which increase enzyme stability. Evolution has created a diversity of protein properties that are encoded in genomic sequences and structural data. Bioinformatics has the power to uncover this evolutionary code and provide a reproducible selection of hotspots - key residues to be mutated in order to produce more stable and functionally diverse proteins and enzymes. Further development of systematic bioinformatic procedures is needed to organize and analyze sequences and structures of proteins within large superfamilies and to link them to function, as well as to provide knowledge-based predictions for experimental evaluation. PMID:25524647

  1. Predicting microbial protein synthesis in beef cattle: relationship to intakes of total digestible nutrients and crude protein.

    Science.gov (United States)

    Galyean, M L; Tedeschi, L O

    2014-11-01

    Prediction of microbial CP (MCP) synthesis in the rumen is an integral part of the MP system. For the NRC beef model, MCP is calculated as 0.13 multiplied by TDN intake (TDNI), with adjustment for physically effective NDF (peNDF) concentrations less than 20%. Despite its application for nearly 2 decades, MCP predictions using this approach have not been extensively evaluated. We assembled a database of 285 treatment means from 66 published papers using beef cattle and dairy or dairy × beef crossbred steers, fed diets with a wide range of TDN, CP, and ether extract (EE) concentrations, in which MCP synthesis was measured. Fat-free TDN (FFTDN) concentration was calculated by subtracting 2.25 × percent EE from the TDN concentration. Based on initial model selection procedures indicating that DMI and concentrations of TDN, FFTDN, and CP were significantly (P CPI) were considered as potential independent variables. Mixed model regression methods were used to fit 1-, 2-, and 3-independent-variable models based on either TDNI or FFTDNI (e.g., TDNI only, TDNI and CPI, and TDNI, CPI, and the quadratic effect of TDNI; or FFTDNI only, FFTDNI and CPI, and FFTDNI, CPI, and the quadratic effect of FFTDNI). True ruminal OM digested (TROMD; g/d) was highly related (r(2) = 0.84 using citation-adjusted data) to MCP synthesis. Similarly, both TDNI and FFTDNI were highly related to citation-adjusted TROMD (r(2) > 0.96) and MCP synthesis (r(2) > 0.89). Models with FFTDNI were slightly more precise with slightly smaller prediction errors than those with TDNI. Randomly dividing the citations into Development (60%) and Evaluation (40%) data sets indicated that models such as those derived from the overall database accounted for 46 to 56% of the variation in MCP synthesis, with neither mean nor linear bias (P ? 0.26). In contrast, calculating MCP as 0.13 × TDNI, with or without adjustment for peNDF concentration, resulted in overprediction of MCP (P < 0.001 for both mean and linear bias). Cross-validation using 5,000 randomly drawn training and testing data sets yielded results similar to the Development/Evaluation approach. Recommended equations are provided, but the errors of prediction associated with these empirical regression equations were on the order of 25 to 30% of the mean MCP. PMID:25253815

  2. Behaviour of digestive enzymes in the pancreatic juice and pancreas of rats fed on a low-protein diet (3 p. 100 of cereal protein) then on a balanced diet (23.5 p. 100 of mixed protein).

    Science.gov (United States)

    Kheroua, O; Belleville, J

    1981-01-01

    The aim of this study in the rat was to determine the effect of a low-protein diet (3 p. 100 cereal protein) and balanced refeeding (23.5 p. 100 mixed protein) on the activity of some pancreatic digestive enzymes and the amount of their secretion. Parallel studies were carried out on the pancreas and its exocrine secretion. 1) With a low-protein diet (21 days), there was a decrease in the amounts of bile and pancreatic juice secreted. During balanced refeeding (18 days), the amount of bile secreted returned to normal and, although that of the juice increased, it was less than the amount secreted by the reference lot. 2) Body and pancreatic weights decreased slightly with the low-protein diet. The protein content and mitotic ability of the pancreas declined. However, during balanced refeeding, the ponderal weight of the pancreas returned to normal more rapidly than that of the overall organism. Pancreatic protein content and mitotic ability also augmented. 3) The low-protein diet produced an overall decrease in enzyme activity in the pancreas and in the juice. However, in the absence of any dietary stimulation, these activities were not affected proportionally in the same way as the pancreas and its exocrine secretion. Nevertheless, no disturbance in the digestion and absorption of the ration was observed. 4) During balanced refeeding, enzyme activities increased to different levels in the juice and the pancreas but after 18 days total enzyme activity had not been entirely recovered. 5) Enzyme activity varied widely, especially during the first 48 hrs of malnutrition and the first 36 hrs of refeeding. PMID:7349559

  3. Disulphide bonds in wheat gluten: cystine peptides derived from gluten proteins following peptic and thermolytic digestion.

    Science.gov (United States)

    Keck, B; Köhler, P; Wieser, H

    1995-06-01

    Gluten from the wheat variety Rektor was extracted with 70% aqueous ethanol. The insoluble portion (whole glutenin) was partially hydrolysed with trypsin at pH 6.5 and separated on a Sephadex G25 column. The high molecular weight fraction 1 was further hydrolysed with pepsin at pH 2.0. To remove low molecular weight proteins, a portion of whole glutenin was extracted with dilute acetic acid. The residue (enriched glutenin), which contained mostly LMW and HMW subunits of glutenin, was hydrolysed with thermolysin at pH 6.5. The peptic and tryptic hydrolysates were separated on a Sephadex G25 column and the peptide fractions with the highest cystine content were separated further by reversed-phase high-performance liquid chromatography (RP-HPLC). Cystine peptides were detected by differential chromatography (RP-HPLC prior to and after reduction of disulphide bonds) and then isolated by preparative RP-HPLC. After reduction, cysteine peptides were alkylated and analysed for their amino acid sequence. Altogether, 19 cystine peptides were characterized and assigned to known sequences of gluten proteins; 16 peptides confirmed the positions of disulphide bonds present in LMW subunits and gamma-gliadins, as described previously. For the first time, a cystine peptide has been isolated, representing an intermolecular disulphide bond between the y-type of HMW and LMW subunits. Furthermore, a cystine peptide was assigned to gamma-gliadins; thus, all cysteine residues of gamma-gliadins are documented by at least one cystine peptide. One peptide analysed came from the alpha-amylase inhibitor CM 16. Altogether the results indicate that the intramolecular linkages of gluten proteins are not formed at random, but are strongly directed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7668061

  4. Effect of Carbohydrate Sources and Levels of Cotton Seed Meal in Concentrate on Feed Intake, Nutrient Digestibility, Rumen Fermentation and Microbial Protein Synthesis in Young Dairy Bulls

    OpenAIRE

    Wanapat, M.; Anantasook, N.; Rowlinson, P.; Pilajun, R.; Gunun, P.

    2013-01-01

    The objective of this study was to investigate the effect of levels of cottonseed meal with various carbohydrate sources in concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in dairy bulls. Four, 6 months old dairy bulls were randomly assigned to receive four dietary treatments according to a 2×2 factorial arrangement in a 4×4 Latin square design. Factor A was carbohydrate source; cassava chip (CC) and cassava chip+rice bran in the ratio ...

  5. Effect of Carbohydrate Source and Cottonseed Meal Level in the Concentrate on Feed Intake, Nutrient Digestibility, Rumen Fermentation and Microbial Protein Synthesis in Swamp Buffaloes

    OpenAIRE

    Wanapat, M.; Pilajun, R.; Polyorach, S.; Cherdthong, A.; Khejornsart, P.; Rowlinson, P.

    2013-01-01

    The objective of this study was to investigate the effect of carbohydrate source and cottonseed meal level in the concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in swamp buffaloes. Four, 4-yr old rumen fistulated swamp buffaloes were randomly assigned to receive four dietary treatments according to a 2×2 factorial arrangement in a 4×4 Latin square design. Factor A was carbohydrate source; cassava chip (CC) and CC+rice bran at a ratio 3...

  6. Influence of Processing on Dietary Fiber, Tannin and <i>in Vitro</i> Protein Digestibility of Pearl Millet

    OpenAIRE

    Florence Suma Pushparaj; Asna Urooj

    2011-01-01

    From the nutritional point of view, data on dietary fiber content, tannin and in vitro protein digestibility of processed millet is of importance, because millets are never eaten raw. Effects of commonly used traditional methods on dietary fiber, tannin content and %IVPD of two locally available pearl millet varieties (Kalukombu and Maharashtra Rabi Bajra) were investigated. The millet was subjected to various processing methods like milling (whole flour, semi refined flour and bran rich frac...

  7. In-line system containing porous polymer monoliths for protein digestion with immobilized pepsin, peptide preconcentration and nano-liquid chromatography separation coupled to electrospray ionization mass spectroscopy

    OpenAIRE

    Geiser, Laurent; Eeltink, Sebastiaan; Svec, Frantisek; Fre?chet, Jean M. J.

    2008-01-01

    The use of two different monoliths located in capillaries for on-line protein digestion, preconcentration of peptides and their separation has been demonstrated. The first monolith was used as support for covalent immobilization of pepsin. This monolith with well defined porous properties was prepared by in situ copolymerization of 2-vinyl-4,4-dimethylazlactone and ethylene dimethacrylate. The second, poly(lauryl methacrylate-co-ethylene dimethacrylate) monolith with a different porous struct...

  8. Componentes antinutricionais e digestibilidade proteica em sementes de abóbora (Cucurbita maxima) submetidas a diferentes processamentos / Antinutritional components and protein digestibility in pumpkin seeds (Cucurbita maxima) submitted to different processing methods

    Scientific Electronic Library Online (English)

    Luciana de Paula, Naves; Angelita Duarte, Corrêa; Custódio Donizete dos, Santos; Celeste Maria Patto de, Abreu.

    2010-05-01

    Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese Subprodutos vegetais têm sido utilizados na dieta com a finalidade de melhorar o estado nutricional de populações desnutridas. Entretanto, os antinutrientes presentes nesses alimentos podem acarretar efeitos indesejáveis. Portanto, os teores de polifenóis, cianeto, saponinas, inibidor de tripsina, a [...] tividade hemaglutinante e a porcentagem da digestibilidade proteica in vitro de sementes de abóbora cruas e tratadas termicamente foram investigados com o objetivo de selecionar o processamento que acarrete maior redução dos antinutrientes e maior digestibilidade proteica. Sementes da abóbora Cucurbita maxima foram, em quatro repetições, submetidas aos seguintes tratamentos: utilizadas na forma crua; cozidas em água em ebulição (AE) por três tempos: 5, 10 e 15 minutos; e cozidas no vapor por 10 minutos. Posteriormente foram liofilizadas, trituradas e armazenadas em temperatura ambiente até a realização das análises. Não houve diferença significativa, entre os tratamentos, quanto aos níveis de polifenóis. As sementes cruas apresentaram o maior teor de cianeto, o menor nível de inibidor de tripsina e a menor digestibilidade proteica. O cozimento em AE por 10 minutos acarretou o menor nível de saponinas e a maior digestibilidade proteica. Não foi detectada atividade hemaglutinante em nenhuma amostra. Conclui-se que o cozimento em AE por 10 minutos foi o que proporcionou melhores resultados. Abstract in english Vegetable subproducts have been used in diets with the purpose of improving the nutritional quality of undernourished populations. However, the antinutrients present in those foods can cause adverse health effects. Therefore, the contents of polyphenols, cyanide, saponins, trypsin inhibitor, hemaglu [...] tinin activity, and the percentage of the in vitro protein digestibility of raw and thermally treated pumpkin seeds were investigated with the objective of selecting the processing that results in major antinutrient reduction and greater protein digestibility. Pumpkin seeds (Cucurbita maxima) were submitted to the following treatments, which were carried out in quadruplicate: used raw; boiled for 5, 10, and 15 minutes; and steamed for 10 minutes. Next, the seeds were freeze-dried, grinded, and stored at room temperature until the analyses were completed. There was no significant difference among the treatments as for the levels of polyphenols. The raw seeds showed the highest content of cyanide, smallest level of trypsin inhibitor, and smallest protein digestibility. The 10-minute boiling resulted in the lowest content of saponins and greatest protein digestibility. No Hemaglutinin activity was detected in the samples. It can be concluded that the 10-minute boiling was the treatment that produced better results.

  9. Componentes antinutricionais e digestibilidade proteica em sementes de abóbora (Cucurbita maxima submetidas a diferentes processamentos Antinutritional components and protein digestibility in pumpkin seeds (Cucurbita maxima submitted to different processing methods

    Directory of Open Access Journals (Sweden)

    Luciana de Paula Naves

    2010-05-01

    Full Text Available Subprodutos vegetais têm sido utilizados na dieta com a finalidade de melhorar o estado nutricional de populações desnutridas. Entretanto, os antinutrientes presentes nesses alimentos podem acarretar efeitos indesejáveis. Portanto, os teores de polifenóis, cianeto, saponinas, inibidor de tripsina, atividade hemaglutinante e a porcentagem da digestibilidade proteica in vitro de sementes de abóbora cruas e tratadas termicamente foram investigados com o objetivo de selecionar o processamento que acarrete maior redução dos antinutrientes e maior digestibilidade proteica. Sementes da abóbora Cucurbita maxima foram, em quatro repetições, submetidas aos seguintes tratamentos: utilizadas na forma crua; cozidas em água em ebulição (AE por três tempos: 5, 10 e 15 minutos; e cozidas no vapor por 10 minutos. Posteriormente foram liofilizadas, trituradas e armazenadas em temperatura ambiente até a realização das análises. Não houve diferença significativa, entre os tratamentos, quanto aos níveis de polifenóis. As sementes cruas apresentaram o maior teor de cianeto, o menor nível de inibidor de tripsina e a menor digestibilidade proteica. O cozimento em AE por 10 minutos acarretou o menor nível de saponinas e a maior digestibilidade proteica. Não foi detectada atividade hemaglutinante em nenhuma amostra. Conclui-se que o cozimento em AE por 10 minutos foi o que proporcionou melhores resultados.Vegetable subproducts have been used in diets with the purpose of improving the nutritional quality of undernourished populations. However, the antinutrients present in those foods can cause adverse health effects. Therefore, the contents of polyphenols, cyanide, saponins, trypsin inhibitor, hemaglutinin activity, and the percentage of the in vitro protein digestibility of raw and thermally treated pumpkin seeds were investigated with the objective of selecting the processing that results in major antinutrient reduction and greater protein digestibility. Pumpkin seeds (Cucurbita maxima were submitted to the following treatments, which were carried out in quadruplicate: used raw; boiled for 5, 10, and 15 minutes; and steamed for 10 minutes. Next, the seeds were freeze-dried, grinded, and stored at room temperature until the analyses were completed. There was no significant difference among the treatments as for the levels of polyphenols. The raw seeds showed the highest content of cyanide, smallest level of trypsin inhibitor, and smallest protein digestibility. The 10-minute boiling resulted in the lowest content of saponins and greatest protein digestibility. No Hemaglutinin activity was detected in the samples. It can be concluded that the 10-minute boiling was the treatment that produced better results.

  10. Digestibility marker and ileal amino acid digestibility in phytase-supplemented soybean or canola meals for growing pigs.

    Science.gov (United States)

    Favero, A; Ragland, D; Vieira, S L; Owusu-Asiedu, A; Adeola, O

    2014-12-01

    Two experiments using soybean meal (SBM) or canola meal (CM) were conducted to investigate whether the choice of digestibility marker influenced the apparent ileal digestibility (AID) or standardized ileal digestibility (SID) of N and AA in diets supplemented with phytase. In each experiment, 18 barrows fitted with T-cannulas at the ileocecal junction were assigned to 3 diets consisting of a N-free diet to determine endogenous losses of N and AA, a semipurified diet (SBM in Exp. 1 or CM in Exp. 2), and the semipurified diet supplemented with phytase at 1,000 phytase units/kg. Three digestibility markers including acid-insoluble ash (AIA), chromic oxide (Cr2O3), and titanium dioxide (TiO2) were added to each diet at 3 g/kg. Each diet was fed for 7 d, consisting of a 5-d adjustment and a 2-d collection of ileal digesta. In both studies, basal ileal endogenous losses determined with Cr2O3 as a digestibility marker were lower (Pphytase and digestibility marker on AID or SID of AA. The AID of N and AA in SBM using AIA as a digestibility marker tended to be lower (PPhytase supplementation increased (PPhytase did not affect the SID of N or any AA in SBM except for Met, for which there was an increase (Pphytase supplementation. Using CM as the protein source in Exp. 2, there were significant interactions between digestibility marker and phytase. Phytase supplementation had effects (Pphytase supplementation increased (Pphytase supplementation of CM when AIA was used as a digestibility marker. In contrast, there were no clear improvements in AA digestibility from phytase supplementation for SBM. Phytase effects on AID or SID of AA were dependent on the digestibility marker used in diets when CM was used as the protein source but not when SBM was used as the protein source. Therefore, AA digestibility response to phytase supplementation may depend on the protein being evaluated as well as the choice of digestibility marker. PMID:25403199

  11. Purification and antioxidant properties of octapeptide from salmon byproduct protein hydrolysate by gastrointestinal digestion.

    Science.gov (United States)

    Ahn, Chang-Bum; Kim, Jeong-Gyun; Je, Jae-Young

    2014-03-15

    Pectoral fin protein from salmon processing byproduct was hydrolyzed using Alcalase, Flavourzyme, Neutrase, pepsin, Protamex, and trypsin, and the peptic hydrolysate showed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity. Antioxidant peptide was purified using consecutive chromatography. The purified antioxidant peptide was identified to be Phe-Leu-Asn-Glu-Phe-Leu-His-Val with molecular weight of 1018.48 Da by time of flight-mass spectrometry/mass spectrometry (TOF-MS) analysis. The IC50 values against DPPH and 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) cation radical scavenging activity were 486 and 152 ?M, respectively, and the octapeptide showed strong ferric reducing power. In addition, the octapeptide showed significant (p<0.05) protection ability against hydroxyl radical-induced DNA damage and hydrogen peroxide-induced hepatic damage in Chang liver cells. Taken together, the pectoral fin protein hydrolysate and/or its active peptides may be useful ingredients in functional food. PMID:24206688

  12. Effect of replacement of fish meal by potato protein concentrate in the diet for rainbow trout on feeding rate, digestibility and growth

    DEFF Research Database (Denmark)

    Xie, Shougi; Jokumsen, Alfred

    1999-01-01

    Six isonitrogenous and isoenergetic diets were composed to investigate the effects of incorporation of potato protein concentrate (PPC) and supplementation of methionine in the diet for rainbow trout (Oncorhynchus mykiss) on feeding rate, digestion, growth, feed utilization and body composition. The control diet contained all Danish L T-fish meal as protein sources. The other experimental diets contained 2.2, 5.6, 8.9 and 11.1% PPC respectively. Diet 6 contained 5.6% PPC and 1.7% methionine. A 4-week trial was conducted at about 12degreeC. The results showed that feeding rate decreased with increased incorporation levels of PPC. Apparent digestibility of dry matter, crude protein and ash increased with increased proportion of dietary PPC, while there was no significant effect on the apparent digestibility of crude fat. The incorporation of 5.6% PPC decreased growth rate and 8.9% PPC decreased both growth and feed efficiency. Supplementation of 1.7% methionine decreased both feeding rateand growth.

  13. Effects of two halophytic plants (kochia and atriplex) on digestibility, fermentation and protein synthesis by ruminal microbes maintained in continuous culture.

    Science.gov (United States)

    Riasi, A; Mesgaran, M Danesh; Stern, M D; Ruiz Moreno, M J

    2012-05-01

    Eight continuous culture fermenters were used in a completely randomized design to evaluate various nutritional values of Kochia (Kochia scoparia) compared with Atriplex (Atriplex dimorphostegia). Dried and pelleted samples (leaves and stems) provided substrate for metabolism by ruminal microbes maintained in a continuous culture fermentation system. Results indicated that there were no differences (p>0.05) in dry matter (DM) and crude protein (CP) digestibility between the two halophytic plants. Atriplex had higher (pAtriplex (411 g/kg) was higher (pAtriplex (406 vs. 234 g/kg). There were no differences (p>0.05) between the two halophytic plants in molar proportion of acetate and propionate, but the concentration of butyrate and valerate in Kochia were about two fold of Atriplex (pAtriplex (5.96 vs. 4.85 g N/kg of OM truly digested). It was concluded that Kochia scoparia and Atriplex dimorphostegia had similar digestibility of DM and CP. It appears that these halophytic plants may not have enough digestible energy for high producing ruminants. PMID:25049608

  14. Amino acid digestibility and energy concentration of high-protein corn dried distillers grains and high-protein sorghum dried distillers grains with solubles for swine.

    Science.gov (United States)

    Jacela, J Y; Frobose, H L; DeRouchey, J M; Tokach, M D; Dritz, S S; Goodband, R D; Nelssen, J L

    2010-11-01

    A study was conducted to determine the AA digestibility and energy concentration of a specialized high-protein corn distillers dried grains (HPC-DDG) product and a high-protein sorghum dried distillers grains with solubles (HPS-DDGS) product. Six growing barrows (BW = 22.7 kg) were surgically fitted with T-cannulas at the terminal ileum and allotted randomly to 3 treatments in a crossover design with 3 periods. The treatment diets were 1) 67% HPC-DDG and 2) 50% HPS-DDGS as the sole protein sources, and 3) an N-free diet for determining basal endogenous AA loss. All diets contained 0.25% chromic oxide as an inert marker. Digesta and fecal samples were collected and analyzed for AA and energy concentrations. After chemical analysis, standardized and apparent ileal digestible (SID and AID, respectively) AA and GE were determined for each coproduct. The DE, ME, and NE values for HPC-DDG and HPS-DDGS also were calculated. The chemical composition of HPC-DDG and HPS-DDGS on a DM basis was 40.8% CP, 5.4% fat, 22.9% ADF, 36.6% NDF, 0.04% Ca, and 0.42% P and 48.2% CP, 3.1% fat, 17.5% ADF, 20.4% NDF, 0.13% Ca, and 0.82% P, respectively. The DM content of HPC-DDG and HPS-DDGS was 89.50 and 91.88%, respectively. Analyzed AA content of HPC-DDG was greater than that of traditional corn DDGS. The Lys content of HPC-DDG was 1.36% (DM basis), resulting in a Lys-to-CP ratio of 3.2%. In HPS-DDGS, most AA were present in greater proportions than in HPC-DDG or conventional sorghum DDGS. The HPS-DDGS Lys content was 1.7% (DM basis), equivalent to a Lys-to-CP ratio of 3.5%. In HPC-DDG, the AID for Lys, Met, Thr, and Trp were 65.9 ± 1.7, 87.0 ± 1.9, 72.8 ± 3.4, and 76.2% ± 3.5, respectively, and SID values were 67.8 ± 1.7, 87.5 ± 1.9, 75.0 ± 3.5, and 78.6 ± 3.7%, respectively. For HPS-DDGS, the AID for Lys, Met, Thr, and Trp were 51.9 ± 5.3, 73.0 ± 3.1, 60.6 ± 5.3, and 71.7 ± 3.4%, respectively, and SID values were 53.7 ± 4.9, 73.8 ± 3.0, 63.0 ± 4.9, and 73.8 ± 3.0%, respectively. The GE, DE, and calculated ME and NE values were 5,293, 3,703 ± 121, 3,426 ± 121, and 2,131 ± 88 kcal/kg of DM, respectively, for HPC-DDG and 5,108, 3,878, 3,549, and 2,256 kcal/kg of DM, respectively, for HPS-DDGS. Results indicate that both coproducts are well suited for use in swine diets and that actual AA digestibility values and calculated energy concentrations can now be used in swine diet formulation. PMID:20656972

  15. Concentration of metabolizable energy and digestibility of energy, phosphorus, and amino acids in lemna protein concentrate fed to growing pigs.

