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Sample records for improves protein digestive

  1. High Hydrostatic Pressure Pretreatment of Whey Protein Isolates Improves Their Digestibility and Antioxidant Capacity

    Directory of Open Access Journals (Sweden)

    Michèle M. Iskandar

    2015-05-01

    Full Text Available Whey proteins have well-established antioxidant and anti-inflammatory bioactivities. High hydrostatic pressure processing of whey protein isolates increases their in vitro digestibility resulting in enhanced antioxidant and anti-inflammatory effects. This study compared the effects of different digestion protocols on the digestibility of pressurized (pWPI and native (nWPI whey protein isolates and the antioxidant and anti-inflammatory properties of the hydrolysates. The pepsin-pancreatin digestion protocol was modified to better simulate human digestion by adjusting temperature and pH conditions, incubation times, enzymes utilized, enzyme-to-substrate ratio and ultrafiltration membrane molecular weight cut-off. pWPI showed a significantly greater proteolysis rate and rate of peptide appearance regardless of digestion protocol. Both digestion methods generated a greater relative abundance of eluting peptides and the appearance of new peptide peaks in association with pWPI digestion in comparison to nWPI hydrolysates. Hydrolysates of pWPI from both digestion conditions showed enhanced ferric-reducing antioxidant power relative to nWPI hydrolysates. Likewise, pWPI hydrolysates from both digestion protocols showed similar enhanced antioxidant and anti-inflammatory effects in a respiratory epithelial cell line as compared to nWPI hydrolysates. These findings indicate that regardless of considerable variations of in vitro digestion protocols, pressurization of WPI leads to more efficient digestion that improves its antioxidant and anti-inflammatory properties.

  2. Improving performance of broilers fed lower digestible protein diets

    NARCIS (Netherlands)

    Qaisrani, S.N.

    2014-01-01

    Background and Problem Statement There is a great interest in recent years to replace soybean meal by cost effective protein sources such as rapeseed meal and maize gluten in animal feed. In poultry, this replacement, however, may result in a poorer performance due to among others i

  3. Antimicrobial treatment reduces intestinal microflora and improves protein digestive capacity without changes in villous structure in weanling pigs

    DEFF Research Database (Denmark)

    Thymann, Thomas; Sørensen, Kristina U.; Hedemann, Mette S.;

    2007-01-01

    -old controls In 8; P amylase, trypsin and small intestinal...... that the beneficial effects of antimicrobials are mediated not only through reduction in intestinal bacteria] load, but also through a stimulation of protein digestive function and goblet cell density....

  4. Steam explosion of Brewer's spent grain improves enzymatic digestibility of carbohydrates and affects solubility and stability of proteins.

    Science.gov (United States)

    Kemppainen, K; Rommi, K; Holopainen, U; Kruus, K

    2016-09-01

    Steam explosion was studied as a means to improve the enzymatic digestibility of carbohydrates in Brewer's spent grain, a protein and lipid-rich lignocellulosic by-product of the brewing industry. Having temperature, treatment time and the presence of acid catalyst as variables, a treatment at 200 °C for 10 min without an acid catalyst was found to be the most efficient, dissolving 12.1 % of the dry matter. Mainly oligomeric non-cellulosic glucan and arabinoxylan were dissolved, and the remaining insoluble carbohydrates could be efficiently hydrolysed by an enzyme cocktail (75 % hydrolysis yield). The process also caused partial protein degradation and dissolved over a third of the total nitrogen. Meanwhile, the insoluble protein appeared to become more strongly associated with acid-insoluble lignin. Compositional changes observed in the proteins and carbohydrates were supported by the results of epifluorescence microscopy. The process yielded three chemically different fractions which could serve as biorefinery products or intermediates.

  5. 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.

  6. Influence of kiwifruit on protein digestion.

    Science.gov (United States)

    Kaur, Lovedeep; Boland, Mike

    2013-01-01

    Green kiwifruit consumption has long been thought to assist in the digestion of food proteins due to the presence of the proteolytic enzyme actinidin. This chapter reviews the recent findings of both in vitro and in vivo studies on the effect of green kiwifruit (which contain the enzyme actinidin) on the gastro- and small-intestinal digestion of a range of common food proteins and protein-rich foods including milk, meat, fish, eggs, legumes, and cereal proteins. Clear evidence is provided that green kiwifruit, and the enzyme actinidin itself, can provide enhanced upper-tract digestion (particularly gastric) of a variety of food proteins, which lends support to a role for dietary kiwifruit as a digestive aid. Kiwifruit influences the digestion patterns of food proteins to varying extents. For some protein sources, digestion in the presence of green kiwifruit resulted in a substantially greater digestion of intact protein, and different peptide patterns were produced compared with those seen after digestion with mammalian digestive enzymes alone. Kiwifruit extract alone (in the absence of other digestive enzymes) has been observed to be capable of digesting some proteins present in foods, particularly those in yoghurt, cheese, fish, and raw eggs. An in vivo (pig) study including a positive control of added actinidin and a negative control where the actinidin in green kiwifruit had been inactivated showed conclusively that actinidin is responsible for the enhanced gastric hydrolysis of food proteins.

  7. 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)

  8. Kafirinas, proteínas clave para conferir digestibilidad y calidad proteica al grano de sorgo Kafirins, key proteins to improve digestibility and proteic quality of sorghum grain

    Directory of Open Access Journals (Sweden)

    Elizabeth Chiquito-Almanza

    2011-04-01

    Full Text Available El sorgo es un alimento básico en varios países de África y Asia. Sin embargo, su grano es deficiente en lisina y su calidad proteica disminuye cuando se cocina. Los intentos para conferir calidad proteica al grano de sorgo, no han satisfecho los requerimientos nutricionales, y las alternativas biotecnológicas se han enfocado a la expresión heteróloga de proteínas, sin prestar atención a incrementar la digestibilidad proteica. El incremento del contenido de lisina en maíz QPM y el silenciamiento génico de las α-zeínas en maíz, sugieren que la modificación de la expresión de las kafirinas, una familia de prolaminas del sorgo homólogas a las zeínas de maíz, permite incrementar el contenido de lisina y la digestibilidad proteica del grano de sorgo. En esta revisión se discuten aspectos básicos de la clasificación de las kafirinas, su homología con las zeínas de maíz, y su contribución en la calidad y digestibilidad proteica del grano de sorgo. El objetivo es sustentar la hipótesis de que la modificación de la expresión de las kafirinas mediante silenciamiento génico es una estrategia clave para mejorar el valor nutritivo del grano del sorgo, el estudio se llevó a cabo durante 2009 y 2010.Sorghum is a basic food in several countries of Africa and Asia. However, its grain is deficient in lysine and its proteic quality diminishes when is cooked. Attempts to confer proteic quality to sorghum grain have not satisfed the nutritional requirements, and biotechnical alternatives have been focused to proteins' heterologous expression, without taking into account to increase proteic digestibility. Increment of lysine content in QPM corn and gene silencing of α-zeins in corn, suggest that modification of expression of kafirins, a prolamin family of sorghum homologous to corn zeins, allows to increase lysin content and the proteic digestibility of sorghum grain. In this revision basic issues of kafirins classification are

  9. Protein enriched pasta: structure and digestibility of its protein network.

    Science.gov (United States)

    Laleg, Karima; Barron, Cécile; Santé-Lhoutellier, Véronique; Walrand, Stéphane; Micard, Valérie

    2016-02-01

    Wheat (W) pasta was enriched in 6% gluten (G), 35% faba (F) or 5% egg (E) to increase its protein content (13% to 17%). The impact of the enrichment on the multiscale structure of the pasta and on in vitro protein digestibility was studied. Increasing the protein content (W- vs. G-pasta) strengthened pasta structure at molecular and macroscopic scales but reduced its protein digestibility by 3% by forming a higher covalently linked protein network. Greater changes in the macroscopic and molecular structure of the pasta were obtained by varying the nature of protein used for enrichment. Proteins in G- and E-pasta were highly covalently linked (28-32%) resulting in a strong pasta structure. Conversely, F-protein (98% SDS-soluble) altered the pasta structure by diluting gluten and formed a weak protein network (18% covalent link). As a result, protein digestibility in F-pasta was significantly higher (46%) than in E- (44%) and G-pasta (39%). The effect of low (55 °C, LT) vs. very high temperature (90 °C, VHT) drying on the protein network structure and digestibility was shown to cause greater molecular changes than pasta formulation. Whatever the pasta, a general strengthening of its structure, a 33% to 47% increase in covalently linked proteins and a higher β-sheet structure were observed. However, these structural differences were evened out after the pasta was cooked, resulting in identical protein digestibility in LT and VHT pasta. Even after VHT drying, F-pasta had the best amino acid profile with the highest protein digestibility, proof of its nutritional interest.

  10. Protein Nitrogen Determination by Kjeldahl Digestion and Ion Chromatography.

    Science.gov (United States)

    Wang, Hsiaoling; Pampati, Nagarani; McCormick, William M; Bhattacharyya, Lokesh

    2016-06-01

    We report development and validation of a simple, rapid, and accurate method for the quantitation of protein nitrogen, which combines Kjeldahl digestion and ion chromatography with suppressed conductivity detection and requires nanomolar amount of nitrogen in samples (≥10 μg protein). The mechanism of suppressed conductivity detection does not permit analysis of samples containing copper (present in Kjeldahl digestion solution) and aluminum (present in many vaccines as adjuvants) due to precipitation of their hydroxides within the suppressor. We overcame this problem by including 10 μM oxalic acid in Kjeldahl digests and in the eluent (30 mM methanesulfonic acid). The chromatography is performed using an IonPac CS-16 cation exchange column by isocratic elution. The method reduces the digestion time to less than 1 h and eliminates the distillation and titration steps of the Kjeldahl method, thereby reducing the analysis time significantly and improving precision and accuracy. To determine protein nitrogen in samples containing non-protein nitrogen, proteins are precipitated by a mixture of deoxycholate and trichloroacetic acid and the precipitates are analyzed after dissolving in KOH. The method is particularly useful for biological samples that are limited and can also be applied to food, environmental, and other materials.

  11. 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.

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

    Science.gov (United States)

    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 ...

  13. The on-bead digestion of protein corona on nanoparticles by trypsin immobilized on the magnetic nanoparticle.

    Science.gov (United States)

    Hu, Zhengyan; Zhao, Liang; Zhang, Hongyan; Zhang, Yi; Wu, Ren'an; Zou, Hanfa

    2014-03-21

    Proteins interacting with nanoparticles would form the protein coronas on the surface of nanoparticles in biological systems, which would critically impact the biological identities of nanoparticles and/or result in the physiological and pathological consequences. The enzymatic digestion of protein corona was the primary step to achieve the identification of protein components of the protein corona for the bottom-up proteomic approaches. In this study, the investigation on the tryptic digestion of protein corona by the immobilized trypsin on a magnetic nanoparticle was carried out for the first time. As a comparison with the usual overnight long-time digestion and the severe self-digestion of free trypsin, the on-bead digestion of protein corona by the immobilized trypsin could be accomplished within 1h, along with the significantly reduced self-digestion of trypsin and the improved reproducibility on the identification of proteins by the mass spectrometry-based proteomic approach. It showed that the number of identified bovine serum (BS) proteins on the commercial Fe3O4 nanoparticles was increased by 13% for the immobilized trypsin with 1h digestion as compared to that of using free trypsin with even overnight digestion. In addition, the on-bead digestion of using the immobilized trypsin was further applied on the identification of human plasma protein corona on the commercial Fe3O4 nanoparticles, which leads the efficient digestion of the human plasma proteins and the identification of 149 human plasma proteins corresponding to putative critical pathways and biological processes.

  14. Non-decoloured In-gel Digestion of Coomassie Blue-stained Proteins

    Institute of Scientific and Technical Information of China (English)

    ZHANG Guo-an; BAO Hui-min; FAN Hui-zhi; YANG Peng-yuan

    2003-01-01

    A simplified method is presented for tryptic digestion of Coomassie brilliant blue(CBB)-stained proteins in polyacrylamide gels. Compared with conventional methods, the proposed method does not require a removal of the dye before digestion, and is thus faster and saves a lot of labor. The resulted digest can be analyzed by either RPLC/ESIMS or MALDI MS for identification of the protein in a conventional way. Model studies with bovine serum albumin(BSA) showed that 50 ng of the protein could be routinely identified. The simplified procedure displays a tendency to produce more incompletely cleaved peptides, which is favorable for improving the sequence coverage.

  15. Effects of Low Protein Diets on Nutrient Digestibility and Excretion in Finishing Pigs

    Institute of Scientific and Technical Information of China (English)

    Hu Qin; Zhu Jianping; Liu Chunxue; Hong Ping; Du Li; Cheng Yufei; Fan Xueliang; Zhou Yanmin; Li Linfeng; Wen Chao

    2015-01-01

    The experiment was conducted to study the effects of low protein diets on nutrient digestibility and excretion of finishing pigs. For the 30-d trial,fortytwo duroc × landrace × yorkshire( DLY) finishing pigs with the initial weight of( 91. 13 ± 1. 64) kg were randomly divided into two groups,with three replicates in each group. The pigs in the two groups were fed with different crude protein( CP) levels( control groups: 15. 49% and low protein groups: 12. 59%). The results showed that:( 1) The digestibility of dry matter and crude fat( CF) in low protein groups were increased significantly( P 0. 05). The results showed that 2. 9% reduction of the CP in diets could improve nutrient digestibility and reduce the excretion of N,but reduced the digestibility of Arg.

  16. Effects of enzymatic dephosphorylation on infant in vitro gastrointestinal digestibility of milk protein concentrate.

    Science.gov (United States)

    Liu, Dasong; Wang, Yuanyuan; Yu, Yun; Hu, Jinhua; Lu, Naiyan; Regenstein, Joe M; Wang, Miao; Zhou, Peng

    2016-04-15

    This study investigated the effects of dephosphorylation extent on infant in vitro gastric clotting property and gastrointestinal digestibility of milk protein concentrate. Dephosphorylation was affected by phosphatase type and incubation pH. A series of milk protein concentrate with 0-69% dephosphorylation were obtained by incubation with calf intestinal alkaline phosphatase at pH 6.5 for 0-420 min. Both β- and αs1-caseins in the modified milk protein concentrate showed multiply dephosphorylated isoforms with different numbers of phosphate groups depending on the extent of dephosphorylation. With increased dephosphorylation of milk protein concentrate, the gastric clotting extent decreased and the gastrointestinal digestibility increased under infant in vitro conditions. These results suggested the potential of developing a dephosphorylated milk protein concentrate, with improved gastric clotting property and gastrointestinal digestibility, to simulate the multiply phosphorylated patterns of human casein and hence to further the humanization of infant formula on a molecular level.

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

    NARCIS (Netherlands)

    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 oc

  18. Gelation of soybean protein and polysaccharides delays digestion.

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    Hu, Bing; Chen, Qing; Cai, Qimeng; Fan, Yun; Wilde, Peter J; Rong, Zhen; Zeng, Xiaoxiong

    2017-04-15

    Xanthan gum and carrageenan, representing the medium and highly negatively charged polysaccharides, were heated respectively together with soybean protein isolate (SPI) at different biopolymer ratios. Upon mixing with simulated stomach juice (SSJ), the xanthan-SPI and carrageenan-SPI at biopolymer ratios higher than 0.01 leads to self-assembled gelation immediately. Stronger gel is formed under higher biopolymer ratios. Highly negatively charged carrageenan forms a stronger gel than that composed with xanthan gum. SDS-PAGE results show the digestibility of SPI is delayed after incorporation with the polysaccharides, which is enhanced with the increase of the biopolymer mass ratios. And the polysaccharide with higher negative charge has stronger potential in delaying the digestion of SPI. Furthermore, the microstructure of the xanthan-SPI and carrageenan-SPI gel before and after simulated stomach digestion was characterized by scanning electron microscope (SEM), which also confirms that the gel delays the digestion of soybean protein.

  19. PROTEIN FRACTIONATION AND IN VITRO DIGESTIBILITY OF AZOLLA IN RUMINANTS

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    S. PARASHURAMULU

    2013-05-01

    Full Text Available A study was undertaken to evaluate the nutritive value and digestibility of Azolla in ruminants by in vitro techniques. The crude protein, crude fibre and ether extract contents were at a level of 21.37%, 12.5% and 2.3%, respectively. The neutral and acid detergent fibre levels were about 35.4 and 23.9%, respectively. The average in vitro dry matter digestibility, in vitro organic matter digestibility and metabolozable energy contents were 79.5%, 63.8 mg/200mg and 7.36 MJ/kg DM (1759 kcal/kg, respectively. The various protein fractions A, B1, B2, B3 and C estimated by Cornell net crude protein solubility system were 18.22, 42.56, 15.15, 7.47 and 16.61% of total protein, respectively. The Azolla contained significantly higher B1 fraction followed by A, B2 and C and lowest fraction of C. Thus in view of above, present study indicated Azolla to be a good source protein supplement with 21.37% crude protein with highest B protein fractions, moderate source of energy (1759 kcal ME/kg, high dry matter and organic matter digestibilities and rich in trace minerals thus could be used as an alternate protein supplement or as supplementary protein supplement to ruminants.

  20. 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

    2014-05-07

    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 α-lactalbumin, β-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.

  1. Tannin contents and protein digestibility of black grams (Vigna mungo) after soaking and cooking.

    Science.gov (United States)

    Zia-Ur-rehman; Shah, W H

    2001-01-01

    The objective of this research was to ascertain the effects of soaking black grams (Cultivar AARI-5732) in different salt solutions at different temperatures and different time periods, and different methods of cooking on the tannin content and protein digestibility. Tannin content of black grams was reduced to various extents by soaking at 30 degrees and 100 degrees C for different time periods. However, soaking at 100 degrees C increased the rate of extraction and reduced the extraction time of tannins. Soaking black grams in water at 100 degrees C reduced tannins by 22.14% in 45 minutes whereas about 2.5 times more tannin was reduced after soaking in sodium bicarbonate solution with or without sodium chloride. Maximum improvement in protein digestibility was also observed after soaking black grams in sodium bicarbonate solution. Tannin contents were further reduced along with improvement in protein digestibility as a result of cooking.

  2. Ultrasonic treatment to improve anaerobic digestibility of dairy waste streams.

    Science.gov (United States)

    Palmowski, L; Simons, L; Brooks, R

    2006-01-01

    The dairy-processing industry generates various types of organic wastes, which are utilised as stock feed, for anaerobic digestion, spread on land or alternatively land-filled at high costs. Owing to the generation of renewable energy, anaerobic digestion is an attractive option for many factories. To enhance the biological degradation process, a mechanical disintegration of various waste dairy streams was undertaken. While the successful application of ultrasonic treatment has been reported for various municipal waste streams, limited information was available for dairy industry applications. The results of this study showed that ultrasonic treatment can improve the digestibility of the more problematic dairy waste streams, such as sludges, by breaking down micro-organisms' cell walls and releasing soluble cell compounds. For more soluble streams, such as dairy factory effluent, an increased gas production was observed and attributed to the reduced particle size of the fat globules.

  3. Effects of dietary protein concentration on performance and nutrient digestibility in Pekin ducks during aflatoxicosis.

    Science.gov (United States)

    Chen, X; Murdoch, R; Zhang, Q; Shafer, D J; Applegate, T J

    2016-04-01

    A 14-d study was conducted to determine the impact of dietary crude protein concentration on performance, serum biochemistry, and nutrient digestive functions in Pekin ducklings during aflatoxicosis. A total of 144 male Pekin ducklings were randomly allotted to 4 dietary treatments arranged in a 2×2 factorial with 2 crude protein (CP) (20 and 24% on an analyzed basis) with or without 0.2 mg/kg aflatoxin B1 (AFB1) (0.21 mg/kg analyzed). The AFB1 reduced BW gain, feed intake, and breast muscle weight by 33 to 43% (Pnutrient digestibility. No statistical interaction of AFB1 by CP was observed for any measures. Results from the current study suggest that AFB1 at low concentration can significantly impair performance of Pekin ducklings primarily through inhibited feed intake, as well as influence nutrient digestion processes (jejunum morphology, digestive enzyme activity, and apparent energy digestibility). Higher dietary CP can improve growth performance of ducklings regardless of AF exposure, but did not interact with dietary AFB1 on performance, serum biochemistry, or nutrient digestion in Pekin ducklings from hatch to 14 d.

  4. Enzymatic Digestion for Improved Bacteria Separation from Leafy Green Vegetables.

    Science.gov (United States)

    Wang, Danhui; Wang, Ziyuan; He, Fei; Kinchla, Amanda J; Nugen, Sam R

    2016-08-01

    An effective and rapid method for the separation of bacteria from food matrix remains a bottleneck for rapid bacteria detection for food safety. Bacteria can strongly attach to a food surface or internalize within the matrix, making their isolation extremely difficult. Traditional methods of separating bacteria from food routinely involve stomaching, blending, and shaking. However, these methods may not be efficient at removing all the bacteria from complex matrices. Here, we investigate the benefits of using enzyme digestion followed by immunomagnetic separation to isolate Salmonella from spinach and lettuce. Enzymatic digestion using pectinase and cellulase was able to break down the structure of the leafy green vegetables, resulting in the detachment and release of Salmonella from the leaves. Immunomagnetic separation of Salmonella from the liquefied sample allowed an additional separation step to achieve a more pure sample without leaf debris that may benefit additional downstream applications. We have investigated the optimal combination of pectinase and cellulase for the digestion of spinach and lettuce to improve sample detection yields. The concentrations of enzymes used to digest the leaves were confirmed to have no significant effect on the viability of the inoculated Salmonella. Results reported that the recovery of the Salmonella from the produce after enzyme digestion of the leaves was significantly higher (P < 0.05) than traditional sample preparation methods to separate bacteria (stomaching and manually shaking). The results demonstrate the potential for use of enzyme digestion prior to separation can improve the efficiency of bacteria separation and increase the likelihood of detecting pathogens in the final detection assay.

  5. Stability and Immunogenicity of Hypoallergenic Peanut Protein-Polyphenol Complexes During In Vitro Pepsin Digestion

    Science.gov (United States)

    Allergenic peanut proteins are relatively resistant to digestion, and if digested, metabolized peptides tend to remain large and immunoreactive, triggering allergic reactions in sensitive individuals. In this study, the stability of hypoallergenic peanut protein-polyphenol complexes was evaluated d...

  6. Relationship between Molecular Structure Characteristics of Feed Proteins and Protein In vitro Digestibility and Solubility.

    Science.gov (United States)

    Bai, Mingmei; Qin, Guixin; Sun, Zewei; Long, Guohui

    2016-08-01

    The nutritional value of feed proteins and their utilization by livestock are related not only to the chemical composition but also to the structure of feed proteins, but few studies thus far have investigated the relationship between the structure of feed proteins and their solubility as well as digestibility in monogastric animals. To address this question we analyzed soybean meal, fish meal, corn distiller's dried grains with solubles, corn gluten meal, and feather meal by Fourier transform infrared (FTIR) spectroscopy to determine the protein molecular spectral band characteristics for amides I and II as well as α-helices and β-sheets and their ratios. Protein solubility and in vitro digestibility were measured with the Kjeldahl method using 0.2% KOH solution and the pepsin-pancreatin two-step enzymatic method, respectively. We found that all measured spectral band intensities (height and area) of feed proteins were correlated with their the in vitro digestibility and solubility (p≤0.003); moreover, the relatively quantitative amounts of α-helices, random coils, and α-helix to β-sheet ratio in protein secondary structures were positively correlated with protein in vitro digestibility and solubility (p≤0.004). On the other hand, the percentage of β-sheet structures was negatively correlated with protein in vitro digestibility (pproteins are closely related to their in vitro digestibility at 28 h and solubility. Furthermore, the α-helix-to-β-sheet ratio can be used to predict the nutritional value of feed proteins.

  7. [Digestibility and protein quality of quinua: comparative study of quinua (Chenopodium Quinoa) seed and flour in children].

    Science.gov (United States)

    López de Romaña, G; Graham, G G; Rojas, M; MacLean, W C

    1981-09-01

    Based on the hypothesis that the digestibility of quinua seed is the limiting factor in the utilization of nutrients from this staple, two quinua-based diets were prepared using quinua seeds and quinua flour. Theses diets were offered to children recovering from malnutrition. The digestibility and protein quality of the quinua diets were compared to those of a casein control diet by analyzing the children's metabolic balance. Results showed that digestibility of the quinua diets were compared to those of a casein control diet by analyzing the children's metabolic balance. Results showed that digestibility of the quinua seed is the limiting factor in the protein and energy utilization, and that milling improves significantly the digestibility of fat and carbohydrates. Findings also confirmed that the protein quality of quinua seeds is adequate for human consumption.

  8. Toxoplasma gondii Ingests and Digests Host Cytosolic Proteins

    Science.gov (United States)

    Dou, Zhicheng; McGovern, Olivia L.; Di Cristina, Manlio

    2014-01-01

    ABSTRACT The protozoan parasite Toxoplasma gondii resides within a nonfusogenic vacuole during intracellular replication. Although the limiting membrane of this vacuole provides a protective barrier to acidification and degradation by lysosomal hydrolases, it also physically segregates the parasite from the host cytosol. Accordingly, it has been suggested that T. gondii acquires material from the host via membrane channels or transporters. The ability of the parasite to internalize macromolecules via endocytosis during intracellular replication has not been tested. Here, we show that Toxoplasma ingests host cytosolic proteins and digests them using cathepsin L and other proteases within its endolysosomal system. Ingestion was reduced in mutant parasites lacking an intravacuolar network of tubular membranes, implicating this apparatus as a possible conduit for trafficking to the parasite. Genetic ablation of proteins involved in the pathway is associated with diminished parasite replication and virulence attenuation. We show that both virulent type I and avirulent type II strain parasites ingest and digest host-derived protein, indicating that the pathway is not restricted to highly virulent strains. The findings provide the first definitive evidence that T. gondii internalizes proteins from the host during intracellular residence and suggest that protein digestion within the endolysosomal system of the parasite contributes to toxoplasmosis. PMID:25028423

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

    DEFF Research Database (Denmark)

    Alstrup, Lene; Weisbjerg, Martin Riis; Hymøller, Lone

    2014-01-01

    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...

  10. Protein digestion and absorption in the blind loop syndrome.

    Science.gov (United States)

    Curtis, K J; Prizont, R; Kim, Y S

    1979-12-01

    Protein digestion and absorption was studied in rats with 6-week-old surgically constructed self-filling intestinal blind loops and steatorrhea, ie, blind-loop animals and controls were fed a 14C-labeled protein meal containing a nonabsorbable marker, 51CrCl3, and sacrificed 1 or 2 hr later. Intestinal contents were analyzed for 14C, 51Cr, protein, trypsin, and the products of digestion. At 1 hr, 14C absorption was greater in controls, but at 2 hr there was no difference in absorption between the two groups. Marker studies showed that blind-loop filling resulted in a delay of the progression of intestinal contents distally. Intraluminal trypsin and porteolysis were similar in the two groups. Endogenous protein was greater in the blind-loop animals. The early stages of the blind-loop syndrome may be characterized by delayed protein absorption secondary to blind-loop filling, which is compensated for by the distal gut resulting in an absence of overall protein malabsorption.

  11. Dry anaerobic digestion of lignocellulosic and protein residues

    Directory of Open Access Journals (Sweden)

    Maryam M Kabir

    2015-12-01

    Full Text Available Utilisation of wheat straw and wool textile waste in dry anaerobic digestion (AD process was investigated. Dry-AD of the individual substrates as well as co-digestion of those were evaluated using different total solid (TS contents ranging between 6 to 30%. Additionally, the effects of the addition of nutrients and cellulose- or protein-degrading enzymes on the performance of the AD process were also investigated. Dry-AD of the wheat straw resulted in methane yields of 0.081 – 0.200 Nm3CH4/kgVS with the lowest and highest values obtained at 30 and 21% TS, respectively. The addition of the cellulolytic enzymes could significantly increase the yield in the reactor containing 13% TS (0.231 Nm3CH4/kg VS. Likewise, degradation of wool textile waste was enhanced significantly at TS of 13% with the addition of the protein-degrading enzyme (0.131 Nm3CH4/kg VS. Furthermore, the co-digestion of these two substrates showed higher methane yields compared with the methane potentials calculated for the individual fractions at all the investigated TS contents due to synergetic effects and better nutritional balance.

  12. On-plate digestion of proteins using novel trypsin-immobilized magnetic nanospheres for MALDI-TOF-MS analysis.

    Science.gov (United States)

    Li, Yan; Yan, Bo; Deng, Chunhui; Tang, Jia; Liu, Junyan; Zhang, Xiangmin

    2007-10-01

    In this study, a novel method of on-plate digestion using trypsin-immobilized magnetic nanospheres was developed followed by MALDI-TOF-MS for rapid and effective analysis and identification of proteins. We utilized a facile one-pot method for the direct preparation of amine-functionalized magnetic nanospheres with highly magnetic properties and the amino groups on the outer surface. Through the reaction of the aldehyde groups with amine groups, trypsin was simply and stably immobilized onto the magnetic nanospheres. The obtained trypsin-linked magnetic nanospheres were then applied for on-plate digestion of sample proteins (myoglobin and Cytochrome c). Moreover, after digestion, the trypsin-linked nanospheres could be easily removed from the plate due to their magnetic property, which would avoid causing contamination on the ion source chamber in MS. The effects of the temperature and incubation time on the digestion efficiency were characterized. Within only 5 min, proteins could be efficiently digested with the peptide sequence coverage higher than or equal to that of the traditional in-solution digestion for 12 h. Furthermore, RPLC fractions of rat liver extract were also successfully processed using this novel method. These results suggested that our improved on-plate digestion protocol for MALDI-MS may find further application in automated analysis of large sets of proteins.

  13. Secondary Structure and Subunit Composition of Soy Protein In Vitro Digested by Pepsin and Its Relation with Digestibility

    Directory of Open Access Journals (Sweden)

    Yong Yang

    2016-01-01

    Full Text Available In the present study, in vitro digestibility and structure of soybean protein isolates (SPIs prepared from five soybean varieties were investigated in simulated gastric fluid (SGF, using FT-IR microspectroscopy and SDS-PAGE. The result indicated that β-conformations were prone to be hydrolyzed by pepsin preferentially and transformed to unordered structure during in vitro digestion, followed by the digestion of α-helix and unordered structure. A negative linear correlation coefficient was found between the β-conformation contents of five SPIs and their in vitro digestibility values. The intensities of the protein bands corresponding to 7S and 11S fractions were decreased and many peptide bands appeared at 11~15 kDa during enzymatic hydrolysis. β-conglycinin was poorly hydrolyzed with pepsin, especially the β-7S subunit. On the other hand, basic polypeptides of glycinin degraded slower than acidic polypeptides and represented a large proportion of the residual protein after digestion. 11S-A3 of all SPIs disappeared after 1 h digestion. Moreover, a significant negative linear correlation coefficient (r=-0.89 was found between the β-7S contents of five SPIs and their in vitro digestibility values. These results are useful for further studies of the functional properties and bioactive properties of these varieties and laid theoretical foundations for the development of the specific functional soy protein isolate.

  14. Effects of Industrial Heating Processes of Milk-Based Enteral Formulas on Site-Specific Protein Modifications and Their Relationship to in Vitro and in Vivo Protein Digestibility.

    Science.gov (United States)

    Wada, Yasuaki; Lönnerdal, Bo

    2015-08-01

    Heat treatments are applied to milk and dairy products to ensure their microbiological safety and shelf lives. Types of heating processes may have different effects on protein modifications, leading to different protein digestibility. In this study, milk-based liquid nutritional formulas (simulating enteral formulas) were subjected to steam injection ultra-high-temperature treatment or in-can sterilization, and the formulas were investigated by proteomic methods and in vitro and in vivo digestion assays. Proteomic analyses revealed that in-can sterilization resulted in higher signals for N(ε)-carboxymethyllysine and dephosphorylation of Ser residues in major milk proteins than in steam-injected formula, reflecting the more severe thermal process of in-can sterilization. In vitro and in vivo digestion assays indicated that steam injection improved protein digestibility, supposedly by denaturation, while the improvement seemed to be overwhelmed by formation of aggregates that showed resistance to digestion in in-can sterilized formula. Adverse effects of heat treatment on protein digestibility are more likely to be manifested in milk-based formulas than in cow's milk. Although the differences might be of limited significance in terms of amino acid bioavailability, these results emphasize the importance of protein quality of raw materials and selection of heating processes.

  15. 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

    2014-02-07

    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.

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

    Directory of Open Access Journals (Sweden)

    Silia Maria de Negreiros Sousa

    2013-08-01

    Full Text Available 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 diets 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.

  17. Digestibility Improvement of Sorted Waste with Alkaline Hydrothermai Pretreatment

    Institute of Scientific and Technical Information of China (English)

    WANG Hao; WANG Hongtao; LU Wenjing; ZHAO Yan

    2009-01-01

    The digestibility of sorted municipal solid waste (MSW) is often limited by the high content of structured green waste. The objectives of this study are to investigate the effect of alkaline hydrothermal pretreatment on the anaerobic digestion of sorted waste and to analyze the biogas production of different parts of the waste. The waste was hydrothermally pretreated in a dilute alkali solution. The hydrolysis product was then incubated in a 500 mL saline bottle to determine the biochemical methane potential (BMP) under mesophilic anaerobic conditions. The optimum hydrothermal condition was 170℃ at 4 g NaOH/100 g solid for one hour. The concentration of chemical oxygen demand (COD) was 13 936 mg/L and the methane yield was 164 mL/g volatile solid (VS) for 6 days incubation at the optimum conditions. The biogas production was increased more than 50% over the control, with the methane conversion ratio on a carbon basis enhanced to 30.6%. The organic part of the sorted waste was mainly kitchen garbage and leaves. Model kitchen garbage completely liquified at 130℃ for one hour had a methane yield of 276 mL/g VS. The alkali addition slightly enhanced the hydrolyzation rate and methane yield. The biogas potential of leaves was improved by pre-treatment at above 150℃ under alkaline conditions.

  18. 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.

  19. Serial CSTR digester configuration for improving biogas production from manure

    DEFF Research Database (Denmark)

    Boe, Kanokwan; Angelidaki, Irini

    2009-01-01

    A new configuration of manure digesters for improving biogas production has been investigated in laboratory scale. A single thermophilic continuous-flow stirred tank reactor (CSTR) operated with a hydraulic retention time (HRT) of 15 days was compared to a serial CSTR configuration with volume...... distribution ratio of 80/20 and 90/10, and total HRT of 15 days. The results showed that the serial CSTR could obtain 11% higher biogas yield compared to the single CSTR. The increased biogas yield in the serial CSTR was mainly from the second reactor, which accounted for 16% and 12% of total biogas yield...... in the 90/10 and 80/20 configuration, respectively. VFA concentration in the serial CSTR was high in the first reactor but very low in the second reactor. The results from organic pulse load test showed that the second reactor in serial CSTR helped utilizing VFA produced from overloading in the first...

  20. Protein identification and in vitro digestion of fractions from Tenebrio molitor

    NARCIS (Netherlands)

    Yi, Liya; Boekel, van M.A.J.S.; Boeren, Sjef; Lakemond, Catriona M.M.

    2016-01-01

    The nutritional value of insect protein is evaluated not only in amino acid composition, but also in protein digestibility. The general amino acid composition of Tenebrio molitor has been reported before, but limited knowledge is available on its digestibility. The objective of this study was to

  1. Improving biogas quality and methane yield via co-digestion of agricultural and urban biomass wastes.

    Science.gov (United States)

    Poulsen, Tjalfe G; Adelard, Laetitia

    2016-08-01

    Impact of co-digestion versus mono-digestion on biogas and CH4 yield for a set of five biomass materials (vegetable food waste, cow dung, pig manure, grass clippings, and chicken manure) was investigated considering 95 different biomass mixes of the five materials under thermophilic conditions in bench-scale batch experiments over a period of 65days. Average biogas and CH4 yields were significantly higher during co-digestion than during mono-digestion of the same materials. This improvement was most significant for co-digestion experiments involving three biomass types, although it was independent of the specific biomasses being co-digested. Improvement in CH4 production was further more prominent early in the digestion process during co-digestion compared to mono-digestion. Co-digestion also appeared to increase the ultimate CH4/CO2 ratio of the gas produced compared to mono-digestion although this tendency was relatively weak and not statistically significant.