    Science.gov (United States)

    Rojas, O J; Liu, Y; Stein, H H

    2014-11-01

    Lemna protein concentrate (LPC; 68.0% CP) is produced by extracting protein from de-oiled and dehydrated biomaterials from plants of the Lemnaceae family and may be used as a protein source for animals. There are, however, no published data on the nutritional value of LPC fed to pigs. Three experiments were, therefore, conducted to determine the concentration of ME, the standardized total tract digestibility (STTD) of P, and the standardized ileal digestibility (SID) of AA in LPC and to compare these values to values for fish meal and soybean meal (SBM). Experiment 1 was conducted to determine the ME of LPC, fish meal, SBM, and corn. Thirty-two barrows (initial BW: 16.8 ± 2.8 kg) were placed in metabolism cages and allotted to a randomized complete block design with 4 diets and 8 replicate pigs per diet. A corn-based diet and 3 diets that contained corn and LPC, fish meal, or SBM were formulated. Feces and urine were collected for 5 d after a 5-d adaptation period, and all samples were analyzed for GE. Results indicated that the concentration of ME was not different among corn, fish meal, and SBM (3,855, 3,904, and 4,184 kcal/kg DM, respectively), but there was a tendency (P = 0.08) for a reduced ME in LPC (3,804 kcal/kg DM) compared with SBM. In Exp. 2, 24 barrows (initial BW: 12.5 ± 2.5 kg) were allotted to a randomized complete block design with 3 diets and 8 replicate pigs per diet and used to determine the STTD of P in LPC, fish meal, and SBM. Three diets that each contained 1 of the 3 test ingredients as the sole source of P were formulated. Pigs were placed in metabolism cages, and feces were collected for 5 d after a 5-d adaptation period. The STTD of P in LPC (72.8%) was not different from the STTD of P in fish meal (65.6%), but tended (P = 0.07) to be greater than in SBM (62.8%). The SID of AA in LPC, SBM, and fish meal was determined in Exp. 3. Eight barrows (initial BW: 21.4 ± 4.0 kg) were equipped with a T-cannula in the distal ileum and randomly allotted to a replicated 4 × 4 Latin square design. A N-free diet and 3 cornstarch-based diets in which SBM, SBM and LPC or SBM and fish meal were the only sources of AA were formulated. The SID of most indispensable AA was greater (P < 0.05) in fish meal than in LPC, but the overall SID of AA was not different between fish meal and LPC. In conclusion, the ME and the STTD of P are not different between LPC and fish meal, but there is a tendency for greater ME in SBM than in LPC, whereas the STTD of P tends to be greater in LPC than in SBM. The SID of the most indispensable AA is greater in fish meal than in LPC. PMID:25349364

  16. Effect of exogenous cellulase enzyme on feed digestibility in lamb

    International Nuclear Information System (INIS)

    The aim of this study was to determine the effect of exogenous enzyme on digestibility and N retention in lamb. Eight lambs were randomly allocated to 2 experiment group in group comparison design trial. Experimental treatments were: 1) CTL (No enzyme) and 2 50NZ (Mixed enzyme with high cellulase at 50g/100kg.feed). The digestibility study showed that Exogenous enzyme increased (P<0.05) dry matter and crude protein digestibility of treated lamb compared to those of control. A similar trend (P=0.11) was observed for the NDF digestibility. Mean values for dry matter digestibility were 57.86 and 69.83% and for protein digestibility were 64.76 and 73.38%, for CTL and 50NZ, respectively). The N intake was similar among treatment, averaging 22.57g/head/day. Percent N retained of 50 NZ treated lambs was higher (P<.05) than those of CTL group (mean value were 47.74 and 59.07 for CTC and 50NZ, respectively). Feed efficiency or feed conversion ratio was numerically improved for enzyme-treated groups. Overall, the results of this study provide evidence that mixed cellulase enzyme can be used to improver performance of lambs as compare to non-enzyme diet.

  17. Altering protein conformation to improve fermentative hydrogen production from protein wastewater.

    Science.gov (United States)

    Xiao, Naidong; Chen, Yinguang; Ren, Hongqiang

    2013-10-01

    One important reason for low hydrogen production from protein wastewater is due to the native folded conformation of protein. In this study the enhancement of bio-hydrogen production from protein wastewater by altering protein conformation via pretreatment was reported. Firstly, the effect of different pretreatment methods (acid, alkali, heat, and ultraviolet) on hydrogen production from synthetic protein wastewater was compared. The hydrogen production from the ultraviolet pretreated wastewater was 111.3 mL/g-protein, which was 3.79-, 3.73-, 3.54-, and 1.36-fold of that from the unpretreated (blank), acid, alkali, and heat pretreated wastewater, respectively. Then, the reasons for ultraviolet pretreatment showing significantly higher hydrogen production than other pretreatments were investigated. It was found that all pretreatments did not cause the cleavage of peptide bond, but the ultraviolet one caused much greater damage of hydrogen bonding networks and unfolding of protein. Thus, during anaerobic fermentation much higher protease activity and protein utilization were observed, which resulted in the bio-hydrogen production being remarkably improved. Further studies indicated that the photo-oxidization of aromatic residues in protein was not the reason for ultraviolet pretreatment remarkably improving bio-hydrogen production. Finally, the application of ultraviolet pretreatment to enhance hydrogen production from real protein wastewater was testified. PMID:23850214

  18. Seasonal dynamics of ruminal crude protein digestion of browse species from baja california sur, mexico

    Scientific Electronic Library Online (English)

    Rafael, Ramírez-Orduña; Roque G., Ramírez-Lozano; Marco V., Gómez-Meza; Juan A., Armenta-Quintana; Juan M., Ramírez-Orduña; Ramón, Cepeda-Palacios; Juan M., Ávila-Sandoval.

    2003-07-01

    Full Text Available Dez arbustos nativos da Baixa Califórnia Sul, México, foram nutricionalmente avaliados em cada estação durante três anos consecutivos, medindo proteína crua (PC), fração protéica na fibra detergente neutro (FPDN) e na fibra detergente ácido (FPDA), e velocidade e grau de degradabilidade de PC. Cinco [...] leguminosas e cinco não leguminosas foram coletadas de 16 transectos lineares de 30 m. Todas as plantas de cada transecto foram identificadas e medidas para estimar o índice de diversidade Shannon, riqueza das espécies, cobertura aérea e freqüência relativa, dominância, densidade e importância. Feno de Medicago sativa L. foi usado como forragem de referência de alta qualidade nutritiva. Para estimar a degradabilidade efetiva de PC (DEPC), se usaram por duplicado sacolas nylon (10 x 15 cm; poro de 50 ìm) contendo 4 g de cada espécie de cada estação e ano, incubadas na parte ventral do rúmen de quatro bezerros de carne canulados. Em geral, o conteúdo de PC, FPDN, FPDA e DEPC foram significativamente maiores em leguminosas. A PC solúvel na fibra detergente neutro foi maior nas não leguminosas na primavera e no verão, e PC e DEPC foram maiores no outono. A DEPC em leguminosas foi significativamente menor no verão, possivelmente por redução na proteína disponível da parede celular. Somente espécies como Opuntia cholla, Prosopis sp. e Cercidium floridium tiveram valores de DEPC comparáveis ou maiores à alfafa. Cyrtocarpa edulis teve a menor DEPC. A PC de leguminosas foi mais degradada no rúmen que as não leguminosas. Distinguem-se dois períodos de qualidade nutritiva, um de alta qualidade no outono e no inverno e outro de baixa na primavera e no verão. Abstract in spanish Diez arbustos nativos de Baja California Sur, México, fueron nutricionalmente evaluados en cada estación durante tres años consecutivos, midiendo proteína cruda (PC), fracción proteica en la fibra detergente neutro (FPDN) y en la fibra detergente ácido (FPDA), y velocidad y grado de degradabilidad d [...] e PC. Cinco leguminosas y cinco no-leguminosas fueron colectadas de 16 transectos lineales de 30m. Todas las plantas de cada transecto fueron identificadas y medidas para estimar el índice de diversidad Shannon, riqueza de las especies, cobertura aérea y frecuencia relativa, dominancia, densidad e importancia. Heno de Medicago sativa L. fue usado como forraje de referencia de alta calidad nutritiva. Para estimar la degradabilidad efectiva de PC (DEPC), se usaron por duplicado bolsas nylon (10x15cm; poro de 50¼m) conteniendo 4g de cada especie de cada estación y año, incubadas en la parte ventral del rumen de cuatro becerros de carne canulados. En general, el contenido de PC, FPDN, FPDA y DEPC fueron significativamente mayores en leguminosas. La PC soluble en la fibra detergente neutro fue mayor en las no-leguminosas en primavera y verano, y PC y DEPC fueron mayores en otoño. La DEPC en leguminosas fue significativamente menor en verano, posiblemente por reducción en la proteína disponible de la pared celular. Solo especies como Opuntia cholla, Prosopis sp. y Cercidium floridium tuvieron valores de DEPC comparables o mayores a la alfalfa. Cyrtocarpa edulis tuvo la menor DEPC. La PC de leguminosas fue más degradada en el rumen que las no-leguminosas. Se distinguen dos períodos de calidad nutritiva, uno de alta calidad en otoño e invierno y otro de baja en primavera y verano. Abstract in english Ten native browse species from Baja California Sur, Mexico, were nutritionally evaluated in each season of three consecutive years measuring crude protein (CP), protein fraction in neutral detergent fiber (NDIP) and in acid detergent fiber (ADIP), and rate and extent of crude protein degradability. [...] Five legumes and five non-legumes were collected from 16 linear 30m transects. All plants in each transect were identified and measured to estimate Shannon’s diversity index, species richness, aerial cover and relative frequency, dominance, density and importance. Medicago sativa L. hay was

  19. Chemical composition and ruminal degradation kinetics of crude protein and amino acids, and intestinal digestibility of amino acids from tropical forages

    Scientific Electronic Library Online (English)

    Lidia Ferreira, Miranda; Norberto Mario, Rodriguez; Elzânia Sales, Pereira; Augusto César de, Queiroz; Roberto Daniel, Sainz; Patrícia Guimarães, Pimentel; Miguel Marques, Gontijo Neto.

    2012-03-01

    Full Text Available The objective of this research was to determine the chemical composition and ruminal degradation of the crude protein (CP), total and individual amino acids of leaves from tropical forages: perennial soybean (Neonotonia wightii), cassava (Manihot esculenta), leucaena (Leucaena leucocephala) and rami [...] e (Boehmeria nivea), and to estimate the intestinal digestibility of the rumen undegradable protein (RUDP) and individual amino acids of leaves from the tropical forages above cited, but including pigeon pea (Cajanus cajan). Three nonlactating Holstein cows were used to determine the in situ ruminal degradability of protein and amino acids from leaves (6, 18 and 48 hours of ruminal incubation). For determination of the intestinal digestibility of RUDP, the residue from ruminal incubation of the materials was used for 18 hours. A larger concentration of total amino acids for ramie and smaller for perennial soybean were observed; however, they were very similar in leucaena and cassava. Leucine was the essential amino acid of greater concentration, with the exception of cassava, which exhibited a leucine concentration 40.45% smaller. Ramie showed 14.35 and 22.31% more lysine and methionine, respectively. The intestinal digestibility of RUDP varied from 23.56; 47.87; 23.48; 25.69 and 10.86% for leucaena, perennial soybean, cassava, ramie and pigeon pea, respectively. The individual amino acids of tropical forage disappeared in different extensions in the rumen. For the correct evaluation of those forages, one should consider their composition of amino acids, degradations and intestinal digestibility, once the amino acid composition of the forage does not reflect the amino acid profiles that arrived in the small intestine. Differences between the degradation curves of CP and amino acids indicate that degradation of amino acids cannot be estimated through the degradation curve of CP, and that amino acids are not degraded in a similar degradation profile.

  20. Efeitos da fonte de proteína da dieta sobre a digestão de amido em bovinos Effects of diet protein source on starch digestion in cattle

    Directory of Open Access Journals (Sweden)

    Kátia Cylene Guimarães

    2001-05-01

    Full Text Available O objetivo deste trabalho foi avaliar o efeito da utilização de três fontes de proteína: uréia (UR, farinha de penas (FPE e farelo de glúten de milho (FGM sobre a digestão ruminal, intestinal e total do amido em bovinos. Utilizaram-se três bovinos da raça Holandês Preto e Branco, castrados, fistulados no rúmen e duodeno, distribuídos em um delineamento experimental quadrado latino 3 x 3. Não houve efeito da fonte protéica (P > 0,05 sobre os parâmetros de digestão avaliados com relação à matéria seca, bem como a composição química microbiana e pH ruminal. A dieta UR apresentou maior produção ruminal de amônia seguida pelas dietas FGM e FPE (P The effect of three protein sources, urea (UR, feather meal (FM and corn gluten meal (CGM on ruminal, intestinal and total starch digestion in cattle was evaluated. Three Holstein steers implanted with ruminal and duodenal cannulae, in a 3x3 Latin square design, were used. Protein source (p > 0.05 did not affect dry matter evaluations, chemical microbial composition and ruminal pH. UR diet showed highest ruminal ammonium production, followed by CGM and FM diets (P < 0.05. The apparent and true microbial efficiency was higher in CGM diet (P < 0.05 and identical to UR and FM diets. Protein source did not affect intestinal digestibility starch. Total digestibility coefficient (% of starch failed to show any significant difference, albeit total digestion of starch (g/day was higher (P < 0.05 in UR and FM diets.

  1. Chemical composition and ruminal degradation kinetics of crude protein and amino acids, and intestinal digestibility of amino acids from tropical forages

    Directory of Open Access Journals (Sweden)

    Lidia Ferreira Miranda

    2012-03-01

    Full Text Available The objective of this research was to determine the chemical composition and ruminal degradation of the crude protein (CP, total and individual amino acids of leaves from tropical forages: perennial soybean (Neonotonia wightii, cassava (Manihot esculenta, leucaena (Leucaena leucocephala and ramie (Boehmeria nivea, and to estimate the intestinal digestibility of the rumen undegradable protein (RUDP and individual amino acids of leaves from the tropical forages above cited, but including pigeon pea (Cajanus cajan. Three nonlactating Holstein cows were used to determine the in situ ruminal degradability of protein and amino acids from leaves (6, 18 and 48 hours of ruminal incubation. For determination of the intestinal digestibility of RUDP, the residue from ruminal incubation of the materials was used for 18 hours. A larger concentration of total amino acids for ramie and smaller for perennial soybean were observed; however, they were very similar in leucaena and cassava. Leucine was the essential amino acid of greater concentration, with the exception of cassava, which exhibited a leucine concentration 40.45% smaller. Ramie showed 14.35 and 22.31% more lysine and methionine, respectively. The intestinal digestibility of RUDP varied from 23.56; 47.87; 23.48; 25.69 and 10.86% for leucaena, perennial soybean, cassava, ramie and pigeon pea, respectively. The individual amino acids of tropical forage disappeared in different extensions in the rumen. For the correct evaluation of those forages, one should consider their composition of amino acids, degradations and intestinal digestibility, once the amino acid composition of the forage does not reflect the amino acid profiles that arrived in the small intestine. Differences between the degradation curves of CP and amino acids indicate that degradation of amino acids cannot be estimated through the degradation curve of CP, and that amino acids are not degraded in a similar degradation profile.

  2. Heat shock response improves heterologous protein secretion in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Hou, Jin; Österlund, Tobias

    2013-01-01

    The yeast Saccharomyces cerevisiae is a widely used platform for the production of heterologous proteins of medical or industrial interest. However, heterologous protein productivity is often low due to limitations of the host strain. Heat shock response (HSR) is an inducible, global, cellular stress response, which facilitates the cell recovery from many forms of stress, e.g., heat stress. In S. cerevisiae, HSR is regulated mainly by the transcription factor heat shock factor (Hsf1p) and many of its targets are genes coding for molecular chaperones that promote protein folding and prevent the accumulation of mis-folded or aggregated proteins. In this work, we over-expressed a mutant HSF1 gene HSF1-R206S which can constitutively activate HSR, so the heat shock response was induced at different levels, and we studied the impact of HSR on heterologous protein secretion. We found that moderate and high level over-expression of HSF1-R206S increased heterologous ?-amylase yield 25 and 70 % when glucose was fully consumed, and 37 and 62 % at the end of the ethanol phase, respectively. Moderate and high level over-expression also improved endogenous invertase yield 118 and 94 %, respectively. However, human insulin precursor was only improved slightly and this only by high level over-expression of HSF1-R206S, supporting our previous findings that the production of this protein in S. cerevisiae is not limited by secretion. Our results provide an effective strategy to improve protein secretion and demonstrated an approach that can induce ER and cytosolic chaperones simultaneously.

  3. Ethanol-based organosolv treatment with trace hydrochloric acid improves the enzymatic digestibility of Japanese cypress (Chamaecyparis obtusa) by exposing nanofibers on the surface.

    Science.gov (United States)

    Hideno, Akihiro; Kawashima, Ayato; Endo, Takashi; Honda, Katsuhisa; Morita, Masatoshi

    2013-03-01

    The effects of adding trace acids in ethanol based organosolv treatment were investigated to increase the enzymatic digestibility of Japanese cypress. A high glucose yield (60%) in the enzymatic hydrolysis was obtained by treating the sample at 170 °C for 45 min in 50% ethanol liquor containing 0.4% hydrochloric acid. Moreover, the enzymatic digestibility of the treated sample was improved to ?70% by changing the enzyme from acremonium cellulase to Accellerase1500. Field emission scanning electron microscopy revealed the presence of lignin droplets and partial cellulose nanofibers on the surface of the treated sample. Simultaneous saccharification and fermentation of the treated samples using thermotolerant yeast (Kluyveromyces marxianus NBRC1777) was tested. A high ethanol concentration (22.1 g/L) was achieved using the EtOH50/W50/HCl0.4-treated sample compared with samples from other treatments. PMID:23395739

  4. Digestibilidade intestinal in vitro da proteína de coprodutos da indústria do biodiesel Intestinal protein digestibility of by-products from biodiesel industry

    Directory of Open Access Journals (Sweden)

    G.S. Couto

    2012-10-01

    Full Text Available Determinou-se a digestibilidade intestinal (DI da proteína de vários coprodutos do biodiesel nas formas de farelo e torta. Foram avaliados oito coprodutos: tortas e farelos de pinhão manso, nabo forrageiro, tremoço, algodão. Os coprodutos foram incubados no rúmen por 16 horas, e os resíduos não degradados no rúmen submetidos à digestão enzimática com solução de pepsina e pancreatina para a determinação da DI. Ainda, nos resíduos da incubação ruminal, foram determinadas: degradabilidade da matéria seca (DR, proteína degradável no rúmen (PDR e proteína não degradável no rúmen (PNDR. A digestibilidade intestinal da proteína para os coprodutos do biodiesel variou de 2,4 a 48,6%. Todos os coprodutos avaliados caracterizaram-se como alimentos de alto teor proteico, sendo considerados de alta PDR, e apresentaram baixa digestibilidade intestinal da proteína. A DI da proteína dos coprodutos do biodiesel na forma de torta foi maior em comparação com a dos farelos. A torta e o farelo de algodão apresentaram os maiores coeficientes de DI.The objective of this research was to determine intestinal protein digestibility (ID of some biodiesel by-products in the form of cakes and the meals. Eight by-products were: cakes and meals of physic nut, turnip, lupine, cotton cake, cottonseed meal. The by-products were incubated in the rumen for 16 hours, were the undegradable rumen residues were submitted to enzymatic digestion with pepsin and pancreatin solution for the determination of ID. In the incubation residues the following was also determined: dry matter degradability (RD, rumen degradable protein (RDP and rumen undegradable protein (RUP. The intestinal protein digestibility of biodiesel by-products ranged from 2.4 to 48.6%. All the by-products evaluated in this study were characterized as high protein sources and were considered high-RDP. The by-products presented low intestinal protein digestibility. The ID protein of biodiesel by-products was higher in the cakes than the meals. The by-products evaluated, the cottonseed cake and meal presented the highest ID coefficients.

  5. Digestibilidade intestinal in vitro da proteína de coprodutos da indústria do biodiesel / Intestinal protein digestibility of by-products from biodiesel industry

    Scientific Electronic Library Online (English)

    G.S., Couto; J.C., Silva Filho; A.D., Corrêa; E.A., Silva; R.M.P., Pardo; C., Esteves.

    1216-12-01

    Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese Determinou-se a digestibilidade intestinal (DI) da proteína de vários coprodutos do biodiesel nas formas de farelo e torta. Foram avaliados oito coprodutos: tortas e farelos de pinhão manso, nabo forrageiro, tremoço, algodão. Os coprodutos foram incubados no rúmen por 16 horas, e os resíduos não deg [...] radados no rúmen submetidos à digestão enzimática com solução de pepsina e pancreatina para a determinação da DI. Ainda, nos resíduos da incubação ruminal, foram determinadas: degradabilidade da matéria seca (DR), proteína degradável no rúmen (PDR) e proteína não degradável no rúmen (PNDR). A digestibilidade intestinal da proteína para os coprodutos do biodiesel variou de 2,4 a 48,6%. Todos os coprodutos avaliados caracterizaram-se como alimentos de alto teor proteico, sendo considerados de alta PDR, e apresentaram baixa digestibilidade intestinal da proteína. A DI da proteína dos coprodutos do biodiesel na forma de torta foi maior em comparação com a dos farelos. A torta e o farelo de algodão apresentaram os maiores coeficientes de DI. Abstract in english The objective of this research was to determine intestinal protein digestibility (ID) of some biodiesel by-products in the form of cakes and the meals. Eight by-products were: cakes and meals of physic nut, turnip, lupine, cotton cake, cottonseed meal. The by-products were incubated in the rumen for [...] 16 hours, were the undegradable rumen residues were submitted to enzymatic digestion with pepsin and pancreatin solution for the determination of ID. In the incubation residues the following was also determined: dry matter degradability (RD), rumen degradable protein (RDP) and rumen undegradable protein (RUP). The intestinal protein digestibility of biodiesel by-products ranged from 2.4 to 48.6%. All the by-products evaluated in this study were characterized as high protein sources and were considered high-RDP. The by-products presented low intestinal protein digestibility. The ID protein of biodiesel by-products was higher in the cakes than the meals. The by-products evaluated, the cottonseed cake and meal presented the highest ID coefficients.

  6. Protein sources and digestive enzyme activities in jundiá (Rhamdia quelen) / Fontes protéicas e atividade de enzimas digestivas em jundiás (Rhamdia quelen)

    Scientific Electronic Library Online (English)

    Rafael, Lazzari; João, Radünz Neto; Fabio de Araújo, Pedron; Vania Lucia, Loro; Alexandra, Pretto; Carolina Rosa, Gioda.

    Full Text Available SciELO Brazil | Language: English Abstract in portuguese As enzimas digestivas influenciam a utilização dos alimentos em peixes, e seu conhecimento é importante para otimizar a formulação de dietas. Este trabalho descreve a atividade de enzimas digestivas em juvenis de jundiá alimentados com fontes protéicas. Os peixes foram alimentados com seis dietas (9 [...] 0 dias): MBY (farinha de carne e ossos + levedura de cana), SY (farelo de soja + levedura de cana), S (farelo de soja), MBS (farinha de carne e ossos + farelo de soja), FY (farinha de peixe + levedura de cana) e FS (farinha de peixe + farelo de soja). A cada 30 dias, foram analisadas as enzimas digestivas (tripsina, quimiotripsina e amilase) no intestino. No estômago, foi mensurada a protease ácida. Foram estimados os índices digestivo e hepato-somático, quociente intestinal, comprimento do trato digestório e ganho em peso. As atividades de tripsina e quimiotripsina foram maiores (p Abstract in english Digestive enzymes activity influence feed utilization by fish, and its understanding is important to optimize diet formulation. This study reports the digestive enzyme activities of jundiá juveniles fed diets with protein sources. Fish were fed six experimental diets for 90 days: MBY (meat and bone [...] meal + sugar cane yeast), SY (soybean meal + sugar cane yeast), S (soybean meal), MBS (meat and bone meal + soybean meal), FY (fish meal + sugar cane yeast) and FS (fish meal + soybean meal), and then sampled every 30 days and assayed in two intestine sections for digestive enzymes - trypsin, chymotrypsin and amylase - activities; gastric protease was assayed in the stomach. Digestive and hepatosomatic index, intestinal quotient, digestive tract length and weight gain were also measured. Trypsin and chymotrypsin activities were higher (p

  7. Effect of carbohydrate source and cottonseed meal level in the concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in swamp buffaloes.

    Science.gov (United States)

    Wanapat, M; Pilajun, R; Polyorach, S; Cherdthong, A; Khejornsart, P; Rowlinson, P

    2013-07-01

    The objective of this study was to investigate the effect of carbohydrate source and cottonseed meal level in the concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in swamp buffaloes. Four, 4-yr old rumen fistulated swamp buffaloes were randomly assigned to receive four dietary treatments according to a 2×2 factorial arrangement in a 4×4 Latin square design. Factor A was carbohydrate source; cassava chip (CC) and CC+rice bran at a ratio 3:1 (CR3:1), and factor B was level of cottonseed meal (CM); 109 g CP/kg (LCM) and 328 g CP/kg (HCM) in isonitrogenous diets (490 g CP/kg). Buffaloes received urea-treated rice straw ad libitum and supplemented with 5 g concentrate/kg BW. It was found that carbohydrate source did not affect feed intake, nutrient intake, digested nutrients, nutrient digestibility, ammonia nitrogen concentration, fungi and bacterial populations, or microbial protein synthesis (p>0.05). Ruminal pH at 6 h after feeding and the population of protozoa at 4 h after feeding were higher when buffalo were fed with CC than in the CR3:1 treatment (pnitrogen supply than the LCM fed group (p0.05). Based on this experiment, concentrate with a low level of cottonseed meal could be fed with cassava chips as an energy source in swamp buffalo receiving rice straw. PMID:25049873

  8. Desempenho de cordeiros e estimativa da digestibilidade do amido de dietas com diferentes fontes protéicas / Lamb performance and estimation of starch digestibility of diets with different protein sources

    Scientific Electronic Library Online (English)

    Mário Adriano Ávila, Queiroz; Ivanete, Susin; Alexandre Vaz, Pires; Clayton Quirino, Mendes; Renato Shinkai, Gentil; Omer Cavalcanti, Almeida; Rafael Camargo do, Amaral; Gerson Barreto, Mourão.

    1193-12-01

    Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese O objetivo deste trabalho foi avaliar a influência de fontes protéicas sobre desempenho, características da carcaça e da carne de cordeiros confinados, e estimar a digestibilidade do amido de rações com alta proporção de grãos. Foram distribuídos 28 cordeiros Santa Inês, em blocos completos ao acaso [...] , de acordo com o peso vivo e a idade, no início do experimento. As fontes protéicas foram os farelos de: soja, amendoim, canola e algodão, em dietas isonitrogenadas com 90% de concentrado e 10% de volumoso (feno de coast-cross). Na determinação da digestibilidade, foram utilizados quatro borregos em delineamento experimental em quadrado latino 4x4, e a digestibilidade do amido foi estimada a partir do teor de amido fecal. Não houve diferenças (p>0,05) quanto ao consumo de matéria seca, ganho de peso vivo médio, conversão alimentar, características da carcaça e da carne, entre as fontes protéicas avaliadas. A digestibilidade do amido apresentou coeficiente de determinação de 93%. Independentemente da fonte protéica utilizada, o teor de amido nas fezes é um indicador eficiente na estimativa da digestibilidade do amido de dietas com alta proporção de concentrado para cordeiros. Abstract in english The objective of this work was to evaluate the influence of protein sources on feedlot lambs performance, carcass and meat characteristics, and to estimate starch digestibility of high grain diets. Twenty eight Santa Inês ram lambs were selected in a complete randomized block design, according to bo [...] dy weight and age in the beginning of the experiment. Protein sources were the following meals: soybean, peanut, canola, and cottonseed in isonitrogenous diets with 90% concentrate and 10% roughage (coast-cross hay). Four ram lambs (48 kg) were used to determine starch digestibility in a 4x4 latin square design, and starch digestibility was estimated by fecal concentration of starch. There were no differences (p>0.05) for dry matter intake, average daily gain, feed conversion, carcass characteristics and meat quality, among the protein sources evaluated. Determination coefficient for starch digestibility was 93%. Fecal starch content is an accurate indicator of the starch digestibility estimation in lambs feeding high grain diets, independently of the protein source used.

  9. Emulsification of algal oil with soy lecithin improved DHA bioaccessibility but did not change overall in vitro digestibility.

    Science.gov (United States)

    Lin, Xinjie; Wang, Qi; Li, Weili; Wright, Amanda J

    2014-11-01

    Oil emulsification facilitates digestive lipolysis and subsequent lipid bioaccessibility. This study aimed to understand the effects of emulsification on DHA-rich algal oil digestion and bioaccessibility. An oil/water emulsion (50?:?44?:?6 oil-water-soy lecithin) was subjected to an in vitro digestion model with gastric pH 1.6 or 4.0 and particle size distributions, duodenal stage lipolysis and DHA bioaccessibility were determined. The emulsion was destabilized at gastric pH 1.6, with subsequent slow duodenal lipolysis. With gastric pH 4.0, the emulsion structure remained intact, initial lipolysis proceeded rapidly and DHA bioaccessibility was higher than for bulk oil, a mixture of oil, water and soy lecithin, and the gastric pH 1.6 destabilized emulsion (p < 0.05). However, the extent of lipolysis was not affected by emulsification or gastric pH. Therefore, the presence of an intact emulsion at the start of duodenal digestion, while not impacting the extent of lipolysis, did impact the initial lipolysis and DHA bioaccessibility. PMID:25208938

  10. Bayesian proteoform modeling improves protein quantification of global proteomic measurements.

    Science.gov (United States)

    Webb-Robertson, Bobbie-Jo M; Matzke, Melissa M; Datta, Susmita; Payne, Samuel H; Kang, Jiyun; Bramer, Lisa M; Nicora, Carrie D; Shukla, Anil K; Metz, Thomas O; Rodland, Karin D; Smith, Richard D; Tardiff, Mark F; McDermott, Jason E; Pounds, Joel G; Waters, Katrina M

    2014-12-01

    As the capability of mass spectrometry-based proteomics has matured, tens of thousands of peptides can be measured simultaneously, which has the benefit of offering a systems view of protein expression. However, a major challenge is that, with an increase in throughput, protein quantification estimation from the native measured peptides has become a computational task. A limitation to existing computationally driven protein quantification methods is that most ignore protein variation, such as alternate splicing of the RNA transcript and post-translational modifications or other possible proteoforms, which will affect a significant fraction of the proteome. The consequence of this assumption is that statistical inference at the protein level, and consequently downstream analyses, such as network and pathway modeling, have only limited power for biomarker discovery. Here, we describe a Bayesian Proteoform Quantification model (BP-Quant)(1) that uses statistically derived peptides signatures to identify peptides that are outside the dominant pattern or the existence of multiple overexpressed patterns to improve relative protein abundance estimates. It is a research-driven approach that utilizes the objectives of the experiment, defined in the context of a standard statistical hypothesis, to identify a set of peptides exhibiting similar statistical behavior relating to a protein. This approach infers that changes in relative protein abundance can be used as a surrogate for changes in function, without necessarily taking into account the effect of differential post-translational modifications, processing, or splicing in altering protein function. We verify the approach using a dilution study from mouse plasma samples and demonstrate that BP-Quant achieves similar accuracy as the current state-of-the-art methods at proteoform identification with significantly better specificity. BP-Quant is available as a MatLab® and R packages. PMID:25433089

  11. Bayesian Proteoform Modeling Improves Protein Quantification of Global Proteomic Measurements

    Energy Technology Data Exchange (ETDEWEB)

    Webb-Robertson, Bobbie-Jo M.; Matzke, Melissa M.; Datta, Susmita; Payne, Samuel H.; Kang, Jiyun; Bramer, Lisa M.; Nicora, Carrie D.; Shukla, Anil K.; Metz, Thomas O.; Rodland, Karin D.; Smith, Richard D.; Tardiff, Mark F.; McDermott, Jason E.; Pounds, Joel G.; Waters, Katrina M.

    2014-12-01

    As the capability of mass spectrometry-based proteomics has matured, tens of thousands of peptides can be measured simultaneously, which has the benefit of offering a systems view of protein expression. However, a major challenge is that with an increase in throughput, protein quantification estimation from the native measured peptides has become a computational task. A limitation to existing computationally-driven protein quantification methods is that most ignore protein variation, such as alternate splicing of the RNA transcript and post-translational modifications or other possible proteoforms, which will affect a significant fraction of the proteome. The consequence of this assumption is that statistical inference at the protein level, and consequently downstream analyses, such as network and pathway modeling, have only limited power for biomarker discovery. Here, we describe a Bayesian model (BP-Quant) that uses statistically derived peptides signatures to identify peptides that are outside the dominant pattern, or the existence of multiple over-expressed patterns to improve relative protein abundance estimates. It is a research-driven approach that utilizes the objectives of the experiment, defined in the context of a standard statistical hypothesis, to identify a set of peptides exhibiting similar statistical behavior relating to a protein. This approach infers that changes in relative protein abundance can be used as a surrogate for changes in function, without necessarily taking into account the effect of differential post-translational modifications, processing, or splicing in altering protein function. We verify the approach using a dilution study from mouse plasma samples and demonstrate that BP-Quant achieves similar accuracy as the current state-of-the-art methods at proteoform identification with significantly better specificity. BP-Quant is available as a MatLab ® and R packages at https://github.com/PNNL-Comp-Mass-Spec/BP-Quant.

  12. Effect of different dietary crude protein levels on performance, N digestibility and some blood parameters in Kivircik lambs

    Directory of Open Access Journals (Sweden)

    Keser Onur

    2008-01-01

    Full Text Available In this study, 45 weaned Kivircik male lambs with an average initial body weight ŠBW¹ of 26.23 kg were randomly assigned to five experimental groups (G10, G12, G14, G16, G18 fed 10, 12, 14, 16, 18 % crude protein (CP, respectively. It was observed that G16 had higher BW than the first three groups (p<0.05, but there were no significant differences between G16 and G18. G16 had higher daily body weight gain ŠBWG¹ than other groups. During the study, when compared with first three groups, G16 had significantly higher BWG (p<0.05, but no significant difference was observed between G16 and G18. The lowest DMI was observed in G10 (p<0.05, however, there were no differences between G14, G16 and G18. The best and the lowest feed efficiency were observed in G16 and G10, respectively. When the analysis results of faeces samples were compared, the lowest nitrogen (N excretion was detected in G10 and G12, and the highest N excretion was detected in G18 (p<0.05. However, there were no significant differences between G14 and G16. Percentages of N digestibility's of G16 and G18 were higher than those of other groups (p<0.05. There were no significant differences between G16 and G18, and between G10, G12 and G14. The analysis results of serum samples obtained from experimental groups showed that, except for serum urea and albumin levels, there were no significant differences between the level of other metabolites. G10 had lowest serum albumin level (p<0.05, and there were no significant differences between the other groups. Serum urea levels of G14, G16, G18 were similar and higher than those of G10 and G12 (p<0.05. The lowest serum urea level was determined in G10 (p<0.05. Consequently, when it was considered that feeding Kivircik lambs with higher protein level than 16 % had no advantage for performance and would be cause of economic loss, it can be said that 16 % CP was optimal.

  13. In vitro starch digestion correlates well with rate and extent of starch digestion in broiler chickens

    OpenAIRE

    Weurding, R. E.; Veldman, R.; Veen, W. A. G.; Aar, P. J.; Verstegen, M. W. A.

    2001-01-01

    Current feed evaluation systems for poultry are based on digested components (fat, protein and nitrogen-free extracts). Digestible starch is the most important energy source in broiler chicken feeds and is part of the nitrogen-free extract fraction. Digestible starch may be predicted using an in vitro method that mimics digestive processes in the gastrointestinal tract of broiler chickens. An experiment was designed to use this method for predicting site, rate and extent of starch digestion i...

  14. Detecting Protein Complexes by an Improved Affinity Propagation Algorithm in Protein-Protein Interaction Networks

    Directory of Open Access Journals (Sweden)

    Yu Wang

    2012-07-01

    Full Text Available Identification of protein complexes in protein-protein interaction (PPI networks is important in understanding cellular processes. In this paper, we propose a computationally efficient algorithm, named by Overlapped Affinity Propagation (OAP, which is based on Affinity Propagation algorithm (AP to detect protein complexes. First, AP algorithm is adopted to obtain a hard partition of the network. Then the candidate overlapping proteins for each community are identified. Finally, a strategy is constructed with an immediate purpose to filter noise in these detected protein complexes. We apply the OAP to the Saccharomyces cerevisiae PPI network, and the experimental results demonstrate that our algorithm can discover protein complexes with high precision by compared with the AP, MCL, CoAch and CPM algorithms. Our proposed method is validated as an effective algorithm in identifying protein complexes and can provide more insights for future biological study.

  15. Evidence for validity of ileal digestibility coefficients in monogastrics.

    Science.gov (United States)

    Columbus, Daniel; de Lange, Cornelis F M

    2012-08-01

    Measures of amino acid (AA) digestibility are used widely to estimate bioavailability of AA in feed and food ingredients for monogastric animals. In principle, the digestibility assay is simpler than in vivo assessments of AA bioavailability and allows for simultaneous estimation of the bioavailability of all AA in an experimental diet. It is generally assumed that absorption of intact AA in the hindgut of monogastrics is minimal, even though colonocytes do contain AA transporters and have been shown to absorb AA. This assumption is supported by the observation that infusion of AA into the hindgut does not improve nitrogen balance in monogastrics. In addition, growth performance of monogastrics is more highly correlated with ileal than faecal AA digestibility. Therefore, ileal digestibility coefficients provide better estimates of AA bioavailability than faecal digestibility coefficients. Measures of apparent ileal digestibility (AID) of AA are confounded with endogenous gut AA losses (EAAL). The curvilinear increase in AID of AA with increasing dietary AA level has been attributed to the relatively large contribution of EAAL to total ileal AA flows at low dietary AA levels. Subtracting basal EAAL from total ileal AA flows yields standardized ileal digestibility (SID) coefficients that appear to be more additive than AID coefficients in mixtures of feed ingredients. An implicit assumption when using SID AA coefficients in diet formulation is that the post-absorptive utilization of AA is not influenced by the dietary protein source. This assumption appears inappropriate when using feed or food ingredients that have been over-heated, induce large amounts of EAAL, or contain substantial amounts of fermentable fibre. Improved understanding of processes that contribute to the discrepancy between bioavailability and ileal digestibility will allow a more effective use of AA digestibility coefficients in diet formulation. PMID:23107537

  16. Improving the Performance of an HMM for Protein Family Modelling

    Directory of Open Access Journals (Sweden)

    Mohamed Hamza El-Sayed

    2007-01-01

    Full Text Available A hidden Markov model for protein modelling consists of sub-models for alpha-helix, beta-sheets, coil and possibly more. It is described how to estimate the model parameters as a whole from labeled sequences instead of estimating the parameters from the individual parts independently from subsequences. It is argued that the standard maximum likelihood ML estimation is not the optimal for training such model. In this study a new method is used where instead of estimating the parameters of model that maximizing the probability of the protein sequences (ML, we maximize the probability of the correct labels prediction, such a criterion is called conditional maximum likelihood CML. The advantage of this method is to optimize recognition of model. We tested our method on some of protein families such as L-asparagines, we noted that the performance of HMM is improved in prediction process.

  17. Breeding for improved plant architecture and high protein yields

    International Nuclear Information System (INIS)

    It has been increasingly recognized that the problem of protein malnutrition in many developing countries is closely correlated with inadequate intake of calories. Breeding for higher grain yields, therefore, continues to be a major objective of agricultural research in these countries. It follows that studies on nutritional improvement of foodgrains must also be planned with the objective of harvesting more protein per hectare combined with high grain yields. As far as cereal crops like wheat and rice are concerned, significant advances have been made in recent years in developing high yielding cultivars, which show an improved plant type marked by resistance to lodging and a high harvest index. Experimental approaches that can be used to increase the protein content of grains in these varieties are reviewed. The position in the case of grain legumes is quite different. The foremost requirement in their case is to develop high-yielding varieties similar to those of the cereal crops and the possibilities that exist in this regard are very great. The concept of improved plant type is even more relevant for increasing their yields. The existing cultivars of these crops have been selected more for adaptation to stress conditions rather than for high yields and in this process they have acquired a bushy, spreading and indeterminate growth habit combined with late maturity. A major requirement is to reconstruct the plant type to improve the grain-straw ratio, generate respprove the grain-straw ratio, generate response to higher plant populations per unit area, and to fit these crops into multiple and inter-cropping patterns. Using these concepts it has been possible to evolve new ideotypes and improve the yields of several species of grain legumes both through hybridization and induced mutations. Some of these results are reviewed. (author)

  18. Bioaugmentation of a Two-Stage Thermophilic (68°C/55°C) Anaerobic Digestion Concept for Improvement of the Methane Yield From Cattle Manure

    DEFF Research Database (Denmark)

    BangsØ Nielsen, Henrik; Mladenovska, Zuzana

    2007-01-01

    The possibility of improving a two-stage (68°C/55°C) anaerobic digestion concept for treatment of cattle manure was studied. In batch experiments, a 10-24% increase of the specific methane yield from cattle manure and its fractions was obtained, when the substrates were inoculated with bacteria of the genus Caldicellusiruptor and Dictyoglomus. In a reactor experiment inoculation of a 68°C pretreatment reactor with Caldicellusiruptor resulted in a 93% increase in the methane yield of the pre-treatment reactor for a period of 18 days, but gave only a slight increase in the overall methane yield of the two-stage setup.

  19. Improving methane production and phosphorus release in anaerobic digestion of particulate saline sludge from a brackish aquaculture recirculation system.

    Science.gov (United States)

    Zhang, Xuedong; Ferreira, Rui B; Hu, Jianmei; Spanjers, Henri; van Lier, Jules B

    2014-06-01

    In this study, batch tests were conducted to examine the effects of trehalose and glycine betaine as well as potassium on the specific methanogenic activity (SMA), acid and alkaline phosphatase activity of anaerobic biomass and phosphorus release in anaerobic digestion of saline sludge from a brackish recirculation aquaculture system. The results of ANOVA and Tukey's HSD (honestly significant difference) tests showed that glycine betaine and trehalose enhanced SMA of anaerobic biomass and reactive phosphorus release from the particulate waste. Moreover, SMA tests revealed that methanogenic sludge, which was long-term acclimatized to a salinity level of 17 g/L was severely affected by the increase in salinity to values exceeding 35 g/L. Addition of compatible solutes, such as glycine betaine and trehalose, could be used to enhance the specific methane production rate and phosphorus release in anaerobic digestion from particulate organic waste produced in marine or brackish aquaculture recirculation systems. PMID:24785791

  20. Glycosylation: impact, control and improvement during therapeutic protein production.

    Science.gov (United States)

    Costa, Ana Rita; Rodrigues, Maria Elisa; Henriques, Mariana; Oliveira, Rosário; Azeredo, Joana

    2014-12-01

    The emergence of the biopharmaceutical industry represented a major revolution for modern medicine, through the development of recombinant therapeutic proteins that brought new hope for many patients with previously untreatable diseases. There is a ever-growing demand for these therapeutics that forces a constant technological evolution to increase product yields while simultaneously reducing costs. However, the process changes made for this purpose may also affect the quality of the product, a factor that was initially overlooked but which is now a major focus of concern. Of the many properties determining product quality, glycosylation is regarded as one of the most important, influencing, for example, the biological activity, serum half-life and immunogenicity of the protein. Consequently, monitoring and control of glycosylation is now critical in biopharmaceutical manufacturing and a requirement of regulatory agencies. A rapid evolution is being observed in this context, concerning the influence of glycosylation in the efficacy of different therapeutic proteins, the impact on glycosylation of a diversity of parameters/processes involved in therapeutic protein production, the analytical methodologies employed for glycosylation monitoring and control, as well as strategies that are being explored to use this property to improve therapeutic protein efficacy (glycoengineering). This work reviews the main findings on these subjects, providing an up-to-date source of information to support further studies. PMID:23919242

  1. Improved method for predicting protein fold patterns with ensemble classifiers.

    Science.gov (United States)

    Chen, W; Liu, X; Huang, Y; Jiang, Y; Zou, Q; Lin, C

    2012-01-01

    Protein folding is recognized as a critical problem in the field of biophysics in the 21st century. Predicting protein-folding patterns is challenging due to the complex structure of proteins. In an attempt to solve this problem, we employed ensemble classifiers to improve prediction accuracy. In our experiments, 188-dimensional features were extracted based on the composition and physical-chemical property of proteins and 20-dimensional features were selected using a coupled position-specific scoring matrix. Compared with traditional prediction methods, these methods were superior in terms of prediction accuracy. The 188-dimensional feature-based method achieved 71.2% accuracy in five cross-validations. The accuracy rose to 77% when we used a 20-dimensional feature vector. These methods were used on recent data, with 54.2% accuracy. Source codes and dataset, together with web server and software tools for prediction, are available at: http://datamining.xmu.edu.cn/main/~cwc/ProteinPredict.html. PMID:22370884

  2. Characterizing the Range of Extracellular Protein Post-Translational Modifications in a Cellulose-Degrading Bacteria Using a Multiple Proteolyic Digestion/Peptide Fragmentation Approach

    Energy Technology Data Exchange (ETDEWEB)

    Dykstra, Andrew B [ORNL; Rodriguez, Jr., Miguel [ORNL; Raman, Babu [Dow Chemical Company, The; Cook, Kelsey [ORNL; Hettich, Robert {Bob} L [ORNL

    2013-01-01

    Post-translational modifications (PTMs) are known to play a significant role in many biological functions. The focus of this study is to characterize the post-translational modifications of the cellulosome protein complex used by the bacterium Clostridium thermocellum to better understand how this protein machine is tuned for enzymatic cellulose solubilization. To enhance comprehensive characterization, the extracellular cellulosome proteins were analyzed using multiple proteolytic digests (trypsin, Lys-C, Glu-C) and multiple fragmentation techniques (collisionally-activated dissociation, electron transfer dissociation, decision tree). As expected, peptide and protein identifications were increased by utilizing alternate proteases and fragmentation methods, in addition to the increase in protein sequence coverage. The complementarity of these experiments also allowed for a global exploration of PTMs associated with the cellulosome based upon a set of defined PTMs that included methylation, oxidation, acetylation, phosphorylation, and signal peptide cleavage. In these experiments, 85 modified peptides corresponding to 28 cellulosome proteins were identified. Many of these modifications were located in active cellulolytic or structural domains of the cellulosome proteins, suggesting a level of possible regulatory control of protein function in various cellulotyic conditions. The use of multiple enzymes and fragmentation technologies allowed for independent verification of PTMs in different experiments, thus leading to increased confidence in PTM identifications.

  3. An improved method for scoring protein-protein interactions using semantic similarity within the gene ontology

    Directory of Open Access Journals (Sweden)

    Jain Shobhit

    2010-11-01

    Full Text Available Abstract Background Semantic similarity measures are useful to assess the physiological relevance of protein-protein interactions (PPIs. They quantify similarity between proteins based on their function using annotation systems like the Gene Ontology (GO. Proteins that interact in the cell are likely to be in similar locations or involved in similar biological processes compared to proteins that do not interact. Thus the more semantically similar the gene function annotations are among the interacting proteins, more likely the interaction is physiologically relevant. However, most semantic similarity measures used for PPI confidence assessment do not consider the unequal depth of term hierarchies in different classes of cellular location, molecular function, and biological process ontologies of GO and thus may over-or under-estimate similarity. Results We describe an improved algorithm, Topological Clustering Semantic Similarity (TCSS, to compute semantic similarity between GO terms annotated to proteins in interaction datasets. Our algorithm, considers unequal depth of biological knowledge representation in different branches of the GO graph. The central idea is to divide the GO graph into sub-graphs and score PPIs higher if participating proteins belong to the same sub-graph as compared to if they belong to different sub-graphs. Conclusions The TCSS algorithm performs better than other semantic similarity measurement techniques that we evaluated in terms of their performance on distinguishing true from false protein interactions, and correlation with gene expression and protein families. We show an average improvement of 4.6 times the F1 score over Resnik, the next best method, on our Saccharomyces cerevisiae PPI dataset and 2 times on our Homo sapiens PPI dataset using cellular component, biological process and molecular function GO annotations.

  4. Effect of pectin extracted from citrus pulp on digesta characteristics and nutrient digestibility in broilers chickens

    Scientific Electronic Library Online (English)

    Vanessa Karla, Silva; Viviane de Souza, Morita; Isabel Cristina, Boleli.