  2. Serial CSTR digester configuration for improving biogas production from manure.

    Science.gov (United States)

    Boe, Kanokwan; Angelidaki, Irini

    2009-01-01

    A new configuration of manure digesters for improving biogas production has been investigated in laboratory scale. A single thermophilic continuous-flow stirred tank reactor (CSTR) operated with a hydraulic retention time (HRT) of 15 days was compared to a serial CSTR configuration with volume distribution ratio of 80/20 and 90/10, and total HRT of 15 days. The results showed that the serial CSTR could obtain 11% higher biogas yield compared to the single CSTR. The increased biogas yield in the serial CSTR was mainly from the second reactor, which accounted for 16% and 12% of total biogas yield in the 90/10 and 80/20 configuration, respectively. VFA concentration in the serial CSTR was high in the first reactor but very low in the second reactor. The results from organic pulse load test showed that the second reactor in serial CSTR helped utilizing VFA produced from overloading in the first reactor, which improved the effluent quality and conversion efficiency of the serial CSTR.

  3. Trypsin immobilization in ordered porous polymer membranes for effective protein digestion

    Energy Technology Data Exchange (ETDEWEB)

    Qiao, Juan [Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, No. 2 Zhongguancun Beiyijie, Beijing 100190 (China); Kim, Jin Yong [Department of Chemistry, Yonsei University, 50 Yonsei-ro, Seoul 120-749 (Korea, Republic of); Wang, Yuan Yuan [Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, No. 2 Zhongguancun Beiyijie, Beijing 100190 (China); Qi, Li, E-mail: qili@iccas.ac.cn [Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, No. 2 Zhongguancun Beiyijie, Beijing 100190 (China); Wang, Fu Yi [Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, No. 2 Zhongguancun Beiyijie, Beijing 100190 (China); Moon, Myeong Hee, E-mail: mhmoon@yonsei.ac.kr [Department of Chemistry, Yonsei University, 50 Yonsei-ro, Seoul 120-749 (Korea, Republic of)

    2016-02-04

    Fast and effective protein digestion is a vital process for mass spectrometry (MS) based protein analysis. This study introduces a porous polymer membrane enzyme reactor (PPMER) coupled to nanoflow liquid chromatography-tandem MS (nLC-ESI-MS/MS) for on-line digestion and analysis of proteins. Poly (styrene-co-maleic anhydride) (PS-co-MAn) was fabricated by the breath figure method to make a porous polymer membrane in which the MAn group was covalently bound to enzyme. Based on this strategy, microscale PPMER (μPPMER) was constructed for on-line connection with the nLC-ESI-MS/MS system. Its capability for enzymatic digestion with bovine serum albumin (BSA) was evaluated with varied digestion periods. The on-line proteolysis of BSA and subsequent analysis with μPPMER-nLC-ESI-MS/MS revealed that peptide sequence coverage increased from 10.3% (digestion time 10 min) to 89.1% (digestion time 30 min). μPPMER can efficiently digest proteins due to the microscopic confinement effect, showing its potential application in fast protein identification and protease immobilization. Applications of on-line digestion using μPPMER with human plasma and urinary proteome samples showed that the developed on-line method yielded equivalent or better performance in protein coverage and identified more membrane proteins than the in-solution method. This may be due to easy accommodation of hydrophobic membrane proteins within membrane pores. - Highlights: • A porous polymer membrane enzyme reactor was developed. • Breath figure method was used for the fabrication of porous polymer membrane. • The enzyme reactor was coupled to nLC-ESI-MS/MS for proteins on-line digestion.

  4. 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)

  5. In vitro digestion of soluble cashew proteins and characterization of surviving IgE-reactive peptides

    Science.gov (United States)

    The stability of food allergens to digestion varies; and the ability of food proteins to cause an allergic reaction may be affected by the susceptibility of the allergen to digestion by proteases, including pepsin and trypsin. Recent studies have demonstrated that cashew nut allergens are often a ca...

  6. Development of continuous microwave-assisted protein digestion with immobilized enzyme.

    Science.gov (United States)

    Chen, Zhengyi; Li, Yongle; Lin, Shuhai; Wei, Meiping; Du, Fuyou; Ruan, Guihua

    2014-03-01

    In this study, an easy and efficiency protein digestion method called continuous microwave-assisted protein digestion (cMAED) with immobilized enzyme was developed and applied for proteome analysis by LC-MS(n). Continuous microwave power outputting was specially designed and applied. Trypsin and bromelain were immobilized onto magnetic micropheres. To evaluate the method of cMAED, bovine serum albumin (BSA) and protein extracted from ginkgo nuts were used as model and real protein sample to verify the digestion efficiency of cMAED. Several conditions including continuous microwave power, the ratio of immobilized trypsin/BSA were optimized according to the analysis of peptide fragments by Tricine SDS-PAGE and LC-MS(n). Subsequently, the ginkgo protein was digested with the protocols of cMAED, MAED and conventional heating enzymatic digestion (HED) respectively and the LC-MS(n) profiles of the hydrolysate was compared. Results showed that cMAED combined with immobilized enzyme was a fast and efficient digestion method for protein digestion and microwave power tentatively affected the peptide producing. The cMAED method will be expanded for large-scale preparation of bioactive peptides and peptide analysis in biological and clinical research.

  7. Rice Protein Extracted by Different Methods Affects Cholesterol Metabolism in Rats Due to Its Lower Digestibility

    Directory of Open Access Journals (Sweden)

    Hongbo Liu

    2011-11-01

    Full Text Available To elucidate whether the digestibility is responsible for the hypocholesterolemic action of rice protein, the effects of rice proteins extracted by alkali (RP-A and α-amylase (RP-E on cholesterol metabolism were investigated in 7-week-old male Wistar rats fed cholesterol-free diets for 3 weeks. The in vitro and in vivo digestibility was significantly reduced by RP-A and RP-E as compared to casein (CAS. The digestibility was lower in RP-E than that of RP-A. Compared with CAS, the significant cholesterol-lowering effects were observed in rats fed by RP-A and RP-E. Fecal excretion of bile acids was significantly stimulated by RP-E, but not by RP-A. The apparent cholesterol absorption was more effectively inhibited by RP-E than RP-A because more fecal neutral sterols were excreted in rats fed RP-E. There was a significant correlation between protein digestibility and cholesterol absorption (r = 0.8662, P < 0.01, resulting in a significant correlation between protein digestibility and plasma cholesterol level (r = 0.7357, P < 0.01 in this study. The present study demonstrates that the digestibility of rice protein affected by extraction method plays a major role in the modulation of cholesterol metabolism. Results suggest that the hypocholesterolemic action induced by rice protein with lower digestibility primarily contribute to the inhibition of cholesterol absorption.

  8. Continual feeding of two types of microalgal biomass affected protein digestion and metabolism in laying hens.

    Science.gov (United States)

    Ekmay, R D; Chou, K; Magnuson, A; Lei, X G

    2015-01-01

    A 14-wk study was conducted to determine the nutritional efficacy and ssmetabolic impact of 2 types of microalgal biomass as alternative protein sources in laying hen diets. Shaver hens (total = 150 and 26 wk old) were fed 1 of 5 diets: a control or a defatted green microalgal biomass (DG; Desmodesmus spp.) at 25% and a full-fatted diatom biomass (FD; Staurosira spp.) at 11.7% inclusion with or without protease. This experiment consisted of 5 replicates per treatment and each replicate contained 6 hens individually reared in cages (1 hen for biochemical data/replicate). Despite decreased ADFI (P = 0.03), hens fed DG or FD had final BW, overall hen-day egg production, and egg quality similar to the controls. Feeding DG or FD did not alter plasma concentrations of insulin, glutamine, and uric acid or alkaline phosphatase activity at wk 8 or 14 but decreased plasma 3-methyhistine concentrations (P = 0.03) and tartrate-resistant acid phosphatase (TRAP) activities (P < 0.001) at wk 14 and improved (P = 0.002) ileal total AA digestibility. Although DG or FD exhibited moderate effects on intestinal brush border protease activities and mRNA levels of duodenal transporters Pept1, Lat1, and Cat1, both substantially enhanced (P < 0.05) phosphorylation of hepatic protein synthesis key regulator S6 ribosomal protein (S6) and the ratio of phospho-S6 to S6 in the liver of hens. However, DG and FD manifested with different impacts on weights of egg and egg albumen, proteolytic activity of jejunal digesta, plasma TRAP activity, ileal total AA digestibility, and several intestinal genes and hepatic proteins. Supplemental protease in the DG and FD diets produced mixed effects on a number of measures. In conclusion, our findings revealed the feasibility of including greater levels of microalgal biomass as a source of feed protein for laying hens and a novel potential of the biomass in improving dietary protein digestion and body protein metabolism than previously perceived.

  9. Protein quality and oil digestibility of Lupinus mutabilis: metabolic studies in children.

    Science.gov (United States)

    Lopez de Romaãna, G; Graham, G G; Morales, E; Massa, E; MacLean, W C

    1983-04-01

    The nutritional quality of lupins (Lupinus mutabilis) for infants and children was evaluated in two sets of balance studies. In the first the digestibility and protein quality of diets based on lupin flour, with and without methionine supplementation, were compared with those of a control diet consisting of casein, sucrose and vegetable oil. Apparent nitrogen absorption from lupin flour (81.8 and 84.3% of intake) was slightly but significantly less than that during casein control periods (87.2 and 86.8% of intake, P less than 0.05 and less than 0.001). Apparent nitrogen retention from unsupplemented lupin (15.6 +/- 5.8% of intake) was significantly less than that from casein in the corresponding control periods (29.8 +/- 4.9%, P less than 0.001); a small but significant (P less than 0.05) increase in nitrogen retention was observed during the control period following the lupin diet when compared with that preceding it. Methionine supplementation of lupin produced a marked improvement in apparent nitrogen retention (to 22.2 +/- 6.9%, P less than 0.05). In the second set of studies the digestibility of lupin oil was compared with that of a blend of soybean and cottonseed oils (50:50). Excretion of fecal fat (9.8 +/- 3.0% of intake) and fecal energy (6.7 +/- 1.2% of intake) with the diet containing lupin oil were similar to those observed with the control diet. Both the protein quality and oil digestibility of Lupinus mutabilis are very similar to those from soybeans processed in a similar manner. For certain countries the lupin could be a valuable source of protein and edible oil for human consumption.

  10. Facile trypsin immobilization in polymeric membranes for rapid, efficient protein digestion.

    Science.gov (United States)

    Xu, Fei; Wang, Wei-Han; Tan, Yu-Jing; Bruening, Merlin L

    2010-12-15

    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 albumin provide 84% sequence coverage, which is higher than the 71% coverage obtained with in-solution digestion for 16 h or the sodium dodecyl sulfate (SDS), whereas in-solution digestion under similar conditions yields no peptide signals in mass spectra even after removal of SDS. These membrane reactors, which can be easily prepared in any laboratory, have a shelf life of several months and continuously digest protein for at least 33 h without significant loss of activity.

  11. 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.

  12. Effect of initial protein concentration and pH on in vitro gastric digestion of heated whey proteins.

    Science.gov (United States)

    Zhang, Sha; Vardhanabhuti, Bongkosh

    2014-02-15

    The in vitro digestion of heated whey protein aggregates having different structure and physicochemical properties was evaluated under simulated gastric conditions. Aggregates were formed by heating whey protein isolates (WPI) at 3-9% w/w initial protein concentration and pH 3.0-7.0. Results showed that high protein concentration led to formation of larger WPI aggregates with fewer remaining monomers. Aggregates formed at high protein concentrations showed slower degradation rate compared to those formed at low protein concentration. The effect of initial protein concentration on peptide release pattern was not apparent. Heating pH was a significant factor affecting digestion pattern. At pH above the isoelectric point, the majority of the proteins involved in the aggregation, and aggregates formed at pH 6.0 were more susceptible to pepsin digestion than at pH 7.0. At acidic conditions, only small amount of proteins was involved in the aggregation and heated aggregates were easily digested by pepsin, while the remaining unaggregated proteins were very resistant to gastric digestion. The potential physiological implication of these results on satiety was discussed.

  13. Improving Adult ESL Learners' Pronunciation Skills. ERIC Digest.

    Science.gov (United States)

    Florez, MaryAnn Cunningham

    This digest reviews the current status of pronunciation instruction in adult English-as-a-Second-Language (ESL) classes. The current focus on communicative approaches to ESL instruction and the concern for building teamwork and communication skills in an increasingly diverse workplace are renewing interest in the role that pronunciation plays in…

  14. 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 ω-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 ω-3 oils.

  15. 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.

  16. 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

  17. Compact structure and proteins of pasta retard in vitro digestive evolution of branched starch molecular structure.

    Science.gov (United States)

    Zou, Wei; Sissons, Mike; Warren, Frederick J; Gidley, Michael J; Gilbert, Robert G

    2016-11-01

    The roles that the compact structure and proteins in pasta play in retarding evolution of starch molecular structure during in vitro digestion are explored, using four types of cooked samples: whole pasta, pasta powder, semolina (with proteins) and extracted starch without proteins. These were subjected to in vitro digestion with porcine α-amylase, collecting samples at different times and characterizing the weight distribution of branched starch molecules using size-exclusion chromatography. Measurement of α-amylase activity showed that a protein (or proteins) from semolina or pasta powder interacted with α-amylase, causing reduced enzymatic activity and retarding digestion of branched starch molecules with hydrodynamic radius (Rh)pasta protects the starch and proteins in the interior of the whole pasta, reducing the enzymatic degradation of starch molecules, especially for molecules with Rh>100nm.

  18. Digestion, absorption and utilization of single-cell protein by the preruminant calf. The true digestibility of milk and bacterial protein and the apparent digestibility and utilization of their constituent amino acids.

    Science.gov (United States)

    Sedgman, C A; Roy, J H; Thomas, J; Stobo, I J; Ganderton, P

    1985-07-01

    Two experiments of Latin square design were made, each with four Friesian bull calves fitted with re-entrant duodenal and ileal cannulas at 4-10 d of age. The calves were used to study the effect of giving milk-substitutes containing 0, 300, 500 and 700 g bacterial protein (Pruteen)/kg total protein on apparent digestibility of nitrogen fractions and amino acids and true digestibility of 3H-labelled milk protein and 35S-labelled bacterial protein in the small intestine. A third experiment of Latin square design with four intact Friesian calves was used to measure apparent digestibility of nutrients throughout the alimentary tract and retention of N, calcium and phosphorus. At the duodenum, volume of outflow, its pH, and outflow of total-N(TN), protein-N (PN) and non-protein-N (NPN) decreased with time after feeding. At the ileum, volume of outflow and TN outflow were unaffected by time after feeding but PN outflow decreased; NPN outflow at the ileum increased to a maximum 6 h after feeding and then declined. Increased inclusion of Pruteen did not affect the volume of outflow at the duodenum or ileum, but duodenal PN outflow increased. At the ileum, pH values were lower and TN, PN and NPN outflows were higher with increasing concentration of Pruteen in the diet. Apparent digestibility in the small intestine tended to decrease with greater amounts of Pruteen, but was only significant for NPN. Apparent digestibility from mouth to ileum significantly decreased for TN and PN as Pruteen inclusion increased. Amino acid concentration in duodenal outflow, with the exception of that of arginine, reflected intake. The total amount of each amino acid in ileal outflow increased and the apparent digestibility of most amino acids decreased with greater amounts of Pruteen in the diet. Apparent digestibility of nucleic acid-N from Pruteen was very high. True digestibility in the small intestine and between mouth and ileum of 3H-labelled milk protein was high and did not differ

  19. Digestible fibre to starch ratio and protein level in diets for growing rabbits

    Directory of Open Access Journals (Sweden)

    Gerolamo Xiccato

    2010-01-01

    Full Text Available To evaluate the effect of digestible fibre (DF to starch ratio (0.8, 1.5, and 2.8 and protein level (15% and 16% on health status, digestive physiology, growth performance, and carcass traits, 246 rabbits weaned at 33 d were fed until slaughter (75 d six diets formulated according to a bi-factorial arrangement (3 DF to starch ratios by 2 protein levels. Growth performance and carcass quality at slaughter were not af- fected by treatments. Increasing DF to starch ratio did not modify dry matter digestibility (62.0% on average, while increased (P<0.001 DF digestibility (52.3 to 68.1%, stimulated caecal fermentation (total VFA: 56.0 vs 67.8 and 67.2 mmol/l; P=0.02 and changed VFA molar proportions. Increasing dietary protein increased digestibility of dry matter (P=0.02, crude protein (P<0.001 and digestible fibre (P<0.001 and increased cae- cal VFA production (P<0.01. The highest mortality (17.1% vs 1.5% average mortality of the other groups, P<0.001 was found in rabbits fed the diet with the lowest DF to starch ratio and the highest protein level.

  20. Co-digestion of press water and food waste in a biowaste digester for improvement of biogas production.

    Science.gov (United States)

    Nayono, Satoto E; Gallert, Claudia; Winter, Josef

    2010-09-01

    Co-digestion of press water from organic municipal wastes and of homogenized food residues with defibered kitchen wastes (food waste) as the main substrate was examined to improve biogas production. Although the biowaste digester was operated already at high organic loading (OLR) of 12.3 kg CODm(-3)d(-1) during the week, addition of co-substrates not only increased biogas production rates but also improved total biogas production. By feeding the two co-substrates up to 20 kg CODm(-3)d(-1) gas production followed the increasing OLR linearly. When the OLR was further increased with food waste, not more gas than for 20 kg CODm(-3)d(-1) OLR was obtained, indicating the maximum metabolic capabilities of the microbes. During weekends (no biowaste available) food waste could substitute for biowaste to maintain biogas production. Addition of press water or food waste to biowaste co-digestion resulted in more buffer capacity, allowing very high loadings without pH control.

  1. Proteolytic Digestion and TiO2 Phosphopeptide Enrichment Microreactor for Fast MS Identification of Proteins

    Science.gov (United States)

    Deng, Jingren; Lazar, Iulia M.

    2016-04-01

    The characterization of phosphorylation state(s) of a protein is best accomplished by using isolated or enriched phosphoprotein samples or their corresponding phosphopeptides. The process is typically time-consuming as, often, a combination of analytical approaches must be used. To facilitate throughput in the study of phosphoproteins, a microreactor that enables a novel strategy for performing fast proteolytic digestion and selective phosphopeptide enrichment was developed. The microreactor was fabricated using 100 μm i.d. fused-silica capillaries packed with 1-2 mm beds of C18 and/or TiO2 particles. Proteolytic digestion-only, phosphopeptide enrichment-only, and sequential proteolytic digestion/phosphopeptide enrichment microreactors were developed and tested with standard protein mixtures. The protein samples were adsorbed on the C18 particles, quickly digested with a proteolytic enzyme infused over the adsorbed proteins, and further eluted onto the TiO2 microreactor for enrichment in phosphopeptides. A number of parameters were optimized to speed up the digestion and enrichments processes, including microreactor dimensions, sample concentrations, digestion time, flow rates, buffer compositions, and pH. The effective time for the steps of proteolytic digestion and enrichment was less than 5 min. For simple samples, such as standard protein mixtures, this approach provided equivalent or better results than conventional bench-top methods, in terms of both enzymatic digestion and selectivity. Analysis times and reagent costs were reduced ~10- to 15-fold. Preliminary analysis of cell extracts and recombinant proteins indicated the feasibility of integration of these microreactors in more advanced workflows amenable for handling real-world biological samples.

  2. Experimental co-digestion of corn stalk and vermicompost to improve biogas production.

    Science.gov (United States)

    Chen, Guangyin; Zheng, Zheng; Yang, Shiguan; Fang, Caixia; Zou, Xingxing; Luo, Yan

    2010-10-01

    Anaerobic co-digestion of corn stalk and vermicompost (VC) as well as mono-digestion of corn stalk were investigated. Batch mono-digestion experiments were performed at 35+/-1 degrees C and initial total solid loading (TSL) ranged from 1.2% to 6.0%. Batch co-digestion experiments were performed at 35+/-1 degrees C and initial TSL of 6% with VC proportions ranged from 20% to 80% of total solid (TS). For mono-digestion of corn stalk, a maximum methane yield of 217.60+/-13.87 mL/g TS(added) was obtained at initial TSL of 4.8%, and acidification was found at initial TSL of 6.0% with the lowest pH value of 5.10 on day 4. Co-digestion improved the methane yields by 4.42-58.61% via enhancing volatile fatty acids (VFAs) concentration and pH value compared with mono-digestion of corn stalk. The maximum biogas yield of 410.30+/-11.01 mL/g TS(added) and methane yield of 259.35+/-13.85 mL/g TS(added) were obtained for 40% VC addition. Structure analysis by X-ray diffractometry (XRD) showed that the lowest crystallinity of 35.04 of digested corn stalk was obtained from co-digestion with 40% VC, which decreased 29.4% compared to 49.6 obtained from un-treated corn stalk. It is concluded that co-digestion with VC is beneficial for improving biodigestibility and methane yield from corn stalk.

  3. Stability and immunogenicity of hypoallergenic peanut protein-polyphenol complexes during in vitro pepsin digestion.

    Science.gov (United States)

    Plundrich, Nathalie J; White, Brittany L; Dean, Lisa L; Davis, Jack P; Foegeding, E Allen; Lila, Mary Ann

    2015-07-01

    Allergenic peanut proteins are relatively resistant to digestion, and if digested, metabolized peptides tend to remain large and immunoreactive, triggering allergic reactions in sensitive individuals. In this study, the stability of hypoallergenic peanut protein-polyphenol complexes was evaluated during simulated in vitro gastric digestion. When digested with pepsin, the basic subunit of the peanut allergen Ara h 3 was more rapidly hydrolyzed in peanut protein-cranberry or green tea polyphenol complexes compared to uncomplexed peanut flour. Ara h 2 was also hydrolyzed more quickly in the peanut protein-cranberry polyphenol complex than in uncomplexed peanut flour. Peptides from peanut protein-cranberry polyphenol complexes and peanut protein-green tea polyphenol complexes were substantially less immunoreactive (based on their capacity to bind to peanut-specific IgE from patient plasma) compared to peptides from uncomplexed peanut flour. These results suggest that peanut protein-polyphenol complexes may be less immunoreactive passing through the digestive tract in vivo, contributing to their attenuated allergenicity.

  4. On-line digestion system for protein characterization and proteome analysis.

    Science.gov (United States)

    López-Ferrer, Daniel; Petritis, Konstantinos; Lourette, Natacha M; Clowers, Brian; Hixson, Kim K; Heibeck, Tyler; Prior, David C; Pasa-Tolić, Ljiljana; Camp, David G; Belov, Mikhail E; Smith, Richard D

    2008-12-01

    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 online pressurized digestion system coupled to an ion mobility time-of-flight mass spectrometer, an ion trap mass spectrometer, or both. The system denatured, digested, and separated product peptides in a manner of minutes, making it amenable to on-line high-throughput applications. In addition to simplifying and expediting sample processing, the system was easy to implement and no cross-contamination was observed among samples. As a result, the online digestion system offers a powerful approach for high-throughput screening of proteins that could prove valuable in biochemical research (rapid screening of protein-based drugs).

  5. 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 α-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.

  6. Improvement of dewatering characteristics by co-digestion of rice straw with sewage sludge.

    Science.gov (United States)

    Gu, Tingting; Yamamoto-Ikemoto, Ryoko; Tsuchiya-Nakakihara, Eri; Watanabe, Haruki; Suetsugu, Yasutaka; Yanai, Atsushi

    2016-12-01

    A continuous mesophilic co-digestion of sewage sludge and softened rice straw was conducted and the dewatering characteristics of digested sludge were evaluated by a dewatering experiment using a belt press. The digestion was operated with solid retention time (SRT) of 25 days, and the feeding ratio of sludge to rice straw was 1:0.5 (total solids base). After 129 days of stable operation, the properties of digested sludge were analysed; then five kinds of cationic coagulants were tested to select the optimal coagulants for dewatering, and two coagulants were selected and used in the dewatering experiment because of lower doses and lower moisture of sludge cakes. Sludge property analysis showed that by the addition of rice straw, the fibrous materials in the digested sludge increased remarkably and the normalized capillary suction time (CST) decreased significantly, indicating that the dewatering properties was improved. The results of dewatering experiment showed that by the addition of rice straw, specific filtration rate of digested sludge increased by 81.2% and 174.6%, respectively; water content of dewatered sludge cakes decreased by 8.2% and 13.4%, respectively. The dewaterability of digested sludge was suggested to be improved due to rice straw addition.

  7. Evaluating the quality of protein from hemp seed (Cannabis sativa L.) products through the use of the protein digestibility-corrected amino acid score method.

    Science.gov (United States)

    House, James D; Neufeld, Jason; Leson, Gero

    2010-11-24

    The macronutrient composition and the quality of protein of hemp seed and products derived from hemp seed grown in Western Canada were determined. Thirty samples of hemp products (minimum 500 g), including whole hemp seed, hemp seed meal from cold-press expelling, dehulled, or shelled, hemp seed and hemp seed hulls, were obtained from commercial sources. Proximate analysis, including crude protein (% CP), crude fat (% fat) and fiber, as well as full amino acid profiles, were determined for all samples. Protein digestibility-corrected amino acid score (PDCAAS) measurements, using a rat bioassay for protein digestibility and the FAO/WHO amino acid requirement of children (2-5 years of age) as reference, were conducted on subsets of hemp products. Mean (±SD) percentage CP and fat were 24.0(2.1) and 30.4(2.7) for whole hemp seed, 40.7(8.8) and 10.2(2.1) for hemp seed meal, and 35.9(3.6) and 46.7(5.0) for dehulled hemp seed. The percentage protein digestibility and PDCAAS values were 84.1-86.2 and 49-53% for whole hemp seed, 90.8-97.5 and 46-51% for hemp seed meal, and 83.5-92.1 and 63-66% for dehulled hemp seed. Lysine was the first limiting amino acid in all products. Removal of the hull fraction improved protein digestibility and the resultant PDCAAS value. The current results provide reference data in support of protein claims for hemp seed products and provide evidence that hemp proteins have a PDCAAS equal to or greater than certain grains, nuts, and some pulses.

  8. Improved corn protein based articles

    Science.gov (United States)

    Developing higher value uses for zein (corn protein), a potential major co-product of the bio-ethanol industry, will improve the economics of this business. Historically, zein was predominantly used in the textile fiber industry. Unfortunately the techniques used at that time to modify the zein cann...

  9. 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...

  10. 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.

  11. Super-atmospheric pressure ionization mass spectrometry and its application to ultrafast online protein digestion analysis.

    Science.gov (United States)

    Chen, Lee Chuin; Ninomiya, Satoshi; Hiraoka, Kenzo

    2016-06-01

    Ion source pressure plays a significant role in the process of ionization and the subsequent ion transmission inside a mass spectrometer. Pressurizing the ion source to a gas pressure greater than atmospheric pressure is a relatively new approach that aims to further improve the performance of atmospheric pressure ionization sources. For example, under a super-atmospheric pressure environment, a stable electrospray can be sustained for liquid with high surface tension such as pure water, because of the suppression of electric discharge. Even for nano-electrospray ionization (nano-ESI), which is known to work with aqueous solution, its stability and sensitivity can also be enhanced, particularly in the negative mode when the ion source is pressurized. A brief review on the development of super-atmospheric pressure ion sources, including high-pressure electrospray, field desorption and superheated ESI, and the strategies to interface these ion sources to a mass spectrometer will be given. Using a recent ESI prototype with an operating temperature at 220 °C under 27 atm, we also demonstrate that it is possible to achieve an online Asp-specific protein digestion analysis in which the whole processes of digestion, ionization and MS acquisition could be completed on the order of a few seconds. This method is fast, and the reaction can even be monitored on a near-real-time basis. Copyright © 2016 John Wiley & Sons, Ltd.

  12. Microwave irradiation induced changes in protein molecular structures of barley grains: relationship to changes in protein chemical profile, protein subfractions, and digestion in dairy cows.

    Science.gov (United States)

    Yan, Xiaogang; Khan, Nazir A; Zhang, Fangyu; Yang, Ling; Yu, Peiqiang

    2014-07-16

    The objectives of this study were to evaluate microwave irradiation (MIR) induced changes in crude protein (CP) subfraction profiles, ruminal CP degradation characteristics and intestinal digestibility of rumen undegraded protein (RUP), and protein molecular structures in barley (Hordeum vulgare) grains. Samples from hulled (n = 1) and hulless cultivars (n = 2) of barley, harvested from four replicate plots in two consecutive years, were evaluated. The samples were either kept as raw or irradiated in a microwave for 3 min (MIR3) or 5 min (MIR5). Compared to raw grains, MIR5 decreased the contents of rapidly degradable CP subfraction (from 45.22 to 6.36% CP) and the ruminal degradation rate (from 8.16 to 3.53%/h) of potentially degradable subfraction. As a consequence, the effective ruminal degradability of CP decreased (from 55.70 to 34.08% CP) and RUP supply (from 43.31 to 65.92% CP) to the postruminal tract increased. The MIR decreased the spectral intensities of amide 1, amide II, α-helix, and β-sheet and increased their ratios. The changes in protein spectral intensities were strongly correlated with the changes in CP subfractions and digestive kinetics. These results show that MIR for a short period (5 min) with a lower energy input can improve the nutritive value and utilization of CP in barely grains.

  13. Fermentation and addition of enzymes to a diet based on high-moisture corn, rapeseed cake, and peas improve digestibility of nonstarch polysaccharides, crude protein, and phosphorus in pigs.

    Science.gov (United States)

    Jakobsen, G V; Jensen, B B; Bach Knudsen, K E; Canibe, N

    2015-05-01

    Fluctuating prices of cereals have led to an interest in alternative ingredients for feed. The aim of this study was to evaluate the effect of fermentation and the addition of nonstarch polysaccharide (NSP)-degrading enzymes on the ileal and total tract digestibility of nutrients of a diet based on locally grown crops. Four diets were fed including a nonfermented liquid standard grower diet (Control) and 3 experimental diets based on high-moisture corn, rapeseed cake, and peas fed as nonfermented liquid feed (nFLF), fermented liquid feed (FLF), or FLF supplemented with an enzyme mixture of β-glucanase + xylanase + pectinase (FLF+Enz). The FLF was prepared by mixing feed and water (1:2.5, wt/wt) and, once daily, replacing 50% of the mixture with an equal amount of fresh feed and water. The diets were fed to 8 ileal cannulated barrows in a double Latin square design. Ileal digesta and feces were collected after an adaption period of 10 d. Results showed microbiologically good-quality fermented diets. The levels of Enterobacteriaceae were 5.1 to 5.4 log cfu/g in FLF and FLF+Enz vs. 6.3 log cfu/g in nFLF in the ileum and 5.1 to 5.2 log cfu/g in FLF and FLF+Enz vs. 6.3 log cfu/g in nFLF in the feces. Apparent total tract digestibility (ATTD) of CP was increased by fermentation (73.2% in FLF vs. 69.0% in nFLF; P = 0.033), and digestibility of P showed a tendency (P = 0.073) toward an increase. Addition of the enzyme mixture resulted in a pronounced reduction of dietary NSP compared with FLF (12.8% total NSP in FLF+Enz vs. 15.9% total NSP in FLF; Pfermentation and enzyme addition brought the digestibility to the level of the Control. In conclusion, fermentation increased the ATTD of CP and the AID of P, with the same tendency (P ≤ 0.07) for the ATTD. Addition of NSP-degrading enzymes resulted in a pronounced reduction in the concentration of NSP in the feed along with increased AID of NSP. Hence, the experimental diet seems to be a possible alternative to a traditional

  14. Effect of high hydrostatic pressure processing on in vitro digestion of milk proteins and fats

    Science.gov (United States)

    The use of high hydrostatic pressure processing (HPP) is increasing in popularity in the food industry. Its ability to modify milk proteins and fats suggests that it may be useful in creating foods that suppress appetite; however, its effect on the digestibility of proteins and fats is unclear. The...

  15. Digestibility of glandless cottonseed protein in diets for pacific white shrimp, litopenaeus vannamei

    Science.gov (United States)

    Accurate digestibility coefficients for protein, energy, and lipid are needed by feed formulators to optimize diets to meet nutritional requirements and to substitute ingredients cost-effectively. Of particular interest is protein, which accounts for the majority of shrimp feed content and expense. ...

  16. Pepsin diffusivity in whey protein gels and its effect on gastric digestion

    NARCIS (Netherlands)

    Luo, Q.; Borst, J.W.; Westphal, A.H.; Boom, R.M.; Janssen, A.E.M.

    2017-01-01

    Protein is essential to human health, but its digestion kinetics in varied structures are not yet well understood. We previously found different kinetics of protein hydrolysis in solution and in gels, and we hypothesized that the difference stemmed from the steric hindrance of gel structure to the d

  17. Accelerated protein digestion and amino acid absorption after Roux-en-Y gastric bypass

    DEFF Research Database (Denmark)

    Bojsen-Møller, Kirstine N; Jacobsen, Siv H; Dirksen, Carsten;

    2015-01-01

    . CONCLUSIONS: RYGB accelerates caseinate digestion and amino acid absorption, resulting in faster and higher but more transient postprandial elevation of plasma amino acids. Changes are likely mediated by accelerated intestinal nutrient entry and clearly demonstrate that protein digestion is not impaired after......BACKGROUND: Roux-en-Y gastric bypass (RYGB) involves exclusion of major parts of the stomach and changes in admixture of gastro-pancreatic enzymes, which could have a major impact on protein digestion and amino acid absorption. OBJECTIVE: We investigated the effect of RYGB on amino acid appearance...... in the systemic circulation from orally ingested protein and from endogenous release. DESIGN: Nine obese glucose-tolerant subjects, with a mean body mass index (in kg/m(2)) of 39.2 (95% CI: 35.2, 43.3) and mean glycated hemoglobin of 5.3% (95% CI: 4.9%, 5.6%), were studied before and 3 mo after RYGB. Leucine...

  18. High pressure processing of meat: effects on ultrastructure and protein digestibility.

    Science.gov (United States)

    Kaur, Lovedeep; Astruc, Thierry; Vénien, Annie; Loison, Olivier; Cui, Jian; Irastorza, Marion; Boland, Mike

    2016-05-18

    The effects of high pressure processing (HPP, at 175 and 600 MPa) on the ultrastructure and in vitro protein digestion of bovine longissimus dorsi muscle meat were studied. HPP caused a significant change in the visual appearance and texture of the meat subjected to HPP at 600 MPa so that it appeared similar to cooked meat, unlike the meat subjected to HPP at 175 MPa that showed no significant visible change in the colour and texture compared to the raw meat. The muscles were subjected to digestion under simulated gastric conditions for 1 h and then under simulated small-intestinal conditions for a further 2 h. The digests were analysed using gel electrophoresis (SDS-PAGE) and ninhydrin assay for amino N. The effect of the acid conditions of the stomach alone was also investigated. Reduced SDS-PAGE results showed that pepsin-digested (60 min) HPP meats showed fewer proteins or peptides of high molecular weight than the pepsin-digested untreated meat, suggesting more breakdown of the parent proteins in HPP-treated meats. This effect was more pronounced in the muscles treated at 600 MPa. These results are in accordance with microscopy results, which showed greater changes in the myofibrillar structure after simulated gastric digestion of the sample processed at 600 MPa than at 175 MPa. Transmission electron microscopy also showed the presence of protein aggregates in the former sample, resulting probably from protein denaturation of sarcoplasmic proteins, in the subcellular space and between myofibrils; along with cell contraction (similar to that caused by heating) in the former.