    2013-08-01

    Full Text Available SciELO Brazil | Language: English Abstract in english The experiment was carried out to evaluate the effects of continuous ingestion of pectin on intestinal viscosity, intestinal transit time, excreta moisture content, nutrient digestibility and energy metabolism of broilers at starter and growth phases. We used 240 one-day-old Cobb male broiler chicks [...] , distributed in a completely randomized experimental design. Treatments consisted of four concentrations of pectin (0, 10, 30 and 50 g kg-1) with six replicates of 10 birds each. The ingestion of pectin supplied in the feed by broilers at the starter phase increased intestinal viscosity and intestinal transit time, reduced excreta moisture, improved the use of apparent metabolizable energy, nitrogen-corrected apparent metabolizable energy, coefficient of apparent metabolizability, coefficient of nitrogen-corrected apparent metabolizability, apparent digestibility coefficient of crude protein and organic matter; worsened calcium utilization and coefficients of apparent digestibility of dry matter; and did not influence the coefficients of apparent digestibility of crude fat, ash and phosphorus. Pectin ingestion during the growth phase increased intestinal viscosity and apparent digestibility coefficients of ash and organic matter, but decreased the dry matter, crude fat, crude protein and calcium. Intestinal transit time, energy metabolism and apparent digestibility coefficient of crude protein showed quadratic behavior according to pectin levels in the feed. Therefore, pectin ingestion by broilers at the starter phase increases intestinal viscosity and intestinal transit time, reduces excreta moisture and improves energy utilization, whereas at the growth phase nutrient digestibility is decreased.

  5. Degradação ruminal e digestibilidade intestinal da proteína bruta de alimentos para bovinos Rumen degradation and intestinal digestibility of crude protein in feeds for cattle

    Directory of Open Access Journals (Sweden)

    Marcos Inácio Marcondes

    2009-11-01

    Full Text Available Este trabalho foi conduzido com os objetivos de determinar as frações A e B e a taxa de degradação ruminal (Kd da matéria seca (MS e da proteína bruta (PB de 27 alimentos e avaliar a digestibilidade intestinal da proteína nãodegradada no rúmen pelas técnicas do saco de náilon móvel e de três estágios. Os alimentos avaliados foram farelos de arroz, babaçu, gérmen de milho e trigo; milho desintegrado com palha e sabugo, milho desintegrado com sabugo, milho, polpa cítrica, sorgo, amireia, farelos de algodão com 28, 38 e 46% de PB, farelos de amendoim, girassol e soja; feijão-bandinha, glúten de milho, grão de soja, levedura, promil, refinazil, cascas de cacau, café e soja e silagens de capim-elefante e milho. Para obtenção da degradabilidade ruminal da MS e PB dos alimentos, utilizaram-se sacos de náilon de 10 × 20 cm e os tempos de incubação de 0, 2, 4, 6, 12, 16, 24, 48 e 72 horas. A digestibilidade intestinal foi determinada pelas técnicas do saco de náilon móvel e de três estágios. Os dados de degradação ruminal da matéria seca e da proteína bruta, em sua maioria, estão de acordo com a literatura. A técnica dos três estágios não estimou de forma satisfatória a digestibilidade interstinal de todos os alimentos estudados em conjunto, mas foi adequada para os alimentos proteicos. A maioria dos alimentos possui aproximadamente 90% de digestibilidade da PB, com exceção das cascas de soja, café e cacau e das silagens de milho e capimelefante. A técnica de três estágios estimou corretamente a digestibilidade intestinal dos alimentos proteicos, mas recomenda-se a utilização da equação DIPB (% = -5,1906 + 1,1053 × X para corrigir a digestibilidade obtida pela técnica dos três estágios para alimentos não-proteicos.The objective of the present study was to determine the A and B fractions and the rumen degradation rate (Kd of dry matter (DM and crude protein (CP in 27 feeds and determine the intestinal digestibility of non-degraded rumen protein by the three steps and mobile nylon bag techniques. Rice meal, babassu meal, wheat and corn germ, ground corn cobs and straw, ground corn cobs, corn, citrus pulp, sorghum, starea and cottonseed meal with 28, 38 and 46% CP, peanut meal, sunflower meal, soybean meal, bean byproduct, corn gluten, soybean grain, dry yeast, corn gluten meal, cacao hulls, coffee hulls, soybean hulls, elephant-grass silage and corn silage were sampled. To obtain the ruminal degradation of DM and CP, 10 × 20 cm nylon bags with incubation times of 0, 2, 4, 6, 12, 16, 24, 48 and 72 hours were used. The intestinal digestibility was determined by the three steps and the mobile nylon bag techniques. The ruminal degradation data of the dry matter and crude protein were similar to the literature. The three step technique did not estimate properly the intestinal digestibility of all the feeds evaluated, excepted for the protein feeds. Most feeds had total CP digestibility close to 90%, excepted for soybean, coffee and cacao hulls and elephant-grass and corn silage. The three step technique correctly estimated the intestinal digestibility of the protein feeds, but the equation IDCP (% = -5,1906 + 1,1053 × X is recommended to correct the digestibility obtained by the three steps technique for non-protein feeds.

  6. Degradação ruminal e digestibilidade intestinal da proteína bruta de alimentos para bovinos / Rumen degradation and intestinal digestibility of crude protein in feeds for cattle

    Scientific Electronic Library Online (English)

    Marcos Inácio, Marcondes; Sebastião de Campos, Valadares Filho; Edenio, Detmann; Rilene Ferreira Diniz, Valadares; Luiz Fernando Costa e, Silva; Mozart Alves, Fonseca.

    2247-22-01

    Full Text Available Este trabalho foi conduzido com os objetivos de determinar as frações A e B e a taxa de degradação ruminal (Kd) da matéria seca (MS) e da proteína bruta (PB) de 27 alimentos e avaliar a digestibilidade intestinal da proteína nãodegradada no rúmen pelas técnicas do saco de náilon móvel e de três está [...] gios. Os alimentos avaliados foram farelos de arroz, babaçu, gérmen de milho e trigo; milho desintegrado com palha e sabugo, milho desintegrado com sabugo, milho, polpa cítrica, sorgo, amireia, farelos de algodão com 28, 38 e 46% de PB, farelos de amendoim, girassol e soja; feijão-bandinha, glúten de milho, grão de soja, levedura, promil, refinazil, cascas de cacau, café e soja e silagens de capim-elefante e milho. Para obtenção da degradabilidade ruminal da MS e PB dos alimentos, utilizaram-se sacos de náilon de 10 × 20 cm e os tempos de incubação de 0, 2, 4, 6, 12, 16, 24, 48 e 72 horas. A digestibilidade intestinal foi determinada pelas técnicas do saco de náilon móvel e de três estágios. Os dados de degradação ruminal da matéria seca e da proteína bruta, em sua maioria, estão de acordo com a literatura. A técnica dos três estágios não estimou de forma satisfatória a digestibilidade interstinal de todos os alimentos estudados em conjunto, mas foi adequada para os alimentos proteicos. A maioria dos alimentos possui aproximadamente 90% de digestibilidade da PB, com exceção das cascas de soja, café e cacau e das silagens de milho e capimelefante. A técnica de três estágios estimou corretamente a digestibilidade intestinal dos alimentos proteicos, mas recomenda-se a utilização da equação DIPB (%) = -5,1906 + 1,1053 × X para corrigir a digestibilidade obtida pela técnica dos três estágios para alimentos não-proteicos. Abstract in english The objective of the present study was to determine the A and B fractions and the rumen degradation rate (Kd) of dry matter (DM) and crude protein (CP) in 27 feeds and determine the intestinal digestibility of non-degraded rumen protein by the three steps and mobile nylon bag techniques. Rice meal, [...] babassu meal, wheat and corn germ, ground corn cobs and straw, ground corn cobs, corn, citrus pulp, sorghum, starea and cottonseed meal with 28, 38 and 46% CP, peanut meal, sunflower meal, soybean meal, bean byproduct, corn gluten, soybean grain, dry yeast, corn gluten meal, cacao hulls, coffee hulls, soybean hulls, elephant-grass silage and corn silage were sampled. To obtain the ruminal degradation of DM and CP, 10 × 20 cm nylon bags with incubation times of 0, 2, 4, 6, 12, 16, 24, 48 and 72 hours were used. The intestinal digestibility was determined by the three steps and the mobile nylon bag techniques. The ruminal degradation data of the dry matter and crude protein were similar to the literature. The three step technique did not estimate properly the intestinal digestibility of all the feeds evaluated, excepted for the protein feeds. Most feeds had total CP digestibility close to 90%, excepted for soybean, coffee and cacao hulls and elephant-grass and corn silage. The three step technique correctly estimated the intestinal digestibility of the protein feeds, but the equation IDCP (%) = -5,1906 + 1,1053 × X is recommended to correct the digestibility obtained by the three steps technique for non-protein feeds.

  7. Improving homology models for protein-ligand binding sites.

    Science.gov (United States)

    Kauffman, Chris; Rangwala, Huzefa; Karypis, George

    2008-01-01

    In order to improve the prediction of protein-ligand binding sites through homology modeling, we incorporate knowledge of the binding residues into the modeling framework. Residues are identified as binding or nonbinding based on their true labels as well as labels predicted from structure and sequence. The sequence predictions were made using a support vector machine framework which employs a sophisticated window-based kernel. Binding labels are used with a very sensitive sequence alignment method to align the target and template. Relevant parameters governing the alignment process are searched for optimal values. Based on our results, homology models of the binding site can be improved if a priori knowledge of the binding residues is available. For target-template pairs with low sequence identity and high structural diversity our sequence-based prediction method provided sufficient information to realize this improvement. PMID:19642282

  8. The Effect of Soybean Hulls Substitution Levels to Corn in The Ration on Energy and Protein Digestibility and on Sheep Performance

    Directory of Open Access Journals (Sweden)

    Ning Iriyanti

    2000-05-01

    Full Text Available An experiment was conducted to investigate the effect of soybean hulls levels substitution in to corn as ration in twenty seven rams. The animal had a mean of live weight of 12.86 ± 1.94 kg. Levels of substitution were 0, 50, and 100% or 0, 25, and 50% ration dry matter basis and rations were iso-nitrogeneous. Analysis of variance were conducted for completely randomized design and polynomial orthogonal test were applied for the statistical analysis. Inclusion of soybean hulls in 50% ration dry matter had no effect on daily gain (99.71 ± 22.97 g, and digestible crude protein (62.43 ± 6.56 %. However, dry matter consumption tended to increase linearly and as soybean hulls level increase in the ration (P<0.01. The digestible energy significantly decrease linearly (P<0.01, whereas intake of the energy had same was 1.27 ± 0.33, 1.44 ± 0.17, and 1.23 ± 0.19 Mcal/day for ration containing 0, 25, 50% of soybean hulls, respectively. It was concluded that the soybean hulls may be substituted for corn. (Animal Production 2(2: 98-103 (2000 Key words : soybean hulls, corn, digestibility, daily gain.

  9. Effect of carbohydrate sources and levels of cotton seed meal in concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in young dairy bulls.

    Science.gov (United States)

    Wanapat, M; Anantasook, N; Rowlinson, P; Pilajun, R; Gunun, P

    2013-04-01

    The objective of this study was to investigate the effect of levels of cottonseed meal with various carbohydrate sources in concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in dairy bulls. Four, 6 months old dairy bulls were randomly assigned to receive four dietary treatments according to a 2×2 factorial arrangement in a 4×4 Latin square design. Factor A was carbohydrate source; cassava chip (CC) and cassava chip+rice bran in the ratio of 3:1 (CR3:1), and factor B was cotton seed meal levels in the concentrate; 109 g CP/kg (LCM) and 328 g CP/kg (HCM) at similar overall CP levels (490 g CP/kg). Bulls received urea-lime treated rice straw ad libitum and were supplemented with 10 g of concentrate/kg BW. It was found that carbohydrate source and level of cotton seed meal did not have significant effects on ruminal pH, ammonia nitrogen concentration, microbial protein synthesis or feed intake. Animals which received CC showed significantly higher BUN concentration, ruminal propionic acid and butyric acid proportions, while dry matter, organic matter digestibility, populations of total viable bacteria and proteolytic bacteria were lower than those in the CR3:1 treatment. The concentration of total volatile fatty acids was higher in HCM than LCM treatments, while the concentration of butyric acid was higher in LCM than HCM treatments. The population of proteolytic bacteria with the LCM treatments was higher than the HCM treatments; however other bacteria groups were similar among the different levels of cotton seed meal. Bulls which received LCM had higher protein digestibility than those receiving HCM. Therefore, using high levels of cassava chip and cotton seed meal might positively impact on energy and nitrogen balance for the microbial population in the rumen of the young dairy bull. PMID:25049819

  10. Effects of ratios of non-fibre carbohydrates to rumen degradable protein in diets of Holstein cows: 1. Feed intake, digestibility and milk production

    Scientific Electronic Library Online (English)

    A., Afzalzadeh; H., Rafiee; A.A., Khadem; A., Asadi.

    Full Text Available To evaluate the effect of different ratios of non-fibre carbohydrates (NFC) to rumen degradable protein (RDP) on lactation responses, digestion and dry matter intake (DMI), nine multiparous mid-lactation Holstein cows, averaging 171 ± 17 days in milk and 24.1 ± 3.3 kg of milk/d were assigned to a 3 [...] × 3 Latin square design. The total mixed rations (TMR) included 45% forage on a dry matter (DM) basis and 55% concentrates. The RDP (g/kg of DM) and NFC : RDP ratios of diets were respectively: 1) 98 and 4.10 (LRDP), 2) 108 and 3.71 (MRDP), 3) 118 and 3.34 (HRDP). Ratios were achieved through altering the RDP content of the diets while the NFC was held constant at 401 g/kg of DM. Urea was supplemented as a source of RDP to decrease the ratio. DMI and organic matter intake (OMI) were affected by treatment and tended to decrease linearly from LRDP to HRDP. Crude protein intake (CPI) increased linearly as the amount of RDP in the diets increased. Apparent organic matter digestion in the total tract decreased significantly in the HRDP diet. Percentage of protein and solid non-fat (SNF) and concentration of milk urea-nitrogen (MUN) in milk increased linearly when cows were fed diets with the higher amounts of RDP. Results showed that low producing cows are less likely to respond to altering NFC : RDP ratios than high producing cows, but digestibility in the MRDP treatment was higher and RDP level equal to 108 (g/kg of DM). A NFC : RDP ratio equal to 3.71 was useful and recommendable for mid-lactation dairy cow.

  11. Urea free and more efficient sample preparation method for mass spectrometry based protein identification via combining the formic acid-assisted chemical cleavage and trypsin digestion.

    Science.gov (United States)

    Wu, Shuaibin; Yang, Kaiguang; Liang, Zhen; Zhang, Lihua; Zhang, Yukui

    2011-10-30

    A formic acid (FA)-assisted sample preparation method was presented for protein identification via mass spectrometry (MS). Detailedly, an aqueous solution containing 2% FA and dithiothreitol was selected to perform protein denaturation, aspartic acid (D) sites cleavage and disulfide linkages reduction simultaneously at 108°C for 2h. Subsequently, FA wiped off via vacuum concentration. Finally, iodoacetamide (IAA) alkylation and trypsin digestion could be performed ordinally. A series of model proteins (BSA, ?-lactoglobulin and apo-Transferrin) were treated respectively using such method, followed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) analysis. The identified peptide number was increased by ? 80% in comparison with the conventional urea-assisted sample preparation method. Moreover, BSA identification was achieved efficiently down to femtomole (25 ± 0 sequence coverage and 16 ± 1 peptides) via such method. In contrast, there were not peptides identified confidently via the urea-assisted method before desalination via the C18 zip tip. The absence of urea in this sample preparation method was an advantage for the more favorable digestion and MALDI-TOF MS analysis. The performances of two methods for the real sample (rat liver proteome) were also compared, followed by a nanoflow reversed-phase liquid chromatography with electrospray ionization tandem mass spectrometry system analysis. As a result, 1335 ± 43 peptides were identified confidently (false discovery rate <1%) via FA-assisted method, corresponding to 295 ± 12 proteins (of top match=1 and requiring 2 unique peptides at least). In contrast, there were only 1107 ± 16 peptides (corresponding to 231 ± 10 proteins) obtained from the conventional urea-assisted method. It was serving as a more efficient protein sample preparation method for researching specific proteomes better, and providing assistance to develop other proteomics analysis methods, such as, peptide quantitative analysis. PMID:22063562

  12. Effect of Plants Containing Secondary Compounds with Palm Oil on Feed Intake, Digestibility, Microbial Protein Synthesis and Microbial Population in Dairy Cows

    OpenAIRE

    Anantasook, N.; Wanapat, M.; Cherdthong, A.; Gunun, P.

    2013-01-01

    The objective of this study was to determine the effect of rain tree pod meal with palm oil supplementation on feed intake, digestibility, microbial protein synthesis and microbial populations in dairy cows. Four, multiparous early-lactation Holstein-Friesian crossbred (75%) lactating dairy cows with an initial body weight (BW) of 405±40 kg and 36±8 DIM were randomly assigned to receive dietary treatments according to a 4×4 Latin square design. The four dietary treatments were un-supplemen...

  13. Effects of Replacement of Fish Meal by Soy Protein Isolate on the Growth, Digestive Enzyme Activity and Serum Biochemical Parameters for Juvenile Amur Sturgeon (Acipenser schrenckii)

    OpenAIRE

    Xu, Q. Y.; Wang, C. A.; Zhao, Z. G.; Luo, L.

    2012-01-01

    An 8-wk experiment was conducted to evaluate the effect of replacing fish meal (FM) with soy protein isolate (SPI) on the growth, digestive enzyme activity and serum biochemical parameters of juvenile Amur sturgeon (Acipenser schrenckii). SPI was used to replace 0, 25, 50, 62.5, 75, 87.5, 100% of dietary FM and 100% replacement supplemented crystalline amino acid. Healthy sturgeon with an average initial weight of 26.38±0.24 g were randomly assigned to 24 aquaria (8 treatments with triplicat...

  14. Effect of Mannanase on Broiler Performance, Ileal and In-vitro Protein Digestibility, Uric Acid and Litter Moisture in Broiler Feeding

    OpenAIRE

    Saki, A. A.; Mazugi, M. T.; Kamyab, A.

    2005-01-01

    Protein digestibility and uric acid excretion may play the vital role in feed efficiency and consequently effect on broiler performance. Five hundred unsexed Arian day old chickens were placed after pre-starter feeding (10 days) in 22 pens with 23 chickens in each. This study was modulated in two levels of Mannanase (Tread mark of Hemicell enzyme) enzyme (0 and 5 kg/ton) and three levels of metabolizable energy (3000, 2900 and 2850 kcal/kg). Six treatments were arranged and statistical status...

  15. Desempenho zootécnico de girinos de rã-touro com diferentes níveis de proteína e energia digestíveis / Zootechnical performance of bullfrog tadpoles under different levels of digestible protein and energy

    Scientific Electronic Library Online (English)

    José Teixeira de, Seixas Filho; Rodrigo Diana, Navarro; Marcelo Maia, Pereira; Sílvia Conceição Reis Pereira, Mello; Eduardo Arruda Teixeira, Lanna; Jorge Luiz Pereira, Lima.

    1112-11-01

    Full Text Available A partir dos valores de digestibilidade de alguns ingredientes foram propostas dietas, com diferentes níveis de proteína e energia digestíveis, em esquema fatorial 3 x 3, destinadas a girinos de rã-touro para avaliar o desempenho zootécnico. O experimento, com duração de 60 dias, foi conduzido com 8 [...] 10 girinos, oriundos da mesma desova, distribuídos em 27 caixas de polipropileno, em uma densidade de 1girino/L. O delineamento experimental foi um esquema fatorial com três níveis de proteína digestível (27; 31 e 35%) e três níveis de energia digestível (2.700; 2.800 e 2.900kcal/kg) com três réplicas. Os parâmetros avaliados aos 60 dias foram ganho de peso, consumo da dieta, consumo em proteína da dieta, conversão alimentar, taxa de eficiência proteica e índice de sobrevivência. Os resultados foram submetidos à análise de variância e ao teste de Duncan (95% de precisão). Os resultados encontrados não apresentaram interação entre proteína e energia digestível para nenhum parâmetro avaliado. Em relação à proteína digestível, foram verificadas diferenças nos parâmetros consumo em proteína da dieta e taxa de eficiência proteica (TEP), o que demonstra ineficiência das dietas com altos valores proteicos. Deve-se oferecer aos girinos de rã-touro dietas com 27% de proteína digestível, por apresentarem melhor resultado para taxa de eficiência proteica e para consumo em proteína da dieta. Abstract in english From the digestibility values of some ingredients, it was proposed diets with different levels of digestible protein and energy, in a 3x3 factorial design for bullfrog tadpoles, aiming at evaluate the zootechnical performance. The experiment was conducted during 60 days with 810 tadpoles from the sa [...] me spawning distributed into 27 polypropylene boxes at a density of 1 tadpole/L. The experimental design consisted of a factorial arrangement with three levels of digestible protein (27; 31 and 35%) and three levels of digestible energy (2700; 2800 and 2900kcal/kg) with three repetitions. Parameters evaluated at 60 days were weight gain, feed intake, dietary protein intake, feed conversion, protein efficiency rate and survival rate. Results were subjected to analysis of variance and Duncan test (95% accuracy), and presented no interaction between digestible protein and digestible energy for any parameter evaluated. As for the digestible protein, significant differences were detected in the dietary protein intake, and protein efficiency rate, evidencing inefficiency of diets with high protein values. Bullfrog tadpoles should be given diets with 27% digestible protein by having the best result for the protein efficiency rate and for the dietary protein intake, with the lowest value and similar to other levels among evaluated parameters.

  16. Improving classification in protein structure databases using text mining

    Directory of Open Access Journals (Sweden)

    Jones David T

    2009-05-01

    Full Text Available Abstract Background The classification of protein domains in the CATH resource is primarily based on structural comparisons, sequence similarity and manual analysis. One of the main bottlenecks in the processing of new entries is the evaluation of 'borderline' cases by human curators with reference to the literature, and better tools for helping both expert and non-expert users quickly identify relevant functional information from text are urgently needed. A text based method for protein classification is presented, which complements the existing sequence and structure-based approaches, especially in cases exhibiting low similarity to existing members and requiring manual intervention. The method is based on the assumption that textual similarity between sets of documents relating to proteins reflects biological function similarities and can be exploited to make classification decisions. Results An optimal strategy for the text comparisons was identified by using an established gold standard enzyme dataset. Filtering of the abstracts using a machine learning approach to discriminate sentences containing functional, structural and classification information that are relevant to the protein classification task improved performance. Testing this classification scheme on a dataset of 'borderline' protein domains that lack significant sequence or structure similarity to classified proteins showed that although, as expected, the structural similarity classifiers perform better on average, there is a significant benefit in incorporating text similarity in logistic regression models, indicating significant orthogonality in this additional information. Coverage was significantly increased especially at low error rates, which is important for routine classification tasks: 15.3% for the combined structure and text classifier compared to 10% for the structural classifier alone, at 10-3 error rate. Finally when only the highest scoring predictions were used to infer classification, an extra 4.2% of correct decisions were made by the combined classifier. Conclusion We have described a simple text based method to classify protein domains that demonstrates an improvement over existing methods. The method is unique in incorporating structural and text based classifiers directly and is particularly useful in cases where inconclusive evidence from sequence or structure similarity requires laborious manual classification.