  19. Effect of Digestible Protein and Sulfur Amino Acids in Starter Diet on Performance and Small Intestinal (Jejunum Morphology of Broilers

    Directory of Open Access Journals (Sweden)

    Avisa Akhavan khaleghi

    2016-04-01

    amount, higher than the recommended levels by Ross in the starter period, had no significant increase in the WG, FI and FCR. Increasing DSAA in the starter diet, although did not improve the FCR, but significantly increased daily weight gain and feed intake at the end of the period. With a decrease of 10 percent in digestible protein of the starter diet and its counter-action with digestible sulfur amino acids had no effect on WG, FI and FCR in any experimental periods. By taking the use of the diets of 20% Crude Protein (CP and 0.641% and 0.926% of Total Sulfur Amino Acids (TSAA or the 22% of CP and 0.705% and 0.926% of TSAA, no weight gain difference was observed from 1 to 7 days of age. Also, a 2% reduction of CP in diet of the starter period was not effective on the WG. The results of the present experiment showed that in the starter diets containing 3025 kCal/kg of metabolism energy, the levels of the digestible protein could be increased 20 gr per every kg of diet, without observing any negative effect on the performance of the broilers and in this situation extra amino acids are added to the diet. Results of the effects of different levels of the digestible protein and digestible sulfur amino acids on the carcass parts and the gastro intestinal tract are presented in the table 4. Increasing the DSAA, had no significant effect on carcass parts and relative weight of the gastro intestinal tract in the day 42, but the abdominal fat decreased (P

  20. Effect of Different Protein Levels On Nutrient Digestion Metabolism and Serum Biochemical Parameters in Calves

    Institute of Scientific and Technical Information of China (English)

    LI Hui; DIAO Qi-yu; ZHANG Nai-feng; TU Yan; WANG Ji-feng

    2008-01-01

    The current study has been performed to examine the effects of different dietary protein levels on the growth,nutrient digestion and absorption,as well as biochemical parameters in calves.Nine healthy newborn calves were selected,randomly divided into 3 groups and fed 3 milk replacers with different protein levels(18,22,and 26%),respectively.Five period-digestion-metabolism trials were taken between 12-20,22-30,32-40,42-50,and 52-60 days after birth.All 3 groups showed a similar growth curve during 11 to 61 experimental days,however,the growth rate of the 22%crude protein(CP) group was 8.89%higher than that of the 18%CP group and 19.48%higher than that of the 26%CP group.respectively. The apparent digestibility of dry material(DM)declined gradually with age,whereas,the apparent digestibility of N,extract ether(EE)rose slightly.Compared to the 22%CP and 26%CP group,calves fed with 18%CP apparently had a lower digestibility than DM,EE,and nitrogen(N).The average apparent digestibilities of N were 69.39,75.36,and 74.55%, respectively.Both the apparent digestibility and retention of calcium(Ca)and phosphorus(P)were steady throughout the experiment,but the average apparent digestibility of P in the 26%CP group was only 63.83%,which was markedly lower than that of the 18%CP group(70.40%)and 22%CP group(69.73%).In addition,the sernm concentrations of total protein(TP),albumin(ALB),and globulin(GLOB)of the 22%CP group were higher than those in the 18%CP and 26%CP groups.The urea N(BUN)content in the 18%CP group,on the other hand,was significantly lower than that of the other two groups.The highest glucose(GLU)concentration was found in the 22%CP group(5.38 mmol L-1),at the end of the trials.The protein levels in the milk replacer affected the digestion metabolism of nutrition and the serum biochemical parameters of calves at different physiological phases.Calves fed with 22%CP milk replacer had a better growth performance than the other groups.

  1. Pea proteins for piglets: effects on digestive processes.

    NARCIS (Netherlands)

    le Guen, M.P.

    1993-01-01

    White-flowered pea ( Pisum sativum ) contains 20 to 25% crude protein (Nx6.25). The pea proteins consist in globulins (60%) and albumins (25%). Because the pea albumin proteins have biological activities due to their metabolic roles in the seed, some of them are called antinutritional factors (ANFs)

  2. 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.

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

    Directory of Open Access Journals (Sweden)

    Joseph G. Usack

    2014-01-01

    Full Text Available 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.

  4. Improvement of anaerobic digestion control from urban sludges; Mejora del control de la digestion anaerobia de fangos urbanos

    Energy Technology Data Exchange (ETDEWEB)

    Minguez Gabina, R.

    2002-07-01

    Due to its many advantages, the anaerobic digestion is a widespread choice in our water treatment plants. Furthermore, the companies demand an optimization of the control of the process. Until today, the practice of the operation has been made modifying the conditions of the digester by controlling the outlet parameters of the digestion, so that a delay in the application of the corrections is enough to decrease the performance. The investigation concludes that the most secure way to drive an anaerobic digester is to mange a strict control of the inlet parameters into digestion process. (Author)

  5. Evaluation of the National Research Council (2001) dairy model and derivation of new prediction equations. 1. Digestibility of fiber, fat, protein, and nonfiber carbohydrate.

    Science.gov (United States)

    White, R R; Roman-Garcia, Y; Firkins, J L; VandeHaar, M J; Armentano, L E; Weiss, W P; McGill, T; Garnett, R; Hanigan, M D

    2017-03-02

    Evaluation of ration balancing systems such as the National Research Council (NRC) Nutrient Requirements series is important for improving predictions of animal nutrient requirements and advancing feeding strategies. This work used a literature data set (n = 550) to evaluate predictions of total-tract digested neutral detergent fiber (NDF), fatty acid (FA), crude protein (CP), and nonfiber carbohydrate (NFC) estimated by the NRC (2001) dairy model. Mean biases suggested that the NRC (2001) lactating cow model overestimated true FA and CP digestibility by 26 and 7%, respectively, and under-predicted NDF digestibility by 16%. All NRC (2001) estimates had notable mean and slope biases and large root mean squared prediction error (RMSPE), and concordance (CCC) ranged from poor to good. Predicting NDF digestibility with independent equations for legumes, corn silage, other forages, and nonforage feeds improved CCC (0.85 vs. 0.76) compared with the re-derived NRC (2001) equation form (NRC equation with parameter estimates re-derived against this data set). Separate FA digestion coefficients were derived for different fat supplements (animal fats, oils, and other fat types) and for the basal diet. This equation returned improved (from 0.76 to 0.94) CCC compared with the re-derived NRC (2001) equation form. Unique CP digestibility equations were derived for forages, animal protein feeds, plant protein feeds, and other feeds, which improved CCC compared with the re-derived NRC (2001) equation form (0.74 to 0.85). New NFC digestibility coefficients were derived for grain-specific starch digestibilities, with residual organic matter assumed to be 98% digestible. A Monte Carlo cross-validation was performed to evaluate repeatability of model fit. In this procedure, data were randomly subsetted 500 times into derivation (60%) and evaluation (40%) data sets, and equations were derived using the derivation data and then evaluated against the independent evaluation data. Models

  6. Effect of germination periods and hydrothermal treatments on in vitro protein and starch digestibility of germinated legumes.

    Science.gov (United States)

    Uppal, Veny; Bains, Kiran

    2012-04-01

    Germination of legumes followed by hydrothermal treatments is an effective mean of improving nutritive value of legumes. The protein content of mungbean, chickpea and cowpea increased by 9-11, 11-16 and 8-11% after germination. A significant (p ≤ 0.05) decrease in protein content was observed on pressure cooking and microwaving in all three legumes. The carbohydrates decreased by 1 to 3% during soaking and 2 to 6% during germination. A significant (p ≤ 0.05) improvement in in vitro protein digestibility (IVPD) was observed after soaking as well as after three germination periods. Germination resulted in an increase in IVPD from 15 to 25% in mungbean, 6 to 17% in chickpea and 6 to 17% in cowpea. A significant (p ≤ 0.05) increase in IVPD was observed when raw sprouts of three legumes were subjected to pressure cooking and microwaving. In vitro starch digestibility (IVSD) increased significantly (p ≤ 0.05) after germination, the percent increase being 8 to 12% in mungbean, 9 to 11% in chickpea and 10 to 13% in cowpea. The duration of germination had significant (p ≤ 0.05) effect on IVSD. A significant (p ≤ 0.05) improvement in IVSD was observed when legume sprouts were subjected to pressure cooking and microwave cooking.

  7. In vitro digestibility, protein composition and techno-functional properties of Saskatchewan grown yellow field peas (Pisum sativum L.) as affected by processing.

    Science.gov (United States)

    Ma, Zhen; Boye, Joyce I; Hu, Xinzhong

    2017-02-01

    Saskatchewan grown yellow field pea was subjected to different processing conditions including dehulling, micronization, roasting, conventional/microwave cooking, germination, and combined germination and conventional cooking/roasting. Their nutritional and antinutritional compositions, functional properties, microstructure, thermal properties, in vitro protein and starch digestibility, and protein composition were studied. Processed field peas including conventional cooked yellow peas (CCYP), microwave cooked yellow peas (MCYP), germinated-conventional cooked yellow peas (GCCYP), and germinated-roasted yellow peas (GRYP) exhibited the significantly higher in vitro protein digestibility (IVPD), which was in accordance with their significantly lower trypsin inhibitor activity and tannin content. The SDS-PAGE and size exclusion HPLC profiles of untreated pea proteins and their hydrolysates also confirmed the IVPD result that these four treatments facilitated the hydrolysis of pea proteins to a greater extent. The CCYP, MCYP, GCCYP, and GRYP also exhibited significantly higher starch digestibility which was supported by their lower onset (To), peak (Tp), and conclusion (Tc) temperatures obtained from DSC thermogram, their lower pasting properties and starch damage results, as well as their distinguished amorphous flakes' configuration observed on the scanning electron microscopic image. LC/ESI-MS/MS analysis following in-gel digests of SDS-PAGE separated proteins allowed detailed compositional characterization of pea proteins. The present study would provide fundamental information to help to better understand the functionality of field peas as ingredients, and particularly in regards to agri-food industry to improve the process efficiency of field peas with enhanced nutritional and techno-functional qualities.

  8. 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.

  9. Thesis Abstract Bean (Phaseolus vulgaris L.) lines: chemical composition and protein digestibility.

    Science.gov (United States)

    Mesquita, F R; Silva, M I A; Corrêa, A D

    2016-05-09

    The bean represents the main source of proteins for the low income populations, although the digestibility of those proteins is relatively low. Consequently, the programs of plant genetic breeding have been working on the search for new lines with higher protein levels. Thus, with the purpose of supplying information to the researchers, in this study, 21 bean (Phaseolus vulgaris L.) lines were analyzed for the centesimal and mineral composition, protein digestibility, phenolic compounds, and trypsin inhibitor. The entirely randomized experimental design was used with 21 treatments (lines) and three repetitions. All values were within the following ranges: 22.34 to 36.28 g crude protein/100 g dry matter (DM); 7.56 to 20.91 g neutral detergent fiber/100 g DM; 0.53 to 2.55 g fat/100 g DM and 2.97 to 4.87 g ashes/100 g DM. The levels of phosphorus, potassium, calcium, magnesium, and sulfur, in g/100 g DM, varied from 0.45 to 0.72; 1.51 to 2.48; 0.03 to 0.28; 0.18 to 0.34 and 0.28 to 0.45, respectively. Regarding copper, manganese, zinc and iron, the levels, in mg/kg DM, varied from 11.37 to 17.73; 14.93 to 28.90; 36.67 to 69.90 and 71.37 to 126.90, respectively. The in vitro protein digestibility varied from 18.03 to 48.32%. The levels of phenolic compounds varied from 0.28 to 1.08 mg acid tanic/100 g DM and the one of trypsin inhibitor from 59.93 to 151.07 trypsin inhibited units/mg DM. Among the lines with higher protein contents, "ESAL 569" (beige with brown stripe) presented the largest protein digestibility and considerable levels of minerals. "P-180" (beige with brown stripe) was one of the lines with higher crude protein contents and digestibilities, and also presented high levels for most of the minerals. No relation between protein digestibility and the contents of phenolic compounds or trypsin inhibitor was observed.

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

    DEFF Research Database (Denmark)

    Bundgaard, Anja Marie; Dalgaard, Randi; Thrane, Mikkel;

    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...

  11. Improving the Digestive Tract Robustness of Probiotic Lactobacilli

    NARCIS (Netherlands)

    Bokhorst-van de Veen, van H.; Bron, P.A.; Kleerebezem, M.

    2015-01-01

    This chapter describes stress responses of probiotic lactobacilli, in relation to gastrointestinal (GI-)tract robustness. An overview is given of some newly developed tools to understand and improve stress responses of the model probiotic L. plantarum WCFS1 in relation to its GI persistence. These i

  12. Zeolite nanoparticle modified microchip reactor for efficient protein digestion.

    Science.gov (United States)

    Huang, Yi; Shan, Wei; Liu, Baohong; Liu, Yun; Zhang, Yahong; Zhao, Yue; Lu, Haojie; Tang, Yi; Yang, Pengyuan

    2006-04-01

    An enzymatic microreactor has been fabricated based on the poly(methyl methacrylate) (PMMA) microchchip surface-modified with zeolite nanoparticles. By introducing the silanol functional groups, the surface of PMMA microchannel has been successfully modified with silicalite-1 nanoparticle for the first time due to its large external surface area and high dispersibility in solutions. Trypsin can be stably immobilized in the microchannel to form a bioreactor using silica sol-gel matrix. The immobilization of enzyme can be realized with a stable gel network through a silicon-oxygen-silicon bridge via tethering to those silanol groups, which has been investigated by scanning electron microscopy and microchip capillary electrophoresis with laser-induced fluorescence detection. The maximum proteolytic rate constant of the immobilized trypsin is measured to be about 6.6 mM s(-1). Using matrix assisted laser desorption and ionization time-of-flight mass spectrometry, the proposed microreactor provides an efficient digestion of cytochrome c and bovine serum albumin at a fast flow rate of 4.0 microL min(-1), which affords a very short reaction time of less than 5 s.

  13. Improved ADM1 model for anaerobic digestion process considering physico-chemical reactions.

    Science.gov (United States)

    Zhang, Yang; Piccard, Sarah; Zhou, Wen

    2015-11-01

    The "Anaerobic Digestion Model No. 1" (ADM1) was modified in the study by improving the bio-chemical framework and integrating a more detailed physico-chemical framework. Inorganic carbon and nitrogen balance terms were introduced to resolve the discrepancies in the original bio-chemical framework between the carbon and nitrogen contents in the degraders and substrates. More inorganic components and solids precipitation processes were included in the physico-chemical framework of ADM1. The modified ADM1 was validated with the experimental data and used to investigate the effects of calcium ions, magnesium ions, inorganic phosphorus and inorganic nitrogen on anaerobic digestion in batch reactor. It was found that the entire anaerobic digestion process might exist an optimal initial concentration of inorganic nitrogen for methane gas production in the presence of calcium ions, magnesium ions and inorganic phosphorus.

  14. Amino acid composition, score and in vitro protein digestibility of foods commonly consumed in Norhwest Mexico

    Directory of Open Access Journals (Sweden)

    Graciela Caire-Juvera

    2013-04-01

    Full Text Available A better knowledge of the amino acid composition of foods commonly consumed in different regions is essential to calculate their scores and, therefore, to predict their protein quality. This paper presents the amino acid composition, amino acid score and in vitro protein digestibility of fifteen foods that are commonly consumed in Northwest Mexico. The foods were prepared by the traditional methods and were analyzed by reverse-phase HPLC. The chemical score for each food was determined using the recommendations for children of 1-2 years of age, and the digestibility was evaluated using a multienzyme technique. Lysine was the limiting amino acid in cereal-based products (scores 15 to 54, and methionine and cysteine were limiting in legume products (scores 41 to 47, boiled beef (score = 75 and hamburger (score = 82. The method of preparation had an effect on the content of certain amino acids, some of them increased and others decreased their content. Meat products and regional cheese provided a high amino acid score (scores 67 to 91 and digestibility (80.7 to 87.8%. Bologna, a processed meat product, had a lower digestibility (75.4%. Data on the amino acid composition of foods commonly consumed in Mexico can be used to provide valuable information on food analysis and protein quality, and to contribute to nutrition and health research and health programs.

  15. Measurement of the postruminal digestibility of crude protein by the bag technique in cows.

    Science.gov (United States)

    Voigt, J; Piatkowski, B; Engelmann, H; Rudolph, E

    1985-08-01

    A new method has been developed which permits the crude protein digestibility of feedstuffs in the intestine of cattle to be measured with little effort in terms of samples and experimental work. It consists of welding 0.4 ... 0.8 g of the feedstuff (particle size: 125 ... 1000 micron) into polyamide fabric bags (25 X 40 mm) which are inserted via cannulae into the digestive tracts of fistulated cows from the abomasum/duodenum to the ileum or from the abomasum/duodenum to the faeces. The mean retention time of the bags in the animal was 8.5 +/- 2.7 h from the abomasum to the end of the ileum and (13.3 +/- 1.9 h from the abomasum to the faeces. Up to 15 bags per day and cow may be used. The random error of the method is 1.3% (absolute) when the measurements are performed on two animals using two bags each. Intestinal digestibilities of over 90% were measured for concentrate proteins (except linseed meal) and of 72 ... 95% for forage proteins. Post-ruminal digestion was virtually finished at the end of the small intestine.

  16. Application of protein misfolding cyclic amplification to detection of prions in anaerobic digestate.

    Science.gov (United States)

    Gilroyed, Brandon H; Braithwaite, Shannon L; Price, Luke M; Reuter, Tim; Czub, Stefanie; Graham, Catherine; Balachandran, Arumuga; McAllister, Tim A; Belosevic, Miodrag; Neumann, Norman F

    2015-11-01

    The exceptional physio-chemical resistance of prions to established decontamination procedures poses a challenge to assessing the suitability of applied inactivation methods. Prion detection is limited by the sensitivity level of Western blotting or by the cost and time factors of bioassays. In addition, prion detection assays can be limited by either the unique or complex nature of matrices associated with environmental samples. To investigate anaerobic digestion (AD) as a practical and economical approach for potential conversion of specified risk materials (SRM) into value added products (i.e., renewable energy), challenges associated with detection of prions in a complex matrix need to be overcome to determine potential inactivation. Protein misfolding cyclic amplification (PMCA) assay, with subsequent Western blot visualization, was used to detect prions within the AD matrix. Anaerobic digestate initially inhibited the PMCA reaction and/or Western blot detection. However, at concentrations of ≤1% of anaerobic digestate, 263K scrapie prions could be amplified and semi-quantitatively detected. Infectious 263K prions were also proven to be bioavailable in the presence of high concentrations of digestate (10-90%). Development of the PMCA application to digestate provides extremely valuable insight into the potential degradation and/or fate of prions in complex biological matrices without requiring expensive and time-consuming bioassays.

  17. Improved method for identification of low abundance proteins using 2D-gel electrophoresis, MALDI-TOF and TOF/TOF

    Science.gov (United States)

    Introduction: Differential protein expression studies have been routinely performed in our laboratory to determine the health effects of environmentally-important chemicals. In this abstract, improvements in the in-gel protein digestion, MALDI plate spotting and data acquisition...

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

    DEFF Research Database (Denmark)

    Leon, Ileana R; Schwämmle, Veit; Jensen, Ole N;

    2013-01-01

    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...... 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...

  19. Surface adsorption alters the susceptibility of whey proteins to pepsin-digestion.

    Science.gov (United States)

    Nik, Amir Malaki; Wright, Amanda J; Corredig, Milena

    2010-04-15

    An in vitro digestion model mimicking the gastric phase of the human gastrointestinal tract coupled with SDS-PAGE and MALDI-TOF mass spectroscopy was employed to study the hydrolysis profiles of whey proteins in solution and adsorbed at the oil-water interface. The objective of this work was to understand the differences in hydrolysis behaviour of whey protein isolates once adsorbed at the interface, and comparisons were carried out with pure beta-lactoglobulin and alpha-lactalbumin fractions. In solution, while beta-lactoglobulin appeared to be resistant to enzymatic treatment, alpha-lactalbumin was fully degraded. Adsorption of both proteins at the oil-water interface affected their conformational structure and susceptibility to peptic hydrolysis. Adsorbed beta-lactoglobulin was hydrolyzed into small polypeptides and in contrast, the resistance of alpha-lactalbumin to pepsin increased upon adsorption at the interface. In addition, changes in the particle size distribution of the droplets during pepsin hydrolysis mainly depended on the original protein concentration. The results suggested that exchanges occur at the interface between adsorbed and non-adsorbed protein, that is to say that either some protein desorb from the interface and does not fully recover its structure in solution, or that hydrolysis of the protein at the interface induces further adsorption and hydrolysis of the protein in solution. These mechanisms have important implications in the digestibility of the proteins.

  20. A cyclic-olefin-copolymer microfluidic immobilized-enzyme reactor for rapid digestion of proteins from dried blood spots.

    Science.gov (United States)

    Wouters, Bert; Dapic, Irena; Valkenburg, Thalassa S E; Wouters, Sam; Niezen, Leon; Eeltink, Sebastiaan; Corthals, Garry L; Schoenmakers, Peter J

    2017-03-31

    A critical step in the bottom-up characterization of proteomes is the conversion of proteins to peptides, by means of endoprotease digestion. Nowadays this method typically uses overnight digestion and as such represents a considerable bottleneck for high-throughput analysis. This report describes protein digestion using an immobilized-enzyme reactor (IMER), which enables accelerated digestion times that are completed within seconds to minutes. For rapid digestion to occur, a cyclic-olefin-copolymer microfluidic reactor was constructed containing trypsin immobilized on a polymer monolithic material through a 2-vinyl-4,4-dimethylazlactone linker. The IMER was applied for the rapid offline digestion of both singular protein standards and a complex protein mixture prior to liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS/MS) analysis. The effects of protein concentration and residence time in the IMER were assessed for protein standards of varying molecular weight between 11 and 240kDa. Compared to traditional in-solution digestion, IMER-facilitated protein digestion at room temperature for 5min yielded similar results in terms of sequence coverage and number of identified peptides. Good repeatability was demonstrated with a relative standard deviation of 6% for protein-sequence coverage. The potential of the IMER was also demonstrated for a complex protein mixture in the analysis of dried blood spots. Compared to a traditional workflow a similar number of proteins could be identified, while reducing the total analysis time from 22.5h to 4h and importantly omitting the sample-pre-treatment steps (denaturation, reduction, and alkylation). The identified proteins from two workflows showed similar distributions in terms of molecular weight and hydrophobic character.

  1. Prediction of crude protein digestibility of animal by-product meals for dogs by the protein solubility in pepsin method.

    Science.gov (United States)

    Kawauchi, Iris M; Sakomura, Nilva K; Pontieri, Cristiana F F; Rebelato, Aline; Putarov, Thaila C; Malheiros, Euclides B; Gomes, Márcia de O S; Castrillo, Carlos; Carciofi, Aulus C

    2014-01-01

    Animal by-product meals have large variability in crude protein (CP) content and digestibility. In vivo digestibility procedures are precise but laborious, and in vitro methods could be an alternative to evaluate and classify these ingredients. The present study reports prediction equations to estimate the CP digestibility of meat and bone meal (MBM) and poultry by-product meal (PM) using the protein solubility in pepsin method (PSP). Total tract CP digestibility of eight MBM and eight PM samples was determined in dogs by the substitution method. A basal diet was formulated for dog maintenance, and sixteen diets were produced by mixing 70 % of the basal diet and 30 % of each tested meal. Six dogs per diet were used to determine ingredient digestibility. In addition, PSP of the MBM and PM samples was determined using three pepsin concentrations: 0·02, 0·002 and 0·0002 %. The CP content of MBM and PM ranged from 39 to 46 % and 57 to 69 %, respectively, and their mean CP digestibility by dogs was 76 (2·4) and 85 (2·6) %, respectively. The pepsin concentration with higher Pearson correlation coefficients with the in vivo results were 0·0002 % for MBM (r 0·380; P = 0·008) and 0·02 % for PM (r 0·482; P = 0·005). The relationship between the in vivo and in vitro results was better explained by the following equations: CP digestibility of MBM = 61·7 + 0·2644 × PSP at 0·0002 % (P = 0·008; R (2) 0·126); and CP digestibility of PM = 54·1 + 0·3833 × PSP at 0·02 % (P = 0·005; R (2) 0·216). Although significant, the coefficients of determination were low, indicating that the models were weak and need to be used with caution.

  2. Improved biogas production from food waste by co-digestion with de-oiled grease trap waste.

    Science.gov (United States)

    Wu, Li-Jie; Kobayashi, Takuro; Kuramochi, Hidetoshi; Li, Yu-You; Xu, Kai-Qin

    2016-02-01

    The objective of this study was to assess the feasibility of co-digesting food waste (FW) and de-oiled grease trap waste (GTW) to improve the biogas production. A lab-scale mesophilic digester (MD), a temperature-phased anaerobic digester (TPAD) and a TPAD with recycling (TPAD-R) were synchronously operated under mono-digestion (FW) and co-digestion (FW+de-oiled GTW). Co-digestion increased the biogas yield by 19% in the MD and TPAD-R, with a biogas yield of 0.60L/g VS added. Specific methanogenic activity in the TPAD-R was much higher than that in the MD. In addition to methane, hydrogen at a yield of approximately 1mol/mol hexose was produced in the TPAD-R. Alkalinity was consumed more in the co-digestion than in mono-digestion. Co-digestion resulted in more lipid accumulation in each digester. The MD favored the degradation of lipid and conversion of long-chain fatty acids more than the TPAD and TPAD-R.

  3. Analysis of Protein Amino Acids in Tobacco Using Microwave Digestion of Plant Material

    Directory of Open Access Journals (Sweden)

    Moldoveanu SC

    2014-12-01

    Full Text Available This paper describes a technique using microwave digestion and gas chromatography-mass spectrometry (GC-MS, which makes possible the analysis of protein amino acids in tobacco. The technique involves first the measurement of free amino acids, a hydrolysis using microwave digestion, and a measurement of total resulting amino acids. The content of protein amino acids is determined from the difference of total and free amino acids. The digestion is performed with aqueous 6 N HCl (with 1% phenol for two hours in a microwave at 120°C in sealed vials. The GC-MS analysis is performed after the amino acids are derivatized with N-methyl-N-(t-butyldimethylsilyltrifluoroacetamide (MTBSTFA. The technique provides reliable results with less than 10% relative standard deviation (RSD for most amino acids. Only the determination of very low level amino acids is affected by larger errors. The method provides results for free amino acids that are in very good agreement with those obtained by high performance liquid chromatography (HPLC, and also results for protein levels in tobacco in agreement with data previously reported in the literature. Results are given for several single grade tobaccos and for tobacco blends from four Kentucky reference cigarettes.

  4. Modified Protein Improves Vitiligo Symptoms in Mice

    Science.gov (United States)

    ... 2013 (historical) Modified Protein Improves Vitiligo Symptoms in Mice Altering a key protein involved in the development ... pigmentation loss associated with the skin disorder in mice, according to recent research funded by the NIH’s ...

  5. 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.

  6. The effect of replacing lactose by starch on protein and fat digestion in milk-fed veal calves.

    Science.gov (United States)

    Pluschke, A M; Gilbert, M S; Williams, B A; van den Borne, J J G C; Schols, H A; Gerrits, W J J

    2016-08-01

    Replacing dairy components from milk replacer (MR) with vegetable products has been previously associated with decreased protein and fat digestibility in milk-fed calves resulting in lower live weight gain. In this experiment, the major carbohydrate source in MR, lactose, was partly replaced with gelatinized corn starch (GCS) to determine the effect on protein and fat digestibility in milk-fed calves. In total, 16 male Holstein-Friesian calves received either MR with lactose as the carbohydrate source (control) or 18% GCS at the expense of lactose. In the adaptation period, calves were exposed to an increasing dose of GCS for 14 weeks. The indigestible marker cobalt ethylenediaminetetraacetic acid was incorporated into the MR for calculating apparent nutrient digestibility, whereas a pulse dose of chromium (Cr) chloride was fed with the last MR meal 4 h before slaughter as an indicator of passage rates. The calves were anesthetized and exsanguinated at 30 weeks of age. The small intestine was divided in three; small intestine 1 and 2 (SI1 and SI2, respectively) and the terminal ileum (last ~100 cm of small intestine) and samples of digesta were collected. Small intestinal digesta was analysed for α-amylase, lipase and trypsin activity. Digestibility of protein was determined for SI1, SI2, ileum and total tract, whereas digestibility of fat was determined for SI1, SI2 and total tract. Apparent protein digestibility in the small intestine did not differ between treatments but was higher in control calves at total tract level. Apparent crude fat digestibility tended to be increased in SI1 and SI2 for GCS calves, but no difference was found at total tract level. Activity of α-amylase in SI2 and lipase in both SI1 and SI2 was higher in GCS calves. Activity of trypsin tended to be higher in control calves and was higher in SI1 compared with SI2. A lower recovery of Cr in SI2 and a higher recovery of Cr in the large intestine suggest an increased rate of passage for GCS

  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 (p0.05), except propionic acid (C3) was increased (pstraw improved feed intake, digestibility, rumen fermentation and efficiency of microbial N synthesis in crossbred dairy steers.

  8. Improving methane production from digested manure biofibers by mechanical and thermal alkaline pretreatment.

    Science.gov (United States)

    Tsapekos, P; Kougias, Panagiotis G; Frison, A; Raga, R; Angelidaki, I

    2016-09-01

    Animal manure digestion is associated with limited methane production, due to the high content in fibers, which are hardly degradable lignocellulosic compounds. In this study, different mechanical and thermal alkaline pretreatment methods were applied to partially degradable fibers, separated from the effluent stream of biogas reactors. Batch and continuous experiments were conducted to evaluate the efficiency of these pretreatments. In batch experiments, the mechanical pretreatment improved the degradability up to 45%. Even higher efficiency was shown by applying thermal alkaline pretreatments, enhancing fibers degradability by more than 4-fold. In continuous experiments, the thermal alkaline pretreatment, using 6% NaOH at 55°C was proven to be the most efficient pretreatment method as the methane production was increased by 26%. The findings demonstrated that the methane production of the biogas plants can be increased by further exploiting the fraction of the digested manure fibers which are discarded in the post-storage tank.

  9. DIGESTIBLE VALINE REQUIREMENTS IN LOW-PROTEIN DIETS FOR BROILERS CHICKS

    Directory of Open Access Journals (Sweden)

    GR Nascimento

    Full Text Available ABSTRACT Three experiments were carried out to evaluate the levels of digestible valine in diets with reduced crude protein on the performance, carcass yield and muscle fiber diameter of male broilers during the pre-starter (1 to 8 d of age, starter (9 to 21 d of age and grower phases (21 to 42 d of age. A total of 1,080 chickens in the pre-starter phase, 900 in the starter phase and 864 in the grower phase were distributed in a completely randomized design with six treatments, consisting of a control positive diet (C+ and diets with 2 percentage points (p.p. reduction of crude protein level and five graded levels of digestible valine (Val, with six replicates of 30, 25 and 24 birds per experimental unit, respectively. The recommended level of TID Val in the low-protein diets for broilers in the pre-starter, starter and grower stages were 1.028; 0.905 and 0.789%, respectively. The reduction of 2 percentage points of the crude protein level in diets based on corn and soybean meal impaired (p ≤0.05 the feed conversion ratio in the starter and grower stages. Likewise, the reduced-protein diets decreased (p ≤0.05 muscle fiber diameter, but did not affect (p> 0.05 carcass and parts yields, or abdominal fat percentage at 42 days.

  10. 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 ( β -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 β -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.

  11. 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.

  12. Nutrient digestion, microbial protein synthesis, and blood metabolites of Jersey heifers fed chitosan and whole raw soybeans

    Directory of Open Access Journals (Sweden)

    Jefferson Rodrigues Gandra

    2016-03-01

    Full Text Available ABSTRACT This study was undertaken to determine the effects of chitosan and whole raw soybean on nutrient intake and total tract digestion, nitrogen utilization, microbial protein synthesis, blood metabolites, and energy balance of dairy heifers. Twelve Jersey heifers (6±0.5 months of age and 139.50±25.56 kg of live weight; mean ± standard deviation were randomly assigned to a replicated Latin square design with a 2 × 2 factorial arrangement. The experimental period consisted of 14 days of adaptation to diets, six days of sampling, and five days of washout. The experimental diets were: control (CO; chitosan (CHI, inclusion of 2.0 g kg−1 DM of chitosan; whole raw soybean (WS, 163.0 g kg−1 of WS on diet DM basis; and chitosan + whole raw soybean (CHI+WS. Chitosan decreased dry matter and neutral detergent fiber intakes; however, CHI increased DM total tract digestion. An interaction effect was observed on retained nitrogen, which increased when animals were fed CHI+WS compared with CO or CHI, but did not differ from that of animals fed WS. Chitosan decreased microbial nitrogen and crude protein flow of heifers. Energy balance was improved when heifers received diets containing WS. Efficiency of energy utilization was not affected by experimental diets. An interaction effect was observed for blood high-density lipoprotein (HDL concentration, which increased with both dietary inclusion of CHI and WS compared with the other diets, and CHI provided the lowest value of HDL cholesterol. Chitosan and whole raw soybean do not alter nutrient intake and total tract digestion; however, they decrease nitrogen urinary excretion and increase blood HDL cholesterol of heifers.

  13. Minor milk constituents are affected by protein concentration and forage digestibility in the feed ration

    DEFF Research Database (Denmark)

    Larsen, Torben; Alstrup, Lene; Weisbjerg, Martin Riis

    2016-01-01

    The present study was conducted in order to investigate if selected minor milk components would be indicative for the nutritional situation of the cow. Forty-eight dairy cows were offered a high digestible ration vs. a lower digestible ration combined with 2 protein levels in a 4 × 4 Latin square...... design. Milk glucose, glucose-6-phosphate, cholesterol, triacylglycerides (TAG), uric acid and β-hydroxybutyrate (BHBA) were measured and correlated mutually and towards other milking parameters (yield, h since last milking, days in milk (DIM), urea, etc). The variation range of the suggested variables...... were broad, a fact that may support their utilisation as predictive parameters. The content of milk metabolites was significantly affected by the change in rations as milk glucose, glucose-6-phosphate, uric acid, and the ratio cholesterol: triacylglycerides increased with higher energy intake while...

  14. Quantification of the degree of biotinylation of proteins using proteinase K digestion and competition ELISA.

    Science.gov (United States)

    Rispens, Theo; Ooijevaar-de Heer, Pleuni

    2016-03-01

    Quantification of the degree of biotinylation of proteins is useful to achieve and maintain a high degree of consistency of reagents used in research and diagnostic setting. Unfortunately, existing protocols and commercial kits suffer from a number of shortcomings that limit their usefulness. Here, we describe a simple protocol that overcomes the limitations of current assays. A robust competition ELISA was developed that is easy to carry out, uses no specialized equipment other than a standard plate reader for absorbance measurements and only reagents that are commonly available. The protocol uses a proteinase K digestion step of a sample of biotinylated protein to eliminate multivalency issues and sterical hindrance from bulky proteins. Furthermore, the use of an anti-biotin antibody instead of streptavidin results in a convenient range of sensitivity, avoiding million-fold dilutions that may impair precision. The resulting assay typically consumes about 1 μg of biotinylated protein.