  17. Development, standardization and validation of nuclear based technologies for estimating microbial protein supply in ruminant livestock for improving productivity

    International Nuclear Information System (INIS)

    The primary constraint to livestock production in developing countries is the scarcity and fluctuating quantity and quality of the year-round feed supply. These countries experience serious shortages of animal feeds and fodders of the conventional type. Natural forages are very variable both in quality and quantity, conventional agro-industrial by-products are scarce and vary seasonal, and grains are required almost exclusively for human consumption. The small farmers in developing countries have limited resources available to them for feeding their ruminant livestock. Poor nutrition results in low rates of reproduction and production as well as increased susceptibility to disease and mortality. Providing adequate good-quality feed to livestock to raise and maintain their productivity is a major challenge to agricultural scientists and policy makers all over the world. Recent advances in ration balancing include manipulation of feed to increase the quantity and quality of protein and energy delivered to the small intestine. Selection of feeds based on high efficiency of microbial protein synthesis in the rumen along with the high dry matter digestibility, and development of feeding strategies based on high efficiency as well as high microbial protein synthesis in the rumen will lead to higher supply of protein post-ruminally. The strategy for improving production has therefore been to maximize the efficiency of utilization of available feed resources in the rumen by pavailable feed resources in the rumen by providing optimum conditions for microbial growth and thereby supplementing dietary nutrients to complement and balance the products of rumen digestion to the animal's requirement

  18. Ammonia treatment of wheat straw. 2. Efficiency of microbial protein synthesis, rumen microbial protein pool size and turnover, and small intestinal protein digestion in sheep.

    OpenAIRE

    Oosting, S. J.; Viets, T. C.; Lammers-wienhoven, S. C. W.; Bruchem, J.

    1993-01-01

    Ammonia-treated wheat straw (AWS) was compared with untreated wheat straw (UWS) and untreated wheat straw supplemented with urea (SWS) in an experiment with 6 wether sheep. Microbial protein synthesis increased after ammonia treatment due to the higher intake of rumen degradable organic matter (OM). The efficiency of microbial protein synthesis was low for all diets, averaging 22.1 g N/kg apparently rumen degradable OM and was not significantly affected by ammonia treatment. Estimates of micr...

  19. Approaches to improving the nutritional quality of barley seed proteins

    International Nuclear Information System (INIS)

    The poor nutritional quality of barley grain is determined by the low level of lysine in the prolamin storage proteins (hordein). These account for between 35 to 50% of the total grain nitrogen, depending on the nutritional status of the plant. There is a reduced proportion of hordein in mutant high lysine lines but these also have reduced synthesis of storage carbohydrates and hence low yields. Three strategies for improvement are discussed. Increases in the lysine content of hordein may be difficult to achieve because of the presence of complex families of structural genes. It would also be necessary to insert a large number of additional lysine residues. Two more promising approaches are to increase the level of expression of genes coding for lysine-rich globulin storage proteins and to increase the pool of free lysine by selecting mutant lines with relaxed feedback regulation of lysine synthesis. (author)

  20. Intake, digestibility, rumen characteristics and microbial protein synthesis efficiency in bovine and bubaline fed sugar cane silage with additives

    Scientific Electronic Library Online (English)

    Emilyn Midori, Maeda; Lucia Maria, Zeoula; Clóves Cabreira, Jobim; Ulysses, Cecato; Luiz Paulo, Rigolon; Ricardo, Kazama; Guido, Jacobi; Acir Felipe Grolli, Carvalho.

    2012-03-01

    Full Text Available Intake and total and ruminal digestibility, ruminal characteristics and microbial synthesis efficiency in bovine and bubaline fed sugar cane silages treated with microbial additive Lactobacillus buchneri (inoculate) and the addition of: cassava byproduct meal at 250 g/kg; soybean hulls at 250 g/kg; [...] and urea at 33 g/kg, in natural matter were evaluated. The forage:concentrate ratio was 600:400 g/kg. Bubaline body weight (BW) was 403±49 kg and bovine BW was 492±10 kg. The experimental design consisted of two 4 × 4 Latin squares in a 4 × 2 factorial arrangement. The average DM intake did not differ between species, and the diet with urea showed lowest intake (13 g/kg BW). Among the diets, the one with soybean hulls addition increased the total digestibility of DM, organic matter (OM) and neutral detergent fiber (NDF), while addition of urea reduced the DM, OM and non-fibrous carbohydrates (NFC) total digestibility in both species. Bovines have higher ruminal digestibility of NFC in the diet with addition of inoculate, inoculate + soybean meal and inoculate + urea diets than bubaline. Ruminal liquid pH was higher for bubaline (6.61) than bovines (6.48). Diets with addition of cassava byproduct and soybean hulls presented higher ruminal short chain fatty acids concentrations. Ruminal ammonia concentration (N-NH3) did not differ between species and the treatment with urea presented the highest value of N-NH3 (17.25 mg/100 mL). Liquid passage rate was higher for bubaline (12.9%/h) than bovines (9.9%/h). Bubaline has higher microbial synthesis efficiency than bovine: 35.1 vs 24.6 g of N-microbial/kg OM of the total rumen digestibility, regardless of the diet.

  1. Intake, digestibility, rumen characteristics and microbial protein synthesis efficiency in bovine and bubaline fed sugar cane silage with additives

    Directory of Open Access Journals (Sweden)

    Emilyn Midori Maeda

    2012-03-01

    Full Text Available Intake and total and ruminal digestibility, ruminal characteristics and microbial synthesis efficiency in bovine and bubaline fed sugar cane silages treated with microbial additive Lactobacillus buchneri (inoculate and the addition of: cassava byproduct meal at 250 g/kg; soybean hulls at 250 g/kg; and urea at 33 g/kg, in natural matter were evaluated. The forage:concentrate ratio was 600:400 g/kg. Bubaline body weight (BW was 403±49 kg and bovine BW was 492±10 kg. The experimental design consisted of two 4 × 4 Latin squares in a 4 × 2 factorial arrangement. The average DM intake did not differ between species, and the diet with urea showed lowest intake (13 g/kg BW. Among the diets, the one with soybean hulls addition increased the total digestibility of DM, organic matter (OM and neutral detergent fiber (NDF, while addition of urea reduced the DM, OM and non-fibrous carbohydrates (NFC total digestibility in both species. Bovines have higher ruminal digestibility of NFC in the diet with addition of inoculate, inoculate + soybean meal and inoculate + urea diets than bubaline. Ruminal liquid pH was higher for bubaline (6.61 than bovines (6.48. Diets with addition of cassava byproduct and soybean hulls presented higher ruminal short chain fatty acids concentrations. Ruminal ammonia concentration (N-NH3 did not differ between species and the treatment with urea presented the highest value of N-NH3 (17.25 mg/100 mL. Liquid passage rate was higher for bubaline (12.9%/h than bovines (9.9%/h. Bubaline has higher microbial synthesis efficiency than bovine: 35.1 vs 24.6 g of N-microbial/kg OM of the total rumen digestibility, regardless of the diet.

  2. Improving accuracy of protein contact prediction using balanced network deconvolution.

    Science.gov (United States)

    Sun, Hai-Ping; Huang, Yan; Wang, Xiao-Fan; Zhang, Yang; Shen, Hong-Bin

    2015-03-01

    Residue contact map is essential for protein three-dimensional structure determination. But most of the current contact prediction methods based on residue co-evolution suffer from high false-positives as introduced by indirect and transitive contacts (i.e., residues A-B and B-C are in contact, but A-C are not). Built on the work by Feizi et al. (Nat Biotechnol 2013; 31:726-733), which demonstrated a general network model to distinguish direct dependencies by network deconvolution, this study presents a new balanced network deconvolution (BND) algorithm to identify optimized dependency matrix without limit on the eigenvalue range in the applied network systems. The algorithm was used to filter contact predictions of five widely used co-evolution methods. On the test of proteins from three benchmark datasets of the 9th critical assessment of protein structure prediction (CASP9), CASP10, and PSICOV (precise structural contact prediction using sparse inverse covariance estimation) database experiments, the BND can improve the medium- and long-range contact predictions at the L/5 cutoff by 55.59% and 47.68%, respectively, without additional central processing unit cost. The improvement is statistically significant, with a P-value?co-evolution-based contact prediction to the three-dimensional structure modeling relies on the number of homologous sequences existing in the sequence databases. BND can be used as a general contact refinement method, which is freely available at: http://www.csbio.sjtu.edu.cn/bioinf/BND/. Proteins 2015; 83:485-496. © 2014 Wiley Periodicals, Inc. PMID:25524593

  3. Improvements in growth performance, bone mineral status and nutrient digestibility in pigs following the dietary inclusion of phytase are accompanied by modifications in intestinal nutrient transporter gene expression.

    Science.gov (United States)

    Vigors, Stafford; Sweeney, Torres; O'Shea, Cormac J; Browne, John A; O'Doherty, John V

    2014-09-14

    Phytase (PHY) improves growth performance, nutrient digestibility and bone structure in pigs; however, little is known about its effects on intestinal nutrient transporter gene expression. In the present study, a 44 d experiment was carried out using forty-eight pigs (11·76 (sem 0·75) kg) assigned to one of three dietary treatment groups to measure growth performance, coefficient of apparent ileal digestibility (CAID), coefficient of apparent total tract nutrient digestibility (CATTD) and intestinal nutrient transporter gene expression. Dietary treatments during the experimental period were as follows: (1) a high-P (HP) diet containing 3·4 g/kg available P and 7·0 g/kg Ca; (2) a low-P (LP) diet containing 1·9 g/kg available P and 5·9 g/kg Ca; (3) a PHY diet containing LP diet ingredients+1000 phytase units (FTU)/kg of PHY. The PHY diet increased the average daily gain (P< 0·05) and final body weight (P< 0·01) and decreased the feed conversion ratio (P< 0·05) compared with the LP diet. Pigs fed the PHY diet had a higher CAID of gross energy compared with those fed the HP and LP diets (P< 0·001). Pigs fed the PHY diet had increased CAID of P (P< 0·01) and CATTD of Ca and P (P< 0·001) compared with those fed the LP diet. The PHY diet increased the gene expression of the peptide transporter 1 (PEPT1/SLC15A1) (P< 0·05) in the ileum compared with the LP diet. The LP diet decreased the gene expression of the sodium-glucose-linked transporter 1 (SGLT1/SLC5A1) and GLUT2/SLC2A2 (P< 0·05) and increased the expression of membrane Ca channel (TRPV6) and calbindin compared with the HP diet (P< 0·001). In conclusion, feeding a diet supplemented with PHY improves growth performance and nutrient digestibility as well as increases the gene expression of the peptide transporter PEPT1. PMID:24998244

  4. Influence of crude protein content, ingredient complexity, feed form, and duration of feeding of the Phase I diets on productive performance and nutrient digestibility of Iberian pigs.

    Science.gov (United States)

    Berrocoso, J D; Saldaña, B; Serrano, M P; Cámara, L; Ibáñez, M A; Mateos, G G

    2013-03-01

    The influence of CP content and ingredient complexity, feed form, and duration of feeding of the Phase I diets on growth performance and total tract apparent digestibility (TTAD) of energy and nutrients was studied in Iberian pigs weaned at 28 d of age. There were 12 dietary treatments with 2 type of feeds (high-quality, HQ; and low-quality, LQ), 2 feed forms (pellets vs. mash), and 3 durations (7, 14, and 21 d) of supply of the Phase I diets. From d 7, 14, or 21 (depending on treatment) to d 35, all pigs received a common diet in mash form. Each treatment was replicated 3 times (6 pigs/pen). For the entire experiment, ADG (P pigs fed the HQ diets than pigs fed the LQ diets and pigs fed pellets than those fed mash (P pigs fed the Phase I diet for 14 or 21 d than those fed the diet for 7 d (P 0.10) by feed quality, but feeding pellets or increasing the duration of feeding the Phase I diets improved G:F (P pigs fed the Phase I diets for 14 or 21 d than for pigs fed the diet for only 7 d (P pigs previously fed the LQ diet had greater ADG than pigs fed the HQ Phase I diets (P pigs fed the Phase I diets for 21 d had decreased ADG (P pigs fed these diets for 7 or 14 d. Organic matter digestibility was greater for pigs fed the HQ Phase I diets than pigs fed the LQ Phase I diets (P Iberian pigs from d 0 to 35. Also, pelleting improved energy and nutrient digestibility and feed efficiency. Increasing the duration of supply of the Phase I diets from 7 to 21 d improved feed efficiency but reduced ADG. Therefore, the use of LQ Phase I diets in pellet form for no more than 7 d after weaning is recommended in Iberian pigs. PMID:23307842

  5. Digestibilidad aparente de proteína cruda y lípidos en la brótola, Urophycis brasiliensis (Kamp, 1858) (Pisces: Gadiformes), alimentada con reemplazos parciales de harina de soja y harina de carne / Apparent digestibility of crude protein and lipids in Brazilian codling, Urophycis brasiliensis (Kamp, 1858) (Pisces: Gadiformes), fed with partial replacements of soybean meal and meat meal diets

    Scientific Electronic Library Online (English)

    Sergio N, Bolasina; Jorge L, Fenucci.

    2005-12-01

    Full Text Available SciELO Chile | Language: Spanish Abstract in spanish El objetivo de este trabajo fue determinar la digestibilidad aparente de dietas (proteína cruda y lípidos) formuladas para brótola. Se investigaron tres dietas (43% proteína cruda): una control con harina de pescado como la principal fuente protéica y otras dos con reemplazo parcial de este ingredie [...] nte por harina de soja desgrasada y harina de carne. Los peces se alimentaron diariamente hasta la saciedad y no hubo mortalidades durante los 14 días que duró el experimento. En los días 7 y 14, los peces fueron anestesiados y se extrajeron las heces por masaje abdominal. La digestibilidad de proteína cruda, al séptimo día, fue significativamente diferente al comparar el grupo control con los grupos donde se reemplazó la harina de pescado. Se encontraron diferencias significativas en la digestibilidad de lípidos entre el grupo alimentado con harina de soja y el grupo alimentado con harina de carne durante todo el experimento. Los valores de digestibilidad más altos y los menores porcentajes de humedad en las heces se encontraron en el grupo control, sugiriendo una mejor digestión de esta dieta. En las dietas con reemplazos, la digestibilidad aparente se incrementó de la primera a la segunda semana. Se puede concluir que esta especie puede ser alimentada con dietas con reemplazos de hasta un 30% de harina de pescado utilizando harina de soja desgrasada o de carne; el tiempo necesario para la adaptación enzimática de los peces a esta dieta es de alrededor de dos semanas. Abstract in english The objective of this study was to determine apparent digestibility of crude protein and lipid in Brazilian codling. Three diets (43% crude protein) were tested: a control diet with fish meal as the main protein source, and two others with partial replacement of fish meal by defatted soybean and mea [...] t meal. Fishes were fed daily to satiation, and no mortality was observed during the 14 day-experiment. On days 7 and 14, fish were anesthetized and faeces were obtained by stripping. Apparent crude protein digestibility at day 7 was significantly different between the control group and the groups in which fish meal was replaced. Lipid digestibility showed significant differences between soybean group and meat meal group throughout the trial. The highest digestibility values and the lowest moisture percentages in faeces were found in the control group, suggesting an improved digestion of this diet. Apparent digestibility increased from first to second week in diets with replacements. Brazilian codling can be reared on diets in which up to 30% replacement of the fish meal protein can be replaced by defatted soybean or meat meal. Fish need about two weeks for enzymatic adaptation to these diets.

  6. Chapter 11. Digestive tube

    International Nuclear Information System (INIS)

    Exploration of the digestive tube by radioactive tracers relates mainly to the functional study of certain digestion or absorption troubles. The tracer absorbed by the digestibe system was followed by counting of the stools, successive measurements of the plasma radioactivity, measurements of urinary elimination or uptake on a storage organ such as the liver in the case of vitamin B12, measurement of whole-body radioactivity for vitamin B12 and iron. The different isotopic techniques used to study intestinal absorption of lipids, proteins and aminoacids, vitamin B 12 and iron were described and their contribution to the detection of exudative enteropathies and digestive haemorrhage was shown. It was pointed out that the stomach is one of the organs most accessible to standard exploration techniques. The role of sup(99m)Tc in both the morphological exploration of stomach and the study of gastric secretion, of 51Cr and 129Cs in the study of gastric evacuation were demonstrated

  7. Fermentation of Sugarcane Sludge to Improve Dry- and and Organic Matters Digestibility and Rumen Parameters In Vitro

    Directory of Open Access Journals (Sweden)

    Yusuf Widodo

    2008-01-01

    Full Text Available The objective of the research was to study the utilization of fungi and yeast on sugarcane sludge fermentation. The treatments of the first experiment were P0= sugarcane sludge without fermentation; P1= fermentation by Saccharomyces cereviceae; P2 = fermentation by Aspergillus oryzae; P3 = fermentation by Aspergillus niger; P4= fermentation by Rhizopus orryzae. Treatments were arranged in a Completely Randomised Design. The treatments of the second experiments were R0 = basic rations, R1 = R0 + 5% fermented of sugarcane sludge, R2 = R0 + 10% fermented of sugarcane sludge, R3 = R0 + 15% fermented of sugarcane sludge, R4 = R0 + 20% fermented of sugarcane sludge. Treatments have significant effects (P<0.05 on VFA and NH3 parameters. Based on these parameters, fermentation of sugarcane sludge using Saccharomyces cereviceae had better results compared to others. The treatments had significant effect (P<0.05 on NH3, and based on polynomial orthogonal test the treatments had linear curve of Y= 4,035 +0,237X. However, treatments had no significant effects on VFA and organic matters digestibility. The dry matters digestibility parameters were significantly affected (P<0.05, and the curve response was linear with equation of Y=45,964 - 0,294X. (Animal Production 10(1: 30-33 (2008 Key Words: Sugarcane sludge processing, fungi, yeast, in vitro

  8. VOLUNTARY INTAKE, APPARENT DIGESTIBILITY AND DIGESTA KINETICS OF THREE PROTEIN-LEVEL FORAGES FED TO SHEEP CONSUMO VOLUNTÁRIO, DIGESTIBILIDADE APARENTE E CINÉTICA DIGESTIVA DE TRÊS FORRAGEIRAS EM OVINOS

    Directory of Open Access Journals (Sweden)

    Adibe Luiz Abdalla

    2007-12-01

    Full Text Available

    The aim of this work was to evaluate the voluntary intake, apparent digestibility and digesta kinetics of three protein-level forages fed to Santa Inês sheep. Six sheep (LW = 40 ± 5.7 kg fitted with rumen cannulas were used. Lucerne (LUC, signalgrass (SIG and Tifton-85 grass (TIF hays were chosen due to their crude protein (CP content (respectively, 191, 29 and 75 g kg-1 DM. Treatments and animals were fitted in a double Latin square (3 treatments, 3 periods, 6 animals. To measure the voluntary intake, animals were kept in individual stall with free access to water and food, during the first phase of each period. The feed consisted exclusively of LUC, SIG or TIF and mineral supplementation. Apparent digestibility was determined by total faeces collection. Digesta kinetics was estimated using Co-EDTA and Cr mordant fibre as markers to liquid and solid phases, respectively. Means were compared by Tukey test. Dry matter voluntary intake of treatment LUC was higher (P < 0.05 than treatments SIG and TIF and it reflected the strong relationship between intake and protein content in the diet. Dry matter and organic matter apparent digestibility coefficients were superior (P < 0.05 for LUC. Crude protein apparent digestibility showed differences (P < 0.05 between the three treatments, being LUC (0.694 the highest observed followed by TIF (0.500. Crude protein apparent digestibility of SIG was practically null (0.001, indicating intense protein deficit. Dry matter voluntary intake and digestibility of tested feeds were influenced by chemical composition and digestion kinetics. Low protein feeds had a prejudicial effect in nutrient apparent digestibi-lity, decreasing until nullity for protein of treatment SIG.

    Key-words: Lucerne, marker, nutrition, ruminant, signalgrass, Tifton.

    O objetivo deste trabalho foi avaliar o consumo voluntário, a digestibilidade aparente e a cinética diges-tiva de três forragens, com teores protéicos distintos, em ovinos. Utilizaram-se  seis ovinos da raça Santa Inês (PV = 40 ± 5,7 kg, machos adultos, castrados e providos de cânulas ruminais. Optou-se pelo uso de fenos de alfafa (ALF, de braquiária (BRA e de capim Tifton-85 (TIF pelos seus teores de proteína bruta (PB, respectivamente, 191, 29 e 75 g kg-1 MS. Alocaram-se os tratamentos e os animais em delineamento de dois quadrados latinos (três tratamentos, três períodos e seis animais. As dietas consistiam exclusivamente de ALF, BRA ou TIF e mistura mineral, oferecida separadamente. O ensaio de consumo voluntário foi realizado em baias individuais e os de digestibilidade e cinética digestiva, em gaiolas de metabolismo. Determinou-se a digestibilidade aparente por coleta total de fezes. Estimou-se a cinética digestiva mediante o uso de Co-EDTA e fibra Cr-mordantada como marcadores das fases líquida e sólida, respectivamente. Compararam-se as médias por teste de Tukey. O consumo voluntário de MS do tratamento ALF foi maior (P < 0,05 que os tratamentos BRA e TIF, o que refletiu a forte relação entre consumo e teor de proteína na dieta. Os coeficientes de digestibilidade aparente da MS e da MO também foram superiores (P < 0,05 para ALF. A digestibilidade aparente da proteína bruta mostrou diferenças (P < 0,05 entre os três tratamentos, sendo ALF (0,694 a mais alta que se observou, seguida por TIF (0,500. Para o tratamento BRA, essa variável foi praticamente nula (0,001, indicando déficit protéico intenso. O consumo voluntário e a digestibilidade aparente dos alimentos testados receberam influência da composição química e da cinética digestiva. Alimentos com baixo teor protéico prejudicaram a digestibilidade aparente de nutrientes, diminuindo, próximo à nulidade, para a proteína do tratamento BRA.

    Palavras-chaves:  Alfafa, braquiária, marcadores, nutrição, ruminantes, Tifton.

  9. Protein sources and digestive enzyme activities in jundiá (Rhamdia quelen Fontes protéicas e atividade de enzimas digestivas em jundiás (Rhamdia quelen

    Directory of Open Access Journals (Sweden)

    Rafael Lazzari

    2010-01-01

    Full Text Available Digestive enzymes activity influence feed utilization by fish, and its understanding is important to optimize diet formulation. This study reports the digestive enzyme activities of jundiá juveniles fed diets with protein sources. Fish were fed six experimental diets for 90 days: MBY (meat and bone meal + sugar cane yeast, SY (soybean meal + sugar cane yeast, S (soybean meal, MBS (meat and bone meal + soybean meal, FY (fish meal + sugar cane yeast and FS (fish meal + soybean meal, and then sampled every 30 days and assayed in two intestine sections for digestive enzymes - trypsin, chymotrypsin and amylase - activities; gastric protease was assayed in the stomach. Digestive and hepatosomatic index, intestinal quotient, digestive tract length and weight gain were also measured. Trypsin and chymotrypsin activities were higher (p As enzimas digestivas influenciam a utilização dos alimentos em peixes, e seu conhecimento é importante para otimizar a formulação de dietas. Este trabalho descreve a atividade de enzimas digestivas em juvenis de jundiá alimentados com fontes protéicas. Os peixes foram alimentados com seis dietas (90 dias: MBY (farinha de carne e ossos + levedura de cana, SY (farelo de soja + levedura de cana, S (farelo de soja, MBS (farinha de carne e ossos + farelo de soja, FY (farinha de peixe + levedura de cana e FS (farinha de peixe + farelo de soja. A cada 30 dias, foram analisadas as enzimas digestivas (tripsina, quimiotripsina e amilase no intestino. No estômago, foi mensurada a protease ácida. Foram estimados os índices digestivo e hepato-somático, quociente intestinal, comprimento do trato digestório e ganho em peso. As atividades de tripsina e quimiotripsina foram maiores (p < 0,0001 nos peixes alimentados com as dietas com fontes de origem animal (MBY, MBS, FY e FS. As proteases alcalinas foram afetadas negativamente pelo farelo de soja nas dietas. A atividade de amilase apresentou grandes variações. Os peixes alimentados com as dietas MBY e MBS apresentaram maior atividade de protease ácida (p < 0,0001. Os ganhos em peso foram maiores nos peixes alimentados com as dietas MBS e FS (p < 0,05. Os maiores valores (p < 0,05 de comprimento do trato digestório foram verificados nos peixes alimentados com as dietas MBS e FY. A composição da dieta influencia na atividade de enzimas digestivas do jundiá.

  10. Food allergen digestibility: The influence on allergenicity

    DEFF Research Database (Denmark)

    BØgh, Katrine Lindholm; Madsen, Charlotte Bernhard

    Food allergy is a major health problem in the Western countries, affecting 3-8% of the population. What makes a dietary protein a food allergen has not yet been established, though several characteristics have been proposed to be shared by the food allergens. One of the features believed to be a general characteristic is resistance to digestion. This is based on studies showing that allergenic dietary proteins in general were more resistant to digestion than dietary proteins with no proven allergenicity, leading to the conclusion, that a correlation between stability to digestion and allergenic potential exist. Resistance to digestion is therefore a test parameter included in the safety assessment of the allergenic potential of novel proteins in genetically modified foods. In recent years, the association between resistance to digestion and allergenic potential has been challenged. When reviewing existing data from digestibility studies on known food allergens, it becomes evident that food allergens do not necessarily resist digestion. However, the choice of assay conditions, the method used for detection of residual intact protein as well as fragments hereof greatly influences the outcome. Studies assessing the allergenicity of digestion products, by either IgE-binding, elicitation or sensitising capacity, shows that digestion may abolish, decrease, have no effect, or even increase the allergenicity of food allergens. However, this dependents on the given allergen. In conclusion, reviewing existing digestibility data shows that no absolute correlation between resistance to digestion and allergenic potential exist. Therefore stability to digestion may not necessarily be a good parameter for assessing the allergenic potential of novel proteins. Even very small peptides from food allergens may retain both IgE-binding, eliciting and sensitising capacity. As a consequence immunological studies should be performed when evaluating the digestibility of protein allergens.

  11. Bulgur processes increase nutrition value: possible role in in-vitro protein digestability, phytic acid, trypsin inhibitor activity and mineral bioavailability.

    Science.gov (United States)

    Erta?, Nilgün; Türker, Selman

    2014-07-01

    Changes in the chemical constituents and nutritive quality of chickpea bulgur process, were studied in seeds that were soaked at different time (2, 8 and 12 h), different soaking water pH (pH 4, 6 and 8). Soaking in pH 8 soaking water and 12 h soaking time significantly (p?chickpea bulgur samples. Compared to the raw material, the protein content and in-vitro protein digestibility increased, but starch, crude fiber, fat and energy values decreased and trypsin inhibitor activity was completely eliminated by bulgur process. As the soaking time increased, the phytic acid content also decreased. The highest total phenolic content was determinated with bulgur samples soaked in pH 4 soaking water. The P, Ca, and K values decreased with increasing soaking time. The HCl-extractability of P, Ca, Mg, Fe and K present in chickpea bulgur samples were significantly higher than the raw chickpea seeds. PMID:24966437

  12. Using support vector machine for improving protein-protein interaction prediction utilizing domain interactions

    Energy Technology Data Exchange (ETDEWEB)

    Singhal, Mudita; Shah, Anuj R.; Brown, Roslyn N.; Adkins, Joshua N.

    2010-10-02

    Understanding protein interactions is essential to gain insights into the biological processes at the whole cell level. The high-throughput experimental techniques for determining protein-protein interactions (PPI) are error prone and expensive with low overlap amongst them. Although several computational methods have been proposed for predicting protein interactions there is definite room for improvement. Here we present DomainSVM, a predictive method for PPI that uses computationally inferred domain-domain interaction values in a Support Vector Machine framework to predict protein interactions. DomainSVM method utilizes evidence of multiple interacting domains to predict a protein interaction. It outperforms existing methods of PPI prediction by achieving very high explanation ratios, precision, specificity, sensitivity and F-measure values in a 10 fold cross-validation study conducted on the positive and negative PPIs in yeast. A Functional comparison study using GO annotations on the positive and the negative test sets is presented in addition to discussing novel PPI predictions in Salmonella Typhimurium.

  13. Integrative Effects of Feeding Aspergillus awamori and fructooligosaccharide on Growth Performance and Digestibility in Broilers: Promotion Muscle Protein Metabolism

    OpenAIRE

    Saleh, Ahmed A.; Khairy Amber; El-magd, Mohammed A.; Atta, Mostafa S.; Mohammed, Ahmed A.; Ragab, Mohamed M.; Hanaa Abd El-Kader

    2014-01-01

    This study was conducted to show the effect of Aspergillus awamori (AA), fructooligosaccharide (FOS), and combined Aspergillus awamori and fructooligosaccharide (AA?+?FOS) on growth, digestibility, blood parameters, and expression of some growth-related genes. A total of 60 broiler chicks at the age of 15?d were divided into a control group (n = 15) and 3 treatment groups. The control group was fed a basal diet, and the treatment groups were fed basal diets supplemented with 0.05% AA, 0...

  14. Use of the mobile nylon bag technique for determination of apparent ileal digestibilities of crude protein and amino acids in feedstuffs for pigs

    DEFF Research Database (Denmark)

    Steiner, T; Bornholdt, U

    2011-01-01

    Three digestibility experiments were conducted to evaluate the potential of determination of apparent ileal digestibilities (AID) of crude protein (CP) and amino acids (AA) by the mobile nylon bag technique (MNBT) using 21 feedstuffs and three mixed diets. In two conventional digestibility experiments (Exp. 1 and 2), AID were determined using in total 10 barrows (BW 35 kg) fitted with simple T-cannulas at the terminal ileum. For the MNBT studies, four pigs were fitted with a simple T-cannula at the proximal duodenum and a Post-Valve T-Caecum (PVTC) cannula at the terminal ileum. The MNBT studies included the feedstuffs (n = 10) from Exp. 1 and 2 as well as 14 further feedstuffs and mixed diets in which AID coefficients had been determined in previous trials. For each feedstuff 60 nylon bags were used. In vitro digestion of the bags was carried out in pepsin-HCl solution with 450 IU pepsin/l at pH 2.0 and 37°C for 4 h. In the 28-day in vivo experiment, 15 nylon bags per pig and day were inserted through the duodenal cannula and collected through the PVTC cannula after passage through the small intestine. Coefficients of AID were calculated based on the disappearance of CP and AA from the nylon bags during the in vitro and in vivo phase. In comparison with AID determined by the conventional method, AID of CP was on average 2.4% lower, whereas AID of lysine was on average 8.5% higher when determined by the MNBT. There was no significant (P > 0.05) correlation between AID coefficients of CP and AA determined by the conventional method and the MNBT, when all feedstuffs were taken into account. However, in cereals (n = 11), the correlation between AID coefficients determined by both methods was significant (P < 0.05) for CP (r = 0.61) and some AA (r ranging between 0.62 and 0.72). In conclusion, the potential of the MNBT to determine AID of CP and AA is rather limited. Differences in coefficients of AID of CP and AA were attributed to several factors such as diffusion of sample particles or endogenous protein through the nylon bags as well as to the presence of anti-nutritional factors (e.g. in legume seeds and oilseed meals).

  15. Double restriction-enzyme digestion improves the coverage and accuracy of genome-wide CpG methylation profiling by reduced representation bisulfite sequencing

    Directory of Open Access Journals (Sweden)

    Wang Junwen

    2013-01-01

    Full Text Available Abstract Background Reduced representation bisulfite sequencing (RRBS was developed to measure DNA methylation of high-CG regions at single base-pair resolution, and has been widely used because of its minimal DNA requirements and cost efficacy; however, the CpG coverage of genomic regions is restricted and important regions with low-CG will be ignored in DNA methylation profiling. This method could be improved to generate a more comprehensive representation. Results Based on in silico simulation of enzyme digestion of human and mouse genomes, we have optimized the current single-enzyme RRBS by applying double enzyme digestion in the library construction to interrogate more representative regions. CpG coverage of genomic regions was considerably increased in both high-CG and low-CG regions using the double-enzyme RRBS method, leading to more accurate detection of their average methylation levels and identification of differential methylation regions between samples. We also applied this double-enzyme RRBS method to comprehensively analyze the CpG methylation profiles of two colorectal cancer cell lines. Conclusion The double-enzyme RRBS increases the CpG coverage of genomic regions considerably over the previous single-enzyme RRBS method, leading to more accurate detection of their average methylation levels. It will facilitate genome-wide DNA methylation studies in multiple and complex clinical samples.

  16. An Improved Fast Photochemical Oxidation of Proteins (FPOP) Platform for Protein Therapeutics

    Science.gov (United States)

    Zhang, Ying; Rempel, Don L.; Zhang, Hao; Gross, Michael L.

    2015-03-01

    Unlike small-molecule drugs, the size and dynamics of protein therapeutics challenge existing methods for assessing their high order structures (HOS). To extend fast photochemical oxidation of proteins (FPOP) to protein therapeutics, we modified its platform by introducing a mixing step prior to laser irradiation to minimize unwanted H2O2-induced oxidation. This improvement plus standardizing each step yield better reproducibility as determined by a fitting process whereby we used a non-FPOP spectrum as a template to report the unmodified level. We also tested different buffer systems for this modified FPOP platform with cytochrome c. The outcome is a standard oxidation profile that can be compared between different laboratories and regulatory agencies that wish to adopt FPOP for quality control purposes.

  17. An Improved Fast Photochemical Oxidation of Proteins (FPOP) Platform for Protein Therapeutics.

    Science.gov (United States)

    Zhang, Ying; Rempel, Don L; Zhang, Hao; Gross, Michael L

    2015-03-01

    Unlike small-molecule drugs, the size and dynamics of protein therapeutics challenge existing methods for assessing their high order structures (HOS). To extend fast photochemical oxidation of proteins (FPOP) to protein therapeutics, we modified its platform by introducing a mixing step prior to laser irradiation to minimize unwanted H2O2-induced oxidation. This improvement plus standardizing each step yield better reproducibility as determined by a fitting process whereby we used a non-FPOP spectrum as a template to report the unmodified level. We also tested different buffer systems for this modified FPOP platform with cytochrome c. The outcome is a standard oxidation profile that can be compared between different laboratories and regulatory agencies that wish to adopt FPOP for quality control purposes. PMID:25519854

  18. Effect of different concentrations of protein on the digestive system of juvenile silver catfish Efeito de diferentes concentrações de proteína sobre o sistema digestivo de juvenis de jundiá

    Directory of Open Access Journals (Sweden)

    J.F.B. Melo

    2012-04-01

    Full Text Available Aspects of digestion in juvenile silver catfish Rhamdia quelen fed with four levels of protein (20, 27, 34 and 41% were studied. The studied traits were digestive enzymes (nonspecific proteases, trypsin, chymotrypsin, amylase and lipase and weight gain. The experiment lasted 60 days; 160 juvenile catfish with initial body weight of 32 grams were transferred to the experimental system of 500L tanks with re-circulated and thermostatized water. The average weight of silver catfish increased with dietary protein and 41% of crude protein (CP was accepted as the best one. Gastric protease activity increases with the CP level. The intestinal alkaline protease reached the peak at 27% CP and the highest activities were observed in the anterior section of the gut (AS. The trypsin peak was observed at 34% CP in the AS and medium section of the gut (MS. Amylase and lipase activities were inverse to the dietary CP and the highest values were observed in AS. In conclusion, the digestive enzymes of catfishes are responsive to the dietary protein level. The dietary CP required for highest weight gain of juvenile silver catfish is 41%.Foram estudados aspectos digestivos de juvenis de jundiá Rhamdia quelen alimentados com quatro níveis de proteína (20, 27, 34 e 41 %. As características estudadas foram enzimas digestivas (proteases inespecíficas, tripsina, quimiotripsina, amilase e lipase e ganho de peso. O período experimental foi de 60 dias; 160 juvenis de jundiá com peso médio inicial de 32 gramas foram transferidos para um sistema experimental de tanques de 500L, com recirculação de água termostatizada. O peso médio dos jundiás aumentou com o teor de proteínas da dieta até 41% de proteína bruta (PB, valor esse considerado o melhor. A atividade de protease gástrica aumentou com o nível de PB. A protease alcalina intestinal atingiu o pico em 27% de PB e as maiores atividades foram observadas na secção anterior do intestino (SA. O pico de tripsina foi observado com 34% de PB na SA e na secção média do intestino (SM. As atividades de amilase e lipase foram inversas ao teor de PB, e os valores mais elevados foram observados na SA. Em conclusão, as enzimas digestivas de jundiá são responsivas ao nível de proteína da dieta. O teor mais alto de PB da dieta necessário ao maior ganho de peso em juvenis de jundiá é 41%.

  19. Effect of different concentrations of protein on the digestive system of juvenile silver catfish / Efeito de diferentes concentrações de proteína sobre o sistema digestivo de juvenis de jundiá

    Scientific Electronic Library Online (English)

    J.F.B., Melo; L.M., Lundstedt; G., Moraes; L.A.K.A., Inoue.

    2012-04-01

    Full Text Available Foram estudados aspectos digestivos de juvenis de jundiá Rhamdia quelen alimentados com quatro níveis de proteína (20, 27, 34 e 41 %). As características estudadas foram enzimas digestivas (proteases inespecíficas, tripsina, quimiotripsina, amilase e lipase) e ganho de peso. O período experimental f [...] oi de 60 dias; 160 juvenis de jundiá com peso médio inicial de 32 gramas foram transferidos para um sistema experimental de tanques de 500L, com recirculação de água termostatizada. O peso médio dos jundiás aumentou com o teor de proteínas da dieta até 41% de proteína bruta (PB), valor esse considerado o melhor. A atividade de protease gástrica aumentou com o nível de PB. A protease alcalina intestinal atingiu o pico em 27% de PB e as maiores atividades foram observadas na secção anterior do intestino (SA). O pico de tripsina foi observado com 34% de PB na SA e na secção média do intestino (SM). As atividades de amilase e lipase foram inversas ao teor de PB, e os valores mais elevados foram observados na SA. Em conclusão, as enzimas digestivas de jundiá são responsivas ao nível de proteína da dieta. O teor mais alto de PB da dieta necessário ao maior ganho de peso em juvenis de jundiá é 41%. Abstract in english Aspects of digestion in juvenile silver catfish Rhamdia quelen fed with four levels of protein (20, 27, 34 and 41%) were studied. The studied traits were digestive enzymes (nonspecific proteases, trypsin, chymotrypsin, amylase and lipase) and weight gain. The experiment lasted 60 days; 160 juvenile [...] catfish with initial body weight of 32 grams were transferred to the experimental system of 500L tanks with re-circulated and thermostatized water. The average weight of silver catfish increased with dietary protein and 41% of crude protein (CP) was accepted as the best one. Gastric protease activity increases with the CP level. The intestinal alkaline protease reached the peak at 27% CP and the highest activities were observed in the anterior section of the gut (AS). The trypsin peak was observed at 34% CP in the AS and medium section of the gut (MS). Amylase and lipase activities were inverse to the dietary CP and the highest values were observed in AS. In conclusion, the digestive enzymes of catfishes are responsive to the dietary protein level. The dietary CP required for highest weight gain of juvenile silver catfish is 41%.

  20. Effects of duration of moderate increases in grain feeding on endotoxins in the digestive tract and acute phase proteins in peripheral blood of yearling calves.

    Science.gov (United States)

    Plaizier, J C; Li, S; Le Sciellour, M; Schurmann, B L; Górka, P; Penner, G B

    2014-11-01

    Effects of duration of grain feeding on the concentration of endotoxic lipopolysaccharide (LPS) in digesta throughout the digestive tract and on acute phase proteins and LPS in peripheral blood were determined in Holstein yearling calves. Twenty-five Holstein yearling steer calves received either a forage-based diet containing 92% hay and 8% of a mineral and vitamin pellet on a dry matter basis (CON) or a moderate-grain diet, obtained by replacing 41.5% of the hay in the forage-based diet with barley grain, for 3 (MG3), 7 (MG7), 14 (MG14), or 21 d (MG21) before slaughter. Immediately before slaughter, blood samples were collected from the jugular vein. Immediately after slaughter, digesta samples were collected from the rumen, jejunum, ileum, cecum, colon, and rectum. Rumen liquid digesta, digesta from the intestines, and peripheral blood plasma were analyzed for LPS. Peripheral blood plasma and serum were analyzed for the acute phase proteins amyloid A, haptoglobin, and LPS-binding protein. Feeding the grain diet increased the LPS concentration in rumen fluid linearly from 15,488 endotoxin units (EU)/mL for CON to 70,146 EU/mL for MG7. Concentrations of LPS in rumen fluid in MG14 and MG21 were 61,944 and 56,234 EU/mL, respectively, and did not differ. The LPS concentrations in jejunal digesta were much lower than that in digesta elsewhere in the digestive tract, which suggests that ruminal LPS is broken down in the abomasum or proximal jejunum. The concentration of digesta LPS in the ileum was higher than that of digesta elsewhere in the intestines and similar to that in rumen fluid. The duration of grain feeding increased the LPS concentration in digesta in the ileum and cecum and tended to increase that in the colon cubically. Concentrations of LPS in this part of the digestive tract were highest in the MG3 and MG21 groups. The highest concentrations of LPS in digesta in the cecum, colon, and rectum were 3.7, 3.8, and 5.6 times higher than that in CON, respectively. Grain feeding and the increase in LPS in digesta were not accompanied by an acute phase response or a detectable concentration of LPS in peripheral blood. The absence of LPS in peripheral blood and the lack of increase in acute phase proteins indicated that the grain feeding protocol used in the current study and the accompanying changes in LPS concentrations of the digesta did not result in systemic inflammation. PMID:25242427

  1. Microbial community structure reveals how microaeration improves fermentation during anaerobic co-digestion of brown water and food waste.

    Science.gov (United States)

    Lim, Jun Wei; Chiam, Jun An; Wang, Jing-Yuan

    2014-11-01

    The purpose of this study was to investigate the impact of microaeration on the fermentation process during anaerobic co-digestion of brown water (BW) and food waste (FW). This was achieved by daily monitoring of reactor performance and the determination of its bacterial consortium towards the end of the study. Molecular cloning and sequencing results revealed that bacteria within phyla Firmicutes and Bacteriodetes represented the dominant phylogenetic group. As compared to anaerobic conditions, the fermentation of BW and FW under microaeration conditions gave rise to a significantly more diverse bacterial population and higher proportion of bacterial clones affiliated to the phylum Firmicutes. The acidogenic reactor was therefore able to metabolize a greater variety of substrates leading to higher hydrolysis rates as compared to the anaerobic reactor. Other than enhanced fermentation, microaeration also led to a shift in fermentation production pattern where acetic acid was metabolized for the synthesis of butyric acid. PMID:25194261

  2. Níveis de proteína na dieta de novilhas da raça Pantaneira: desempenho e digestibilidade aparente / Increasing levels of protein in the diet of heifers of Pantaneira Breed: performance and digestibility

    Scientific Electronic Library Online (English)

    A.L., Teodoro; M.V.M., Oliveira; F.M., Vargas Junior; R.S., Juliano; M.L., Longo; L.O., Seno; C.A.L., Oliveira.

    2013-09-01

    Full Text Available SciELO Spain | Language: Portuguese Abstract in portuguese Objetivou-se com este trabalho verificar a influência de níveis crescentes de proteína dietética sobre o desempenho, digestibilidade aparente e concentrações de uréia plasmática e urinária, em bovinos da raça Pantaneira. Os dados foram obtidos de 15 novilhas, com 18 meses de idade e peso corporal in [...] icial de 146 kg, recebendo dietas com 11; 13; 15 e 17 % de proteína bruta (PB). Os animais permaneceram confinados por 114 dias e em intervalos de 28 dias foram pesados e realizadas coletas de sangue, de fezes na ampola retal e de urina na forma spot. O aumento do teor de PB não influenciou o consumo de matéria seca (MS) e o ganho de peso médio diário (GMD); porém melhorou a digestibilidade dos nutrientes, elevou os níveis de nitrogênio no sangue e promoveu maior excreção de uréia urinária. Observou-se, em média, consumos de 5,13 kg MS/animal/dia; GMD de 570 g; coeficientes de digestibilidade aparente de 40,9; 59,6; 39,2; 41,3 e 34,0 % para MS, PB, carboidrato total (CT), fibra detergente neutro (FDN) e ácido (FDA), respectivamente; concentração de uréia plasmática de 49,6 mg/dL; e perda de uréia urinária de 110,0 g/animal/dia. Abstract in english The objective of this experiment was to verify the influence of increasing levels of dietary protein on performance, apparent digestibility and plasma and urinary urea in Pantaneira breed. The data were obtained from 15 heifers, 18 months old and 146 kg initial body weight, fed diets containing 11, [...] 13, 15 and 17 % of crude protein (CP). The animals were confined for 114 days. At intervals of 28 days they were weighed and blood samples and spot samples of feces from the rectum and of urine were taken. The increase in dietary CP content did not influenced dry matter (DM) intake and average daily weight gain (ADG), but improved nutrient digestibility, increased blood nitrogen levels and increased excretion of urinary urea. A mean intake of 5.13 kg DM/animal/day and an ADG of 570 g were observed. Apparent digestibility values of 40.9, 59.6, 39.2, 41.3 and 34,0 % for DM, CP, total carbohydrate, neutral detergent fiber and acid detergent fiber, respectively, were found. Plasma urea concentration was 49.6 mg/dL; and an urinary urea loss of 110,0 g/animal/day was observed.

  3. Métodos de coleta de fezes e determinação dos coeficientes de digestibilidade da fração protéica e da energia de alimentos para o pacu, Piaractus mesopotamicus (Holmberg, 1887) / Fecal collection methods and determination of crude protein and of gross energy digestibility coefficients of feedstuffs for pacu, Piaractus mesopotamicus (Holmberg, 1887)

    Scientific Electronic Library Online (English)

    Eduardo Gianini, Abimorad; Dalton José, Carneiro.