  15. Feeding Value of Silybum marianum for Sheep and its Effect on Fiber and Protein Digestion

    Directory of Open Access Journals (Sweden)

    Ali Mojadam

    2016-04-01

    Full Text Available Introduction Due to scarcity of forage and water resources and high feed prices in Iran, it seems that the utilization of native and cheap forage resources for ruminants is important. The Milk thistle (Silybum marianum is distributed in different regions of Iran included: Gonbad Kavoos, Gorgan, Nodeh Kalardasht, Dareh Hezareh, Dasht Mugan, Poshtkoh, Mollassani, Shush, Hamidieh, Ramhormuz, Izeh and Kazeroon. The Silybum marianum seed extract and whole plant contain many compounds including Silybin A and B, Silychristin, Apigenin, deoxy silydianin (flavonolignans. Seeds of this plant has about 20 to 25 percent oil, which oleic acids (31.85 %, linoleic acid (45.36 % percent and palmitic acid (8.25 % are the major fatty acids of Silybum marianum seed. Also, the Silybum marianum contains flavonoids and anti-nutritional compounds such as tannins and nitrates. Tannins, unsaturated oils and other anti-nutritional compounds may have negative effects on the digestion of fiber and protein fractions. Tannins can make complexes with large number of nutrients, such as carbohydrates, proteins, bacterial cell membrane protein and carbohydrates, and even digestive enzymes. The Silybum marianum plants are native to northern areas of Ahvaz and abundantly growth as self-propelled. The farm animals in these area (sheep, goats, camels, water buffalo and native cows, etc. graze this plant as well as the manual feeding, but there is little known about its effects on the health, performance and digestibility of nutrients (in particular on the fiber and protein, due to existing the tannin and unsaturated oils in it in those animals, and no research work has been done about Silybum marianum. Therefore, this experiment was conducted to determine the feeding value of Silybum marianum and to measure its effects on rumen fermentation and digestion of fiber (straw and protein constituents (soybean meal feedstuff in Arabi sheep. Materials and Methods Twelve male Arabi sheep

  16. The Effects of Fortification of Legumes and Extrusion on the Protein Digestibility of Wheat Based Snack

    Directory of Open Access Journals (Sweden)

    Swapnil S. Patil

    2016-04-01

    Full Text Available Cereal food products are an important part of the human diet with wheat being the most commonly consumed cereal in many parts of the world. Extruded snack products are increasing in consumer interest due to their texture and ease of use. However, wheat based foods are rich in starch and are associated with high glycaemic impact products. Although legume materials are generally rich in fibre and protein and may be of high nutritive value, there is a paucity of research regarding their use in extruded snack food products. The aim of this study was to prepare wheat-based extrudates using four different legume flours: lentil, chickpea, green pea, and yellow pea flour. The effects of adding legumes to wheat-based snacks at different levels (0%, 5%, 10%, and 15% during extrusion were investigated in terms of protein digestibility. It was observed that fortification of snacks with legumes caused a slight increase in the protein content by 1%–1.5% w/w, and the extrusion technique increased the protein digestibility by 37%–62% w/v. The product developed by extrusion was found to be low in fat and moisture content.

  17. Culturing Schwann Cells from Neonatal Rats by Improved Enzyme Digestion Combined with Explants-culture Method.

    Science.gov (United States)

    Liu, Di; Liang, Xiao-Chun; Zhang, Hong

    2016-08-01

    Objective To develop an improved method for culturing Schwann cells(SCs) by using both enzyme digestion and explants-culture approaches and compared with traditional explants-culture method and general hemi-explants-culture method. Methods Bilaterally sciatic nerves and brachial plexus nerves were dissected from 3 to 5-day-old neonatal SD rats and explants-culture method,general hemi-explants-culture method,and improved enzyme digestion combined with explants-culture method were adopted to culture SCs,respectively. SCs were digested and passaged after 7 days in culture and counted under the microscope. The purity of SCs was identified by S-100 immunofluorescence staining. Results The SCs of improved method group grew fastest and the total number of cells obtained was(1.85±0.13)×10(6);the SCs of the hemi-explants-culture method group grew slower than the improved method group and the total number of cells obtained was (1.10±0.10)×10(6);the SCs of the explants-culture method group grew slowest and the total number of cells obtained was (0.77±0.03)×10(6).The total number of cells obtained showed significant difference among the three groups(Pculture method group,and (74.50±4.23)% in the explants-culture method group(Pculture method can obtain sufficient amount of high-purity SCs in a short time and thus may be applied in further research on peripheral nerve regeneration.

  18. The Addition of Hatchery Liquid Waste to Dairy Manure Improves Anaerobic Digestion

    Directory of Open Access Journals (Sweden)

    WRT Lopes

    Full Text Available ABSTRACT The objective of this study was to determine the optimal inclusion level of liquid egg hatchery waste for the anaerobic co-digestion of dairy cattle manure. A completely randomized experimental was applied, with seven treatments (liquid hatchery waste to cattle manure ratios of0: 100, 5:95, 10:90, 15:85, 20:80, 25:75 and 30:70, with five replicates (batch digester model each. The evaluated variables were disappearance of total solids (TS, volatile solids (VS, and neutral detergent fiber (NDF, and specific production of biogas and of methane. Maximum TS and VS disappearance of 41.3% and 49.6%, were obtained at 15.5% and 16.0% liquid hatchery waste inclusion levels. The addition of 22.3% liquid hatchery considerably reduced NDF substrate content (53.2%. Maximum specific biogas production was obtained with 17% liquid hatchery waste, with the addition of 181.7 and 229.5 L kg-1TS and VS, respectively. The highest methane production, at 120.1 and 151.8 L CH4 kg-1TS and VS, was obtained with the inclusion of 17.5 and 18.0% liquid hatchery waste, respectively. The addition of liquid hatchery waste atratios of up to 15.5%in co-digestion with cattle manure reduced solid and fiber levels in the effluent, and improved biogas and methane production.

  19. Stable Isotope Labeling for Improved Comparative Analysis of RNA Digests by Mass Spectrometry

    Science.gov (United States)

    Paulines, Mellie June; Limbach, Patrick A.

    2017-01-01

    Even with the advent of high throughput methods to detect modified ribonucleic acids (RNAs), mass spectrometry remains a reliable method to detect, characterize, and place post-transcriptional modifications within an RNA sequence. Here we have developed a stable isotope labeling comparative analysis of RNA digests (SIL-CARD) approach, which improves upon the original 18O/16O labeling CARD method. Like the original, SIL-CARD allows sequence or modification information from a previously uncharacterized in vivo RNA sample to be obtained by direct comparison with a reference RNA, the sequence of which is known. This reference is in vitro transcribed using a 13C/15N isotopically enriched nucleoside triphosphate (NTP). The two RNAs are digested with an endonuclease, the specificity of which matches the labeled NTP used for transcription. As proof of concept, several transfer RNAs (tRNAs) were characterized by SIL-CARD, where labeled guanosine triphosphate was used for the reference in vitro transcription. RNase T1 digestion products from the in vitro transcript will be 15 Da higher in mass than the same digestion products from the in vivo tRNA that are unmodified, leading to a doublet in the mass spectrum. Singlets, rather than doublets, arise if a sequence variation or a post-transcriptional modification is present that results in a relative mass shift different from 15 Da. Moreover, the use of the in vitro synthesized tRNA transcript allows for quantitative measurement of RNA abundance. Overall, SIL-CARD simplifies data analysis and enhances quantitative RNA modification mapping by mass spectrometry.

  20. Bioavailability of the digestible lysine and total sulfur amino acids in meat and bone meals varying in protein quality.

    Science.gov (United States)

    Wang, X; Parsons, C M

    1998-07-01

    Experiments were conducted to determine whether the digestible Lys, Met, and TSAA in a high and low quality meat and bone meal (MBM) were totally bioavailable for protein synthesis in chicks. True digestibility of amino acids (AA) in the two MBM was determined by the precision-fed cecectomized rooster assay. Bioavailability of the digestible AA was then assessed in three slope-ratio chick growth assays using Lys-, Met-, or TSAA-deficient crystalline AA basal diets that were supplemented with varying levels of the test AA, high or low quality MBM, or AA mixtures simulating the mean digestible AA composition of the high and low quality MBM. Response parameters were weight gain, feed efficiency, body N gain, and body Lys gain in the Lys assay and weight gain and feed efficiency in the Met and TSAA assays. Bioavailability values for the digestible Lys, Met, and TSAA in the MBM and AA mixtures simulating MBM varied depending on response parameter, with values based on feed efficiency generally being highest. No consistent differences in bioavailability of the digestible AA were observed between the two MBM when all AA were considered; however, the bioavailability of the digestible Lys in the low quality MBM was lower than that in the high quality MBM for two of four performance criteria. When considering all response parameters and the AA mixture results, bioavailability of the digestible Lys and Met in the two MBM was generally 90% or greater, whereas bioavailability of the digestible TSAA was only 80% or less. The results of this study indicated that essentially all of the digestible Lys and Met in MBM were bioavailable for protein synthesis and metabolism but suggested that a significant amount of the TSAA, particularly Cys, was not bioavailable.

  1. Pepsin immobilized in dextran-modified fused-silica capillaries for on-line protein digestion and peptide mapping

    Energy Technology Data Exchange (ETDEWEB)

    Stigter, E.C.A. [Division of Biomedical Analysis, Department of Pharmaceutical Sciences, Faculty of Science, Utrecht University, Sorbonnelaan 16, 3584 CA Utrecht (Netherlands)], E-mail: e.c.a.stigter@uu.nl; Jong, G.J. de; Bennekom, W.P. van [Division of Biomedical Analysis, Department of Pharmaceutical Sciences, Faculty of Science, Utrecht University, Sorbonnelaan 16, 3584 CA Utrecht (Netherlands)

    2008-07-07

    On-line digestion of proteins under acidic conditions was studied using micro-reactors consisting of dextran-modified fused-silica capillaries with covalently immobilized pepsin. The proteins used in this study differed in molecular weight, isoelectric point and sample composition. The injected protein samples were completely digested in 3 min and the digest was analyzed with micro-high performance liquid chromatography (HPLC) and tandem mass spectrometry (MS/MS). The different proteins present in the samples could be identified with a Mascot database search on the basis of auto-MS/MS data. It proved also to be possible to digest and analyze protein mixtures with a sequence coverage of 55% and 97% for the haemoglobin {beta}- and {alpha}-chain, respectively, and 35-55% for the various casein variants. Protease auto-digestion, sample carry-over and loss of signal due to adsorption of the injected proteins were not observed. The backpressure of the reactor is low which makes coupling to systems such as Surface Plasmon Resonance biosensors, which do not tolerate too high pressure, possible. The reactor was stable for at least 40 days when used continuously.

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

    Science.gov (United States)

    El-Niely, Hania F. G.

    2007-06-01

    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 ( pphytic 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.

  3. The Protein Digestibility-Corrected Amino Acid Score (PDCAAS) - A concept for describing protein quality in foods and food ingredients: A critical review

    NARCIS (Netherlands)

    Schaafsma, G.

    2005-01-01

    Protein Digestibility-Corrected Amino Score (PDCAAS) is discussed. PDCAAS is now widely used as a routine assay for protein quality evaluation, replacing the more traditional biological methods [e.g., measurement of the Protein Efficiency Ratio (PER) in rats]. PDCAAS is based on comparison of the es

  4. Struvite Precipitation as a Means of Recovering Nutrients and Mitigating Ammonia Toxicity in a Two-Stage Anaerobic Digester Treating Protein-Rich Feedstocks

    Directory of Open Access Journals (Sweden)

    Shunli Wang

    2016-08-01

    Full Text Available Accumulation of ammonia, measured as total ammonia nitrogen (TAN, a product of protein decomposition in slaughterhouse wastes, inhibits the anaerobic digestion process, reducing digester productivity and leading to failure. Struvite precipitation (SP is an effective means to remove TAN and enhance the buffering of substrates. Different Mg and P sources were evaluated as reactants in SP in acidogenic digester effluents to reduce its TAN levels. In order to measure impact of TAN removal, a standard biochemical methane potential (BMP test was conducted to measure methane yield from treatments that had the highest TAN reductions. SP results showed 6 of 9 reagent combinations resulted in greater than 70% TAN removal. The BMP results indicated that SP treatment by adding Mg(OH2 and H3PO4 resulted in 57.6% nitrogen recovery and 41.7% increase in methane yield relative to the substrate without SP. SP is an effective technology to improve nutrient recovery and methane production from the anaerobic digestion of protein-rich feedstocks.

  5. Effects of protein and energy supplementation on growth, forage intake, forage digestion and nitrogen balance in meat goat kids.

    Science.gov (United States)

    Patterson, J M; Lambert, B D; Muir, J P; Foote, A P

    2009-08-01

    The objective of this study was to further the understanding of the effects of dietary protein and energy supplements on growth, performance, feed intake and grass forage digestibility in growing meat goat wethers. In Experiment 1, an 18% CP complete goat pellet was offered alone (control diet, C) or added (+), or not, as supplement to three grass hays (coastal bermudagrass, CB; Tifton 85 bermudagrass, T; and sorghum-Sudan grass hay, SS), to Boer-cross wethers (n = 72). The resulting seven diets were offered ad libitum. In Experiment 2, four wether goats in metabolism crates were used in a 4 × 4 Latin square design and fed a SS basal diet ad libitum with treatments consisting of no supplement, supplemental urea (200 mg/kg BW daily), supplemental dextrose (0.2% BW daily), or urea + dextrose (200 mg/kg BW daily and 0.2% BW daily, respectively). In Experiment 1, average daily gain (ADG) were -3.8, -5.0 and -6.6 g/day for goats consuming CB, T and SS, respectively, and 69.2, 61.6 and 58.1 g/day for supplemented CB (CB+), T (T+) and SS (SS+), respectively, as compared to 245.8 g/day for ad libitum access to C. Supplementation in Experiment 1 increased (P diets. In Experiment 2, protein and energy supplementation increased (P diet digestibility. The beneficial effects of supplements in Experiment 1 and the increase in nitrogen retention in Experiment 2 cannot be explained by improvements in ruminal fiber utilization, but could be due to post-ruminal nutrient supply and/or increased ruminal microbial protein synthesis.

  6. In vitro crude protein digestibility of Tenebrio molitor and Hermetia illucens insect meals and its correlation with chemical composition traits

    Directory of Open Access Journals (Sweden)

    Stefania Marono

    2015-07-01

    Full Text Available The aims of this study were to evaluate the correlation between in vitro crude protein digestibility coefficients of insect meals from Tenebrio molitor (TI and Hermetia illucens (HI and their chemical composition traits as well as to develop regression equations able to estimate the in vitro crude protein digestibility (CPd from proximate analysis of insect meals. Twelve samples of insect meals (6 from TM larvae, TM 1-6 and 6 from HI larvae, HI 1-6 were obtained from different producers and analysed for chemical composition and in vitro crude protein digestibility by a two-step enzymatic method (digestion with pepsin and trypsin-enriched pancreatin. For both insect meal samples, CPd was negatively correlated to ADF and chitin contents, while just for HI there was a positive correlation (P<0.01 between CP percentage of the samples and CPd. For both insect meals the former variable chosen in the stepwise analysis was the chitin, explaining the 79.45% of CPd variability for Tenebrio molitor samples and the 98.30% for Hermetia illucens. In the second step, the amount of protein linked to ADF was added in the model for T. molitor and CP for H. illucens samples. The coefficients chitin is the main constituent of insect body able to affect the crude protein digestibility of Tenebrio molitor and Hermetia illucens larvae meals estimated by an in vitro enzymatic method.

  7. Denaturation and in Vitro Gastric Digestion of Heat-Treated Quinoa Protein Isolates Obtained at Various Extraction pH

    NARCIS (Netherlands)

    Ruiz, Geraldine Avila; Opazo-Navarrete, Mauricio; Meurs, Marlon; Minor, Marcel; Sala, Guido; Boekel, van Martinus; Stieger, Markus; Janssen, Anja E.M.

    2016-01-01

    The aim of this study was to determine the influence of heat processing on denaturation and digestibility properties of protein isolates obtained from sweet quinoa (Chenopodium quinoa Willd) at various extraction pH values (8, 9, 10 and 11). Pretreatment of suspensions of protein isolates at 60,

  8. Combining protein extraction and anaerobic digestion to produce feed, fuel and fertilizer from green biomass – An organic biorefinery concept

    DEFF Research Database (Denmark)

    Fernandez, Maria Santamaria; Salces, Beatriz Molinuevo; Lübeck, Mette

    Organically grown green biomass (red clover, clover grass) was investigated as a resource for organic feed and organic fertilizer by combination of proteins extraction and anaerobic digestion of the residues. Extraction of proteins from both crops revealed very favourable amino acid composition...

  9. Nitrite addition to acidified sludge significantly improves digestibility, toxic metal removal, dewaterability and pathogen reduction

    Science.gov (United States)

    Du, Fangzhou; Keller, Jürg; Yuan, Zhiguo; Batstone, Damien J.; Freguia, Stefano; Pikaar, Ilje

    2016-12-01

    Sludge management is a major issue for water utilities globally. Poor digestibility and dewaterability are the main factors determining the cost for sludge management, whereas pathogen and toxic metal concentrations limit beneficial reuse. In this study, the effects of low level nitrite addition to acidified sludge to simultaneously enhance digestibility, toxic metal removal, dewaterability and pathogen reduction were investigated. Waste activated sludge (WAS) from a full-scale waste water treatment plant was treated at pH 2 with 10 mg NO2‑-N/L for 5 h. Biochemical methane potential tests showed an increase in the methane production of 28%, corresponding to an improvement from 247 ± 8 L CH4/kg VS to 317 ± 1 L CH4/kg VS. The enhanced removal of toxic metals further increased the methane production by another 18% to 360 ± 6 L CH4/kg VS (a total increase of 46%). The solids content of dewatered sludge increased from 14.6 ± 1.4% in the control to 18.2 ± 0.8%. A 4-log reduction for both total coliforms and E. coli was achieved. Overall, this study highlights the potential of acidification with low level nitrite addition as an effective and simple method achieving multiple improvements in terms of sludge management.

  10. Improving anaerobic digestion of easy-acidification substrates by promoting buffering capacity using biochar derived from vermicompost.

    Science.gov (United States)

    Wang, Dou; Ai, Jing; Shen, Fei; Yang, Gang; Zhang, Yanzong; Deng, Shihuai; Zhang, Jing; Zeng, Yongmei; Song, Chun

    2017-03-01

    Acid-buffering of VCBC and VC was evaluated using 4 VFAs, and their application on anaerobic digestion of CM and KW was investigated. Results indicated acid-buffering capacity of VCBC to acetic, propionic, butyric, and valeric acid was 2.5, 1.1, 1.9 and 1.6-fold higher comparing with VC. CM digestion was not initiated at higher organic loading of 50gTS/kg, while it worked well with 5.0% VCBC or VC. KW was not digested even though VC or VCBC was increased to 15% and 20%. However, KW digestion can be alleviated with increasing VCBC or VC proportion, in which the alleviation by VCBC was better than VC. Average VFAs concentration during CM digestion with VC was 4077.7mg/L comparing with 2835.8mg/L of VCBC, and biogas release was delayed for 10-days accompanying rapid pH decrease in CM digestion with VC, which reflected acid-buffering of biochar played a crucial role on improving anaerobic digestion.

  11. Enrichment of Extracellular Matrix Proteins from Tissues and Digestion into Peptides for Mass Spectrometry Analysis.

    Science.gov (United States)

    Naba, Alexandra; Clauser, Karl R; Hynes, Richard O

    2015-07-23

    The extracellular matrix (ECM) is a complex meshwork of cross-linked proteins that provides biophysical and biochemical cues that are major regulators of cell proliferation, survival, migration, etc. The ECM plays important roles in development and in diverse pathologies including cardio-vascular and musculo-skeletal diseases, fibrosis, and cancer. Thus, characterizing the composition of ECMs of normal and diseased tissues could lead to the identification of novel prognostic and diagnostic biomarkers and potential novel therapeutic targets. However, the very nature of ECM proteins (large in size, cross-linked and covalently bound, heavily glycosylated) has rendered biochemical analyses of ECMs challenging. To overcome this challenge, we developed a method to enrich ECMs from fresh or frozen tissues and tumors that takes advantage of the insolubility of ECM proteins. We describe here in detail the decellularization procedure that consists of sequential incubations in buffers of different pH and salt and detergent concentrations and that results in 1) the extraction of intracellular (cytosolic, nuclear, membrane and cytoskeletal) proteins and 2) the enrichment of ECM proteins. We then describe how to deglycosylate and digest ECM-enriched protein preparations into peptides for subsequent analysis by mass spectrometry.

  12. Improving the energy balance of grass-based anaerobic digestion through combined harvesting and pretreatment

    DEFF Research Database (Denmark)

    Tsapekos, Panagiotis; Kougias, Panagiotis; Egelund, H.

    2017-01-01

    An important challenge that has to be addressed to achieve sustainable anaerobic digestion of lignocellulosic substrates is the development of energy and cost efficient pretreatment methods. Technologies orientated to simultaneously harvest and mechanically pretreat the biomass at the field could...... meet these criteria as they can potentially reduce the energy losses. The objective of this study was to elucidate the effect of two full-scale harvesting machines to enhance the biogas production and subsequently, improve energy balance. The performances of Disc-mower and Excoriator were assessed...... technology. More specifically, Excoriator, which cuts and subsequently applies shearing forces on harvested biomass, enhanced the methane production up to 10% and the overall energy budget was improved proportionally to the driving speed increase....

  13. 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; Hvelplund, Torben

    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...

  14. Improving the stability of thermophilic anaerobic digesters treating SS-OFMSW through enrichment with compost and leachate seeds

    KAUST Repository

    Ghanimeh, Sophia A.

    2013-03-01

    This paper examines the potential of improving the stability of thermophilic anaerobic digestion of source-sorted organic fraction of municipal solid waste (SS-OFMSW) by adding leachate and compost during inoculation. For this purpose, two stable thermophilic digesters, A (control) and B (with added leachate and compost), were subjected to a sustained substrate shock by doubling the organic loading rate for one week. Feeding was suspended then gradually resumed to reach the pre-shock loading rate (2. gVS/l/d). Digester A failed, exhibiting excessive increase in acetate and a corresponding decrease in pH and methane generation, and lower COD and solids removal efficiencies. In contrast, digester B was able to restore its functionality with 90% recovery of pre-shock methane generation rate at stable pH, lower hydrogen levels, and reduced VFAs and ammonia accumulation. © 2012 Elsevier Ltd.

  15. Improving the mixing performances of rice straw anaerobic digestion for higher biogas production by computational fluid dynamics (CFD) simulation.

    Science.gov (United States)

    Shen, Fei; Tian, Libin; Yuan, Hairong; Pang, Yunzhi; Chen, Shulin; Zou, Dexun; Zhu, Baoning; Liu, Yanping; Li, Xiujin

    2013-10-01

    As a lignocellulose-based substrate for anaerobic digestion, rice straw is characterized by low density, high water absorbability, and poor fluidity. Its mixing performances in digestion are completely different from traditional substrates such as animal manures. Computational fluid dynamics (CFD) simulation was employed to investigate mixing performances and determine suitable stirring parameters for efficient biogas production from rice straw. The results from CFD simulation were applied in the anaerobic digestion tests to further investigate their reliability. The results indicated that the mixing performances could be improved by triple impellers with pitched blade, and complete mixing was easily achieved at the stirring rate of 80 rpm, as compared to 20-60 rpm. However, mixing could not be significantly improved when the stirring rate was further increased from 80 to 160 rpm. The simulation results agreed well with the experimental results. The determined mixing parameters could achieve the highest biogas yield of 370 mL (g TS)(-1) (729 mL (g TS(digested))(-1)) and 431 mL (g TS)(-1) (632 mL (g TS(digested))(-1)) with the shortest technical digestion time (T 80) of 46 days. The results obtained in this work could provide useful guides for the design and operation of biogas plants using rice straw as substrates.

  16. Improved diffusion methods for nitrogen and 15nitrogen analysis of Kjeldahl digests.

    Science.gov (United States)

    Stevens, W B; Mulvaney, R L; Khan, S A; Hoeft, R G

    2000-01-01

    Simple methods are described that permit the use of either H3BO3 indicator solution or acidified filter disks to collect NH3 liberated by treatment of Kjeldahl digests with NaOH. These methods incorporate modifications to improve reliability, analytical capacity, and convenience. A semimicro digest was diluted to 25 mL with deionized water, and a 10 mL aliquot, containing up to 4 mg N (150 microg N for diffusions into acidified disks), was transferred to a shell vial, which was placed inside a 473 mL (1 pint) Mason jar containing 10 mL 10N NaOH. The NH3 liberated by overturning the vial was collected after 12 to 48 h at ambient temperature, or after 4 h at 45 to 50 degrees C on a hotplate, for quantitative and/or isotope-ratio analyses. With either H3BO3 indicator solution or acidified filter disks, recovery of diffused N was quantitative. Isotope-ratio analyses of diffused N from 15N-labeled chemical, plant, and soil samples were within 3% of analyses using steam distillation.

  17. Amino acid profiles and digestible indispensable amino acid scores of proteins from the prioritized key foods in Bangladesh.

    Science.gov (United States)

    Shaheen, Nazma; Islam, Saiful; Munmun, Sarah; Mohiduzzaman, Md; Longvah, Thingnganing

    2016-12-15

    Concentrations of standard amino acids were determined in the composite samples (representing 30 agro-ecological zones of Bangladesh) of six prioritized key dietary protein sources: Oryza sativa (rice), Triticum aestivum (wheat flour), Lens culinaris (lentils), Pangusius pangusius (pangas), Labeo rohita (rohu) and Oreochromis mossambicus (tilapia). Digestible indispensable amino acid scores (DIAAS) was calculated using published data on amino acids' digestibility to evaluate the protein quality of these foods. Indispensable amino acid (IAA) contents (mg IAA/g protein), found to be highest in pangas (430) and lowest in wheat (336), of all these analyzed foods exceeded the FAO recommended daily allowance (277mg IAA/g protein) and contributed on average 40% to total amino acid contents. Untruncated DIAAS values ranged from 51% (lysine) in wheat to 106% (histidine) in pangas and distinguished pangas, rohu, and tilapia containing 'excellent quality' protein (DIAAS>100%) with potential to complement lower quality protein of cereals, fruits, and vegetables.

  18. Effect of various domestic processing methods on antinutrients and in vitro protein and starch digestibility of two indigenous varieties of Indian tribal pulse, Mucuna pruriens Var. utilis.

    Science.gov (United States)

    Siddhuraju, P; Becker, K

    2001-06-01

    The effect of various domestic processing methods on antinutrients and starch fractions and in vitro protein and starch digestibilities of white and black varieties of Mucuna pruriens var. utilis was studied. Cooking or autoclaving of both raw seeds and presoaked seeds in different solutions (water, tamarind extract, sodium bicarbonate, and citric acid) significantly (p < 0.05) reduced the content of total phenolics, phytic acid, trypsin inhibitor and chymotrypsin inhibitor activities, and L-dopa compared to soaking or dry heating techniques. The germination processes (24 and 48 h) were also effective in the reduction of various antinutrients, although this reduction appeared to be more pronounced in a prolonged period of germination (72 h). Water soaking followed by dehusking was found to be ineffective in the reduction of trypsin and chymotrypsin inhibitor activities in both varieties. All of the treatments were effective in significantly (p < 0.05) reducing the resistant starch content in the presently investigated samples. Cooking as well as autoclaving brought about a more significant (p < 0.05) improvement in the digestibility of protein and starch compared to germination and dry heat treatment. Moreover, among the different processing techniques, soaking in sodium bicarbonate solution followed by cooking (29.6-34.8%) or autoclaving (33.0-37.2%) seemed to be the best method for improving starch digestibility.

  19. Enhanced protein digestion through the confinement of nanozeolite-assembled microchip reactors.

    Science.gov (United States)

    Ji, Ji; Zhang, Yahong; Zhou, Xiaoqin; Kong, Jilie; Tang, Yi; Liu, Baohong

    2008-04-01

    An on-chip microreactor was proposed toward the acceleration of protein digestion through the construction of a nanozeolite-assembled network. The nanozeolite microstructure was assembled using a layer-by-layer technique based on poly(diallyldimethylammonium chloride) and zeolite nanocrystals. The adsorption of trypsin in the nanozeolite network was theoretically studied based on the Langmuir adsorption isotherm model. It was found that the controlled trypsin-containing nanozeolite networks assembled within a microchannel could act as a stationary phase with a large surface-to-volume ratio for the highly efficient proteolysis of both proteins at low levels and with complex extracts. The maximum proteolytic rate of the adsorbed trypsin was measured to be 350 mM min-1 microg-1, much faster than that in solution. Moreover, due the large surface-to-volume ratio and biocompatible microenvironment provided by the nanozeolite-assembled films as well as the microfluidic confinement effect, the low-level proteins down to 16 fmol per analysis were confidently identified using the as-prepared microreactor within a very short residence time coupled to matrix-assisted laser desorption-time-of-flight mass spectrometry. The on-chip approach was further demonstrated in the identification of the complex extracts from mouse macrophages integrated with two-dimensional liquid chromatography-electrospray ionization-tandem mass spectrometry. This microchip reactor is promising for the development of a facile means for protein identification.

  20. 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 0.05) by pasteurization, whereas sterilization initially accelerated WP digestion but did not change its overall digestibility.

  1. Improving methane production from digested manure biofibers by mechanical and thermal alkaline pretreatment

    DEFF Research Database (Denmark)

    Tsapekos, Panagiotis; Kougias, Panagiotis; Frison, A.

    2016-01-01

    Animal manure digestion is associated with limited methane production, due to the high content in fibers, which are hardly degradable lignocellulosic compounds. In this study, different mechanical and thermal alkaline pretreatment methods were applied to partially degradable fibers, separated from...... the effluent stream of biogas reactors. Batch and continuous experiments were conducted to evaluate the efficiency of these pretreatments. In batch experiments, the mechanical pretreatment improved the degradability up to 45%. Even higher efficiency was shown by applying thermal alkaline pretreatments......, enhancing fibers degradability by more than 4-fold. In continuous experiments, the thermal alkaline pretreatment, using 6% NaOH at 55 °C was proven to be the most efficient pretreatment method as the methane production was increased by 26%. The findings demonstrated that the methane production of the biogas...

  2. Towards automation in protein digestion: Development of a monolithic trypsin immobilized reactor for highly efficient on-line digestion and analysis.

    Science.gov (United States)

    Naldi, Marina; Černigoj, Urh; Štrancar, Ales; Bartolini, Manuela

    2017-05-15

    Reducing experimental variability, limiting contamination and increasing automation are essential goals in the development of reliable analytical platforms for mass spectrometry (MS)-based proteomics. In this work novel trypsin-based monolithic immobilized enzyme reactors (tryp-IMERs), obtained by covalent immobilization on convective interaction media (CIMac™) analytical columns (5mm×5.2mm I.D.), were developed. Notwithstanding the small dimensions, column format allowed the insertion in common high performance liquid chromatography (HPLC) systems, thus avoiding the use of expensive micro- or nano-platforms. Monolith pore diameter and surface chemistry were optimized to achieve high digestion efficiency even with high molecular weight proteins and to avoid protein/peptide adsorption, peak broadening and sample loss. A full characterization of the tryp-IMERs was undertaken to select the best protocol for preparation and type of trypsin. Optimization of the operational and storage conditions was carried out by an off-line approach. On-line studies were performed by setting a multidimensional analytical platform, which included the tryp-IMER, a trapping column, an analytical C4 column and a high resolution hybrid mass spectrometer (ESI-Q-TOF). In the optimized conditions rapid protein digestion (90±9s), high protein coverage (≥60%) and high score values were achieved for five selected sample proteins (cytochrome c, myoglobin and albumins from different sources) differing in molecular size, isoelectric point and accessibility to cleavage sites as well as for a protein mixture of 200ng. The best performing tryp-IMERs showed high sensitivity down to the pmole level. The platform also resulted suitable for the analysis of high-molecular weight proteins such as a pool of human immunoglobulins G (hIgG) and for the high molecular weight fraction of human plasma proteins, which were digested in less than two minutes to an extent similar to that achieved by overnight

  3. 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.

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

    Science.gov (United States)

    Vaštag, Zužana; Popović, Ljiljana; Popović, Senka; Peričin-Starčević, Ivana; Krimer-Malešević, 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 α-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.

  5. Effects of diets containing vegetable protein concentrates on performance and activity of digestive enzymes in silver catfish (Rhamdia quelen

    Directory of Open Access Journals (Sweden)

    Naglezi de Menezes Lovatto

    2014-02-01

    Full Text Available The purpose of study was to evaluate the effect of using protein concentrates crambe and sunflower meal in the diet of silver catfish juveniles, as substitute for animal protein source. A total of 300 silver catfish had been separate in 15 experimental units of 280 L, totaling five treatments with three replications. We evaluated two levels (25% and 50% replacement of the meat and bone meal by protein concentrates of crambe and sunflower meals. Evaluated growth parameters, biological index and digestive enzymes in fish. There was no statistical difference for mass (g and standard length (cm, but the fish diet CPFCr-25% had greater total length (cm. No difference in dry matter, crude protein and total protein deposited (calculated. However, there was a higher concentration of ash in the carcass of the animals fed the control diet and CPFCr-50% in relation to diet CPFG- 50%, in addition, higher levels of lipids in fish fed diet CPFG-50%. No significant differences for hepatosomatic index, digestive somatic index and intestinal quotient of animals subjected to different treatments. The activity of digestive enzymes trypsin and chymotrypsin did not change. There was increased activity of acid protease. The quantitative and qualitative increase in protein concentration from this fraction allows the use of bran protein concentrates crambe and sunflower as substitutes for animal protein source.

  6. 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

    NARCIS (Netherlands)

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

    2013-01-01

    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 undige

  7. 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

  8. Ruminal degradability and intestinal digestibility of protein and amino acids in soybean and corn distillers grains products.

    Science.gov (United States)

    Mjoun, K; Kalscheur, K F; Hippen, A R; Schingoethe, D J

    2010-09-01

    New fractionation and fermentation technologies in the ethanol industry have resulted in the production of different forms of distillers grains (DG). Such products are reduced-fat, high-protein, and "modified" wet feeds. Characterization of protein fractions of these co-products and other commonly used feedstuffs is important for the formulation of dairy cattle diets. In situ and in vitro techniques were conducted to compare crude protein (CP) availability in 4 DG products with commonly used soybean proteins. Soybean protein products included solvent-extracted soybean meal (SBM; 44% CP), expeller soybean meal (ESBM), and extruded soybeans (ES). The DG products were conventional distillers dried grains with solubles, reduced-fat distillers dried grains with solubles (RFDGS), high-protein distillers dried grains, and modified wet distillers grains with solubles (MWDGS). Nylon bags containing 5 g of each feed were incubated in the rumen of 3 cannulated lactating cows for 2, 4, 8, 16, 24, and 48 h. The rapidly degradable CP fraction varied from 8.1 to 37.2% for SBM and MWDGS, respectively. The slowly degradable CP fraction was greatest for SBM, ES, and high-protein distillers dried grains (88.0%+/-3.7), followed by ESBM, distillers dried grains with solubles, and RFDGS (76.8+/-4.1%). The MWDGS had the lowest slowly degradable CP fraction (61.1%). The rate of degradation of the slowly degradable CP fraction ranged from 11.8 for SBM to 2.7%/h for RFDGS. Rumen-undegradable protein varied widely (32.3 to 60.4%), with RFDGS having the greatest and SBM the lowest concentrations. Intestinal digestibility of rumen-undegradable protein (IDP) was estimated by pepsin-pancreatin digestion of ruminally preincubated (16 h) samples. The IDP was greatest for SBM, ESBM, and ES (97.7%+/-0.75), whereas IDP of DG products was 92.4%+/-0.87. Similarly, total digestible protein was greatest (99.0%) for soybean products, whereas DG products had a total digestible protein of 96.0%. Intestinal

  9. Methylated-antibody affinity purification to improve proteomic identification of plant RNA polymerase Pol V complex and the interacting proteins

    Science.gov (United States)

    Qin, Guochen; Ma, Jun; Chen, Xiaomei; Chu, Zhaoqing; She, Yi-Min

    2017-01-01

    Affinity purification followed by enzymatic digestion and mass spectrometry has been widely utilized for the sensitive detection of interacting proteins and protein complexes in various organisms. In plants, the method is technically challenging due to the low abundance proteins, non-specific binding and difficulties of eluting interacting proteins from antibody beads. In this report, we describe a strategy to modify antibodies by reductive methylation of lysines without affecting their binding properties, followed by on-bead digestion of bound proteins with endoproteinase Lys-C. By this method, the antibody remains intact and does not interfere with the downstream identification of interacting proteins. Non-specific binding proteins were excluded using 14N/15N-metabolic labeling of wild-type and the transgenic plant counterparts. The method was employed to identify 12 co-immunoprecipitated protein subunits in Pol V complex and to discover 17 potential interacting protein targets in Arabidopsis. Our results demonstrated that the modification of antibodies by reductive dimethylation can improve the reliability and sensitivity of identifying low-abundance proteins through on-bead digestion and mass spectrometry. We also show that coupling this technique with chemical crosslinking enables in-depth characterization of endogenous protein complexes and the protein-protein interaction networks including mapping the surface topology and post-translational modifications of interacting proteins. PMID:28224978

  10. Quantification of the main digestive processes in ruminants: the equations involved in the renewed energy and protein feed evaluation systems.

    Science.gov (United States)

    Sauvant, D; Nozière, P

    2016-05-01

    The evolution of feeding systems for ruminants towards evaluation of diets in terms of multiple responses requires the updating of the calculation of nutrient supply to the animals to make it more accurate on aggregated units (feed unit, or UF, for energy and protein digestible in the intestine, or PDI, for metabolizable protein) and to allow prediction of absorbed nutrients. The present update of the French system is based on the building and interpretation through meta-analysis of large databases on digestion and nutrition of ruminants. Equations involved in the calculation of UF and PDI have been updated, allowing: (1) prediction of the out flow rate of particles and liquid depending on the level of intake and the proportion of concentrate, and the use of this in the calculation of ruminal digestion of protein and starch from in situ data; (2) the system to take into account the effects of the main factors of digestive interactions (level of intake, proportion of concentrate, rumen protein balance) on organic matter digestibility, energy losses in methane and in urine; (3) more accurate calculation of the energy available in the rumen and the efficiency of its use for the microbial protein synthesis. In this renewed model UF and PDI values of feedstuffs vary depending on diet composition, and intake level. Consequently, standard feed table values can be considered as being only indicative. It is thus possible to predict the nutrient supply on a wider range of diets more accurately and in particular to better integrate energy×protein interactions occurring in the gut.

  11. Nutrient recovery and improvement of anaerobic digestion process by low grade magnesium oxide application

    OpenAIRE

    Romero Güiza, Maycoll Stiven

    2015-01-01

    Anaerobic digestion is a worldwide technology to treat organic waste streams, primarily due to its capacity to produce methane as renewable energy. However, there is an increasing interest on nutrient recovery (N and P), which from both environmental and economic reasons have been identified as key feature in anaerobic digestion plants. Controlled struvite formation has been attracting increasing attention as a near mature technology to recover nutrients from anaerobic digestion. However, str...

  12. Comparative studies of peak intensities and chromatographic separation of proteolytic digests, PTMs, and intact proteins obtained by nanoLC-ESI MS analysis at room and elevated temperatures.

    Science.gov (United States)

    Moskovets, Eugene V; Ivanov, Alexander R

    2016-06-01

    This work demonstrates that the chromatographic separation performed at highly stabilized elevated temperature results in significant improvements in sensitivity, quantitative accuracy, chromatographic resolution, and run-to-run reproducibility of nanoLC-MS analysis of complex peptides mixtures. A newly developed platform was shown to provide conditions for accurate temperature stabilization and temperature homogeneity when performing nanoLC-ESI MS analysis. We quantitatively assessed and compared the recovery of peptides and small proteins from nanoLC columns at room and elevated temperatures. We found that analyses performed at highly stabilized elevated temperatures led to improved detection sensitivity, reproducibility, and chromatographic resolution in reversed-phase LC separation of unmodified peptides (both hydrophilic and hydrophobic), post-translationally modified peptides (O-phosphorylated), and small intact proteins. The analytical benefits of elevated temperatures for qualitative and quantitative proteomic LC-MS profiling were demonstrated using mixtures of synthetic peptides, tryptic digests of mixtures of model proteins, and digested total lysates of isolated rat kidney mitochondria. The effect of elevated temperature on the ion suppression was also demonstrated. Graphical Abstract A fragment of overlaid LC retention time-m/z planar views demonstrates the improved separation performance in the analysis of a complex peptide mixture at elevated temperature. Retention time-m/z 2D peptide features detected at 60 °C (magenta) were matched and aligned with features detected at room temperature (green).

  13. Chemical composition and standardized ileal digestibility of protein and amino acids from blue mussel, starfish, and fish silage in pigs

    DEFF Research Database (Denmark)

    Nørgaard, Jens Vinther; Petersen, Jens Kjerulf; Tørring, Ditte Bruunshøj

    2015-01-01

    fish silage from farmed salmon was also included in theexperiment. The standardized ileal digestibility (SID) of crude protein (CP) and amino acids(AA) was evaluated in a Latin square design with pigs (initial weight 39.3 kg) fitted with asimple T-cannula in the terminal ileum. Diets contained 131...

  14. In vitro availability of iron and zinc: Effects of the type, concentration and fractions of digestion products of the protein

    NARCIS (Netherlands)

    Pérez-Llamas, F.; Diepenmaat-Wolters, M.G.E.; Zamora, S.

    1996-01-01

    An in vitro dialysis method was employed to determine the effect on the Fe and Zn absorption of the type (beef, pork and soyabean) and the amount (10 and 30 g/kg) of protein present. In addition, the effects of low- and high-molecular-weight (LMW and HMW respectively) digestion products were investi

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

    NARCIS (Netherlands)

    Hendriks, W.H.; Baal, van 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 low

  16. A novel protein mixture containing vegetable proteins renders enteral nutrition products non-coagulating after in vitro gastric digestion

    NARCIS (Netherlands)

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

    2013-01-01

    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

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

    Energy Technology Data Exchange (ETDEWEB)

    El-Niely, Hania F.G. [Food Irradiation Research Department, National Center for Radiation Research and Technology, Atomic Energy Authority, P.O. Box 29, Nasr City, Cairo (Egypt)]. E-mail: elniely@hotmail.com

    2007-06-15

    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.

  18. Dual Role (Anti- and Pro-oxidant) of Gallic Acid in Mediating Myofibrillar Protein Gelation and Gel in Vitro Digestion.

    Science.gov (United States)

    Cao, Yungang; True, Alma D; Chen, Jie; Xiong, Youling L

    2016-04-20

    The dose-dependent effects of gallic acid (GA; at 0, 6, 30, and 150 μmol/g protein) on chemical changes and gelling properties of oxidatively stressed porcine myofibrillar protein (MP) and in vitro digestibility of the gels were investigated. The incorporation of GA suppressed lipid oxidation and protein carbonyl formation but promoted the loss of thiol and amine groups, destabilization of the tertiary structure, aggregation, and cross-linking. The gelling potential (storage modulus) of MP was increased by nearly 50% with 6 and 30 μmol/g of GA, corresponding to enhanced protein unfolding and aggregation and formation of disulfide-dominant covalent bonds. However, GA at 150 μmol/g induced macroscopic aggregations and insolubility of MP, resulting in poorly structured gels. Despite the oxidative changes, MP gels did not show reduced susceptibility to digestive enzymes in vitro.

  19. Virginiamycin improves phosphorus digestibility and utilization by growing-finishing pigs fed a phosphorus-deficient, corn-soybean meal diet.

    Science.gov (United States)

    Agudelo, J H; Lindemann, M D; Cromwell, G L; Newman, M C; Nimmo, R D

    2007-09-01

    Evaluations of the nutritional effect of antibiotics have largely centered on effects related to the digestibility and utilization of protein and energy. The current study evaluated the potential effect of virginiamycin (VIR) on P digestibility in swine. A total of 70 barrows (mean initial BW = 51 to 64 kg) were used in 4 nutrient-balance experiments. A basal, corn-soybean meal diet that was not supplemented with any inorganic source of P was used in each experiment. In Exp. 1, two diets were tested: basal vs. basal plus 11 mg/kg of VIR. In Exp. 2, four diets were used with a 2 x 2 factorial arrangement of 0 and 11 mg/kg of VIR and 0 and 750 phytase (PHY) units/kg of diet (PU/kg). Experiments 3 and 4 were the same as Exp. 2, except PHY was reduced to 300 PU/kg. For all experiments, VIR improved P digestibility (32.71 to 37.72%, P < 0.001) and Ca digestibility (54.99 to 58.30%, P = 0.002). The addition of PHY improved both P and Ca digestibility (P < 0.001); 750 PU/kg increased P digestibility 27.3% (from 34.6 to 61.9%, P < 0.001), whereas 300 PU increased it 13.8% (from 33.4 to 47.2%, P < 0.001). In an experiment conducted to evaluate the long-term effects of VIR on gut microbial profile, pigs (24 gilts and 8 barrows; mean BW = 29.1 +/- 0.50 kg) were fed a simple corn-soybean meal diet for 16 wk with a 2 x 2 factorial arrangement of VIR (0 and 11 mg/kg) addition and 0.15% dicalcium phosphate deletion. The long-term feeding of VIR in both the control diet and the diet with a marginally reduced P level resulted in a change in ileal microbial profile. A positive numerical increment in the number of phytate-utilizing bacteria was observed in both the normal and P-deleted diets (log unit increments of 12.4 and 17.2% over the respective controls, P = 0.13) when VIR was added. The addition of VIR also tended to affect lactobacilli populations (main effect, P = 0.11; interaction, P = 0.02); VIR decreased lactobacilli in the normal-P diet but did not affect this bacterial

  20. 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

  1. Improving decoy databases for protein folding algorithms

    KAUST Repository

    Lindsey, Aaron

    2014-01-01

    Copyright © 2014 ACM. Predicting protein structures and simulating protein folding are two of the most important problems in computational biology today. Simulation methods rely on a scoring function to distinguish the native structure (the most energetically stable) from non-native structures. Decoy databases are collections of non-native structures used to test and verify these functions. We present a method to evaluate and improve the quality of decoy databases by adding novel structures and removing redundant structures. We test our approach on 17 different decoy databases of varying size and type and show significant improvement across a variety of metrics. We also test our improved databases on a popular modern scoring function and show that they contain a greater number of native-like structures than the original databases, thereby producing a more rigorous database for testing scoring functions.

  2. The sfe corn mutant: a new approach to improve fiber digestibility and milk production

    Science.gov (United States)

    Cross linking of lignin to hemicellulose by ferulates limits in vitro rumen fiber digestibility of grasses. Impact of ferulate cross linking on feed intake, milk production, and in vivo digestibility was investigated in a dairy cow feeding study using the low-ferulate sfe corn mutant. Silages of fiv...

  3. Minor milk constituents are affected by protein concentration and forage digestibility in the feed ration.

    Science.gov (United States)

    Larsen, Torben; Alstrup, Lene; Weisbjerg, Martin Riis

    2016-02-01

    The present study was conducted in order to investigate if selected minor milk components would be indicative for the nutritional situation of the cow. Forty-eight dairy cows were offered a high digestible ration vs. a lower digestible ration combined with 2 protein levels in a 4 × 4 Latin square design. Milk glucose, glucose-6-phosphate, cholesterol, triacylglycerides (TAG), uric acid and β-hydroxybutyrate (BHBA) were measured and correlated mutually and towards other milking parameters (yield, h since last milking, days in milk (DIM), urea, etc). The variation range of the suggested variables were broad, a fact that may support their utilisation as predictive parameters. The content of milk metabolites was significantly affected by the change in rations as milk glucose, glucose-6-phosphate, uric acid, and the ratio cholesterol: triacylglycerides increased with higher energy intake while BHBA and TAG decreased. The content of some of the milk metabolites changed during 24 h day/night periods: BHBA, cholesterol, uric acid and TAG increased whereas free glucose decreased in the night period. Certain associations between milk metabolites and calculated energy parameters like ECM, body condition score (BCS), and body weight gain were found, however, these associations were to some extent explained by an interaction with DIM, just as changes in milk metabolites during a 24 h period seems to interfere. It is concluded that the practical use of the suggested milk variables should be based on more than one metabolite and that stage of lactation and possibly time of the day where the milk is collected should be incorporated in predictive models.

  4. In vivo and in vitro protein digestibility of formulated feeds for Artemesia longinaris (Crustacea, Penaeidae

    Directory of Open Access Journals (Sweden)

    Analía Verónica Fernández Gimenez

    2009-12-01

    Full Text Available This study was undertaken to determine the in vivo crude protein apparent digestibility in the prawn Artemesia longinaris, using feeds with 0.25% of chromic oxide and animal (fish meal, meat and bone meal and squid protein concentrate and plant (soybean meal ingredients. Three replicate groups of prawn were fed and the feces were collected. The rate of protein hydrolysis was measured in vitro using midgut gland enzyme extract from the prawns fed the respective feeds and was compared with those found with enzyme extract of wild prawn. The in vivo apparent digestibility coefficients showed significant differences among the feeds (PO objetivo do presente trabalho foi determinar a digestibilidade aparente in vivo da proteína bruta de ingredientes de origem animal (farinhas de peixe, osso e carne e concentrado de proteína de lula e ingredientes vegetais (farinha de soja em camarões Artemesia longinaris utilizando rações contendo 0,25% de óxido de cromo. Três grupos de camarões, utilizados como replicatas, foram alimentados e as fezes coletadas. A velocidade de hidrólise da proteína de cada ração foi medida in vitro utilizando extrato enzimático da glândula do intestino médio dos camarões alimentados com a ração correspondente e foi comparado com aqueles obtidos com o extrato enzimático de camarões selvagens. Os coeficientes de digestibilidade aparente in vivo mostraram diferenças significativas entre as rações testadas (P<0,05. A farinha de peixe apresentou a maior digestibilidade (92%, enquanto valores intermediários de digestibilidade (83% foram encontrados para a farinha de carne e ossos. A ração contendo farinha de soja e concentrado de proteína de lula resultou em menor digestibilidade (63%. Não houve diferença significativa entre os valores de digestibilidade in vitro para as rações testadas. Estes resultados indicam a limitação inerente dos ensaios enzimáticos in vitro, os quais poderiam ser complementados com

  5. Review of feedstock pretreatment strategies for improved anaerobic digestion: From lab-scale research to full-scale application.

    Science.gov (United States)

    Carrere, Hélène; Antonopoulou, Georgia; Affes, Rim; Passos, Fabiana; Battimelli, Audrey; Lyberatos, Gerasimos; Ferrer, Ivet

    2016-01-01

    When properly designed, pretreatments may enhance the methane potential and/or anaerobic digestion rate, improving digester performance. This paper aims at providing some guidelines on the most appropriate pretreatments for the main feedstocks of biogas plants. Waste activated sludge was firstly investigated and implemented at full-scale, its thermal pretreatment with steam explosion being most recommended as it increases the methane potential and digestion rate, ensures sludge sanitation and the heat needed is produced on-site. Regarding fatty residues, saponification is preferred for enhancing their solubilisation and bioavailability. In the case of animal by-products, this pretreatment can be optimised to ensure sterilisation, solubilisation and to reduce inhibition linked to long chain fatty acids. With regards to lignocellulosic biomass, the first goal should be delignification, followed by hemicellulose and cellulose hydrolysis, alkali or biological (fungi) pretreatments being most promising. As far as microalgae are concerned, thermal pretreatment seems the most promising technique so far.

  6. The supplementation of low-P diets with microbial 6-phytase expressed in Aspergillus oryzae improves P digestibility in sows.

    Science.gov (United States)

    Torrallardona, D; Llauradó, L; Broz, J

    2012-12-01

    Two trials were conducted to evaluate a novel microbial 6-phytase expressed in Aspergillus oryzae (Ronozyme HiPhos; DSM Nutritional Products, Basel, Switzerland) in gestating and lactating sows. In the first trial, 24 sows (Duroc × Landrace; 223 kg BW) were offered, at 16 d of gestation, a low-P control diet (formulated to provide 4.0 g total P/kg; 1.5 g digestible P/kg) supplemented with 0, 500, or 1000 phytase activity (FYT)/kg of phytase. Two weeks later, fresh feces were sampled from all sows and the apparent total tract digestibility of P was measured using TiO(2) as indigestible marker. Phytase supplementation did not (P > 0.10) affect the total tract digestibility of P but reduced (P phytase. After 2 wk, fresh feces were sampled from all sows and the apparent total tract digestibility of P was measured using TiO(2) as indigestible marker. Phytase supplementation improved (P phytase tested increased the apparent total tract digestibility of P in sows and reduced P excretion in feces.

  7. The effects of condensed tannins from Desmodium intortum and Calliandra calothyrsus on protein and carbohydrate digestion in sheep and goats.

    Science.gov (United States)

    Perez-Maldonado, R A; Norton, B W

    1996-10-01

    A factorial experiment was conducted to study the effects of condensed tannins (CT) from the tropical legumes Desmodium intortum and Calliandra calothyrsus on the digestion and utilization of protein and carbohydrate in sheep and goats. CT-free Centrosema pubescens was also fed for comparison with the CT legumes, and each legume was included (300 g/kg DM) in a basal diet of pangola grass (Digitaria decumbens). Pangola grass alone was used as a control diet. There were no significant (P > 0.05) differences between sheep and goats for the efficiency of digestion of N (0.574, SE 0.013), organic matter (OM; 0.519, SE 0.010), neutral-detergent fibre (NDF; 0.524, SE 0.011) and acid-detergent fibre (ADF; 0.407, SE 0.016). Diets containing desmodium and calliandra were digested less well in the rumen (64 and 62% of total OM digested) when compared with the pangola and centrosema diets (74 and 73% of total OM digested in rumen). There was an apparent net gain of 30% in ADF across the digestive tract of sheep and goats given calliandra, and this gain was ascribed to the formation of 'artifact' fibre as a result of fibre-tannin interaction. Overall, inclusion of legume at 300 g/kg in the diet significantly increased (P desmodium (50%) and calliandra (56%) when compared with animals given the centrosema and pangola diets (35%). Sheep and goats given the CT diets also had higher excretions of faecal N. This increment of faecal N (14%) did not affect post-rumen N digestion (P > 0.05) since animals given CT diets absorbed more N (19%) per kg total OM digested than those given the control diets. It was concluded that whilst the low levels of CT provided in desmodium (1.0%) and calliandra (2.3%) diets protected dietary protein from degradation in the rumen, there were no overall beneficial or detrimental effects of CT in these diets for sheep or goats. A method was developed to categorize CT into fractions representative of their forms (free, protein-bound, and fibre-bound) during

  8. Pre-Sleep Protein Ingestion to Improve the Skeletal Muscle Adaptive Response to Exercise Training.

    Science.gov (United States)

    Trommelen, Jorn; van Loon, Luc J C

    2016-11-28

    Protein ingestion following resistance-type exercise stimulates muscle protein synthesis rates, and enhances the skeletal muscle adaptive response to prolonged resistance-type exercise training. As the adaptive response to a single bout of resistance exercise extends well beyond the first couple of hours of post-exercise recovery, recent studies have begun to investigate the impact of the timing and distribution of protein ingestion during more prolonged recovery periods. Recent work has shown that overnight muscle protein synthesis rates are restricted by the level of amino acid availability. Protein ingested prior to sleep is effectively digested and absorbed, and thereby stimulates muscle protein synthesis rates during overnight recovery. When applied during a prolonged period of resistance-type exercise training, protein supplementation prior to sleep can further augment gains in muscle mass and strength. Recent studies investigating the impact of pre-sleep protein ingestion suggest that at least 40 g of protein is required to display a robust increase in muscle protein synthesis rates throughout overnight sleep. Furthermore, prior exercise allows more of the pre-sleep protein-derived amino acids to be utilized for de novo muscle protein synthesis during sleep. In short, pre-sleep protein ingestion represents an effective dietary strategy to improve overnight muscle protein synthesis, thereby improving the skeletal muscle adaptive response to exercise training.

  9. 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

  10. 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 diets increased. Goats fed the T2, T3 and T4 diets gained 10.2, 34.1 and 52.5 g/head/day, respectively, while the control group (T1) lost weight (-12.7 g/head/day). The apparent digestibilities of DM, OM and CP were similar (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.

  11. Use of whey protein beads as a new carrier system for recombinant yeasts in human digestive tract.

    Science.gov (United States)

    Hébrard, Géraldine; Blanquet, Stéphanie; Beyssac, Eric; Remondetto, Gabriel; Subirade, Muriel; Alric, Monique

    2006-12-15

    A new immobilizing protocol using whey protein isolates was developed to entrap recombinant Saccharomyces cerevisiae. The model yeast strain expresses the heterologous P45073A1 that converts trans-cinnamic acid into p-coumaric acid. Beads resulted from a cold-induced gelation of a whey protein solution (10%) containing yeasts (7.5 x 10(7)cells ml(-1)) into 0.1M CaCl(2). The viability and growth capability of yeasts were not altered by our entrapment process. The release and activity of immobilized yeasts were studied in simulated human gastric conditions. During the first 60 min of digestion, 2.2+/-0.9% (n=3) of initial entrapped yeasts were recovered in the gastric medium suggesting that beads should cross the gastric barrier in human. The P45073A1 activity of entrapped yeasts remained significantly higher (pwhey protein beads. The main potential medical applications include biodetoxication or the correction of digestive enzyme deficiencies.

  12. The expression of proteins involved in digestion and detoxification are regulated in Helicoverpa armigera to cope up with chlorpyrifos insecticide.

    Science.gov (United States)

    Dawkar, Vishal V; Chikate, Yojana R; More, Tushar H; Gupta, Vidya S; Giri, Ashok P

    2016-02-01

    Helicoverpa armigera is a key pest in many vital crops, which is mainly controlled by chemical strategies. To manage this pest is becoming challenging due to its ability and evolution of resistance against insecticides. Further, its subsequent spread on nonhost plant is remarkable in recent times. Hence, decoding resistance mechanism against phytochemicals and synthetic insecticides is a major challenge. The present work describes that the digestion, defense and immunity related enzymes are associated with chlorpyrifos resistance in H. armigera. Proteomic analysis of H. armigera gut tissue upon feeding on chlorpyrifos containing diet (CH) and artificial diet (AD) using nano-liquid chromatography-mass spectrometry identified upregulated 23-proteins in CH fed larvae. Database searches combined with gene ontology analysis revealed that the identified gut proteins engrossed in digestion, proteins crucial for immunity, adaptive responses to stress, and detoxification. Biochemical and quantitative real-time polymerase chain reaction analysis of candidate proteins indicated that insects were struggling to get nutrients and energy in presence of CH, while at the same time endeavoring to metabolize chlorpyrifos. Moreover, we proposed a potential processing pathway of chlorpyrifos in H. armigera gut by examining the metabolites using gas chromatography-mass spectrometry. H. armigera exhibit a range of intriguing behavioral, morphological adaptations and resistance to insecticides by regulating expression of proteins involved in digestion and detoxification mechanisms to cope up with chlorpyrifos. In these contexts, as gut is a rich repository of biological information; profound analysis of gut tissues can give clues of detoxification and resistance mechanism in insects.

  13. 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

  14. Effects of processing moisture on the physical properties and in vitro digestibility of starch and protein in extruded brown rice and pinto bean composite flours.

    Science.gov (United States)

    Sumargo, Franklin; Gulati, Paridhi; Weier, Steven A; Clarke, Jennifer; Rose, Devin J

    2016-11-15

    The influence of pinto bean flour and processing moisture on the physical properties and in vitro digestibility of rice-bean extrudates has been investigated. Brown rice: pinto bean flour (0%, 15%, 30%, and 45% bean flour) were extruded under 5 moisture conditions (17.2%, 18.1%, 18.3%, 19.5%, and 20.1%). Physical properties [bulk density, unit density, radial expansion, axial expansion, overall expansion, specific volume, hardness, color, water solubility index, and water absorption index] and in vitro starch and protein digestibilities were determined. Increasing bean flour and processing moisture increased density and hardness while decreasing expansion. Rapidly digestible starch decreased and resistant starch increased as bean substitution and processing moisture increased. In vitro protein digestibility increased with increasing bean flour or with decreasing processing moisture. Incorporating bean flour into extruded snacks can negatively affect physical attributes (hardness, density, and expansion) while positively affecting in vitro starch (decrease) and protein (increase) digestibilities.

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

    Energy Technology Data Exchange (ETDEWEB)

    Taghinejad-Roudbaneh, M., E-mail: mtaghinejad@iaut.ac.i [Department of Animal Science, Faculty of Agriculture, Islamic Azad University, Tabriz Branch, P.O. Box 51589, Tabriz (Iran, Islamic Republic of); Ebrahimi, S.R. [Department of Animal Science, Faculty of Agriculture, Shahr-e-Qods Branch, Islamic Azad University, P.O. Box 37515-374, Shahr-e-Qods (Iran, Islamic Republic of); Azizi, S. [Department of Clinical Sciences, Faculty of Veterinary Medicine, Urmia University, P.O. Box 57155-1177, Urmia (Iran, Islamic Republic of); Shawrang, P. [Nuclear Science and Technology Research Institute, Agricultural, Medical and Industrial Research School, Atomic Energy Organization of Iran, P.O. Box 31485-498, Karaj (Iran, Islamic Republic of)

    2010-12-15

    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.

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

    Science.gov (United States)

    Taghinejad-Roudbaneh, M.; Ebrahimi, S. R.; Azizi, S.; Shawrang, P.

    2010-12-01

    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.

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

    DEFF Research Database (Denmark)

    Lissens, G.; Thomsen, Anne Belinda; De Baere, L.;

    2004-01-01

    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...

  18. Comparison of cake compositions, pepsin digestibility and amino acids concentration of proteins isolated from black mustard and yellow mustard cakes.

    Science.gov (United States)

    Sarker, Ashish Kumar; Saha, Dipti; Begum, Hasina; Zaman, Asaduz; Rahman, Md Mashiar

    2015-01-01

    As a byproduct of oil production, black and yellow mustard cakes protein are considered as potential source of plant protein for feed applications to poultry, fish and swine industries. The protein contents in black and yellow mustard cakes were 38.17% and 28.80% and their pepsin digestibility was 80.33% and 77.43%, respectively. The proteins were extracted at different pH and maximum proteins (89.13% of 38.17% and 87.76% of 28.80% respectively) isolated from black and yellow mustard cakes at pH 12. The purity of isolated proteins of black and yellow mustard cakes was 89.83% and 91.12% respectively and their pepsin digestibility was 89.67% and 90.17% respectively which assigned the absence of antinutritional compounds. It was found that essential amino acids isoleucine, lysine, methionine, threonine and tryptophan and non essential amino acids arginine and tyrosine were present in greater concentration in black mustard cake protein whereas other amino acids were higher in yellow mustard cake protein.

  19. Recent developments on polyphenol–protein interactions: effects on tea and coffee taste, antioxidant properties and the digestive system.

    Science.gov (United States)

    Bandyopadhyay, Prasun; Ghosh, Amit K; Ghosh, Chandrasekhar

    2012-06-01

    Tea and coffee are widely consumed beverages across the world and they are rich sources of various polyphenols. Polyphenols are responsible for the bitterness and astringency of beverages and are also well known to impart antioxidant properties which is beneficial against several oxidative stress related diseases like cancer, cardiovascular diseases, and aging. On the other hand, proteins are also known to display many important roles in several physiological activities. Polyphenols can interact with proteins through hydrophobic or hydrophilic interactions, leading to the formation of soluble or insoluble complexes. According to recent studies, this complex formation can affect the bioavailability and beneficiary properties of both the individual components, in either way. For example, polyphenol-protein complex formation can reduce or enhance the antioxidant activity of polyphenols; similarly it can also affect the digestion ability of several digestive enzymes present in our body. Surprisingly, no review article has been published recently which has focused on the progress in this area, despite numerous articles having appeared in this field. This review summarizes the recent trends and patterns (2005 onwards) in polyphenol-protein interaction studies focusing on the characterization of the complex, the effect of this complex formation on tea and coffee taste, antioxidant properties and the digestive system.

  20. 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.

  1. Effects of gastrointestinal digestion and heating on the allergenicity of the kiwi allergens Act d 1, actinidin, and Act d 2, a thaumatin-like protein

    NARCIS (Netherlands)

    Bublin, Merima; Radauer, Christian; Knulst, Andre; Wagner, Stefan; Scheiner, Otto; Mackie, Alan R.; Mills, E. N. Clare; Breiteneder, Heimo

    2008-01-01

    Kiwifruit is a significant elicitor of allergy both in children and adults. Digestibility of two kiwifruit allergens, actinidin (Act d 1) and thaumatin-like protein (Act d 2), was assessed using an in vitro digestion system that approximates physiological conditions with respect to the passage of fo

  2. 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)

  3. Improvement through low cost biofilm carrier in anaerobic tubular digestion in cold climate regions.

    Science.gov (United States)

    Martí-Herrero, J; Alvarez, R; Rojas, M R; Aliaga, L; Céspedes, R; Carbonell, J

    2014-09-01

    The aim of this research is to evaluate the increase of biogas production with low cost tubular digesters in cold climates using PET rings inside the reactor. Two similar digesters have been operated and monitored in cold weather conditions and have been fed with cow manure. Digester 1 was filled with PET - rings as a biofilm carrier, Digester 2 was kept as a reference. Through the PET - rings the functional surface could be increased by a factor 4.2. The results show that 44% more biogas per Kg SV has been produced with the biofilm carrier in use (0.33 m(3)/kg SV) (reference digester -0.23 m(3)/kg SV), at an organic load rate of 0.26 kg SV/m(3)/d. The thermal performance shows that with an adaptation of the low cost tubular digester the slurry temperature can be raised up to 16.6°C (average) by surrounding temperature of 6.1°C (average) without using any active heating system.

  4. Super-atmospheric pressure ionization mass spectrometry and its application to ultrafast online protein digestion analysis.

    Science.gov (United States)

    Chen, L C; Ninomiya, S; Hiraoka, K

    2016-06-01

    Pressure is a key parameter for an ionization source. In this Special Feature article, Lee Chuin Chen and colleagues review super-atmospheric pressure ionization MS with electrospray, corona-discharge-based chemical ionization, and field desorption. They routinely run their mass spectrometer with ion source pressures ranging from several to several tens of atmospheres. A number of strategies have been used to preserve the high vacuum of the instrument while working with a high-pressure (HP) ion source. A recent prototype uses a booster pump with variable pumping speed added to the first pumping stage of the mass spectrometer to regulate a constant vacuum pressure. Further, a new HP-ESI source allowing rapid (a few seconds) online protein digestion MS is also reported. Dr. Lee Chuin Chen is Associate Professor in the Department of Interdisciplinary Research at the University of Yamanashi (Yamanashi, Japan). His main research interest is the development of novel mass spectrometric methods for in-situ medical diagnosis.

  5. Effects of fasting and refeeding on the digestive tract of zebrafish (Danio rerio) fed with Spirulina (Arthrospira platensis), a high protein feed source.

    Science.gov (United States)

    Lo Cascio, Patrizia; Calabrò, Concetta; Bertuccio, Clara; Paterniti, Irene; Palombieri, Deborah; Calò, Margherita; Albergamo, Ambrogina; Salvo, Andrea; Gabriella Denaro, Maria

    2017-01-03

    In the present work, morphological and molecular effects of short-term feed deprivation and refeeding with Spirulina (Arthrospira platensis) on zebrafish digestive tract were determined. Once elucidated the proximate composition of Spirulina feed, immunohistochemical and western blot analyses of peptide transporter (PepT1) and cholecystokinin (CCK8) were carried out in the gastrointestinal tract of zebrafish, previously morphologically investigated. Two and five fasting days caused not only morphostructural alterations, but also the downregulation of PepT1 and CCK8 proteins. Conversely, the recovery of normal morphological conditions, along with an increased PepT1 and CCK8 expression, were observed after refeeding with Spirulina. The increase of PepT1 expression in zebrafish may be responsible for the enhanced CCK8 secretion, so that both proteins may contribute to an improved digestion process during refeeding. These observations could be supported not only by compensatory mechanisms induced by fasting and refeeding but also by an higher protein quality of Spirulina-based diet.

  6. Starch digestibility and molecular weight distribution of proteins in rice grains subjected to heat-moisture treatment.

    Science.gov (United States)

    Silva, Wyller Max Ferreira; Biduski, Bárbara; Lima, Karina Oliveira; Pinto, Vânia Zanella; Hoffmann, Jéssica Fernanda; Vanier, Nathan Levien; Dias, Alvaro Renato Guerra

    2017-03-15

    The objective of this study was to evaluate the effects of the heat-moisture treatment (HMT) applied to paddy rice grains on the physicochemical properties, in vitro starch digestibility, and molecular weight distribution of proteins in rice flour. The paddy rice grains were adjusted to 13%, 16%, and 18% moisture and autoclaved at 121°C for 30 and 60min. The HMT promoted a reduction of the amylose content, the swelling power, and the solubility of the rice flour. Changes in the relative crystallinity and molecular weight of the proteins extracted with a sodium phosphate buffer containing 2.0% sodium dodecyl sulfate (SDDS) were observed in HMT samples. The different methods for the quantification of resistant starch resulted in distinct resistant starch contents. The HMT (18%-60min) promoted an increase in resistant starch content and the HMT (16%-60min) caused an increase in the slowly digestible starch in the rice flour.

  7. 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 Matić

    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.

  8. Effects of digestible protein to energy ratios on growth and carcass chemical composition of siamese fighting fish (Betta splendens)

    OpenAIRE

    Marcos Vinicius Antunes de Lemos; Thiago Quirino Arantes; Cristielle Nunes Souto; Graciela Pessoa Martins; Janaína Gomes Araújo; Igo Gomes Guimarães

    2014-01-01

    The expansion in the market of ornamental fish depends on the intensification of production systems and the generation of appropriate technologies. Therefore, it is necessary to know the nutritional requirements of each species for the formulation of nutritionally balanced diets. A 30-day feeding trial was undertaken to evaluate the effects of different dietary digestible protein to energy ratio in diets for Betta splendens fingerlings. Fifty-six all-male B. splendens with 1.03 ± 0.11 g avera...

  9. 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; Déchaux, Claire

    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....

  10. Comparison of liquid hot water and alkaline pretreatments of giant reed for improved enzymatic digestibility and biogas energy production.

    Science.gov (United States)

    Jiang, Danping; Ge, Xumeng; Zhang, Quanguo; Li, Yebo

    2016-09-01

    Liquid hot water (LHW) and alkaline pretreatments of giant reed biomass were compared in terms of digestibility, methane production, and cost-benefit efficiency for electricity generation via anaerobic digestion with a combined heat and power system. Compared to LHW pretreatment, alkaline pretreatment retained more of the dry matter in giant reed biomass solids due to less severe conditions. Under their optimal conditions, LHW pretreatment (190°C, 15min) and alkaline pretreatment (20g/L of NaOH, 24h) improved glucose yield from giant reed by more than 2-fold, while only the alkaline pretreatment significantly (pAlkaline pretreatment achieved 27% higher net electrical energy production than that of non-pretreatment (3859kJ/kg initial total solids), but alkaline liquor reuse is needed for improved net benefit.

  11. 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 μmol Trolox eq/g) than control tortilla (7.8 μmol 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

  12. Hydrocolloids Decrease the Digestibility of Corn Starch, Soy Protein, and Skim Milk and the Antioxidant Capacity of Grape Juice.

    Science.gov (United States)

    Yi, Yue; Jeon, Hyeong-Ju; Yoon, Sun; Lee, Seung-Min

    2015-12-01

    Hydrocolloids have many applications in foods including their use in dysphagia diets. We aimed to evaluate whether hydrocolloids in foods affect the digestibility of starch and protein, and their effects on antioxidant capacity. The thickening hydrocolloids: locust bean gum and carboxymethyl cellulose, and the gel-forming agents: agar agar, konjac-glucomannan, and Hot & Soft Plus were blended with corn starch and soy protein, skim milk, or grape juice and were examined for their in vitro-digestability by comparing the reducing sugar and trichloroacetic acid (TCA)-soluble peptide, for antioxidant capacity by total polyphenol contents and the 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity. The hydrocolloids resulted in a decrease in starch digestibility with the gel-forming agents. Hydrocolloids diminished TCA-soluble peptides in skim milk compared to soy protein with the exception of locust bean gum and decreased free radical scavenging capacities and total phenolic contents in grape juice. Our findings may provide evidence for the use of hydro-colloids for people at risk of nutritional deficiencies such as dysphagia patients.

  13. 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 μmol Trolox eq/g) than control tortilla (7.8 μmol Trolox eq/g). The addition of bean to tortilla modified the starch digestibility and antioxidant characteristics of tortilla, obtaining a product with nutraceutical characteristics.