    1101-11-01

    Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese Para melhorar a alimentação do pacu (Piaracatus mesopotamicus), é necessário conhecer o nível de aproveitamento dos ingredientes utilizados em suas dietas. Neste trabalho, inicialmente, foram analisados quatro métodos de coleta de fezes, em função da precisão e da versatilidade (dissecação, extrusão [...] , Guelph e Guelph modificado), e o tempo entre as coletas de fezes nos sistemas de Guelph, por intermédio do coeficiente de digestibilidade da proteína bruta de uma dieta-referência para o pacu. Não houve diferença entre os métodos de coleta de fezes; já os intervalos de tempo das coletas apresentaram diferenças, podendo interferir nos resultados de digestibilidade da proteína. Em uma segunda etapa, para determinação dos coeficientes de digestibilidade da proteína e da energia de cinco concentrados protéicos de origem animal, cinco de origem vegetal e quatro energéticos, foram utilizadas rações elaboradas com 69,5% de uma dieta-referência, 30% do ingrediente teste e 0,5% de Cr2O3, utilizando o método de extrusão manual para a coleta de fezes. Os valores de digestibilidade da proteína da maioria dos alimentos apresentaram-se altos e não variaram estatisticamente, de 93,89 a 75,73% para o farelo de trigo e a farinha de penas, respectivamente. Entretanto, a farinha de sangue e a levedura apresentaram valores de digestibilidade da fração protéica significativamente inferiores (57,72 e 68,86%, respectivamente). Os coeficientes de digestibilidade da energia dos alimentos variaram consideravelmente, apresentando altos valores (acima de 90%) para o sorgo, o farelo de arroz e as sojas crua e tostada, e baixos valores (menores que 70%) para as farinhas de vísceras e de sangue, os farelos de soja e de algodão e a levedura. Quanto aos métodos de coleta de fezes, conclui-se que qualquer um pode ser adotado para a determinação de digestibilidade, desde que sejam rigorosamente aplicados. De maneira geral, a maioria dos concentrados protéicos pode ser utilizada como ótimas fontes de proteína, com exceção da farinha de sangue e a levedura, enquanto os alimentos energéticos confirmaram-se como excelentes fontes de energia em dietas para o pacu. Abstract in english To improve the pacu (Piaracatus mesopotamicus) feeding in the growth phase, it's necessary to know the digestibility of the ingredients used in his diets. Initially, four fecal collection methods were analyzed (dissection, stripping, Guelph and modified Guelph), based on precision and versatility an [...] d also the most efficient time among fecal collection of the Guelph sedimentation systems, through crude protein digestibility coefficient of pacu basic diet. There was no difference among the fecal collection methods while the intervals of time among the collections were significantly different, which may interfere in the protein digestibility results. In a second stage, five proteins concentrated of animal origin, five of vegetable origin and four energetic were studied for estimating protein and energy digestibility coefficients using the stripping method. The rations contained 69.5% of a reference diet, 30.0% of the ingredient test and 0.5% of Cr2O3. In a general way, crude protein digestibility coefficients were high and did not differ from the wheat meal (93.89%) and feather flour (75.73%), respectively. However, blood and yeast showed lower crude protein digestibility (57.72% and 68.86%), respectively. Gross energy digestibility coefficients varied considerably among the feedstuffs, showing high values (above 90%) for the sorghum, rice meal and raw and toasted soy and lower values (smaller than 70%) for poultry by-product and blood meal, soybean meal, cotton and yeast. However, with regard to the fecal collection methods, the results showed that anyone can be adopted with safety for digestibility determination, if used accurately. In a general way, most of protein concentrate can be used as a very good protein source, except for blood meal and t

  4. EFECTO DEL SEXO DE LA RATA (Sprague Dawley SOBRE LA DIGESTIBILIDAD Y RAZÓN NETA DE PROTEÍNA EN ALIMENTOS DE DISTINTA CALIDAD PROTEICA EFFECT OF RAT (Sprague Dawley SEX ON DIGESTIBILITY AND NET PROTEIN RATIO IN FOODS OF DIFFERENT PROTEIN QUALITY

    Directory of Open Access Journals (Sweden)

    María del Refugio Falcón-Villa

    2006-12-01

    Full Text Available La influencia del agrupamiento por sexo de las ratas Sprague Dawley fue evaluada al determinar la calidad proteica en tres alimentos. Los bioensayos realizados fueron digestibilidad de materia seca (DMsec, digestibilidad de nitrógeno aparente y verdadera (DNap y DNver y razón neta de proteína (NPR. Las muestras empleadas fueron alimentos de distinta calidad proteica: leche en polvo, soya texturizada y cereal, obtenidas del comercio local y caseína como control. Se utilizó un diseño experimental por grupos: mixto, hembras y machos. Se colectaron datos cada tercer día: alimento consumido, el total de heces y el incremento en peso de cada individuo. Se concluye de manera general, que el agrupamiento por sexo de la rata no tiene efecto significativo sobre la respuesta de estos bioensayos evaluados en los alimentos probados, a excepción del cereal que presentó diferencias significativas entre machos y hembras en el parámetro de DNver, sugiriendo que es factible utilizar ratas de diferente sexo en estas evaluaciones de calidad proteicaThe effect of grouping by sex Sprague Dawley rats on protein quality evaluation of three foods was studied. Bioassays performed were dry matter digestibility (DMD, apparent and true nitrogen digestibility (AND y TND and net protein ratio (NPR. Foods of different protein quality used were dry whole milk, texturized soybean and high fiber breakfast cereal obtained from the local market. Casein was used as control. An experimental block design where the blocks were the groups of rats classified as mixed, female and male was used. Data of consumed diet, total feces and weight gain of each rat was recorded every third day. It was concluded that with the exception of the high fiber breakfast cereal, which showed significant differences between males and females in the true nitrogen digestibility parameter, in general grouping by sex did not have a significant effect on dry matter, AND, TND and NPR responses. These results suggest the possibility of using rats from either sex in these protein quality evaluation tests

  5. Preparation of shrimp waste as aqua feedstuff: a study of physicochemical properties and in vitro digestibility

    Directory of Open Access Journals (Sweden)

    Karun Thongprajukeaw

    2014-12-01

    Full Text Available The effects of different preparation methods (boiling, microwave irradiation, oven-drying, soaking, and sun-drying for improving the protein quality of shrimp waste (SW when used as aqua feedstuff were observed. The findings from this study indicate that microwave irradiation maintained both the chemical composition and total carotenoid concentration of the SW. This method controlled the appropriate physicochemical characteristics thus promoting proteolytic digestion, as indicated by measurement of changes in pH, water solubility, microstructures, and thermal transition. The protein digestibility based on the use of digestive enzymes from Nile tilapia (Oreochromis niloticus also increased after preparation with microwave irradiation. Based on this study, improvement in the protein quality in SW may be achieved by microwave irradiation and this method may be used as an alternative method for preparing aqua feedstuff.

  6. A new approach to protein enzymatic digestion for fast protein identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Czech Academy of Sciences Publication Activity Database

    Dy?ka, Filip; Laštovi?ková, Markéta; Bobálová, Janette

    2008-01-01

    Ro?. 102, ?. 15 (2008), s967-s968. ISSN 1803-2389. [Meeting on Chemistry and Life /4./. Brno, 09.09.2008-11.09.2008] R&D Projects: GA AV ?R IAA600040701 Institutional research plan: CEZ:AV0Z40310501 Keywords : enzymatic digestion * fast approach Subject RIV: CB - Analytical Chemistry, Separation

  7. Elucidating the temporal dynamics of chromatin-associated protein release upon DNA digestion by quantitative proteomic approach.

    Science.gov (United States)

    Dutta, Bamaprasad; Adav, Sunil S; Koh, Cheng Gee; Lim, Sai Kiang; Meshorer, Eran; Sze, Siu Kwan

    2012-09-18

    Chromatin is a highly dynamic well organized nucleoprotein complex of DNA and proteins that controls DNA-dependent processes such as transcription, replication, repair and many others. Chromatin structure is regulated by various chromatin associated proteins, post-translational modifications of histones and DNA methylation, but a complete picture of structural changes in chromatin architecture is unclear due to the lack of comprehensive data of chromatin-associated proteins and their bindings to different chromatin regions. This study temporally released chromatin-associated proteins by DNase I and MNase treatment and profiled them by exponentially modified protein abundance index (emPAI) based label-free quantitative proteomics. We identified 694 high confidence proteins, with 160 known chromatin-associated proteins. Identified proteins were functionally classified into histones, non-histones involved in architectural maintenance, proteins involved in DNA replication and repair, transcription machinery, transcription regulation, other chromatin proteins, cell cycle proteins and several novel proteins. Numerous proteins presumed to be chromatin associated were identified and their chromatin interactions were explored. The comprehensive differential chromatin bound proteome might expand our knowledge of the proteins that were associated with different chromatin regions, which could be very useful in elucidating chromatin biology. PMID:22796351

  8. Steam explosion pretreatment of wheat straw to improve methane yields: investigation of the degradation kinetics of structural compounds during anaerobic digestion.

    Science.gov (United States)

    Theuretzbacher, Franz; Lizasoain, Javier; Lefever, Christopher; Saylor, Molly K; Enguidanos, Ramon; Weran, Nikolaus; Gronauer, Andreas; Bauer, Alexander

    2015-03-01

    Wheat straw can serve as a low-cost substrate for energy production without competing with food or feed production. This study investigated the effect of steam explosion pretreatment on the biological methane potential and the degradation kinetics of wheat straw during anaerobic digestion. It was observed that the biological methane potential of the non steam exploded, ground wheat straw (276 l(N) kg VS(-1)) did not significantly differ from the best steam explosion treated sample (286 l(N) kg VS(-1)) which was achieved at a pretreatment temperature of 140°C and a retention time of 60 min. Nevertheless degradation speed was improved by the pretreatment. Furthermore it was observed that compounds resulting from chemical reactions during the pretreatment and classified as pseudo-lignin were also degraded during the anaerobic batch experiments. Based on the rumen simulation technique, a model was developed to characterise the degradation process. PMID:25549903

  9. Composição, digestibilidade protéica e desaminação em cultivares brasileiras de soja submetidas à radiação gama / Composition, protein digestibility and deamidation in Brazilian soy cultivars subjected to gamma radiation

    Scientific Electronic Library Online (English)

    Taís Carolina Franqueira de, Toledo; Solange Guidolin Canniatti, Brazaca; Valter, Arthur; Sônia Maria de Stefano, Piedade.

    2007-12-01

    Full Text Available A soja é reconhecidamente um alimento rico em proteínas e lipídios e sua inclusão na alimentação humana no Brasil tem se tornado cada vez maior. Devido à importância comercial e nutricional da soja, possíveis alterações promovidas por métodos de conservação aplicados a ela devem ser estudadas. O pre [...] sente trabalho visa avaliar a composição química, a digestibilidade in vitro de proteínas e a porcentagem de desaminação de grãos de cinco diferentes cultivares de soja (BRS 212, BRS 213, BRS 214, BRS 231 e E48) submetidas à irradiação (doses de 2, 4 e 8 kGy). Dentro da composição química foram encontrados valores para cinza que variaram de 4,90 a 6,08%, para proteína de 21,23 a 36,99%, para lipídeos de 19,22 a 24,84%, para fibras solúveis de 1,37 a 4,03% e para fibras insolúveis de 15,97 a 18,87%. A porcentagem de desaminação nas diferentes amostras variou de 17,34 a 57,79% e a digestibilidade in vitro de 84,45 a 89,11%. O tratamento de irradiação nas doses utilizadas não promoveu alterações na composição centesimal e na digestibilidade, mas promoveu aumento na porcentagem de desaminação das amostras conforme aumento das doses. Abstract in english Soy is known to be rich in proteins and lipids and its inclusion in the diet of Brazil's population is increasing. Due to the commercial and nutritional importance of soy, possible alterations resulting from the methods of conservation to which they are subjected must be studied. The present work ev [...] aluated the chemical composition, the in vitro digestibility of proteins and the percentage of deamidation of grains of five different soy cultivars (BRS 212, BRS 213, BRS 214, 231 BRS and E48) subjected to irradiation (doses of 2, 4 and 8 kGy). The chemical composition showed ashes varying from 4.90 to 6.08%, proteins from 21.23 to 36.99%, lipids from 19.22 to 24.84%, soluble fibers from 1.37 to 4.03%, and insoluble fibers from 15.97 to 18.87%. The percentage of deamidation in the various samples varied from 17.34 to 57.79% and the in vitro digestibility from 84.45 to 89.11%. Irradiation treatment in the doses applied here did not change the chemical composition or digestibility, but increased the percentage of deamidation of the samples as the dosage increased.

  10. Resolution Improvement in Multidimensional Nuclear Magnetic Resonance Spectroscopy of Proteins

    International Nuclear Information System (INIS)

    The work presented in this thesis is concerned with both liquid-state and solid-state nuclear magnetic resonance (NMR) spectroscopy. Most of this work is devoted to the investigation by solid-state NMR of C13-enriched compounds with the principal aim of presenting techniques devised for further improving the spectral resolution in multidimensional NMR of microcrystalline proteins. In fully C13-labelled compounds, the J-coupling induces a broadening of the carbon lineshapes. We show that spin-state-selective technique called IPAP can be successfully combined with standard polarisation transfer schemes in order to remove the J-broadening in multidimensional solid-state NMR correlation experiments of fully C13-enriched proteins. We present subsequently two techniques tailored for liquid-state NMR spectroscopy. The carbon directly detected techniques provide chemical shift information for all backbone hetero-nuclei. They are very attracting for the study of large bio-molecular systems or for the investigation of paramagnetic proteins. In the last part of this thesis, we study the spin-echo J-modulation for homonuclear two-spin 1/2 systems. Under magic-angle spinning, the theory of J-induced spin-echo modulation allows to derive a set of modulation regimes which give a spin-echo modulation exactly equal to the J-coupling. We show that the chemical-shift anisotropy and the dipolar interaction tend to stabilize the spin-echo J-modulation. The theoretical conclusions are supported by numerical simulations and experimental results obtained for three representative samples containing C13 spin pairs. (author)

  11. The effects of basic substances and acidic ionophores on the digestion of exogenous and endogenous proteins in mouse peritoneal macrophages

    OpenAIRE

    1986-01-01

    Basic substances and acidic ionophores that increase the lysosomal pH in cultured macrophages (Ohkuma, S., and B. Poole, 1978, Proc. Natl. Acad. Sci. USA., 75:3327-3331; Poole, B., and S. Ohkuma, 1981, J. Cell Biol., 90:665-669) inhibited the digestion of heat-denatured acetylated bovine serum albumin (BSA) taken up by the cells. For several substances, the shift in pH sufficed to explain the inhibition of proteolysis. Additional effects, presumably on enzyme activities, have to be postulated...

  12. Effects of Replacement of Fish Meal by Soy Protein Isolate on the Growth, Digestive Enzyme Activity and Serum Biochemical Parameters for Juvenile Amur Sturgeon (Acipenser schrenckii).

    Science.gov (United States)

    Xu, Q Y; Wang, C A; Zhao, Z G; Luo, L

    2012-11-01

    An 8-wk experiment was conducted to evaluate the effect of replacing fish meal (FM) with soy protein isolate (SPI) on the growth, digestive enzyme activity and serum biochemical parameters of juvenile Amur sturgeon (Acipenser schrenckii). SPI was used to replace 0, 25, 50, 62.5, 75, 87.5, 100% of dietary FM and 100% replacement supplemented crystalline amino acid. Healthy sturgeon with an average initial weight of 26.38±0.24 g were randomly assigned to 24 aquaria (8 treatments with triplicates each) at an initial stocking density of 11 fish per aquarium and cultured for 8 wks. The results showed that 75.00% or more substitution resulted in a poor weight gain rate, feed conversion ratio and survival rate compared to that of fish fed the control diet (pgrowth performance, intestine digestive enzyme activities and serum biochemical parameters. In conclusion, the results from this study showed adverse effects of inclusion of SPI in diets on growth performance, feed utilization and serum biochemical parameters in juvenile Amur sturgeon. Based on WGR and replacement ratio presented in this report, a 57.64% replacement level was recommended. PMID:25049521

  13. Determination of hidden hazelnut oil proteins in extra virgin olive oil by cold acetone precipitation followed by in-solution tryptic digestion and MALDI-TOF-MS analysis.

    Science.gov (United States)

    De Ceglie, Cristina; Calvano, Cosima Damiana; Zambonin, Carlo Giorgio

    2014-10-01

    Adulteration of extra-virgin olive oil (EVOO) with hazelnut oil (HO) is an illegal practice that could have severe health consequences for consumers due to the possible exposure to hidden hazelnut allergens. Here, matrix-assisted laser-desorption/ionization (MALDI) mass spectrometry (MS) was used as a rapid and sensitive technique for the detection of a low concentration of hazelnut proteins in oil samples. Different protocols were tested for protein extraction, and the most efficient (cold acetone) was applied to HO and EVOO adulterated with HO. The subsequent in-solution tryptic digestion of protein extracts and MALDI-MS analysis, using ?-cyano-4-chlorocinnamic acid as matrix, allowed the detection of stable hazelnut peptide markers (i.e., the m/z ions 1002.52, 1356.71, 1394.70, 1440.81, 1453.85, 1555.76, 1629.83, 1363.73, and 1528.67) attributable to the main hazelnut proteins Cor a 9, Cor a 11, and Cor a 1. Thus, the approach might allow the direct detection of specific hazelnut allergens in EVOO at low concentration without time-consuming pretreatments. PMID:25209075

  14. Directed evolution methods for improving polypeptide folding and solubility and superfolder fluorescent proteins generated thereby

    Science.gov (United States)

    Waldo, Geoffrey S. (Santa Fe, NM)

    2007-09-18

    The current invention provides methods of improving folding of polypeptides using a poorly folding domain as a component of a fusion protein comprising the poorly folding domain and a polypeptide of interest to be improved. The invention also provides novel green fluorescent proteins (GFPs) and red fluorescent proteins that have enhanced folding properties.

  15. Fed-batch cultivation of Desmodesmus sp. in anaerobic digestion wastewater for improved nutrient removal and biodiesel production.

    Science.gov (United States)

    Ji, Fang; Zhou, Yuguang; Pang, Aiping; Ning, Li; Rodgers, Kibet; Liu, Ying; Dong, Renjie

    2015-05-01

    Desmodesmus sp. was used in anaerobically digested wastewater (ADW) for nutrients removal and the biodiesel production was measured and compared using fed-batch cultivation was investigated and compared with batch cultivation. The Desmodesmus sp. was able to remove 236.143, 268.238 and 6.427mg/L of TN, NH4-N and PO4-P respectively after 40d of fed-batch cultivation, while in batch cultivation the quantities of TN, NH4-N and PO4-P removed were 33.331, 37.227 and 1.323mg/L. Biomass production of Desmodesmus sp. was also enhanced in fed-batch cultivation, when ADW loading was carried out every 2days; the biomass concentration peaked at 1.039g/L, which was three times higher than that obtained in batch cultivation (0.385g/L). The highest lipid production (261.8mg/L) was also recorded in fed-batch cultivation as compared to batch cultivation (83.3mg/L). Fed-batch cultivation of Desmodesmus sp. could provide effective control of nutrients limitation and/or ammonia inhibition on microalgae cultivation. PMID:25451775

  16. Effects of protein concentration and heat treatment on concentration of digestible and metabolizable energy and on amino acid digestibility in four sources of canola meal fed to growing pigs.

    Science.gov (United States)

    Liu, Y; Song, M; Maison, T; Stein, H H

    2014-10-01

    Two experiments were conducted to determine DE and ME and the apparent ileal digestibility (AID) and the standardized ileal digestibility (SID) of CP and AA in 4 sources of canola meal (high-protein [CM-HP], high-temperature-processed [CM-HT], low-temperature-processed [CM-LT], and conventional [CM-CV] canola meal) and in conventional soybean meal (SBM) fed to growing pigs. In Exp. 1, 48 growing barrows (initial BW: 39.7 ± 1.58 kg) were individually housed in metabolism cages and randomly assigned to 6 treatments in a randomized complete block design with 2 blocks of 24 pigs and 8 replicate pigs per treatment. The 6 diets included a corn-based basal diet and 5 diets that were formulated by mixing corn and 1 of the sources of canola meal (39.0% inclusion) or SBM (28.5% inclusion). Feces and urine were collected for 5 d following a 5-d adaptation period. The DE and ME in each source of canola meal and in SBM were calculated using the difference procedure. The DE and ME in the 4 sources of canola meal were less (P ileal digesta were collected on d 6 and 7 of each period. The AID and SID of CP and all AA in SBM were greater (P < 0.05) than in the 4 sources of canola meal. Compared with CM-CV, CM-HP had greater (P < 0.05) AID of Ile, Lys, Asp, Cys, and Pro and greater (P < 0.05) SID of Lys and Cys. However, no differences between CM-HT and CM-LT were observed. In conclusion, regardless of the concentration of CP and the processing used, canola meal provides less DE and ME to pigs than corn and SBM, and the SID of AA in canola meal is less than in SBM. The processing temperature used in this experiment did not affect DE and ME or SID of AA in canola meal. The SID of Lys and Cys was greater in CM-HP than in CM-CV. PMID:25149331

  17. Digestion Simulation

    Science.gov (United States)

    Integrated Teaching and Learning Program,

    To reinforce students' understanding of the human digestion process, the functions of several stomach and small intestine fluids are analyzed, and the concept of simulation is introduced through a short, introductory demonstration of how these fluids work. Students learn what simulation means and how it relates to the engineering process, particularly in biomedical engineering. The teacher demo requires vinegar, baking soda, water and aspirin.

  18. Effects of conifer release with vision(r) (glyphosate) herbicide on moose forage quality (digestible protein). Forest research report No. 139

    Energy Technology Data Exchange (ETDEWEB)

    Cumming, H.G.

    1996-12-01

    Forest managers commonly use herbicides to release young conifers from associated non-crop vegetation. Concern has been raised over the potential effects on wildlife of conifer release with herbicides. The most commonly used herbicide in Canadian forestry is Vision, whose active ingredient is glyphosate. Although glyphosate is considered practically non-toxic to mammals, indirect effects (reduced forage) concern both biologists and hunters. This report examines the effects of Vision on moose browse quantity and moose area use near Thunder Bay, Ontario, to study the effects of conifer release with Vision on nutritional browse quality four and eight years after treatment. Both winter (twigs) and summer (leaves) forages were analysed from four plant species commonly eaten by moose in early successional forests: Hazel, aspen, raspberry, and willow. Results are presented only for the more important nutrient, digestible protein.

  19. Cadmium versus copper toxicity: Insights from an integrated dissection of protein synthesis pathway in the digestive glands of mussel Mytilus galloprovincialis

    International Nuclear Information System (INIS)

    Highlights: • Cu2+-exposure of mussels results in genotoxicity, without affecting MTs production. •Cd2+-exposure of mussels causes low genotoxicity, but induces MTs production. • Both metals induce oxidative stress in mussels, with Cd being the strongest inducer. • Translation is suppressed by both metals, mainly at the initiation and elongation steps. • MTs abrogate translational defects caused by Cd2+, by trapping the toxic metal. -- Abstract: The main purpose of this study was to investigate the impact of metal-mediated stress on the protein-synthesis pathway in mussels. To this end, mussels (Mytilus galloprovincialis) underwent a 15 days exposure to 100 ?g/L Cu2+ or Cd2+. Both metals, in particular Cd2+, accumulated in mussel digestive glands and generated a specific status of oxidative-stress. Exposure of mussels to each metal resulted in 40% decrease of the tRNA-aminoacylation efficiency, at the end of exposure. Cu2+ also caused a progressive loss in the capability of 40S-ribosomal subunits to form 48S pre-initiation complex, which reached 34% of the control at the end of exposure. Other steps of translation underwent less pronounced, but measurable damages. Mussels exposed to Cd2+ for 5 days presented a similar pattern of translational dysfunctions in digestive glands, but during the following days of exposure the ribosomal efficiency was gradually restored. Meanwhile, metallothionein levels significantly increased, suggesting that upon Cd2+-mediated stress the protein-synthesizing activity was reorganized both quantitatively and qualitatively. Conclusively, Cd2+ and Cu2+ affect translation at several levels. However, the pattern of translational responses differs, largely depending on the capability of each metal to affect cytotoxic pathways in the tissues, such as induction of antioxidant defense and specific repair mechanisms

  20. Cadmium versus copper toxicity: Insights from an integrated dissection of protein synthesis pathway in the digestive glands of mussel Mytilus galloprovincialis

    Energy Technology Data Exchange (ETDEWEB)

    Pytharopoulou, S.; Kournoutou, G.G. [Laboratory of Biochemistry, School of Medicine, University of Patras, 26504 Patras (Greece); Leotsinidis, M. [Laboratory of Public Health, School of Medicine, University of Patras, 26504 Patras (Greece); Georgiou, C.D. [Department of Biology, Section of Genetics, Cell Biology and Development, University of Patras, 26504 Patras (Greece); Kalpaxis, D.L., E-mail: dimkal@med.upatras.gr [Laboratory of Biochemistry, School of Medicine, University of Patras, 26504 Patras (Greece)

    2013-09-15

    Highlights: • Cu{sup 2+}-exposure of mussels results in genotoxicity, without affecting MTs production. •Cd{sup 2+}-exposure of mussels causes low genotoxicity, but induces MTs production. • Both metals induce oxidative stress in mussels, with Cd being the strongest inducer. • Translation is suppressed by both metals, mainly at the initiation and elongation steps. • MTs abrogate translational defects caused by Cd{sup 2+}, by trapping the toxic metal. -- Abstract: The main purpose of this study was to investigate the impact of metal-mediated stress on the protein-synthesis pathway in mussels. To this end, mussels (Mytilus galloprovincialis) underwent a 15 days exposure to 100 ?g/L Cu{sup 2+} or Cd{sup 2+}. Both metals, in particular Cd{sup 2+}, accumulated in mussel digestive glands and generated a specific status of oxidative-stress. Exposure of mussels to each metal resulted in 40% decrease of the tRNA-aminoacylation efficiency, at the end of exposure. Cu{sup 2+} also caused a progressive loss in the capability of 40S-ribosomal subunits to form 48S pre-initiation complex, which reached 34% of the control at the end of exposure. Other steps of translation underwent less pronounced, but measurable damages. Mussels exposed to Cd{sup 2+} for 5 days presented a similar pattern of translational dysfunctions in digestive glands, but during the following days of exposure the ribosomal efficiency was gradually restored. Meanwhile, metallothionein levels significantly increased, suggesting that upon Cd{sup 2+}-mediated stress the protein-synthesizing activity was reorganized both quantitatively and qualitatively. Conclusively, Cd{sup 2+} and Cu{sup 2+} affect translation at several levels. However, the pattern of translational responses differs, largely depending on the capability of each metal to affect cytotoxic pathways in the tissues, such as induction of antioxidant defense and specific repair mechanisms.

  1. The MalK protein of the ATP-binding cassette transporter for maltose of Escherichia coli is accessible to protease digestion from the periplasmic side of the membrane.