  14. 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 μmol Trolox eq/g than control tortilla (7.8 μmol Trolox eq/g. The addition of bean to tortilla modified the starch digestibility and antioxidant characteristics of tortilla, obtaining a product with nutraceutical characteristics.

  15. Discovery of salt taste enhancing arginyl dipeptides in protein digests and fermented fish sauces by means of a sensomics approach.

    Science.gov (United States)

    Schindler, Alexander; Dunkel, Andreas; Stähler, Frauke; Backes, Michael; Ley, Jakob; Meyerhof, Wolfgang; Hofmann, Thomas

    2011-12-14

    As enzymatic digests of fish proteins were recently reported to enhance salt taste, the fish protein protamine was digested by chymotrypsin and trypsin and subsequently screened for candidate salt taste modulating (STM) peptides. To achieve this, first, a two-step sensory assay was developed and demonstrated to be a rather suitable tool for the detection of salt taste enhancers and the "quantitation" of their salt taste enhancing activity on the basis of isointensities with reference solutions. By means of activity-guided fractionation using ultrafiltration, gel permeation chromatography, and hydrophilic liquid interaction chromatography in combination with the sensory assay for STM activity assessment, a series of arginyl dipeptides, with RP, RA, AR, RG, RS, RV, VR, and RM being the most active, as well as l-arginine were found as salt taste enhancing molecules in fish protamine digests. For the first time, HPLC-MS/MS analysis on a PFP and a HILIC stationary phase, respectively, enabled the quantitative analysis of the arginyl peptides in a series of commercial and laboratory-made protein hydrolysates as well as fermented fish sauces.

  16. Apparent digestibility coefficients for dry matter, protein, lipid, and organic matter using various animal and plant protein sources by sunshine bass Morone chrysops X Morone saxatilis

    Science.gov (United States)

    Limited information is available on the digestibility of nutrients in various practical ingredients used in diets for commercially-important finfish species. This information is especially needed for sunshine bass, Morone chrysops X M. saxatilis, to improve least-cost diet formulations and to allow ...

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

    DEFF Research Database (Denmark)

    Lu, Jingquan; Gavala, Hariklia N.; Skiadas, Ioannis V.;

    2008-01-01

    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 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....... 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...

  18. Crude protein levels in diets containing pelleted concentrate for lactating goats: intake, digestibility, milk production and composition

    Directory of Open Access Journals (Sweden)

    Edileusa de Jesus dos Santos

    2015-08-01

    Full Text Available This study aimed to evaluate the effects of crude protein (100, 130, 160 and 190 g kg-1 of dry matter of diets composed of 200 g kg-1 of Tifton 85 grass hay and 800 g kg-1 of pelleted concentrate on intake, nutrient digestibility, production and composition of milk in lactating goats. Eight female Saanen goats with 42.7 ± 1.43 kg and 57.7 ± 7.37 days of lactation and milk production of 2 ± 0.22 kg at the beginning of the experiment were housed in individual 1.32 × 3.10 m stalls and distributed into two 4 × 4-balanced Latin squares. Intake of dry matter, organic matter, crude protein, neutral detergent fiber corrected for ash and protein, ether extract and total digestible nutrients showed a quadratic effect, with maximum intake of 2.030; 2.000; 305; 769; 55 and 1.574 g day-1 at the levels of 140.7; 140.8; 189.2; 140.9; 144.9 e 142.7 g kg-1 DM, respectively. Digestibility of dry matter, organic matter, crude protein, non-fibrous carbohydrates, ether extract and total digestible nutrient level varied linearly, with increases estimated at 0.54; 0.50, 2.02, 0.49, 0.80 and 0.63 g/100g for each percentage unit of protein added to the diet, respectively. Milk production was affected, with increase of 0.54 g for each 1% crude protein added to the diet. Milk lactose level decreased linearly, unlike the fat level, which increased linearly. Protein level showed a quadratic behavior, with a maximum of 36.7 g per kg of milk at the level of 160.5 g per kg of DM. It is recommended to use crude protein between 135 g and 150 g per kg of dry matter of diets consisting of 800 g of pelleted concentrate (composed of soybean meal replacing the alfalfa hay as protein source per kg of DM for lactating goats producing 2 kg of milk per day.

  19. Digestibility, chemical compound and protein quality of amaranthus forage at two harvested cut

    Directory of Open Access Journals (Sweden)

    pooria ehsani

    2016-04-01

    Full Text Available Introduction Amaranth, genus Amaranthus, belongs to the Amaranthaceae family and includes more than 60 species (28. Amaranth forage, is distinguished by high yield performance of up to 70 t/ha (30. Plant maturity was found to affect neutral detergent fiber (NDF, acid detergent fiber (ADF, lignin, CP and nitrate content of amaranth forage (27. Several studies (21, 27, 29 have shown that the nutritive value of amaranth as a ruminant feed is equal to, or better than, commonly used forages such as alfalfa. Its favorable chemical analysis (e.g., high crude protein (CP and low lignin ranges from 80 to 285 and from 17 to 73 g/kg DM, respectively and its high dry matter (DM digestibility (590–790 g/kg DM low nitrate and oxalic acid concentrations (below toxic levels suggest that it has potential value as a ruminant feedstuff (21, 23, 27. Materials and Methods The forage was planted in spring at one field (10 ha near Karaj city (Iran. The area is at an altitude of 1215 m above sea level, with a mean annual rainfall and temperature is 305.8 mm and 15C◦, respectively. In the early autumn, samples were harvested by hand from at least 10 locations within the field and pooled to five samples. The dry samples were analyzed for DM (method 930.15, ash (method 924.05 and CP (method 9 84.13 of AOAC (6. Neutral detergent fibre (NDF and acid detergent fibre (ADF according to the method of Robertson and Van Soest (35 with NDF and ADF assayed sequentially without a heat stable amylase, and expressed inclusive of residual ash (25. Insoluble CP, soluble CP (SP, true protein (TP, AD insoluble CP (ADICP and ND insoluble CP (NDICP were determined according to Licitra method (15. Nitrate was determined by a colorimetric method (8, and oxalic acid was determined according to Abaza method (2. Ca and Mg were determined by atomic absorption, P was analyzed by spectrophotometer and K by flame emission spectrometer (6. DMD, OMD and DOMD were determined by a two stage

  20. Key bacterial families (Clostridiaceae, Erysipelotrichaceae and Bacteroidaceae are related to the digestion of protein and energy in dogs

    Directory of Open Access Journals (Sweden)

    Emma N. Bermingham

    2017-03-01

    Full Text Available Background Much of the recent research in companion animal nutrition has focussed on understanding the role of diet on faecal microbiota composition. To date, diet-induced changes in faecal microbiota observed in humans and rodents have been extrapolated to pets in spite of their very different dietary and metabolic requirements. This lack of direct evidence means that the mechanisms by which microbiota influences health in dogs are poorly understood. We hypothesised that changes in faecal microbiota correlate with physiological parameters including apparent macronutrient digestibility. Methods Fifteen adult dogs were assigned to two diet groups, exclusively fed either a premium kibbled diet (kibble; K; n = 8 or a raw red meat diet (meat; M; n = 7 for nine weeks. Apparent digestibility of macronutrients (protein, fat, gross energy and dry matter, faecal weight, faecal health scores, faecal VFA concentrations and faecal microbial composition were determined. Datasets were integrated using mixOmics in R. Results Faecal weight and VFA levels were lower and the apparent digestibility of protein and energy were higher in dogs on the meat diet. Diet significantly affected 27 microbial families and 53 genera in the faeces. In particular, the abundances of Bacteriodes, Prevotella, Peptostreptococcus and Faecalibacterium were lower in dogs fed the meat diet, whereas Fusobacterium, Lactobacillus and Clostridium were all more abundant. Discussion Our results show clear associations of specific microbial taxa with diet composition. For example, Clostridiaceae, Erysipelotrichaceae and Bacteroidaceae were highly correlated to parameters such as protein and fat digestibility in the dog. By understanding the relationship between faecal microbiota and physiological parameters we will gain better insights into the effects of diet on the nutrition of our pets.

  1. Key bacterial families (Clostridiaceae, Erysipelotrichaceae and Bacteroidaceae) are related to the digestion of protein and energy in dogs

    Science.gov (United States)

    Maclean, Paul; Thomas, David G.; Cave, Nicholas J.; Young, Wayne

    2017-01-01

    Background Much of the recent research in companion animal nutrition has focussed on understanding the role of diet on faecal microbiota composition. To date, diet-induced changes in faecal microbiota observed in humans and rodents have been extrapolated to pets in spite of their very different dietary and metabolic requirements. This lack of direct evidence means that the mechanisms by which microbiota influences health in dogs are poorly understood. We hypothesised that changes in faecal microbiota correlate with physiological parameters including apparent macronutrient digestibility. Methods Fifteen adult dogs were assigned to two diet groups, exclusively fed either a premium kibbled diet (kibble; K; n = 8) or a raw red meat diet (meat; M; n = 7) for nine weeks. Apparent digestibility of macronutrients (protein, fat, gross energy and dry matter), faecal weight, faecal health scores, faecal VFA concentrations and faecal microbial composition were determined. Datasets were integrated using mixOmics in R. Results Faecal weight and VFA levels were lower and the apparent digestibility of protein and energy were higher in dogs on the meat diet. Diet significantly affected 27 microbial families and 53 genera in the faeces. In particular, the abundances of Bacteriodes, Prevotella, Peptostreptococcus and Faecalibacterium were lower in dogs fed the meat diet, whereas Fusobacterium, Lactobacillus and Clostridium were all more abundant. Discussion Our results show clear associations of specific microbial taxa with diet composition. For example, Clostridiaceae, Erysipelotrichaceae and Bacteroidaceae were highly correlated to parameters such as protein and fat digestibility in the dog. By understanding the relationship between faecal microbiota and physiological parameters we will gain better insights into the effects of diet on the nutrition of our pets. PMID:28265505

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

    Energy Technology Data Exchange (ETDEWEB)

    Nardiello, Donatella; Palermo, Carmen, E-mail: carmen.palermo@unifg.it; Natale, Anna; Quinto, Maurizio; Centonze, Diego

    2015-01-07

    Highlights: • Multi-enzyme digestion for protein sequencing and characterization by CID/ETD. • Simultaneous use of trypsin/chymotrypsin for the maximization of sequence. • Identification of PTMs, sequence variants and species-specific residues. • Increase of accuracy in sequence assignments by orthogonal fragmentation techniques. - Abstract: 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.

  3. Mejora de la escuela mediante informacion (Data-Driven School Improvement). ERIC Digest.

    Science.gov (United States)

    Johnson, James H.

    With accurate information, educators are better able to make effective decisions. Besides being used to assess student performance, a complete program of data collection and use can also show educators how management skills, individual learning styles, and other elements affect student achievement. This digest in Spanish provides an overview of…

  4. Selection for reduced ferulate esters in seedlings improved maize silage fiber digestibility

    Science.gov (United States)

    Ferulate cross linkage of lignin to arabinoxylans in grasses impedes cell wall digestibility. Formation of cross links requires ferulate esters on arabinoxylan. Subsequent lignin deposition is initiated by formation of cross-linked ferulate/monolignol complexes. We hypothesized that selection for se...

  5. Digestion of high rate activated sludge coupled to biochar formation for soil improvement in the tropics.

    Science.gov (United States)

    Nansubuga, Irene; Banadda, Noble; Ronsse, Frederik; Verstraete, Willy; Rabaey, Korneel

    2015-09-15

    High rate activated sludge (HRAS) is well-biodegradable sludge enabling energy neutrality of wastewater treatment plants via anaerobic digestion. However, even through successful digestion a notable residue still remains. Here we investigated whether this residue can be converted to biochar, for its use as a fertilizer or as a solid fuel, and assessed its characteristics and overall process efficiency. In a first phase, HRAS was anaerobicaly digested under mesophilic conditions at a sludge retention time of 20 days. HRAS digested well (57.9 ± 6.2% VS degradation) producing on average 0.23 ± 0.04 L CH4 per gram VS fed. The digestate particulates were partially air-dried to mimic conditions used in developing countries, and subsequently converted to biochar by fixed-bed slow pyrolysis at a residence time of 15 min and at highest heating temperatures (HHT) of 300 °C, 400 °C and 600 °C. Subsequently, the produced chars were characterized by proximate analysis, CHN-elemental analysis, pH in solution and bomb calorimetry for higher heating value. The yield and volatile matter decreased with increasing HHT while ash content and fixed carbon increased with increasing HHT. The produced biochar showed properties optimal towards soil amendment when produced at a temperature of 600 °C with values of 5.91 wt%, 23.75 wt%, 70.35% on dry basis (db) and 0.44 for volatile matter, fixed carbon, ash content and H/C ratio, respectively. With regard to its use for energy purposes, the biochar represented a lower calorific value than the dried HRAS digestate likely due to high ash content. Based on these findings, it can be concluded that anaerobic digestion of HRAS and its subsequent biochar formation at HHT of 600 °C represents an attractive route for sludge management in tropic settings like in Uganda, coupling carbon capture to energy generation, carbon sequestration and nutrient recovery.

  6. The anaerobic co-digestion of sheep bedding and ⩾ 50% cattle manure increases biogas production and improves biofertilizer quality.

    Science.gov (United States)

    Cestonaro, Taiana; Costa, Mônica Sarolli Silva de Mendonça; Costa, Luiz Antônio de Mendonça; Rozatti, Marcos Antonio Teofilo; Pereira, Dercio Ceri; Lorin, Higor Eisten Francisconi; Carneiro, Leocir José

    2015-12-01

    Sheep manure pellets are peculiarly shaped as small 'capsules' of limited permeability and thus are difficult to degrade. Fragmentation of manure pellets into a homogeneous mass is important for decomposition by microorganisms, and occurs naturally by physical shearing due to animal trampling, when sheep bedding is used. However, the high lignocellulose content of sheep bedding may limit decomposition of sheep manure. Here, we evaluated if co-digestion of sheep bedding with cattle manure would improve the yield and quality of the useful products of anaerobic digestion of sheep bedding--biogas and biofertilizer--by providing a source of nutrients and readily available carbon. Mixtures of sheep bedding and cattle manure in varying proportions (0%, 25%, 50%, 75%, or 100% cattle manure) were added to 6-L digesters, used in a batch system, and analyzed by uni and multivariate statistical tools. PC1, which explained 64.96% of data variability, can be referred to as 'organic fraction/productivity', because higher rates of organic fraction consumption (COD, cellulose and hemicellulose contents) led to higher digester productivity (biogas production, nutrient concentration, and sample stability changes). Therefore, productivity and organic fraction variables were most influenced by manure mixtures with higher (⩾ 50%) or lower (⩽ 25%) ratios of cattle manure, respectively. Increasing the amount of cattle manure up to 50% enhanced the biogas potential production from 142 L kg(-1)TS (0% of cattle manure) to 165, 171, 160 L biogas kg(-1)TS for the mixtures containing 100%, 75% and 50% of cattle manure, respectively. Our results show that the addition of ⩾ 50% cattle manure to the mixture increases biogas production and improves the quality of the final biofertilizer.

  7. Estimation of the True Digestibility of Rumen Undegraded Dietary Protein in the Small Intestine of Ruminants by the Mobile Bag Technique

    DEFF Research Database (Denmark)

    Hvelplund, Torben; Weisbjerg, Martin Riis; Andersen, L. S.

    1992-01-01

    Dietary protein degraded to various extents by varying the time of rumen incubation was prepared from eight concentrates and four roughages. Intestinal digestibility was obtained using the mobile bag technique on intact protein and on the samples of undegraded dietary protein from each feed...... introduced in recent years....

  8. 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; Holm, Jørgen;

    2013-01-01

    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...

  9. Standardized ileal digestibility of proteins and amino acids in sesame expeller and soya bean meal in weaning piglets.

    Science.gov (United States)

    Aguilera, A; Reis de Souza, T C; Mariscal-Landín, G; Escobar, K; Montaño, S; Bernal, M G

    2015-08-01

    Apparent ileal digestibility (AID) of diets containing sesame expeller (SE) and soya bean meal (SBM) was determined using 15 piglets (Genetiporc(®)), weaned at 17 ± 0.4 days with average body weight of 6.4 ± 0.7 kg (Fertilis 20 × G Performance, Genetiporc(®), PIC México, Querétaro, México). Piglets were randomly assigned to three treatments: (i) a reference diet with casein as the sole protein source; (ii) a mixed diet of casein-SE; and (iii) a mixed diet of casein-SBM. The chemical composition of SE and SBM was determined, and AID and standardized ileal digestibility (SID) of crude protein (CP) and amino acids (AAs) were determined for each protein source. SE contained greater quantities of ether extract, neutral detergent fibre, phytic acid, methionine and arginine than SBM. Lysine and proline contents and trypsin inhibitor activity were higher in SBM than in SE. The AID and SID of CP and AA (except for lysine and proline) were similar in SE and SBM. The AID of lysine and proline was higher in SBM than in SE (p < 0.05), and the SID of proline was higher in SE than in SBM (p < 0.05). These findings indicate that SE is an appropriate alternative protein source for early weaned pigs.

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

    Directory of Open Access Journals (Sweden)

    Nivea Regina de Oliveira Felisberto

    2011-10-01

    Full Text Available 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.

  11. Improved biogas production from rice straw by co-digestion with kitchen waste and pig manure

    Energy Technology Data Exchange (ETDEWEB)

    Ye, Jingqing [Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, Guangzhou 510640 (China); School of Environmental Sciences and Engineering, Sun Yat-sen University, Guangzhou 510275 (China); Li, Dong; Sun, Yongming [Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, Guangzhou 510640 (China); Wang, Guohui [School of Environmental Sciences and Engineering, Sun Yat-sen University, Guangzhou 510275 (China); Yuan, Zhenhong, E-mail: yuanzh@ms.giec.ac.cn [Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, Guangzhou 510640 (China); Zhen, Feng; Wang, Yao [Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, Guangzhou 510640 (China)

    2013-12-15

    Highlights: • Biogas production was enhanced by co-digestion of rice straw with other materials. • The optimal ratio of kitchen waste, pig manure and rice straw is 0.4:1.6:1. • The maximum biogas yield of 674.4 L/kg VS was obtained. • VFA inhibition occurred when kitchen waste content was more than 26%. • The dominant VFA were propionate and acetate in successful reactors. - Abstract: In order to investigate the effect of feedstock ratios in biogas production, anaerobic co-digestions of rice straw with kitchen waste and pig manure were carried out. A series of single-stage batch mesophilic (37 ± 1 °C) anaerobic digestions were performed at a substrate concentration of 54 g/L based on volatile solids (VS). The results showed that the optimal ratio of kitchen waste, pig manure, and rice straw was 0.4:1.6:1, for which the C/N ratio was 21.7. The methane content was 45.9–70.0% and rate of VS reduction was 55.8%. The biogas yield of 674.4 L/kg VS was higher than that of the digestion of rice straw or pig manure alone by 71.67% and 10.41%, respectively. Inhibition of biogas production by volatile fatty acids (VFA) occurred when the addition of kitchen waste was greater than 26%. The VFA analysis showed that, in the reactors that successfully produced biogas, the dominant intermediate metabolites were propionate and acetate, while they were lactic acid, acetate, and propionate in the others.

  12. Eliminating methanogenic activity in hydrogen reactor to improve biogas production in a two-stage anaerobic digestion process co-digesting municipal food waste and sewage sludge.

    Science.gov (United States)

    Zhu, Heguang; Parker, Wayne; Conidi, Daniela; Basnar, Robert; Seto, Peter

    2011-07-01

    Laboratory scale two-stage anaerobic digestion process model was operated for 280 days to investigate the feasibility to produce both hydrogen and methane from a mixture feedstock (1:1 (v/v)) of municipal food waste and sewage sludge. The maximum hydrogen and methane yields obtained in the two stages were 0.93 and 9.5 mL/mL feedstock. To eliminate methanogenic activity and obtain substantial hydrogen production in the hydrogen reactor, both feedstock and mixed liquor required treatment. The heat treatment (100°C, 10 min) for feedstock and a periodical treatment (every 2-5 weeks, either heating, removal of biomass particles or flushing with air) for mixed liquor were effective in different extent. The methane production in the second stage was significantly improved by the hydrogen production in the first stage. The maximum methane production obtained in the period of high hydrogen production was more than 2-fold of that observed in the low hydrogen production period.

  13. Hydrolysis by Alcalase Improves Hypoallergenic Properties of Goat Milk Protein

    Science.gov (United States)

    Yun, Sung-Seob; Lee, Won-Jae; Kim, Jin-Wook; Ha, Ho-Kyung; Yoo, Michelle

    2016-01-01

    Goat milk is highly nutritious and is consumed in many countries, but the development of functional foods from goat milk has been slow compared to that for other types of milk. The aim of this study was to develop a goat milk protein hydrolysate (GMPH) with enhanced digestibility and better hypoallergenic properties in comparison with other protein sources such as ovalbumin and soy protein. Goat milk protein was digested with four commercial food-grade proteases (separately) under various conditions to achieve the best hydrolysis of αs -casein and β-lactoglobulin. It was shown that treatment with alcalase (0.4%, 60℃ for 30 min) effectively degraded these two proteins, as determined by SDS-PAGE, measurement of nonprotein nitrogen content, and reverse-phase high-performance liquid chromatography. Hydrolysis with alcalase resulted in a significant decrease in β-lactoglobulin concentration (almost to nil) and a ~40% reduction in the level of αs-casein. Quantification of histamine and TNF-α released from HMC-1 cells (human mast cell line) showed that the GMPH did not induce an allergic response when compared to the control. Hence, the GMPH may be useful for development of novel foods for infants, the elderly, and convalescent patients, to replace cow milk. PMID:27621693

  14. Hydrolysis by Alcalase Improves Hypoallergenic Properties of Goat Milk Protein.

    Science.gov (United States)

    Jung, Tae-Hwan; Yun, Sung-Seob; Lee, Won-Jae; Kim, Jin-Wook; Ha, Ho-Kyung; Yoo, Michelle; Hwang, Hyo-Jeong; Jeon, Woo-Min; Han, Kyoung-Sik

    2016-01-01

    Goat milk is highly nutritious and is consumed in many countries, but the development of functional foods from goat milk has been slow compared to that for other types of milk. The aim of this study was to develop a goat milk protein hydrolysate (GMPH) with enhanced digestibility and better hypoallergenic properties in comparison with other protein sources such as ovalbumin and soy protein. Goat milk protein was digested with four commercial food-grade proteases (separately) under various conditions to achieve the best hydrolysis of αs -casein and β-lactoglobulin. It was shown that treatment with alcalase (0.4%, 60℃ for 30 min) effectively degraded these two proteins, as determined by SDS-PAGE, measurement of nonprotein nitrogen content, and reverse-phase high-performance liquid chromatography. Hydrolysis with alcalase resulted in a significant decrease in β-lactoglobulin concentration (almost to nil) and a ~40% reduction in the level of αs-casein. Quantification of histamine and TNF-α released from HMC-1 cells (human mast cell line) showed that the GMPH did not induce an allergic response when compared to the control. Hence, the GMPH may be useful for development of novel foods for infants, the elderly, and convalescent patients, to replace cow milk.

  15. Improved biogas production from rice straw by co-digestion with kitchen waste and pig manure.

    Science.gov (United States)

    Ye, Jingqing; Li, Dong; Sun, Yongming; Wang, Guohui; Yuan, Zhenhong; Zhen, Feng; Wang, Yao

    2013-12-01

    In order to investigate the effect of feedstock ratios in biogas production, anaerobic co-digestions of rice straw with kitchen waste and pig manure were carried out. A series of single-stage batch mesophilic (37±1 °C) anaerobic digestions were performed at a substrate concentration of 54 g/L based on volatile solids (VS). The results showed that the optimal ratio of kitchen waste, pig manure, and rice straw was 0.4:1.6:1, for which the C/N ratio was 21.7. The methane content was 45.9-70.0% and rate of VS reduction was 55.8%. The biogas yield of 674.4 L/kg VS was higher than that of the digestion of rice straw or pig manure alone by 71.67% and 10.41%, respectively. Inhibition of biogas production by volatile fatty acids (VFA) occurred when the addition of kitchen waste was greater than 26%. The VFA analysis showed that, in the reactors that successfully produced biogas, the dominant intermediate metabolites were propionate and acetate, while they were lactic acid, acetate, and propionate in the others.

  16. Pancreatic lipase-related protein 2 digests fats in human milk and formula in concert with gastric lipase and carboxyl ester lipase

    Science.gov (United States)

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

    2013-01-01

    INTRODUCTION Dietary fats must be digested into fatty acids and monoacylglycerols prior to absorption. In adults, colipase-dependent pancreatic triglyceride lipase (PTL) contributes significantly to fat digestion. In newborn rodents and humans, the pancreas expresses low levels of PTL. In rodents, a homologue of PTL, pancreatic lipase related protein 2 (PLRP2) and carboxyl ester lipase (CEL) compensate for the lack of PTL. In human newborns, the role for PLRP2 in dietary fat digestion is unclear. To clarify the potential of human PLRP2 to influence dietary fat digestion in newborns, we determined PLRP2 activity against human milk and infant formula. METHODS The activity of purified recombinant PLRP2, gastric lipase and CEL against fats in human milk and formula was measured with each lipase alone and in combination with a standard pH-stat assay. RESULTS Colipase added to human milk stimulated fat digestion. PLRP2 and CEL had activity against human milk and formula. Pre-digestion with gastric lipase increased PLRP2 activity against both substrates. Together, CEL and PLRP2 activity was additive with formula and synergistic with human milk. CONCLUSIONS PLRP2 can digest fats in human milk and formula. PLRP2 acts in concert with CEL and gastric lipase to digest fats in human milk in vitro. PMID:23732775

  17. Proximate composition, fatty acid analysis and protein digestibility-corrected amino acid score of three Mediterranean cephalopods.

    Science.gov (United States)

    Zlatanos, Spiros; Laskaridis, Kostas; Feist, Christian; Sagredos, Angelos

    2006-10-01

    Proximate composition, fatty acid analysis and protein digestibility-corrected amino acid score (PDCAAS) in three commercially important cephalopods of the Mediterranean sea (cuttlefish, octopus and squid) were determined. The results of the proximate analysis showed that these species had very high protein:fat ratios similar to lean beef. Docosahexaenoic, palmitic and eicosipentaenoic acid were the most abundant fatty acids among analyzed species. The amount of n-3 fatty acids was higher than that of saturated, monounsaturated and n-6 fatty acids. Despite the fact that cephalopods contain small amounts of fat they were found quite rich in n-3 fatty acids. Finally, PDCAAS indicated that these organisms had a very good protein quality.

  18. 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 β-casein. The numbers of peptides from β-casein, lactoferrin, α-lactalbumin, lactadherin, κ-casein, serum albumin, bile salt-associated lipase, and xanthine dehydrogenase/oxidase were significantly higher in the gastric samples than in the milk samples (P milk and gastric samples (P milk peptides with immunomodulatory and antibacterial properties of clinical relevance to the proximal intestinal tract. Data are available via ProteomeXchange (identifier PXD000688).

  19. 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

  20. Infogest: an international network for improving health properties of food by sharing our knowledge on the digestive process

    OpenAIRE

    Léonil, Joelle

    2012-01-01

    Establishing the links between food and human health was a top research priority for Europe. EU legislation, as advised by EFSA, now demands proper scientific data in nutrition and health claims. There is a lot of data being generated on the link between the food digestion and human health and a significant effort continues to be expended separately in each EU country on optimizing food for preventing the development of food-related diseases. The main objective of the COST Action “Improving...

  1. Use of proteinase K nonspecific digestion for selective and comprehensive identification of interpeptide cross-links: application to prion proteins.

    Science.gov (United States)

    Petrotchenko, Evgeniy V; Serpa, Jason J; Hardie, Darryl B; Berjanskii, Mark; Suriyamongkol, Bow P; Wishart, David S; Borchers, Christoph H

    2012-07-01

    Chemical cross-linking combined with mass spectrometry is a rapidly developing technique for structural proteomics. Cross-linked proteins are usually digested with trypsin to generate cross-linked peptides, which are then analyzed by mass spectrometry. The most informative cross-links, the interpeptide cross-links, are often large in size, because they consist of two peptides that are connected by a cross-linker. In addition, trypsin targets the same residues as amino-reactive cross-linkers, and cleavage will not occur at these cross-linker-modified residues. This produces high molecular weight cross-linked peptides, which complicates their mass spectrometric analysis and identification. In this paper, we examine a nonspecific protease, proteinase K, as an alternative to trypsin for cross-linking studies. Initial tests on a model peptide that was digested by proteinase K resulted in a "family" of related cross-linked peptides, all of which contained the same cross-linking sites, thus providing additional verification of the cross-linking results, as was previously noted for other post-translational modification studies. The procedure was next applied to the native (PrP(C)) and oligomeric form of prion protein (PrPβ). Using proteinase K, the affinity-purifiable CID-cleavable and isotopically coded cross-linker cyanurbiotindipropionylsuccinimide and MALDI-MS cross-links were found for all of the possible cross-linking sites. After digestion with proteinase K, we obtained a mass distribution of the cross-linked peptides that is very suitable for MALDI-MS analysis. Using this new method, we were able to detect over 60 interpeptide cross-links in the native PrP(C) and PrPβ prion protein. The set of cross-links for the native form was used as distance constraints in developing a model of the native prion protein structure, which includes the 90-124-amino acid N-terminal portion of the protein. Several cross-links were unique to each form of the prion protein, including

  2. Helichrysum and grapefruit extracts inhibit carbohydrate digestion and absorption, improving postprandial glucose levels and hyperinsulinemia in rats.

    Science.gov (United States)

    de la Garza, Ana Laura; Etxeberria, Usune; Lostao, María Pilar; San Román, Belén; Barrenetxe, Jaione; Martínez, J Alfredo; Milagro, Fermín I

    2013-12-11

    Several plant extracts rich in flavonoids have been reported to improve hyperglycemia by inhibiting digestive enzyme activities and SGLT1-mediated glucose uptake. In this study, helichrysum ( Helichrysum italicum ) and grapefruit ( Citrus × paradisi ) extracts inhibited in vitro enzyme activities. The helichrysum extract showed higher inhibitory activity of α-glucosidase (IC50 = 0.19 mg/mL) than α-amylase (IC50 = 0.83 mg/mL), whereas the grapefruit extract presented similar α-amylase and α-glucosidase inhibitory activities (IC50 = 0.42 mg/mL and IC50 = 0.41 mg/mL, respectively). Both extracts reduced maltose digestion in noneverted intestinal sacs (57% with helichrysum and 46% with grapefruit). Likewise, both extracts inhibited SGLT1-mediated methylglucoside uptake in Caco-2 cells in the presence of Na(+) (56% of inhibition with helichrysum and 54% with grapefruit). In vivo studies demonstrated that helichrysum decreased blood glucose levels after an oral maltose tolerance test (OMTT), and both extracts reduced postprandial glucose levels after the oral starch tolerance test (OSTT). Finally, both extracts improved hyperinsulinemia (31% with helichrysum and 50% with grapefruit) and HOMA index (47% with helichrysum and 54% with grapefruit) in a dietary model of insulin resistance in rats. In summary, helichrysum and grapefruit extracts improve postprandial glycemic control in rats, possibly by inhibiting α-glucosidase and α-amylase enzyme activities and decreasing SGLT1-mediated glucose uptake.

  3. Corn germ with pericarp in relation to whole corn: nutrient contents, food and protein efficiency, and protein digestibility-corrected amino acid score

    Directory of Open Access Journals (Sweden)

    Maria Margareth Veloso Naves

    2011-03-01

    Full Text Available The germ fraction with pericarp (bran is generated in the industrial processing of corn kernel, and it is used for oil extraction and animal feed. This study evaluated the nutritional and protein quality of this fraction in relation to whole corn. The proximate composition, mineral contents, and amino acid profile of the germ fraction with pericarp and of whole corn were determined. A 4-week experiment was conducted using 36 weanling male Wistar rats, and three 10%-protein diets (reference, germ with 15% lipids and casein with 15% lipids, two 6%-protein diets (whole corn and casein, and a protein-free diet were prepared. The germ showed higher contents of proteins, lipids, dietary fiber (27.8 g.100 g-1, ash, minerals (Fe and Zn- approximately 5 mg.100 g-1, and lysine (57.2 mg.g-1 protein than those of corn. The germ presented good quality protein (Relative Protein Efficiency Ratio-RPER = 80%; Protein Digestibility-Corrected Amino Acid Score-PDCAAS = 86%, higher than that of corn (RPER = 49%; PDCAAS = 60%. The corn germ fraction with pericarp is rich in dietary fiber, and it is a source of good quality protein as well as of iron and zinc, and its use as nutritive raw material is indicated in food products for human consumption.

  4. 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

  5. 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.

  6. DEAD-box protein Ddx46 is required for the development of the digestive organs and brain in zebrafish.

    Directory of Open Access Journals (Sweden)

    Shunya Hozumi

    Full Text Available Spatially and temporally controlled gene expression, including transcription, several mRNA processing steps, and the export of mature mRNA to the cytoplasm, is essential for developmental processes. It is well known that RNA helicases of the DExD/H-box protein family are involved in these gene expression processes, including transcription, pre-mRNA splicing, and rRNA biogenesis. Although one DExD/H-box protein, Prp5, a homologue of vertebrate Ddx46, has been shown to play important roles in pre-mRNA splicing in yeast, the in vivo function of Ddx46 remains to be fully elucidated in metazoans. In this study, we isolated zebrafish morendo (mor, a mutant that shows developmental defects in the digestive organs and brain, and found that it encodes Ddx46. The Ddx46 transcript is maternally supplied, and as development proceeds in zebrafish larvae, its ubiquitous expression gradually becomes restricted to those organs. The results of whole-mount in situ hybridization showed that the expression of various molecular markers in these organs is considerably reduced in the Ddx46 mutant. Furthermore, splicing status analysis with RT-PCR revealed unspliced forms of mRNAs in the digestive organ and brain tissues of the Ddx46 mutant, suggesting that Ddx46 may be required for pre-mRNA splicing during zebrafish development. Therefore, our results suggest a model in which zebrafish Ddx46 is required for the development of the digestive organs and brain, possibly through the control of pre-mRNA splicing.

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

    NARCIS (Netherlands)

    Carol, J.; Gorseling, M.C.J.K.; Jong, C.F. de; Lingeman, H.; Kientz, C.E.; Baar, B.L.M. van; Irth, H.

    2005-01-01

    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

  8. Improving Biomethane Production and Mass Bioconversion of Corn Stover Anaerobic Digestion by Adding NaOH Pretreatment and Trace Elements

    Directory of Open Access Journals (Sweden)

    ChunMei Liu

    2015-01-01

    Full Text Available This research applied sodium hydroxide (NaOH pretreatment and trace elements to improve biomethane production when using corn stover for anaerobic digestion. Full-factor experimental tests identified the best combination of trace elements with the NaOH pretreatment, indicating that the best combination was with 1.0, 0.4, and 0.4 mg·L−1·d−1 of elements Fe, Co, and Ni, respectively. The cumulative biomethane production adding NaOH pretreatment and trace elements was 11,367 mL; total solid bioconversion rate was 55.7%, which was 41.8%–62.2% higher than with NaOH-pretreatment alone and 22.2%–56.3% higher than with untreated corn stover. The best combination was obtained 5–9 days shorter than T90 and maintained good system operation stability. Only a fraction of the trace elements in the best combination was present in the resulting solution; more than 85% of the total amounts added were transferred into the solid fraction. Adding 0.897 g of Fe, 0.389 g of Co, and 0.349 g of Ni satisfied anaerobic digestion needs and enhanced biological activity at the beginning of the operation. The results showed that NaOH pretreatment and adding trace elements improve corn stover biodegradability and enhance biomethane production.

  9. Improving Biomethane Production and Mass Bioconversion of Corn Stover Anaerobic Digestion by Adding NaOH Pretreatment and Trace Elements.

    Science.gov (United States)

    Liu, ChunMei; Yuan, HaiRong; Zou, DeXun; Liu, YanPing; Zhu, BaoNing; Li, XiuJin

    2015-01-01

    This research applied sodium hydroxide (NaOH) pretreatment and trace elements to improve biomethane production when using corn stover for anaerobic digestion. Full-factor experimental tests identified the best combination of trace elements with the NaOH pretreatment, indicating that the best combination was with 1.0, 0.4, and 0.4 mg·L(-1)·d(-1) of elements Fe, Co, and Ni, respectively. The cumulative biomethane production adding NaOH pretreatment and trace elements was 11,367 mL; total solid bioconversion rate was 55.7%, which was 41.8%-62.2% higher than with NaOH-pretreatment alone and 22.2%-56.3% higher than with untreated corn stover. The best combination was obtained 5-9 days shorter than T90 and maintained good system operation stability. Only a fraction of the trace elements in the best combination was present in the resulting solution; more than 85% of the total amounts added were transferred into the solid fraction. Adding 0.897 g of Fe, 0.389 g of Co, and 0.349 g of Ni satisfied anaerobic digestion needs and enhanced biological activity at the beginning of the operation. The results showed that NaOH pretreatment and adding trace elements improve corn stover biodegradability and enhance biomethane production.

  10. Protein switches identified from diverse insertion libraries created using S1 nuclease digestion of supercoiled-form plasmid DNA.

    Science.gov (United States)

    Tullman, Jennifer; Guntas, Gurkan; Dumont, Matthew; Ostermeier, Marc

    2011-11-01

    We demonstrate that S1 nuclease converts supercoiled plasmid DNA to unit-length, linear dsDNA through the creation of a single, double-stranded break in a plasmid molecule. These double-stranded breaks occur not only in the origin of replication near inverted repeats but also at a wide variety of locations throughout the plasmid. S1 nuclease exhibits this activity under conditions typically employed for the nuclease's single-stranded nuclease activity. Thus, S1 nuclease digestion of plasmid DNA, unlike analogous digestion with DNaseI, effectively halts after the first double-stranded break. This property makes easier the construction of large domain insertion libraries in which the goal is to insert linear DNA at a variety of locations throughout a plasmid. We used this property to create a library in which a circularly permuted TEM1 β-lactamase gene was inserted throughout a plasmid containing the gene encoding Escherichia coli ribose binding protein. Gene fusions that encode allosteric switch proteins in which ribose modulates β-lactamase catalytic activity were isolated from this library using a combination of a genetic selection and a screen.

  11. Fermented milk containing Bifidobacterium lactis DN-173 010 improves gastrointestinal well-being and digestive symptoms in women reporting minor digestive symptoms: a randomised, double-blind, parallel, controlled study.

    Science.gov (United States)

    Guyonnet, Denis; Schlumberger, Armelle; Mhamdi, Leila; Jakob, Stefan; Chassany, Olivier

    2009-12-01

    The ability of probiotics to improve bowel habits or transit time has been shown in healthy populations. Additional data are required to support the use of specific probiotics to improve gastrointestinal (GI) well-being. The present study was designed to investigate the effect of consuming fermented milk (FM) on GI well-being, digestive symptoms and health-related quality of life (HRQoL) amongst women without diagnosed GI disorders. In this double-blind, controlled, parallel-design study, subjects were randomised to ingest daily either 2 x 125 g FM containing Bifidobacterium lactis DN-173 010 and yoghurt strains or a control non-fermented dairy product for 4 weeks followed by a 4-week wash-out period. GI well-being and digestive symptoms were assessed weekly. HRQoL was measured every 4 weeks. Data were analysed using analysis of covariance and logistic regression, correcting for baseline values on the full analysis set population of 197 women (aged 18-60 years). The percentage of women reporting an improvement in their GI well-being was significantly (P < 0.01) higher in the FM group v. the control group (OR 1.69; 95 % CI 1.17, 2.45). A significantly (P < 0.05) more pronounced decrease in the composite score of digestive symptoms was observed in the FM group when comparing with the control group (least squares mean - 0.57; 95 % CI - 1.12, - 0.02). Among HRQoL dimensions, the digestive comfort score was significantly (P < 0.05) improved in the FM group compared with the control group. The present study showed that the daily consumption of a specific FM is able to improve GI well-being and digestive symptoms in adult women without GI disorders.

  12. 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.

  13. Effect of Grape Pomace Powder, Mangosteen Peel Powder and Monensin on Nutrient Digestibility, Rumen Fermentation, Nitrogen Balance and Microbial Protein Synthesis in Dairy Steers

    Science.gov (United States)

    Foiklang, S.; Wanapat, M.; Norrapoke, T.

    2016-01-01

    This study was designed to investigate the effect of grape pomace powder (GPP), mangosteen peel powder (MPP) and monensin on feed intake, nutrients digestibility, microorganisms, rumen fermentation characteristic, microbial protein synthesis and nitrogen balance in dairy steers. Four, rumen fistulated dairy steers with initial body weight (BW) of 220±15 kg were randomly assigned according to a 4×4 Latin square design to receive four treatments. The treatments were as follows: T1 = control, T2 = supplementation with monensin at 33 mg/kg diet, T3 = supplementation with GPP at 2% of dry matter intake, and T4 = supplementation with MPP at 30 g/kg diet. The steers were offered the concentrate diet at 0.2% BW and 3% urea treated rice straw (UTRS) was fed ad libitum. It was found that GPP supplemented group had higher UTRS intake and nutrient digestibility in terms of neutral detergent fiber and acid detergent fiber than those in control group (p<0.05). Ammonia nitrogen (NH3-N) and blood urea-nitrogen concentration were higher in monensin, GPP and MPP supplemented groups (p<0.05). Total volatile fatty acids and propionate in the GPP group were higher than those in the control group (p<0.05) while acetate concentration, and acetate to propionate ratio were decreased (p<0.01) when steers were supplemented with GPP, monensin, and MPP, respectively. Moreover, protozoal populations in GPP, MPP, and monensin supplementation were significantly lower than those in the control group (p<0.05), while cellulolytic bacterial population was significantly higher in the control group (p<0.05). Nitrogen retention, microbial crude protein and efficiency of microbial nitrogen synthesis were found significantly higher in steers that received GPP (p<0.05). Based on this study it could be concluded that the GPP has potential as an alternative feed supplement in concentrate diets which can result in improved rumen fermentation efficiency, digestibility and microbial protein synthesis in

  14. Tandem SUMO fusion vectors for improving soluble protein expression and purification.

    Science.gov (United States)

    Guerrero, Fernando; Ciragan, Annika; Iwaï, Hideo

    2015-12-01

    Availability of highly purified proteins in quantity is crucial for detailed biochemical and structural investigations. Fusion tags are versatile tools to facilitate efficient protein purification and to improve soluble overexpression of proteins. Various purification and fusion tags have been widely used for overexpression in Escherichia coli. However, these tags might interfere with biological functions and/or structural investigations of the protein of interest. Therefore, an additional purification step to remove fusion tags by proteolytic digestion might be required. Here, we describe a set of new vectors in which yeast SUMO (SMT3) was used as the highly specific recognition sequence of ubiquitin-like protease 1, together with other commonly used solubility enhancing proteins, such as glutathione S-transferase, maltose binding protein, thioredoxin and trigger factor for optimizing soluble expression of protein of interest. This tandem SUMO (T-SUMO) fusion system was tested for soluble expression of the C-terminal domain of TonB from different organisms and for the antiviral protein scytovirin.

  15. Fat digestion in veal calves fed milk replacers low or high in calcium and containing either casein or soy protein isolate.

    Science.gov (United States)

    Yuangklang, C; Wensing, Th; Van den Broek, L; Jittakhot, S; Beynen, A C

    2004-04-01

    The hypothesis tested was that the inhibitory effect of dietary soy protein versus casein on fat digestion in veal calves would be smaller when diets were fed with high instead of low calcium content. Male calves, 1 wk of age, were fed 1 of 4 experimental milk replacers in a 2 x 2 factorial design. There were 19 animals per dietary group. The milk replacers contained either casein or soy protein isolate as variable protein source and were either low or high in calcium. Body weight gain was not significantly affected by the experimental diets. Soy protein isolate versus casein significantly reduced apparent fat digestibility. High versus low calcium intake also depressed fat digestion. The protein effect was smaller (2.9% units) for the high than the low calcium diets (3.6% units), but the interaction did not reach statistical significance. Soy protein isolate versus casein raised fecal bile acid excretion and so did high versus low calcium intake. The difference in bile acid excretion between the soy and casein containing diets was significantly greater for the high than low calcium diets. The absorption of phosphorus, calcium, and magnesium was higher for the casein diets than for the soy-containing diets. This study shows for the first time that soy protein isolate versus casein depressed fat digestion and raised fecal bile acid excretion in veal calves.

  16. A comparative in vitro study of the digestibility of heat- and high pressure-induced gels prepared from industrial milk whey proteins

    Science.gov (United States)

    He, Jin-Song; Mu, Tai-Hua; Wang, Juan

    2013-06-01

    We undertook this study to compare the digestibility of heat- and high pressure-induced gels produced from whey protein isolate (WPI). To simulate in vivo gastrointestinal digestion of WPI gels, a pepsin-trypsin digestion system was used. The in vitro protein digestibility of WPI gels induced by high pressure (400 MPa and 30 min; P-gel) and those induced by heat (80°C and 30 min; H-gel) was compared using a protein concentration of 0.14 g mL-1. The in vitro protein digestibility of P-gels was significantly greater than that of H-gels (p<0.05). The size-exclusion chromatography profiles of the hydrolysates showed that the P-gel generated more and smaller peptides than natural WPI and H-gels. Furthermore, Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed some soluble disulfide-mediated aggregation in the P-gel, while there was more insoluble aggregation in the H-gel than the P-gel. The P-gel was more sensitive to proteinase than the H-gel, which was related to the content of S-S bonds, and this in turn could be attributed to the differences in the gelation mechanism between the H-gel and P-gel.

  17. Nanoliter-volume protein enrichment, tryptic digestion, and partial separation based on isoelectric points by CE for MALDI mass spectral analysis.

    Science.gov (United States)

    Nesbitt, Chandra A; Jurcic, Kristina; Yeung, Ken K-C

    2008-01-01

    Sequence-specific proteolysis is an important part of protein identification by MS. Digestion of protein is commonly performed in-solution, in sample vials with volumes ranging from milli- to microliters. When digestion is performed with a sample volume below 1 microL, handling of solution and potential sample loss via adsorption become significant issues. In this report, a proof of concept for the digestion of a small volume protein solution inside a capillary was demonstrated using a discontinuous buffer system previously studied (Nesbitt, C. A., et al. J. Chromatogr. A 2005, 1073, 175-180). Upon voltage application, a pH junction was created by the discontinuous buffer. Using myoglobin as an example, the protein molecules were enriched at the junction with an estimated volume of a few nanoliters. A protease, trypsin, was then introduced to myoglobin at the junction by coenrichment to induce in-capillary digestion. The voltage application was then suspended to provide the necessary time (2 h) for the proteolysis to proceed. When completed, voltage application was resumed, and the discontinuous buffer reconcentrated the peptides formed from digestion. Importantly, the refocused peptides appeared to roughly elute according to their pIs, resulting in a partial separation. Direct sample deposition from capillary was performed to facilitate mass spectral analysis by MALDI. The partial separation, according to pI, offered the potential benefits of MALDI MS signal enhancement and provided supplementary pI information for peptide identity assignment.

  18. An improved Bradford protein assay for collagen proteins.

    Science.gov (United States)

    López, J M; Imperial, S; Valderrama, R; Navarro, S

    1993-10-29

    A modification of the protein determination method of Bradford adapted for collagen-rich samples is described. The use of Coomassie-based protein determination methods is limited by the great variation in colour yield obtained for different proteins. This is especially important in samples containing significant amounts of collagen where direct application of the methods of Lowry and Bradford results in underestimated values. Addition of small amounts of sodium dodecyl sulphate (SDS) (0.0035%) to the diluted solutions of Coomassie Brilliant Blue G used as dye reagent in the Bradford colorimetric assay caused a 4-fold increase in the colour response of three collagen proteins (Col I, III and IV) and a decrease in absorbance for various non-collagen proteins. The presence of SDS in the reagent did not result in a significant metachromatic shift of the collagen-dye complexes. This simple modification in the preparation of the reagent for the Bradford assay allows similar response curves to be obtained for collagen and non-collagen proteins, making the modified assay of potential use for protein determination in collagen-rich samples such as pancreatic extracts.

  19. Buffalo Cheese Whey Proteins, Identification of a 24 kDa Protein and Characterization of Their Hydrolysates: In Vitro Gastrointestinal Digestion.

    Science.gov (United States)

    Bassan, Juliana C; Goulart, Antonio J; Nasser, Ana L M; Bezerra, Thaís M S; Garrido, Saulo S; Rustiguel, Cynthia B; Guimarães, Luis H S; Monti, Rubens

    2015-01-01

    Milk whey proteins are well known for their high biological value and versatile functional properties, characteristics that allow its wide use in the food and pharmaceutical industries. In this work, a 24 kDa protein from buffalo cheese whey was analyzed by mass spectrometry and presented homology with Bos taurus beta-lactoglobulin. In addition, the proteins present in buffalo cheese whey were hydrolyzed with pepsin and with different combinations of trypsin, chymotrypsin and carboxypeptidase-A. When the TNBS method was used the obtained hydrolysates presented DH of 55 and 62% for H1 and H2, respectively. Otherwise for the OPA method the DH was 27 and 43% for H1 and H2, respectively. The total antioxidant activities of the H1 and H2 samples with and without previous enzymatic hydrolysis, determined by DPPH using diphenyl-p-picrylhydrazyl radical, was 4.9 and 12 mM of Trolox equivalents (TE) for H2 and H2Dint, respectively. The increased concentrations for H1 and H2 samples were approximately 99% and 75%, respectively. The in vitro gastrointestinal digestion efficiency for the samples that were first hydrolyzed was higher compared with samples not submitted to previous hydrolysis. After in vitro gastrointestinal digestion, several amino acids were released in higher concentrations, and most of which were essential amino acids. These results suggest that buffalo cheese whey is a better source of bioavailable amino acids than bovine cheese whey.

  20. Buffalo Cheese Whey Proteins, Identification of a 24 kDa Protein and Characterization of Their Hydrolysates: In Vitro Gastrointestinal Digestion.

    Directory of Open Access Journals (Sweden)

    Juliana C Bassan

    Full Text Available Milk whey proteins are well known for their high biological value and versatile functional properties, characteristics that allow its wide use in the food and pharmaceutical industries. In this work, a 24 kDa protein from buffalo cheese whey was analyzed by mass spectrometry and presented homology with Bos taurus beta-lactoglobulin. In addition, the proteins present in buffalo cheese whey were hydrolyzed with pepsin and with different combinations of trypsin, chymotrypsin and carboxypeptidase-A. When the TNBS method was used the obtained hydrolysates presented DH of 55 and 62% for H1 and H2, respectively. Otherwise for the OPA method the DH was 27 and 43% for H1 and H2, respectively. The total antioxidant activities of the H1 and H2 samples with and without previous enzymatic hydrolysis, determined by DPPH using diphenyl-p-picrylhydrazyl radical, was 4.9 and 12 mM of Trolox equivalents (TE for H2 and H2Dint, respectively. The increased concentrations for H1 and H2 samples were approximately 99% and 75%, respectively. The in vitro gastrointestinal digestion efficiency for the samples that were first hydrolyzed was higher compared with samples not submitted to previous hydrolysis. After in vitro gastrointestinal digestion, several amino acids were released in higher concentrations, and most of which were essential amino acids. These results suggest that buffalo cheese whey is a better source of bioavailable amino acids than bovine cheese whey.

  1. Comparative study of mechanical, hydrothermal, chemical and enzymatic treatments of digested biofibers to improve biogas production.

    Science.gov (United States)

    Bruni, Emiliano; Jensen, Anders Peter; Angelidaki, Irini

    2010-11-01

    Organic waste such as manure is an important resource for biogas production. The biodegradability of manures is however limited because of the recalcitrant nature of the biofibers it contains. To increase the biogas potential of the biofibers in digested manure, we investigated physical treatment (milling), chemical treatment (CaO), biological treatment (enzymatic and partial aerobic microbial conversion), steam treatment with catalyst (H(3)PO(4) or NaOH) and combination of biological and steam treatments (biofibers steam-treated with catalyst were treated with laccase enzyme). We obtained the highest methane yield increase through the chemical treatment that resulted in 66% higher methane production compared to untreated biofibers. The combination of steam treatment with NaOH and subsequent enzymatic treatment increased the methane yield by 34%. To choose the optimal treatment, the energy requirements relative to the energy gain as extra biogas production have to be taken into account, as well as the costs of chemicals or enzymes.

  2. Long-term thermophilic mono-digestion of rendering wastes and co-digestion with potato pulp

    Energy Technology Data Exchange (ETDEWEB)

    Bayr, S., E-mail: suvi.bayr@jyu.fi; Ojanperä, M.; Kaparaju, P.; Rintala, J.

    2014-10-15

    Highlights: • Rendering wastes’ mono-digestion and co-digestion with potato pulp were studied. • CSTR process with OLR of 1.5 kg VS/m{sup 3} d, HRT of 50 d was unstable in mono-digestion. • Free NH{sub 3} inhibited mono-digestion of rendering wastes. • CSTR process with OLR of 1.5 kg VS/m{sup 3} d, HRT of 50 d was stable in co-digestion. • Co-digestion increased methane yield somewhat compared to mono-digestion. - Abstract: In this study, mono-digestion of rendering wastes and co-digestion of rendering wastes with potato pulp were studied for the first time in continuous stirred tank reactor (CSTR) experiments at 55 °C. Rendering wastes have high protein and lipid contents and are considered good substrates for methane production. However, accumulation of digestion intermediate products viz., volatile fatty acids (VFAs), long chain fatty acids (LCFAs) and ammonia nitrogen (NH{sub 4}-N and/or free NH{sub 3}) can cause process imbalance during the digestion. Mono-digestion of rendering wastes at an organic loading rate (OLR) of 1.5 kg volatile solids (VS)/m{sup 3} d and hydraulic retention time (HRT) of 50 d was unstable and resulted in methane yields of 450 dm{sup 3}/kg VS{sub fed}. On the other hand, co-digestion of rendering wastes with potato pulp (60% wet weight, WW) at the same OLR and HRT improved the process stability and increased methane yields (500–680 dm{sup 3}/kg VS{sub fed}). Thus, it can be concluded that co-digestion of rendering wastes with potato pulp could improve the process stability and methane yields from these difficult to treat industrial waste materials.

  3. Protein supplementation of ruminants consuming low-quality cool- or warm-season forage: differences in intake and digestibility.

    Science.gov (United States)

    Bohnert, D W; DelCurto, T; Clark, A A; Merrill, M L; Falck, S J; Harmon, D L

    2011-11-01

    An in situ study (Exp. 1) using 4 ruminally cannulated steers (343 ± 11 kg of BW) in a completely randomized design was used to compare ruminal degradation characteristics of low-quality cool-season (C3; Kentucky bluegrass straw; Poa pratensis; 6.3% CP; DM basis) and warm-season (C4; tallgrass prairie; 5.7% CP; DM basis) forage. Four ruminally cannulated steers (252 ± 8 kg of BW; Exp. 2) and 4 wethers (38 ± 1 kg of BW; Exp. 3) were used in two 2 × 2 factorial arrangements of treatments to determine the influence of supplemental CP (CPSupp; soybean meal; 0.09 and 0.19% of BW, CP basis, for steers and lambs, respectively) on nutrient intake and digestion of C3 and C4 forages. Steers and wethers were allotted to separate 4 × 4 Latin squares that ran simultaneously with 20-d periods. In Exp. 1, C3 had a greater A fraction (fraction of total pool disappearing at a rate too rapid to measure) and effective degradability of DM and NDF compared with C4 (P ruminal liquid retention time (P = 0.02; CPSupp decreased retention by 3.6 h with C4 and by only 0.6 h with C3 forage) and particulate passage rate (P = 0.02; CPSupp increased passage by 46% with C4 and by 10% with C3 forage). As in Exp. 2, a CPSupp × forage interaction (P = 0.01; CPSupp increased digestibility by 18% with C4 and by 7% with C3 forage) was observed with DM digestibility in Exp. 3. In contrast, only N balance (P ruminants are not similar and, more important, that the physiological response of ruminants to protein supplementation of low-quality forage is dependent on forage type.

  4. Protein fraction and digestibility of marandu, xaraes and campo grande grasses in monocropping and intercropping systems under different sowing methods - doi: 10.4025/actascianimsci.v35i1.15134

    Directory of Open Access Journals (Sweden)

    Welma Santos Crunivel

    2013-01-01

    Full Text Available A study was carried out to evaluate the protein fraction and in vitro dry matter digestibility of marandu, xaraes grasses and campo grande in monocropping and intercropping systems under different planting methods, for a period of two years. The experimental design was a complete randomized block with four replications. The treatments consisted of the following crop systems: campo grande in monocropping; xaraés grass in monocropping; marandu grass in monocropping; xaraés intercropped with campo grande in rows; xaraés intercropped with campo grande, broadcast; marandu grass intercropped with campo grande in rows; and marandu intercropped with campo grand, broadcast. The evaluations were conducted for two years, consisting of seasonal evaluations (autumn, winter, spring and summer in the same plots, with repeated measurements over time. The results showed that xaraes and marandu grasses were similar between crop systems, indicating that both can be intercropped with campo grande. The intercropping of campo grande with Brachiaria brizantha cultivars improved the protein fraction and digestibility. The row method of planting provided better protein fractions and in vitro dry matter digestibility.

  5. Níveis de proteína e de arginina digestível na ração pré-inicial de frangos de corte Protein and digestible arginine levels in pre-starter broiler rations

    Directory of Open Access Journals (Sweden)

    Mônica Schaitl Thon

    2010-05-01

    Full Text Available O experimento foi conduzido para avaliar níveis de proteína bruta e arginina digestível na ração pré-inicial de frangos de corte e seus efeitos no desempenho das aves dos 7 aos 21 dias de idade. Foram utilizados 600 pintos da linhagem Cobb 500, distribuídos em delineamento de blocos casualizados em esquema fatorial 4 × 2, composto de quatro níveis de arginina digestível (1,363; 1,463; 1,563 e 1,663% e dois níveis de proteína bruta (20 e 22%, totalizando oito tratamentos, cada um com cinco repetições de 15 aves. Avaliaram-se o ganho de peso, o consumo de ração, o índice de conversão alimentar, a biometria dos órgãos do trato gastrintestinal e a digestibilidade e retenção de matéria seca e nitrogênio. O maior ganho de peso na fase de 1 a 14 dias de idade foi obtido com a ração com 22% de proteína bruta. Os níveis de arginina digestível tiveram efeito quadrático na conversão alimentar na fase de 1 a 10 dias de idade. O peso do esôfago e inglúvio foi maior nas aves alimentadas com a ração com 20% de proteína bruta, no entanto, houve efeito quadrático dos níveis de arginina digestível sobre o comprimento do intestino aos 10 dias de idade e sobre o peso do esôfago + inglúvio aos 3 dias de idade. Houve interação entre os níveis de proteína bruta e arginina digestível para o peso relativo do fígado aos 14 dias, que respondeu de forma quadrática ao nível de 20% de proteína bruta, e para o comprimento de intestino, cujo maior valor foi obtido com os níveis de 22% de proteína bruta e 1,603% de arginina digestível. O balanço e a retenção de nitrogênio foram maiores no nível de 22% de proteína bruta. O nível de 1,363% de arginina digestível atende às exigências nutricionais dos frangos de corte na fase pré-inicial.This experiment was carried out to evaluate levels of crude protein and digestible arginine in pre-starter broiler ration and their effects on the performance of the broilers from 7 to

  6. Study on the In Vitro Simulated Digestion Process of Whey Protein and Effects of Heat Treatment%乳清蛋白的体外模拟消化过程及热处理的影响

    Institute of Scientific and Technical Information of China (English)

    何光华; 黄俊; 尤玉如; 肖功年; 刘士旺; 储小军; 魏培莲; 王仁风

    2012-01-01

    The in vitro pepsin and trypsin digestion process of whey protein and the effects of heat treatments were investigated in this paper. SDS-PAGE analytical results indicated that the effects of pepsin and trypsin on the digestion of whey protein was not very obvious. The dry heat-treatment (75-100 ℃, 1 h) did nearly not affect the in vitro digestion process by trypsin, while undcr the same conditions, the wet heat-treatment could remarkably improve the digestive effect of trypsin on the whey protein. After wet heat-treatment(75 ℃ and 100 ℃, 30 min) and trypsin digestion (8 h) of whey protein, its nitrogen releasing rate reached to 82.1% and 91.3%, respectively. However, the nitrogen releasing rate without wet heat-treatment was only 66.7%. This result suggested that the in vitro digestion process of whey protein was affected by different heat treatments. Wet heat treatment can effectively improve the digestion rate of whey protein, the higher the temperature of wet heat treatment, the faster the in vitro digestion rate of whey protein.%探讨了乳清蛋白体外胃蛋白酶和胰蛋白酶的消化过程,以及热处理对该消化过程的影响.SDS-PAGE分析结果表明,胃蛋白酶和胰蛋白酶对乳清蛋白的体外消化作用的影响不明显.干热处理(75℃和100C,1h)几乎不影响乳清蛋白的体外胰蛋白酶消化过程,而在同样条件下,湿热处理能显著提高胰蛋白酶对乳清蛋白的消化作用.乳清蛋白分别经75℃和100℃湿热处理30 min后,再由胰蛋白酶消化8h,其氮释放率分别达到82.1%和91.3%.而未湿热处理的氮释放率仅为66.7%,表明不同热处理方式对乳清蛋白的体外消化过程产生不同的影响.湿热处理能有效地提高乳清蛋白的消化速率,且湿热处理温度越高,乳清蛋白的体外消化速率越快.

  7. Effect of four processed animal proteins in the diet on digestibility and performance in laying hens.

    NARCIS (Netherlands)

    Krimpen, van M.M.; Veldkamp, T.; Binnendijk, G.P.; Veer, de R.

    2010-01-01

    An experiment was performed to investigate the effect of animal vs. vegetable protein sources in the diet of laying hens on the development of hen performance. A diet containing protein sources of only vegetable origin was compared with 4 diets, each containing 1 of 4 processed animal proteins (PAP)

  8. Effect of homogenization and heat treatment on the behavior of protein and fat globules during gastric digestion of milk.

    Science.gov (United States)

    Ye, Aiqian; Cui, Jian; Dalgleish, Douglas; Singh, Harjinder

    2017-01-01

    The effects of homogenization and heat treatment on the formation and the breakdown of clots during gastric digestion of whole milk were investigated using a human gastric simulator. Homogenization and heat treatment led to formation of coagula with fragmented and crumbled structures compared with the coagulum formed from raw whole milk, but a larger fraction of the protein and more fat globules were incorporated into the coagula induced by action of the milk-clotting enzyme pepsin. The fat globules in the whole milk appeared to be embedded in the clots as they formed. After formation of the clot, the greater numbers of pores in the structures of the clots formed with homogenized milk and heated whole milk led to greater rates of protein hydrolysis by pepsin, which resulted in faster release of fat globules from the clots into the digesta. Coalescence of fat globules occurred both in the digesta and within the protein clots no matter whether they were in homogenized or heated milk samples. The formation of clots with different structures and hence the changes in the rates of protein hydrolysis and the release of milk fat into the digesta in the stomach provide important information for understanding the gastric emptying of milk and the potential to use this knowledge to manipulate the bioavailability of fat and other fat-soluble nutrients in dairy products.

  9. Reducing agitation energy-consumption by improving rheological properties of corn stover substrate in anaerobic digestion.

    Science.gov (United States)

    Tian, Libin; Shen, Fei; Yuan, Hairong; Zou, Dexun; Liu, Yanping; Zhu, Baoning; Li, Xiujin

    2014-09-01

    Rheological properties of corn stover substrate were investigated to explore agitation energy reduction potential for different total solid (TS) in anaerobic digestion. The effects of particle size and temperature on rheological properties and corresponding energy reduction were studied. The results indicated that corn stover slurry exhibited pseudo-plastic flow behavior at TS of 4.23-7.32%, and was well described by Power-law model. At TS of 4.23%, rheological properties were not obviously affected by particle size and temperature. However, when TS was increased to 7.32%, there was 10.37% shear stress reduction by size-reduction from 20 to 80-mesh, and 11.73% shear stress reduction by temperature-increase from 25 to 55 °C. PTS was advanced as variations of power consumption by TS-increase from 4.23% to 7.32%. There was 9.2% PTS-reduction by size-reduction from 20 to 80-mesh at 35 °C. Moreover, PTS-reduction of 10.3%/10 °C was achieved at 20-mesh compared with 9.0%/10 °C at 80-mesh.

  10. ASSESSMENT OF OZONE AS A PRETREATMENT TO IMPROVE ANAEROBIC DIGESTION OF VINASSE

    Directory of Open Access Journals (Sweden)

    S. Aquino

    Full Text Available Abstract This paper presents an assessment of ozone oxidative effects on the biodegradability of sugar cane vinasse, aiming at increasing the methane yield by anaerobic digestion of this effluent. Furthermore, as a new approach, an economic balance of this process was made. Using a bench scale reactor, ozone was applied at 60, 120, 180, 240 mgO3.gCOD-1 doses in raw vinasse and at three initial pH values (4.8, 7 and 9. Applying 60 mgO3.gCOD-1, the biodegradability of vinasse was increased by 22.7% at the initial pH value of 4.8. The application of the two-way ANOVA test indicated a significant statistical interaction between the pH value and ozone. However, a preliminary energy assessment showed that the amount of electricity consumed in a full-scale ozonation plant would be almost 6 times higher than the energy recovered from the combustion of the additional methane produced (13.6%. These results indicate that ozonation of raw vinasse to increase the methane production in a subsequent anaerobic process is economically unfeasible.

  11. Hydrolysis of soybean protein improves iron bioavailability

    Science.gov (United States)

    Iron is an important trace metal element in human body. Iron deficiency affects human health, especially pregnant women and children. Soybean protein is a popular food in Asia and can contain a high amount of iron (145.70±0.74 ug/g); however, it is usually reported as an inhibitor of iron absorption...

  12. Improve biogas production from low-organic-content sludge through high-solids anaerobic co-digestion with food waste.

    Science.gov (United States)

    Liu, Chuanyang; Li, Huan; Zhang, Yuyao; Liu, Can

    2016-11-01

    Anaerobic co-digestion of sewage sludge and food waste was tested at two different total solid (TS) concentrations. In the low-solids group with TS 4.8%, the biogas production increased linearly as the ratio of food waste in substrate increased from 0 to 100%, but no synergetic effect was found between the two substrates. Moreover, the additive food waste resulted in the accumulation of volatile fatty acids and decelerated biogas production. Thus, the blend ratio of food waste should be lower than 50%. While in the high-solids group with TS 14%, the weak alkaline environment with pH 7.5-8.5 avoided excessive acidification but high concentration of free ammonia was a potential risk. However, good synergetic effect was found between the two substrates because the added food waste improved mass transfer in sludge cake. Thus, 50% was recommended as the optimum ratio of food waste in substrate because of the best synergetic effect.

  13. Co-inoculating ruminal content neither provides active hydrolytic microbes nor improves methanization of ¹³C-cellulose in batch digesters.

    Science.gov (United States)

    Chapleur, Olivier; Bize, Ariane; Serain, Thibaut; Mazéas, Laurent; Bouchez, Théodore

    2014-03-01

    Cellulose hydrolysis often limits the kinetics and efficiency of anaerobic degradation in industrial digesters. In animal digestive systems, specialized microorganisms enable cellulose biodegradation at significantly higher rates. This study aims to assess the potential of ruminal microbial communities to settle and to express their cellulolytic properties in anaerobic digesters. Cellulose-degrading batch incubations were co-inoculated with municipal solid waste digester sludge and ruminal content. ¹³C-labeled cellulose degradation was described over time with Gas Chromatography-Combustion-Isotope Ratio Mass Spectrometry. Results were linked to the identification of the microorganisms assimilating ¹³C and to the monitoring of their relative dynamics. Cellulose degradation in co-inoculated incubations was efficient but not significantly improved. Transient disturbances in degradation pathways occurred, as revealed by propionate accumulation. Automated Ribosomal Intergenic Spacer Analysis dynamics and pyrosequencing revealed that expected classes of Bacteria and Archaea were active and degraded cellulose. However, despite the favorable co-inoculation conditions, molecular tools also revealed that no ruminal species settled in the bioreactors. Other specific parameters were probably needed for this to happen. This study shows that exploiting the rumen's cellulolytic properties in anaerobic digesters is not straightforward. Co-inoculation can only be successful if ruminal microorganisms manage to thrive in the anaerobic digester and outcompete native microorganisms, which requires specific nutritional and environmental parameters, and a meticulous reproduction of the selection pressure encountered in the rumen.

  14. 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

    OpenAIRE

    Lawton, Clare; Walton, Jenny; Hoyland, Alexa; Howarth, Elaine; Allan, Peter; Chesters, David; Dye, Louise

    2013-01-01

    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 subjectiv...

  15. Rice protein improves adiposity, body weight and reduces lipids level in rats through modification of triglyceride metabolism

    Directory of Open Access Journals (Sweden)

    Yang Lin

    2012-02-01

    Full Text Available Abstract Background To elucidate whether rice protein can possess a vital function in improving lipids level and adiposity, the effects of rice proteins extracted by alkaline (RP-A and α-amylase (RP-E on triglyceride metabolism were investigated in 7-week-old male Wistar rats fed cholesterol-enriched diets for 2 weeks, as compared with casein (CAS. Results Compared with CAS, plasma concentrations of glucose and lipids were significantly reduced by RP-feeding (P P P P P > 0.05. There was a significant positive correlation between protein digestibility and deposit fat (r = 0.8567, P P Conclusions The present study demonstrates that rice protein can modify triglyceride metabolism, leading to an improvement of body weight and adiposity. Results suggest that the triglyceride-lowering action as well as the potential of anti-adiposity induced by rice protein is attributed to upregulation of lipolysis and downregulation of lipogenesis, and the lower digestibility of rice protein may be the main modulator responsible for the lipid-lowering action.

  16. CuSO4-TiO2 as Kjeldahl digestion catalyst in manual determination of crude protein in animal feeds.

    Science.gov (United States)

    Kane, P F

    1986-01-01

    The official AOAC manual Kjeldahl methods for determining crude protein in animal feeds have several disadvantages. For the HgO catalyst method, there are environmental concerns and a lengthy digestion. For the CuSO4 catalyst method, the digestion period is shorter, but still 90 min. A different catalyst combination, CuSO4-TiO2, makes 40 min digestion feasible. Comparison of these catalysts on a group of representative feeds resulted in a mean difference, Cu-Ti minus HgO, of 0.034% protein. Standard deviation of the differences was 0.36. A Student's t-test showed no significant difference. The method will be collaboratively studied.

  17. Improved Functional Characteristics of Whey Protein Hydrolysates in Food Industry

    OpenAIRE

    Jeewanthi, Renda Kankanamge Chaturika; Lee, Na-Kyoung; Paik, Hyun-Dong

    2015-01-01

    This review focuses on the enhanced functional characteristics of enzymatic hydrolysates of whey proteins (WPHs) in food applications compared to intact whey proteins (WPs). WPs are applied in foods as whey protein concentrates (WPCs), whey protein isolates (WPIs), and WPHs. WPs are byproducts of cheese production, used in a wide range of food applications due to their nutritional validity, functional activities, and cost effectiveness. Enzymatic hydrolysis yields improved functional and nutr...

  18. Stability of Trans-Resveratrol Encapsulated in a Protein Matrix Produced Using Spray Drying to UV Light Stress and Simulated Gastro-Intestinal Digestion.