    OpenAIRE

    Schneider, E.; Hunke, S.; Tebbe, S.

    1995-01-01

    The ATP-hydrolyzing subunit MalK of the ATP-binding cassette transporter for maltose of Escherichia coli is demonstrated to be accessible to digestion by proteinase K in right-side-out membrane vesicles. This finding suggests a partial transmembrane orientation of the protein.

  2. Tryptic digestion coupled with ambient desorption electrospray ionization and liquid extraction surface analysis mass spectrometry enabling identification of skeletal muscle proteins in mixtures and distinguishing between beef, pork, horse, chicken, and turkey meat.

    Science.gov (United States)

    Montowska, Magdalena; Rao, Wei; Alexander, Morgan R; Tucker, Gregory A; Barrett, David A

    2014-05-01

    The use of ambient desorption electrospray ionization mass spectrometry (DESI-MS) and liquid extraction surface analysis mass spectrometry (LESA-MS) is explored for the first time to analyze skeletal muscle proteins obtained from a mixture of standard proteins and raw meat. Single proteins and mixtures of up to five proteins (myoglobin, troponin C, actin, bovine serum albumin (BSA), tropomyosin) were deposited onto a polymer surface, followed by in situ tryptic digestion and comparative analysis using DESI-MS and LESA-MS using tandem electrospray MS. Peptide peaks specific to individual proteins were readily distinguishable with good signal-to-noise ratio in the five-component mixture. LESA-MS gave a more stable analysis and greater sensitivity compared with DESI-MS. Meat tryptic digests were subjected to peptidomics analysis by DESI-MS and LESA-MS. Bovine, horse, pig, chicken, and turkey muscle digests were clearly discriminated using multivariate data analysis (MVA) of the peptidomic data sets. The most abundant skeletal muscle proteins were identified and correctly classified according to the species following MS/MS analysis. The study shows, for the first time, that ambient ionization techniques such as DESI-MS and LESA-MS have great potential for species-specific analysis and differentiation of skeletal muscle proteins by direct surface desorption. PMID:24673366

  3. Determination of additivity of apparent and standardized ileal digestibility of amino acids in diets containing multiple protein sources fed to growing pigs.

    Science.gov (United States)

    Xue, P C; Ragland, D; Adeola, O

    2014-09-01

    An experiment was conducted in growing pigs to investigate the additivity of apparent ileal digestibility (AID) or standardized ileal digestibility (SID) of CP and AA in mixed diets containing multiple protein sources. Using the determined AID or SID for CP and AA in corn, soybean meal (SBM), corn distillers' dried grains with solubles (DDGS), or canola meal (CM), the AID or SID for 4 mixed diets based on corn-SBM, corn-SBM-DDGS, corn-SBM-CM, or corn-SBM-DDGS-CM were predicted and compared with determined AID or SID, respectively. Eighteen growing pigs (initial BW = 61.3 ± 5.5 kg) were surgically fitted with T-cannulas and assigned to a duplicated 9 × 4 incomplete Latin square design with 9 diets and 4 periods. The 9 experimental diets consisted of a nitrogen-free diet (NFD) to estimate basal ileal endogenous loss (BEL) of AA, 4 semipurified diets to determine the AID and SID of CP and AA in the 4 ingredients, and 4 mixed diets to test the additivity of AID and SID. Chromic oxide was added as an indigestible marker. Pigs were fed 1 of the 9 diets during each 7-d period, and ileal digesta were collected on d 6 and 7, from 0800 to 1800 h. The analyzed AA levels for the mixed diets were close to the calculated values based on the AA composition of each ingredient. The results revealed that the predicted SID were consistent with determined values, except for Leu, Thr, Asp, Cys, Pro, and Ser in the corn-SBM diet and Met and Cys in the corn-SBM-DDGS diet. The determined AID for total AA and Arg, His, Trp, Gly, and Pro in the corn-SBM diet were greater (P ileal digestibility of AA in mixed diets containing multiple protein sources. In addition, the lack of additivity of AID in mixed diets could be attributed to the intrinsic characteristics of the feed ingredient, especially its AA content. PMID:25057030

  4. Protein fraction, degradability and digestibility of pearl millet silage at different cutting ages / Fracionamento de proteína, degradabilidade e digestibilidade da silagem de milheto em diferentes idades de corte

    Scientific Electronic Library Online (English)

    Katia Cylene, Guimarães; Kátia Aparecida de Pinho, Costa; Alexsandra, Paludo; Nayara Fernandes dos, Santos; Robson Marcelo, Rossi; Welma Santos, Cruvinel.

    2014-03-01

    Full Text Available SciELO Brazil | Language: English Abstract in portuguese As determinações das frações proteicas e características de fermentação ruminal de genótipos de milheto são de fundamental importância, pois geram informações relativas ao valor nutritivo do alimento e direcionam os programas de melhoramento genético a serem utilizadas na dieta de ruminantes. Diante [...] disso, objetivou-se determinar as taxas de digestão das frações de proteína, degradabilidade ruminal da matéria seca e digestibilidade "in vitro" da matéria seca das silagens dos genótipos de milheto produzidas em diferentes épocas de corte. O experimento foi conduzido no Campus da Faculdade de Agronomia da Universidade de Rio Verde e Instituto Federal Goiano, Campus Rio Verde. O delineamento experimental utilizado foi de blocos ao acaso, com quatro repetições, em esquema fatorial 5 x 3, sendo, cinco cultivares de milheto: ARD 500, ADR 7010, LAB 0730, LAB 0731 e LAB 0732 e três idades de cortes: 57, 65 e 73 dias após a semeadura (DAS). Os resultados mostraram que as silagens produzidas por milheto, independente do genótipo, podem ser consideradas de boa qualidade. Por se tratar de materiais precoces a melhor idade que proporcionou melhor qualidade da silagem desses materiais, foi quando os materiais foram colhidos aos 57 DAS, em que proporcionaram maiores frações A, menores frações C e elevadas degradabilidade e digestibilidade. Abstract in english The determination of protein fraction and rumen fermentation characteristics of pearl millet genotypes is important, since generate information about food nutritional value and also guide the breeding programs of genotypes to be used in diets of ruminants. The objective of this research was to deter [...] mine the digestion rates of protein fractions, dry matter degradability and in vitro dry matter digestibility of silages from pearl millet genotypes produced at different cutting ages. The experiment was carried out on the Agronomy campus of the Rio Verde University and on the Rio Verde campus of Goiano Federal Institute. The experimental design was randomized blocks with four replications in a 5 x 3 factorial arrangement, with five genotypes of millet: ARD 500, ADR 7010, LAB 0730, LAB 0731 and LAB 0732 and three cutting ages: 57, 65 and 73 days after sowing (DAS). The silage produced from pearl millet, regardless of genotype can be considered of good quality. The evaluated genotypes are considered early, so that the age that provided the best silage quality was 57 DAS. This age provided higher fractions A, lower fraction C and high degradability and digestibility.

  5. Antioxidants reduce endoplasmic reticulum stress and improve protein secretion

    OpenAIRE

    Malhotra, Jyoti D.; Miao, Hongzhi; Zhang, Kezhong; Wolfson, Anna; Pennathur, Subramaniam; Pipe, Steven W.; Kaufman, Randal J.

    2008-01-01

    Protein misfolding in the endoplasmic reticulum (ER) contributes to the pathogenesis of many diseases. Although oxidative stress can disrupt protein folding, how protein misfolding and oxidative stress impact each other has not been explored. We have analyzed expression of coagulation factor VIII (FVIII), the protein deficient in hemophilia A, to elucidate the relationship between protein misfolding and oxidative stress. Newly synthesized FVIII misfolds in the ER lumen, activates the unfolded...

  6. Relação lisina digestível: proteína bruta em dietas para frangos de corte no período de 1 a 21 dias de idade. Desempenho e metabolismo Digestible lysine: crude protein ratios on initial broiler diets - performance and metabolism

    Directory of Open Access Journals (Sweden)

    Kênia Ferreira Rodrigues

    2008-03-01

    Full Text Available Objetivou-se neste experimento testar diferentes relações lisina digestível:proteína bruta (PB em dietas para frangos de corte na fase inicial de criação. Utilizou-se um lote misto de 1.800 aves da linhagem Cobb, com um dia de idade. O delineamento foi inteiramente casualizado com um modelo de classificação hierárquica com três níveis de PB e quatro relações lisina digestível:PB. No ensaio de desempenho, as rações foram fornecidas a 30 aves em cada uma das cinco repetições. Aos 21 dias de idade, avaliaram-se consumo de ração, ganho de peso e conversão alimentar. Não houve efeito significativo das diferentes relações dentro dos três níveis de PB. Observou-se comportamento linear do ganho de peso nas relações de lisina dentro dos níveis de 18,5 e 21,5% de PB. A conversão alimentar melhorou linearmente segundo as relações de lisina dentro dos três níveis de PB estudados. O rendimento de carcaça elevou-se linearmente com o aumento das relações no nível de 15,5% de PB, não sendo significativo nos demais níveis. No ensaio de metabolismo, as relações lisina digestível:proteína bruta apresentaram efeito linear sobre a EMAn (21,5% de PB e excreção de nitrogênio (18,5% de PB e efeito quadrático sobre a EMAn (15,5 e 18,5% de PB e o coeficiente de retenção de N (15,5% de PB. Considerando os resultados de ganho de peso, a relação ideal de lisina digestível:proteína bruta na fase inicial foi de 6,8% (18,5% de PB e 5,9% (21,5% de PB ou 1,26 e 1,27% de lisina digestível. Os teores de PB das rações para frangos de corte na fase inicial podem ser reduzidos para 18,5%, sem afetar o desempenho das aves, mas reduzindo a excreção de nitrogênio em 24%, desde que as rações sejam suplementadas com aminoácidos sintéticos.The experiment aimed to test different ratios digestible lysine:crude protein (CP in diets for broiler until 21 days. One thousand and eight hundred 1-d birds of Cobb strain were allotted to a completely randomized design with a hierarchical rank model with three levels of CP and four ratios digestible lysine:CP was used. In the performance trial, the diets were fed to 30 birds in each of the five replicates. At 21 days of age, feed intake, weight gain and feed conversion were evaluated. A linear increase on weight gain was observed in the 18.5 and 21.5% of CP levels. Feed conversion improved linearly according to the ratios of lysine within the three levels of CP studied. Carcass yield increased linearly with increasing rations at the level of 15.5% of CP, it not being significant at the other levels of CP studied. In the metabolism trial, the ratios digestible lysine:CP showed linear effect on EMAn (21.5% of CP and nitrogen excretion (18.5% of CP, quadratic effect on EMAn (15.5 and 18.5% of CP and N retention coefficient (15.5% of CP. Based on the results of weight gain, the ideal digestible lysine:CP ratio in the initial phase was of 6.8% (18.5% of CP and 5.9% (21.5% of CP, or 1.26 and 1.27% of digestible lysine. The dietary CP contents fed to broilers in the initial phase can be reduced to 18.5%, with no effect on bird performance, but by decreasing nitrogen excretion to 24%, since the diets are supplemented with synthetic aminoacids.

  7. Relação lisina digestível: proteína bruta em dietas para frangos de corte no período de 1 a 21 dias de idade. Desempenho e metabolismo / Digestible lysine: crude protein ratios on initial broiler diets - performance and metabolism

    Scientific Electronic Library Online (English)

    Kênia Ferreira, Rodrigues; Paulo Borges, Rodrigues; Rilke Tadeu Fonseca de, Freitas; Antonio Gilberto, Bertechini; Luiz Fernando Teixeira, Albino; Édison José, Fassani.

    2008-03-01

    Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese Objetivou-se neste experimento testar diferentes relações lisina digestível:proteína bruta (PB) em dietas para frangos de corte na fase inicial de criação. Utilizou-se um lote misto de 1.800 aves da linhagem Cobb, com um dia de idade. O delineamento foi inteiramente casualizado com um modelo de clas [...] sificação hierárquica com três níveis de PB e quatro relações lisina digestível:PB. No ensaio de desempenho, as rações foram fornecidas a 30 aves em cada uma das cinco repetições. Aos 21 dias de idade, avaliaram-se consumo de ração, ganho de peso e conversão alimentar. Não houve efeito significativo das diferentes relações dentro dos três níveis de PB. Observou-se comportamento linear do ganho de peso nas relações de lisina dentro dos níveis de 18,5 e 21,5% de PB. A conversão alimentar melhorou linearmente segundo as relações de lisina dentro dos três níveis de PB estudados. O rendimento de carcaça elevou-se linearmente com o aumento das relações no nível de 15,5% de PB, não sendo significativo nos demais níveis. No ensaio de metabolismo, as relações lisina digestível:proteína bruta apresentaram efeito linear sobre a EMAn (21,5% de PB) e excreção de nitrogênio (18,5% de PB) e efeito quadrático sobre a EMAn (15,5 e 18,5% de PB) e o coeficiente de retenção de N (15,5% de PB). Considerando os resultados de ganho de peso, a relação ideal de lisina digestível:proteína bruta na fase inicial foi de 6,8% (18,5% de PB) e 5,9% (21,5% de PB) ou 1,26 e 1,27% de lisina digestível. Os teores de PB das rações para frangos de corte na fase inicial podem ser reduzidos para 18,5%, sem afetar o desempenho das aves, mas reduzindo a excreção de nitrogênio em 24%, desde que as rações sejam suplementadas com aminoácidos sintéticos. Abstract in english The experiment aimed to test different ratios digestible lysine:crude protein (CP) in diets for broiler until 21 days. One thousand and eight hundred 1-d birds of Cobb strain were allotted to a completely randomized design with a hierarchical rank model with three levels of CP and four ratios digest [...] ible lysine:CP was used. In the performance trial, the diets were fed to 30 birds in each of the five replicates. At 21 days of age, feed intake, weight gain and feed conversion were evaluated. A linear increase on weight gain was observed in the 18.5 and 21.5% of CP levels. Feed conversion improved linearly according to the ratios of lysine within the three levels of CP studied. Carcass yield increased linearly with increasing rations at the level of 15.5% of CP, it not being significant at the other levels of CP studied. In the metabolism trial, the ratios digestible lysine:CP showed linear effect on EMAn (21.5% of CP) and nitrogen excretion (18.5% of CP), quadratic effect on EMAn (15.5 and 18.5% of CP) and N retention coefficient (15.5% of CP). Based on the results of weight gain, the ideal digestible lysine:CP ratio in the initial phase was of 6.8% (18.5% of CP) and 5.9% (21.5% of CP), or 1.26 and 1.27% of digestible lysine. The dietary CP contents fed to broilers in the initial phase can be reduced to 18.5%, with no effect on bird performance, but by decreasing nitrogen excretion to 24%, since the diets are supplemented with synthetic aminoacids.

  8. Improved protein structure selection using decoy-dependent discriminatory functions

    Directory of Open Access Journals (Sweden)

    Levitt Michael

    2004-06-01

    Full Text Available Abstract Background A key component in protein structure prediction is a scoring or discriminatory function that can distinguish near-native conformations from misfolded ones. Various types of scoring functions have been developed to accomplish this goal, but their performance is not adequate to solve the structure selection problem. In addition, there is poor correlation between the scores and the accuracy of the generated conformations. Results We present a simple and nonparametric formula to estimate the accuracy of predicted conformations (or decoys. This scoring function, called the density score function, evaluates decoy conformations by performing an all-against-all C? RMSD (Root Mean Square Deviation calculation in a given decoy set. We tested the density score function on 83 decoy sets grouped by their generation methods (4state_reduced, fisa, fisa_casp3, lmds, lattice_ssfit, semfold and Rosetta. The density scores have correlations as high as 0.9 with the C? RMSDs of the decoy conformations, measured relative to the experimental conformation for each decoy. We previously developed a residue-specific all-atom probability discriminatory function (RAPDF, which compiles statistics from a database of experimentally determined conformations, to aid in structure selection. Here, we present a decoy-dependent discriminatory function called self-RAPDF, where we compiled the atom-atom contact probabilities from all the conformations in a decoy set instead of using an ensemble of native conformations, with a weighting scheme based on the density scores. The self-RAPDF has a higher correlation with C? RMSD than RAPDF for 76/83 decoy sets, and selects better near-native conformations for 62/83 decoy sets. Self-RAPDF may be useful not only for selecting near-native conformations from decoy sets, but also for fold simulations and protein structure refinement. Conclusions Both the density score and the self-RAPDF functions are decoy-dependent scoring functions for improved protein structure selection. Their success indicates that information from the ensemble of decoy conformations can be used to derive statistical probabilities and facilitate the identification of near-native structures.

  9. Effects of single cell protein replacing fish meal in diet on growth performance, nutrient digestibility and intestinal morphology in weaned pigs.

    Science.gov (United States)

    Zhang, H Y; Piao, X S; Li, P; Yi, J Q; Zhang, Q; Li, Q Y; Liu, J D; Wang, G Q

    2013-09-01

    Three experiments were conducted to evaluate the ME value, standardized ileal digestibility (SID) of amino acids (AA) of fish meal, and the effects of single cell protein (Prosin and Protide) replacing fish meal in diet on growth performance, nutrient digestibility and intestinal morphology in weaned piglets. In Exp. 1, twenty-four barrows with initial BW of 30.8±2.6 kg were allotted to one of four dietary treatments. Diet 1 contained corn as the only energy source. The other three diets replaced 20% of the corn in diet 1 with one of the three protein feeds (fish meal, Prosin and Protide), and the DE and ME contents were determined by difference. In Exp. 2, eight barrows (initial BW of 25.6±3.2 kg) were fitted with ileal T-cannulas and allotted to a replicated 4×4 Latin square design. Three cornstarch-based diets were formulated using each of the protein feeds as the sole source of AA. A nitrogen-free diet was also formulated to measure endogenous losses of AA. In Exp. 3, one hundred and eighty piglets (initial BW of 7.95±1.59 kg) weaned at 28±2 d were blocked by weight and assigned to one of five treatments for a 28-d growth performance study, each treatment was fed to six pens with six pigs (three barrows and three gilts) per pen. The five treatments consisted of the control group (CON), which was a corn-soybean meal diet containing 5% fish meal, and the other four treatments, which replaced a set amount of fish meal with either Prosin (2.5% or 5%) or Protide (2.5% or 5%). The diets were formulated to provide same nutrient levels. The results showed that on a DM basis, both of the DE and ME contents were lower in Prosin and Protide than that of fish meal (p<0.05). The SID of CP and all essential AA were greater in fish meal than in Prosin and Protide (p<0.05). The pigs fed CON diet had greater weight gain and lower feed conversion rate (FCR) than pigs fed 5% Prosin and 5% Protide diets (p<0.05). The digestibility of CP was greater in pigs fed CON, 2.5% Prosin and 2.5% Protide diets than the pigs fed 5% Prosin and 5% Protide diets (p<0.05). Villus height in jejunum and ileum, and villus height to crypt depth ratio in the jejunum were higher (p<0.05) in pigs fed CON, 2.5% Prosin and 2.5% Protide diets compared with the 5% Prosin and 5% Protide diets. Pigs fed CON diet had greater villus height to crypt depth ratio in the ileum than the pigs fed 5% Prosin and 5% Protide diets (p<0.05). In conclusion, although Prosin and Protide contained lower ME content and SID of AA than fish meal, Prosin and Protide replacing 50% of fish meal in diet with identical nutrient levels could obtain similar performance, nutrient digestibility and intestinal morphology in weaned pigs. PMID:25049915

  10. Wood ash treatment, a cost-effective way to deactivate tannins in Acacia cyanophylla Lindl. foliage and to improve digestion by Barbarine sheep

    International Nuclear Information System (INIS)

    Three in vitro experiments and one in vivo experiment were carried out to study the effect of wood ash sources (6 L wood ash solution/kg fresh plant leaves) and levels and treatment duration on the nutritive value of acacia leaves. In Experiment 1, samples of fresh (F), dried (D), or dried and ground (DG) acacia were soaked for 6 h in water or acacia wood ash solution (120 g of wood ash dry matter/L of water). Soaking acacia in water decreased total extractable phenols (TP), total extractable tannins (TT) and extractable condensed tannins (CT). Wood ash treatment led to a further decrease of these phenolic compounds and was highest with DG acacia. Experiment 2 investigated different levels of acacia wood ash (0, 120, 180 and 240 g wood ash dry matter/L of water) and treatment duration (1, 2 and 3 days). The higher the level of wood ash, the lower proportion of TP and CT in acacia was noted. In Experiment 3, two sources of wood ash (i.e., acacia and Aleppo pine) and the same solution of each source of wood ash were used eight times. The two sources of wood ash had similar deactivating effect on TP and CT. The rate of decrease of TP and CT was highest when the same wood ash solution was used four consecutive times and decreased progressively thereafter. In these three experiments, water and wood ash treatment reduced organic matter and crude protein content but substantially increased the neutral detergent fibre (NDFom) content of treated acacia. In the fourth experimenof treated acacia. In the fourth experiment, we treated acacia with acacia wood ash (180 g/L of water for 2 days) and the same solution was used five times. Treated and untreated acacia were air-dried and fed ad libitum to two groups, each of four Barbarine rams together with 300 g of concentrate. Wood ash treatment did not affect intake and OM digestibility of the diet but increased crude protein and NDFom digestibility (P < 0.05). Feeding untreated acacia resulted in negative N balances but with wood ash treatment, N balance was positive. Microbial N supply was not affected by wood ash treatment. It is concluded that wood ash treatment is a cost-effective way to deactivate tannins in acacia leaves, although additional energy is needed to ensure utilisation of the available N. (author)

  11. Wood ash treatment, a cost-effective way to deactivate tannins in Acacia cyanophylla Lindl. foliage and to improve digestion by Barbarine sheep

    Energy Technology Data Exchange (ETDEWEB)

    Ben Salem, H. [Institut National de la Recherche Agronomique de Tunisie (INRAT), Laboratoire des Productions Animales et Fourrageres, Ariana (Tunisia)]. E-mail: bensalem.hichem@iresa.agrinet.tn; Abidi, S. [Institut National de la Recherche Agronomique de Tunisie (INRAT), Laboratoire des Productions Animales et Fourrageres, Ariana (Tunisia); Ecole Superieure d' Agriculture de Mateur, Mateur (Tunisia); Makkar, H.P.S. [Animal Production and Health Section, Joint FAO/IAEA Division, International Atomic Energy Agency, Vienna (Austria); Nefzaoui, A. [Institut National de la Recherche Agronomique de Tunisie (INRAT), Laboratoire des Productions Animales et Fourrageres, Ariana (Tunisia)

    2005-08-19

    Three in vitro experiments and one in vivo experiment were carried out to study the effect of wood ash sources (6 L wood ash solution/kg fresh plant leaves) and levels and treatment duration on the nutritive value of acacia leaves. In Experiment 1, samples of fresh (F), dried (D), or dried and ground (DG) acacia were soaked for 6 h in water or acacia wood ash solution (120 g of wood ash dry matter/L of water). Soaking acacia in water decreased total extractable phenols (TP), total extractable tannins (TT) and extractable condensed tannins (CT). Wood ash treatment led to a further decrease of these phenolic compounds and was highest with DG acacia. Experiment 2 investigated different levels of acacia wood ash (0, 120, 180 and 240 g wood ash dry matter/L of water) and treatment duration (1, 2 and 3 days). The higher the level of wood ash, the lower proportion of TP and CT in acacia was noted. In Experiment 3, two sources of wood ash (i.e., acacia and Aleppo pine) and the same solution of each source of wood ash were used eight times. The two sources of wood ash had similar deactivating effect on TP and CT. The rate of decrease of TP and CT was highest when the same wood ash solution was used four consecutive times and decreased progressively thereafter. In these three experiments, water and wood ash treatment reduced organic matter and crude protein content but substantially increased the neutral detergent fibre (NDFom) content of treated acacia. In the fourth experiment, we treated acacia with acacia wood ash (180 g/L of water for 2 days) and the same solution was used five times. Treated and untreated acacia were air-dried and fed ad libitum to two groups, each of four Barbarine rams together with 300 g of concentrate. Wood ash treatment did not affect intake and OM digestibility of the diet but increased crude protein and NDFom digestibility (P < 0.05). Feeding untreated acacia resulted in negative N balances but with wood ash treatment, N balance was pos