    Science.gov (United States)

    Koga, Clarissa C; Andrade, Juan E; Ferruzzi, Mario G; Lee, Youngsoo

    2016-02-01

    Trans-resveratrol has demonstrated the potential to provide both therapeutic and preventive activities against chronic diseases such as heart disease and cancer. The incorporation of trans-resveratrol into food products would allow for broader access of this bioactive compound to a larger population. However, this strategy is limited by instability of trans-resveratrol under environmental conditions and within the digestive system leading to isomerization of trans-resveratrol (bioactive form) to cis-resveratrol (bio-inactive form). Studies in the stabilization of trans-resveratrol into protein microparticles are presented. Trans-resveratrol was encapsulated using whey protein concentrate (WPC) or sodium caseinate (SC), with or without anhydrous milk fat (AMF). Binding of resveratrol and aromatic residues in protein was estimated utilizing the Stern-Volmer equation and the number of tryptophan residues. The stability of encapsulated resveratrol was evaluated after exposure to ultraviolet A (UVA) light and 3-stage in vitro digestion. After UVA light exposure, SC-based microcapsules maintained a higher trans:cis resveratrol ratio (0.63, P < 0.05) than WPC-based microcapsules (0.43) and unencapsulated resveratrol (0.49). In addition, encapsulation of resveratrol in both protein microparticles led to an increased digestive stability and bioaccessibility in comparison to unencapsulated resveratrol (47% and 23%, respectively, P < 0.05). SC-based microcapsules provided a higher digestive stability and bioaccessibility (86% and 81%; P < 0.05) compared to WPC-based microcapsules (71% and 68%). The addition of AMF to the microcapsules did not significantly change the in vitro digestion values. In conclusion, SC-based microencapsulation increased the stability of trans-resveratrol to UVA light exposure and simulated digestion conditions. This encapsulation-system-approach can be extended to other labile, bioactive polyphenols.

  19. Apparent and true digestibility of protein and amino acid in feedstuffs used in Nile Tilapia feed as determined by the technique of dissection

    Directory of Open Access Journals (Sweden)

    Felipe Barbosa Ribeiro

    2012-05-01

    Full Text Available The objective of this study was to determine the coefficients of apparent and true digestibility of protein and amino acids of five feedstuffs (corn, wheat bran, soybean meal, corn gluten meal and fish meal in 900 sex-reversed Nile tilapia (Oreochromis niloticus of the Thai strain during the growth phase, with an initial weight of 315±8.45 g. A total of 750 fish were distributed according to a randomized block design (repetitions in time into five treatments with six replicates of 25 fish each. The remaining 150 fish were fed a protein-free diet to measure endogenous protein and amino acid losses in order to determine the true digestibility of these components. Each tested diet contained a single protein source, which consisted of one of the evaluated feedstuffs. Digestibility was indirectly estimated using chromic oxide at 0.50% as marker. Digesta was collected from the last 5 cm of the distal intestine (between the ileal-rectal valve and the anus using the dissection technique. Apparent digestibility coefficients of protein and amino acids were, on average, 74.69 and 73.62% for corn, 73.74 and 72.81% for wheat bran, 86.01 and 84.66% for soybean meal, 85.19 and 84.29% for corn gluten meal, 76.74 and 75.56% for fish meal, respectively. True digestibility coefficients of protein and amino acids were, on average, 85.21 and 83.97% for corn, 84.41 and 83.74% for wheat bran, 87.22 and 87.51% for soybean meal, 87.97 and 87.34% for corn gluten meal, and finally 79.58 and 78.44% for fish meal, respectively.

  20. Ileal and total tract apparent crude protein and amino acid digestibility of ensiled and dried cassava leaves and sweet potato vines in growing pigs

    NARCIS (Netherlands)

    Nguyen, T.H.L.; Ngoan, L.D.; Bosch, G.; Verstegen, M.W.A.; Hendriks, W.H.

    2012-01-01

    The present study was conducted to determine the ileal and total tract apparent digestibility of crude protein (CP) and amino acids (AA) in ensiled and dried cassava (Manihot esculenta) leaves (CL) and sweet potato (Ipomoea batatas) vines (SPV) as a single ingredient or in a 50:50 mixture of CL + SP

  1. A new calibrant for MALDI-TOF-TOF-PSD-MS/MS of non-digested proteins for top-down proteomic analysis

    Science.gov (United States)

    RATIONALE: Matrix-assisted laser desorption/ionization (MALDI) time-of-flight-time-of-flight (TOF-TOF) tandem mass spectrometry (MS/MS) has seen increasing use for post-source decay (PSD)-MS/MS analysis of non-digested protein ions for top-down proteomic identification. However, there is no commonl...

  2. Development and use of a monoclonal antibody to detect semi-digested proteins of the English grain aphid, Sitobion avenae, in the guts of ladybird beetle predators

    NARCIS (Netherlands)

    Gao, S.J.; Zhou, X.R.; Pang, B.P.; Loon, van J.J.A.; Zhao, G.Q.

    2009-01-01

    A monoclonal antibody (McAb), EGA-4A9, was developed to detect the semi-digested proteins of the English grain aphid, Sitobion avenae (Fabricius) (Hemiptera: Aphididae), in predatory ladybird beetles (species of the genera Adonia, Coccinella, Hippodamia, and Propylea) using the gut homogenate of Ado

  3. Buffalo cheese whey proteins, identification of a 24 kda protein and characterization of their hydrolysates: in vitro gastrointestinal digestion

    OpenAIRE

    Juliana C Bassan; Goulart,Antonio J.; Nasser, Ana L. M.; Bezerra, Thaís M. S. [UNESP; Garrido, Saulo S.; Rustiguel, Cynthia B.; Guimarães, Luis H. S.; Rubens Monti

    2015-01-01

    Milk whey proteins are well known for their high biological value and versatile functional properties, characteristics that allow its wide use in the food and pharmaceutical industries. In this work, a 24 kDa protein from buffalo cheese whey was analyzed by mass spectrometry and presented homology with Bos taurus beta-lactoglobulin. In addition, the proteins present in buffalo cheese whey were hydrolyzed with pepsin and with different combinations of trypsin, chymotrypsin and carboxypeptidase...

  4. Monolithic liquid-chromatography columns for protein analysisprotein digest separation and integrated systems

    NARCIS (Netherlands)

    van de Meent, M.H.M.

    2010-01-01

    The objectives of the research described in this thesis are to evaluate the applicability of both silica-based and polymeric monolithic columns for protein analysis. The first part describes investigations into the effects of column length and stationary-phase chemistry on the separation of protein

  5. 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.

  6. Effect of germination periods and hydrothermal treatments on in vitro protein and starch digestibility of germinated legumes

    OpenAIRE

    Uppal, Veny; Bains, Kiran

    2011-01-01

    Germination of legumes followed by hydrothermal treatments is an effective mean of improving nutritive value of legumes. The protein content of mungbean, chickpea and cowpea increased by 9–11, 11–16 and 8–11% after germination. A significant (p ≤ 0.05) decrease in protein content was observed on pressure cooking and microwaving in all three legumes. The carbohydrates decreased by 1 to 3% during soaking and 2 to 6% during germination. A significant (p ≤ 0.05) improvement in in vitro protein di...

  7. Simplified sample preparation method for protein identification by matrix-assisted laser desorption/ionization mass spectrometry: in-gel digestion on the probe surface

    DEFF Research Database (Denmark)

    Stensballe, A; Jensen, Ole Nørregaard

    2001-01-01

    Identification and detailed characterization of complex mixtures of proteins separated by polyacrylamide gel electrophoresis (PAGE) require optimized and robust methods for interfacing electrophoretic techniques to mass spectrometry. Peptide mapping by matrix-assisted laser desorption/ionization-......Identification and detailed characterization of complex mixtures of proteins separated by polyacrylamide gel electrophoresis (PAGE) require optimized and robust methods for interfacing electrophoretic techniques to mass spectrometry. Peptide mapping by matrix-assisted laser desorption...... for protein identification similar to that obtained by the traditional protocols for in-gel digestion and MALDI peptide mass mapping of human proteins, i.e. approximately 60%. The overall performance of the novel on-probe digestion method is comparable with that of the standard in-gel sample preparation...

  8. Effect of trypsin inhibitor activity in soya bean on growth performance, protein digestibility and incidence of sub-clinical necrotic enteritis in broiler chicken flocks.

    Science.gov (United States)

    Palliyeguru, M W C D; Rose, S P; Mackenzie, A M

    2011-06-01

    1. The effect of three different levels of dietary trypsin inhibitor activity (achieved by varying the amount of non-toasted full fat soya bean in replacement for toasted full fat soya bean) on the incidence of spontaneously-occurring sub-clinical necrotic enteritis (NE) in broiler chickens was compared. A fourth dietary treatment compared the effect of a diet that used potato protein concentrate as the major protein source. The determined trypsin inhibitor activity increased with the increasing content of non-toasted soya bean: 1·90, 6·21, 8·46 and 3·72 mg/g for the three soya bean diets (0, 100 and 200 g of non-toasted soya bean/kg) and the potato protein diet respectively. 2. Although increasing amounts of the non-toasted full-fat soya bean increased the feed intakes of the birds, there was a marked reduction in protein digestibility, weight gain and feed conversion efficiency. 3. There was a linear increase in sub-clinical NE lesions in the duodenum, jejunum, mid small intestine and ileum with increasing non-toasted soya bean. Caecal Clostridium perfringens counts increased with the increasing dietary content of non-toasted soya bean. Serum α-toxin antibodies were higher in the birds fed the 200 g non-toasted soya bean/kg diet compared with the other diets. 4. The results demonstrated that variation in the amount of non-toasted dietary soya bean not only affects growth performance of broilers but also affects the incidence of sub-clinical necrotic enteritis in the flock. Ensuring the lowest possible trypsin-inhibitor activity in soya bean samples is a valuable tool to improve the health and welfare of birds and in reducing the financial losses from this disease.

  9. 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

    OpenAIRE

    Louise Dye; David Chesters; Peter Allan; Elaine Howarth; Jenny Walton; Alexa Hoyland; Clare L Lawton

    2013-01-01

    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 (

  10. 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

    OpenAIRE

    Lawton, CL; Walton, J; Hoyland, A; Howarth, E.; Allan, P; Chesters, D; Dye, L.

    2013-01-01

    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 (

  11. 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.

    Science.gov (United States)

    Lawton, Clare L; Walton, Jenny; Hoyland, Alexa; Howarth, Elaine; Allan, Peter; Chesters, David; Dye, Louise

    2013-04-22

    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 (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.

  12. 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.

  13. Protein breakdown and release of β-casomorphins during in vitro gastro-intestinal digestion of sterilised model systems of liquid infant formula.

    Science.gov (United States)

    Cattaneo, Stefano; Stuknytė, Milda; Masotti, Fabio; De Noni, Ivano

    2017-02-15

    Protein modifications occurring during sterilisation of infant formulas can affect protein digestibility and release of bioactive peptides. The effect of glycation and cross-linking on protein breakdown and release of β-casomorphins was evaluated during in vitro gastro-intestinal digestion (GID) of six sterilised model systems of infant formula. Protein degradation during in vitro GID was evaluated by SDS-PAGE and by measuring the nitrogen content of ultrafiltration (3kDa) permeates before and after in vitro GID of model IFs. Glycation strongly hindered protein breakdown, whereas cross-linking resulting from β-elimination reactions had a negligible effect. Only β-casomorphin 7 (β-CM7) was detected (0.187-0.858mgL(-1)) at the end of the intestinal digestion in all untreated IF model systems. The level of β-CM7 in the sterilised model systems prepared without addition of sugars ranged from 0.256 to 0.655mgL(-1). The release of this peptide during GID was hindered by protein glycation.

  14. DIGEST MATERIALS FOR IMPROVING AND EXTENDING THE JUNIOR HIGH SCHOOL ORCHESTRA REPERTORY. VOLUME 1, BAROQUE.

    Science.gov (United States)

    MOORE, JUNE

    PREPARED AS PART OF "PROJECT IMPROVING AND EXTENDING THE JUNIOR HIGH SCHOOL ORCHESTRA REPERTORY," THIS VOLUME CONTAINS CURRICULAR MATERIALS REPRESENTING THE BAROQUE PERIOD. A MUSICAL HISTORY OF THE PERIOD IS GIVEN, AS WELL AS HISTORIES OF THE COMPOSERS AND THEIR INDIVIDUAL COMPOSITIONS. THE MATERIALS ARE PREPARED FOR THREE DEGREES OF TECHNICAL…

  15. Milk protein tailoring to improve functional and biological properties

    Directory of Open Access Journals (Sweden)

    JEAN-MARC CHOBERT

    2012-01-01

    Full Text Available Proteins are involved in every aspects of life: structure, motion, catalysis, recognition and regulation. Today's highly sophisticated science of the modifications of proteins has ancient roots. The tailoring of proteins for food and medical uses precedes the beginning of what is called biochemistry. Chemical modification of proteins was pursued early in the twentieth century as an analytical procedure for side-chain amino acids. Later, methods were developed for specific inactivation of biologically active proteins and titration of their essential groups. Enzymatic modifications were mainly developed in the seventies when many more enzymes became economically available. Protein engineering has become a valuable tool for creating or improving proteins for practical use and has provided new insights into protein structure and function. The actual and potential use of milk proteins as food ingredients has been a popular topic for research over the past 40 years. With today's sophisticated analytical, biochemical and biological research tools, the presence of compounds with biological activity has been demonstrated. Improvements in separation techniques and enzyme technology have enabled efficient and economic isolation and modification of milk proteins, which has made possible their use as functional foods, dietary supplements, nutraceuticals and medical foods. In this review, some chemical and enzymatic modifications of milk proteins are described, with particular focus on their functional and biological properties.

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

    Directory of Open Access Journals (Sweden)

    Takahiro Ogura

    Full Text Available 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 δconglutin, 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.

  17. Identification of a Low Digestibility δ-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 δconglutin, 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

  18. [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.

  19. In situ identification and quantification of protein-hydrolyzing ruminal bacteria associated with the digestion of barley and corn grain.

    Science.gov (United States)

    Xia, Yun; Kong, Yunhong; Huang, Heping; Yang, Hee Eun; Forster, Robert; McAllister, Tim A

    2016-12-01

    In this study, BODIPY FL DQ™ casein staining combined with fluorescence in situ hybridization (FISH) was used to detect and identify protein-hydrolyzing bacteria within biofilms that produced active cell-surface-associated serine- and metallo-proteases during the ruminal digestion of barley and corn grain in cows fed barley-based diets at 2 different levels. A doublet coccoid bacterial morphotype associated with barley and corn grain particles fluoresced after BODIPY FL DQ™ casein staining. Bacteria with this morphotype accounted for 3%-10% of the total bacteria attached to surface of cereal grain particles, possibly indicative of an important role in the hydrolysis of the protein matrix within the endosperm. However, the identity of these predominant proteolytic bacteria could not be determined using FISH. Quantitative FISH revealed that known proteolytic species, Prevotella ruminicola, Ruminobacter amylophilus, and Butyrivibrio fibrisolvens, were attached to particles of various cultivars of barley grain and corn, confirming their role in the proteolysis of cereal grains. Differences in chemical composition among different barley cultivars did not affect the composition of proteolytic bacterial populations. However, the concentrate level in the basal diet did have an impact on the relative abundance of proteolytic bacteria and thus possibly their overall contribution to the proteolysis of cereal grains.

  20. Wild Raspberry Subjected to Simulated Gastrointestinal Digestion Improves the Protective Capacity against Ethyl Carbamate-Induced Oxidative Damage in Caco-2 Cells.

    Science.gov (United States)

    Chen, Wei; Xu, Yang; Zhang, Lingxia; Li, Ya; Zheng, Xiaodong

    2016-01-01

    Ethyl carbamate (EC), a probable human carcinogen, occurs widely in many fermented foods. Previous studies indicated that EC-induced cytotoxicity was associated with oxidative stress. Wild raspberries are rich in polyphenolic compounds, which possess potent antioxidant activity. This study was conducted to investigate the protective effect of wild raspberry extracts produced before (RE) and after in vitro simulated gastrointestinal digestion (RD) on EC-induced oxidative damage in Caco-2 cells. Our primary data showed that ethyl carbamate could result in cytotoxicity and genotoxicity in Caco-2 cells and raspberry extract after digestion (RD) may be more effective than that before digestion (RE) in attenuating toxicity caused by ethyl carbamate. Further investigation by fluorescence microscope revealed that RD may significantly ameliorate EC-induced oxidative damage by scavenging the overproduction of intracellular reactive oxygen species (ROS), maintaining mitochondrial function and preventing glutathione (GSH) depletion. In addition, HPLC-ESI-MS results showed that the contents of identified polyphenolic compounds (esculin, kaempferol O-hexoside, and pelargonidin O-hexoside) were remarkably increased after digestion, which might be related to the better protective effect of RD. Overall, our results demonstrated that raspberry extract undergoing simulated gastrointestinal digestion may improve the protective effect against EC-induced oxidative damage in Caco-2 cells.

  1. Gluten-free pasta incorporating chia (Salvia hispanica L.) as thickening agent: An approach to naturally improve the nutritional profile and the in vitro carbohydrate digestibility.

    Science.gov (United States)

    Menga, Valeria; Amato, Mariana; Phillips, Tim D; Angelino, Donato; Morreale, Federico; Fares, Clara

    2017-04-15

    A gluten-free pasta was prepared adding chia at rice flour for testing the thickening and nutritional properties of this specie. Chemical analysis showed chia is a source of protein (19.52% and 15.81%, seeds and mucilage respectively), insoluble/soluble dietary fiber ratio (4.3 and 1.79 seeds and mucilage respectively), fat and ash content. The total phenolic acids content ranged from 734.5μg/g to 923.9μg/g for seeds and mucilage respectively. Chia was a good thickening agent and, improved the nutritional profile of enriched samples compared to CGF. After cooking TPAs increased in all samples, ranging from 5.3% in DW to 52.8% in CM5. The addition of chia seeds also increased the slowly digestible starch fraction of rice flour, commonly known to have a high glycemic index. Results suggest that chia should be added as thickening agent in the formulation of GF pasta for conferring healthier characteristics.

  2. Milk protein for improved metabolic health: a review of the evidence

    Science.gov (United States)

    2013-01-01

    Epidemiological evidence shows that consumption of dairy products is associated with decreased prevalence of metabolic related disorders, whilst evidence from experimental studies points towards dairy protein as a dietary component which may aid prevention of type 2 diabetes (T2DM). Poor metabolic health is a common characteristic of overweight, obesity and aging, and is the forerunner of T2DM and cardiovascular disease (CVD), and an ever increasing global health issue. Progressive loss of metabolic control is evident from a blunting of carbohydrate, fat and protein metabolism, which is commonly manifested through decreased insulin sensitivity, inadequate glucose and lipid control, accompanied by a pro-inflammatory environment and hypertension. Adverse physiological changes such as excess visceral adipose tissue deposition and expansion, lipid overspill and infiltration into liver, muscle and other organs, and sarcopaenia or degenerative loss of skeletal muscle mass and function all underpin this adverse profile. ‘Sarcobesity’ and sarcopaenic diabetes are rapidly growing health issues. As well as through direct mechanisms, dairy protein may indirectly improve metabolic health by aiding loss of body weight and fat mass through enhanced satiety, whilst promoting skeletal muscle growth and function through anabolic effects of dairy protein-derived branch chain amino acids (BCAAs). BCAAs enhance muscle protein synthesis, lean body mass and skeletal muscle metabolic function. The composition and processing of dairy protein has an impact on digestion, absorption, BCAA kinetics and function, hence the optimisation of dairy protein composition through selection and combination of specific protein components in milk may provide a way to maximize benefits for metabolic health. PMID:23822206

  3. A continuum of research projects to improve extraction of oil and proteins in oilseed plants

    Directory of Open Access Journals (Sweden)

    Miquel Martine

    2011-05-01

    Full Text Available A key challenge in the actual context of fossil sources rarefaction, global warming, and of increase of the world global population, is to promote the use of molecules derived from renewable sources such as plants. Among these molecules, lipids and proteins are targets of interest. Plant lipids from oilseeds are attractive substitutes to the use of fossil oil. Till the beginning of the 20th century, numerous products used in the daily life were derived from natural renewable products. For instance, plant oil was commonly used as fuel for vehicles and was entering in the composition of paintings, lubricants etc. Unfortunately, natural oils have been progressively replaced by cheaper fossil oil in the fabrication of these products. Nowadays, fossil oils are becoming increasingly expensive being a finite comodity. It is thus important to reduce our dependence from fossil oil and develop substitution industries. Oilseeds contain important amounts of proteins which are mainly used in feed. As several kilograms of plant protein are needed to obtain one kilogram of animal protein, the interest toward using plant protein in food is reinforced. The developments of the use of plant lipids, as well as proteins are a major stakes for the competitiveness of European agriculture and industry, as well as for sustainable development. Extraction of oil and proteins from rapeseed has a significant cost, in term of energy and solvent uses, and finally affects the ultimate quality of the products (protein digestibility. In order to quantitatively extract seed reserves under mild conditions, it will be necessary to limit the amount of energy needed, and avoid any use of solvents. Ideally, seeds should be processed in a bio refinery. In this paper, we will describe how oilseeds store their reserves, and roadblocks for improving actual oilseed extraction processes. A continuum of research projects aimed at answering targeted questions will be presented, with selected

  4. Influence of stage of maturity of grass silages on protein digestion and microbial protein synthesis in the rumen.

    NARCIS (Netherlands)

    Bosch, M.W.; Bruchem, van J.; Bongers, L.J.G.M.; Tamminga, S.

    1994-01-01

    In four change-over experiments, wilted grass silages, differing in growth stage at harvesting and as a consequence in cell wall content, were fed ad libitum to dairy cows in early and late lactation. Ruminal degradation rate of the crude protein fraction of the silages was investigated using nylon

  5. Improving the energy balance of grass-based anaerobic digestion through harvesting optimization

    DEFF Research Database (Denmark)

    Tsapekos, Panagiotis; Kougias, Panagiotis; Egelund, H.

    to determine the treatment with the best energy balance, the energy demand during harvesting was compared to the practical methane yield. The biomethanation process was evaluated using triplicate batch assays under thermophilic conditions following the guidelines of the biochemical methane potential (BMP...... that the appropriate harvester can improve the energy output by approximately 2.4 GJ/ha under optimal conditions and subsequently, the overall sustainability of grass-based AD....... efficiency is still doubted and thus, they are not widely used in full-scale biogas plants. In addition, these methods are typically associated with increased costs or energy demands. Hence, there is a need to find effective and cost-efficient solutions that boost biomass decomposition. In this concept, one...

  6. Development and Optimization of a Flocculation Procedure for Improved Solid-Liquid Separation of Digested Biomass

    Energy Technology Data Exchange (ETDEWEB)

    Patton, Caroline; Lischeske, James J.; Sievers, David A.

    2015-11-03

    One viable treatment method for conversion of lignocellulosic biomass to biofuels begins with saccharification (thermochemical pretreatment and enzymatic hydrolysis), followed by fermentation or catalytic upgrading to fuels such as ethanol, butanol, or other hydrocarbons. The post-hydrolysis slurry is typically 4-8 percent insoluble solids, predominantly consisting of lignin. Suspended solids are known to inhibit fermentation as well as poison catalysts and obstruct flow in catalyst beds. Thus a solid-liquid separation following enzymatic hydrolysis would be highly favorable for process economics, however the material is not easily separated by filtration or gravimetric methods. Use of a polyacrylamide flocculant to bind the suspended particles in a corn stover hydrolyzate slurry into larger flocs (1-2mm diameter) has been found to be extremely helpful in improving separation. Recent and ongoing research on novel pretreatment methods yields hydrolyzate material with diverse characteristics. Therefore, we need a thorough understanding of rapid and successful flocculation design in order to quickly achieve process design goals. In this study potential indicators of flocculation performance were investigated in order to develop a rapid analysis method for flocculation procedure in the context of a novel hydrolyzate material. Flocculation conditions were optimized on flocculant type and loading, pH, and mixing time. Filtration flux of the hydrolyzate slurry was improved 170-fold using a cationic polyacrylamide flocculant with a dosing of approximately 22 mg flocculant/g insoluble solids at an approximate pH of 3. With cake washing, sugar recovery exceeded 90 percent with asymptotic yield at 15 L wash water/kg insoluble solids.

  7. 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 μmolGly/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.

  8. 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.

  9. Identification and Characterization of a 25 kDa Protein That Is Indispensable for the Efficient Saccharification of Eisenia bicyclis in the Digestive Fluid of Aplysia kurodai

    Science.gov (United States)

    Tsuji, Akihiko; Kuwamura, Shuji; Shirai, Akihiro; Yuasa, Keizo

    2017-01-01

    The digestive fluid of the sea hare Aplysia kurodai can liberate approximately 2.5 mg of glucose from 10 mg of dried Eisenia bicyclis powder. Although laminaran, a major storage polysaccharide in E. bicyclis, is easily digested to glucose by the synergistic action of the 110 and 210 kDa A. kurodai β-glucosidases (BGLs), glucose is not liberated from E. bicyclis by direct incubation with these BGLs. To clarify this discrepancy, we searched for an Eisenia hydrolysis enhancing protein (EHEP) in the digestive fluid of A. kurodai. A novel 25 kDa protein that enhances E. bicyclis saccharification by β-glucosidases was purified to a homogeneous state from the digestive fluid of A. kurodai, and its cDNA was cloned from total cDNAs reverse-transcribed from hepatopancreas total RNA. The E. bicyclis extract strongly inhibited BGLs, suggesting some compound within this brown alga functioned as a feeding deterrent. However, when E. bicyclis was incubated with BGLs in the presence of EHEP, glucose production was markedly increased. As E. bicyclis is rich in phlorotannin, which are only found in brown algae, our study suggested that these compounds are the main BGL inhibitors in E. bicyclis extract. EHEP protects BGLs from phlorotannin inhibition by binding to phlorotannins and forming an insoluble complex with phloroglucinol and phlorotannins. These findings indicated that EHEP plays a key role in the saccharification of brown seaweeds containing phlorotannins in the digestive fluid of A. kurodai. This is the first report of EHEP as a phlorotannin-binding protein that protects BGLs from inhibition. PMID:28129373

  10. Full-scale application of focused-pulsed pre-treatment for improving biosolids digestion and conversion to methane.

    Science.gov (United States)

    Rittmann, Bruce E; Lee, Hyung-sool; Zhang, Husen; Alder, Jared; Banaszak, James E; Lopez, Ronald

    2008-01-01

    We tested at full-scale the innovative Focused Pulsed (FP) technology for pre-treating waste sludge in order to improve methane gas production and biosolids reduction in sludge digestion, but without incurring problems of odors, toxicity, and high costs for chemical or energy consumption. FP pre-treatment of a mixture of primary and secondary sludge increased the soluble COD by 160% and DOC 120% over the control. FP pre-treatment of 63% of the input waste sludge increased biogas production by over 40% and reduced biosolids requiring disposal by 30% when compared to the plant baseline. FP pre-treatment also correlated with a shift of the bacterial and archaeal communities. The most significant change was that the acetate-cleaving Methanosaeta became the dominant methanogen. Full FP pre-treatment should increase biogas production and biosolids removal by 60% and 40%, respectively. Full FP pre-treatment should generate energy benefits of at least 2.7 times and as high as 18 times the FP energy input, depending on heat recovery from FP treatment. For a plant treating 76,000 m3/d of wastewater (380 m3-sludge/d), FP treatment should generate an annual economic benefit of approximately $540,000 net of electricity and other operating and maintenance costs. This represents a payback period of three years or less.

  11. Improving protein-protein interactions prediction accuracy using protein evolutionary information and relevance vector machine model.

    Science.gov (United States)

    An, Ji-Yong; Meng, Fan-Rong; You, Zhu-Hong; Chen, Xing; Yan, Gui-Ying; Hu, Ji-Pu

    2016-10-01

    Predicting protein-protein interactions (PPIs) is a challenging task and essential to construct the protein interaction networks, which is important for facilitating our understanding of the mechanisms of biological systems. Although a number of high-throughput technologies have been proposed to predict PPIs, there are unavoidable shortcomings, including high cost, time intensity, and inherently high false positive rates. For these reasons, many computational methods have been proposed for predicting PPIs. However, the problem is still far from being solved. In this article, we propose a novel computational method called RVM-BiGP that combines the relevance vector machine (RVM) model and Bi-gram Probabilities (BiGP) for PPIs detection from protein sequences. The major improvement includes (1) Protein sequences are represented using the Bi-gram probabilities (BiGP) feature representation on a Position Specific Scoring Matrix (PSSM), in which the protein evolutionary information is contained; (2) For reducing the influence of noise, the Principal Component Analysis (PCA) method is used to reduce the dimension of BiGP vector; (3) The powerful and robust Relevance Vector Machine (RVM) algorithm is used for classification. Five-fold cross-validation experiments executed on yeast and Helicobacter pylori datasets, which achieved very high accuracies of 94.57 and 90.57%, respectively. Experimental results are significantly better than previous methods. To further evaluate the proposed method, we compare it with the state-of-the-art support vector machine (SVM) classifier on the yeast dataset. The experimental results demonstrate that our RVM-BiGP method is significantly better than the SVM-based method. In addition, we achieved 97.15% accuracy on imbalance yeast dataset, which is higher than that of balance yeast dataset. The promising experimental results show the efficiency and robust of the proposed method, which can be an automatic decision support tool for future

  12. 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-1β and TNF-α and up-regulated IFN-γ, 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

  13. 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.

    2012-01-01

    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 (p<0.05) and an increase in intestinal digestibility that could be explained both by a linear and a quadratic model for total AA and most individual AA (p<0.05). The 130°C treatment decreased ruminal degradability 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

  14. potentially pathogenic gastro-intestinal microorganisms (ID 1030, 2956, 2958, 2961, 2963, 2966, 2970), improved lactose digestion (ID 1030, 2956, 2958, 2961, 2963, 2966, 2970), “intestinal flora/digestive health” (ID 4231), defence against vaginal pathogens (ID 2950, 2957, 2967) and increasing IL-10

    DEFF Research Database (Denmark)

    Tetens, Inge

    and reduction of gastro-intestinal discomfort, decreasing potentially pathogenic gastro-intestinal microorganisms, improved lactose digestion, “intestinal flora/digestive health”, defence against vaginal pathogens and increasing IL-10 production and/or enhancing the activity of natural killer cells. The food...

  15. 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.

  16. Isolation and Identification of a Bacterium from Marine Shrimp Digestive Tract: A New Degrader of Starch and Protein

    Institute of Scientific and Technical Information of China (English)

    LI Jiqiu; TAN Beiping; MAI Kangsen

    2011-01-01

    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-sporeforming,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.

  17. Effects of a sweetpotato protein digest on lipid metabolism in mice administered a high-fat diet

    Directory of Open Access Journals (Sweden)

    Koji Ishiguro

    2016-12-01

    Full Text Available Sweetpotato peptide (SPP was prepared by enzyme digestion of sweetpotato protein from starch wastewater. Animal experiments assessed the effect of SPP on body weight, abdominal adipose tissue mass, serum lipids and adipocytokines. Body and liver weight and epididymal and mesenteric fat of mice fed a high-fat diet containing 0.5% or 5% SPP for 28 days were significantly lower than control mice. Triglyceride and cholesterol in VLDL and LDL and leptin levels were significantly lower in the serum of SPP-administered mice compared to control mice. Biomarker arrays showed that adiponectin, melanocyte-stimulating-hormone-alpha and neuromedin U were more than 1.5 times higher, while TNF-alpha was about 1.5 times lower in the livers of SPP-administered mice compared to control mice. These results suggest SPP mitigated leptin resistance in mice administered a high-fat diet, and maintained anorexigenic peptide levels. SPP administration may suppress lipogenesis by increasing adiponectin levels and decreasing TNF-alpha levels in adipocytes.

  18. Improved microwave-assisted wet digestion procedures for accurate Se determination in fish and shellfish by flow injection-hydride generation-atomic absorption spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Lavilla, I. [Departamento de Quimica Analitica y Alimentaria, Area de Quimica Analitica, Universidad de Vigo, Facultad de Ciencias (Quimica), As Lagoas - Marcosende s/n, 36310 Vigo (Spain); Gonzalez-Costas, J.M. [Departamento de Quimica Analitica y Alimentaria, Area de Quimica Analitica, Universidad de Vigo, Facultad de Ciencias (Quimica), As Lagoas - Marcosende s/n, 36310 Vigo (Spain); Bendicho, C. [Departamento de Quimica Analitica y Alimentaria, Area de Quimica Analitica, Universidad de Vigo, Facultad de Ciencias (Quimica), As Lagoas - Marcosende s/n, 36310 Vigo (Spain)]. E-mail: bendicho@uvigo.es

    2007-05-22

    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 HNO{sub 3} or the mixture HNO{sub 3}/H{sub 2}O{sub 2} and further thermal reduction of the Se(VI) formed to Se(IV) were evaluated. These procedures were as follows: (I) without H{sub 2}O{sub 2} and without heating to dryness; (II) without H{sub 2}O{sub 2} and with heating to dryness; (III) with H{sub 2}O{sub 2} and without heating to dryness; (IV) with H{sub 2}O{sub 2} 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 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

  19. Improvement of microwave-assisted digestion of milk powder with diluted nitric acid using oxygen as auxiliary reagent

    Energy Technology Data Exchange (ETDEWEB)

    Bizzi, Cezar A. [Departamento de Quimica, Universidade Federal de Santa Maria, 97105-900, Santa Maria, RS (Brazil); Instituto Nacional de Ciencia e Tecnologia de Bioanalitica, Campinas, SP (Brazil); Barin, Juliano S. [Departamento de Tecnologia e Ciencia dos Alimentos, Universidade Federal de Santa Maria, 97105-900, Santa Maria, RS (Brazil); Garcia, Edivaldo E. [Departamento de Quimica, Universidade Estadual de Maringa, 87100-900, Maringa, PR (Brazil); Nobrega, Joaquim A. [Departamento de Quimica, Universidade Federal de Sao Carlos, 13565-905, Sao Carlos, SP (Brazil); Dressler, Valderi L. [Departamento de Quimica, Universidade Federal de Santa Maria, 97105-900, Santa Maria, RS (Brazil); Instituto Nacional de Ciencia e Tecnologia de Bioanalitica, Campinas, SP (Brazil); Flores, Erico M.M., E-mail: ericommf@gmail.com [Departamento de Quimica, Universidade Federal de Santa Maria, 97105-900, Santa Maria, RS (Brazil); Instituto Nacional de Ciencia e Tecnologia de Bioanalitica, Campinas, SP (Brazil)

    2011-05-15

    The feasibility of using diluted HNO{sub 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{sub 3} solutions (1 to 14 mol L{sup -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{sup -1} HNO{sub 3} with oxygen pressure ranging from 7.5 to 20 bar with resultant carbon content in digests lower than 1700 mg L{sup -1}. Using optimized conditions, less than 0.86 mL of concentrated nitric acid (14 mol L{sup -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.

  20. Identification of protein SUMOylation sites by mass spectrometry using combined microwave-assisted aspartic acid cleavage and tryptic digestion

    Science.gov (United States)

    Osula, Omoruyi; Swatkoski, Stephen; Cotter, Robert J.

    2012-01-01

    SUMO (Small-Ubiquitin-like MOdifier) is a post-translational modifier of protein substrates at lysine residues that conjugates to proteins in response to various changes in the cell. As a result of SUMO modification, marked changes in transcription regulation, DNA repair, subcellular localization, and mitosis, among other cellular processes, are known to occur. However, while the identification of ubiquitylation sites by mass spectrometry is aided in part by the presence of a small di-amino acid GlyGly “tag” that remains on lysine residues following tryptic digestion, SUMOylation poses a particular challenge as the absence of a basic residue near to the SUMO C-terminus results in a significant 27 or 32 amino acid sequence branch conjugated to the substrate peptide. MS/MS analyses of these branch peptides generally reveal abundant fragment ions resulting from cleavage of the SUMO tail, but which obscure those needed for characterizing the target peptide sequence. Other approaches for identifying SUMO substrates exist and include overexpression of the SUMO isoforms using an N-terminal histidine tag, as well as site-directed mutagenesis of the C-terminal end of the SUMO sequence. Here, we employ combined enzymatic/chemical approaches which serve to shorten the SUMO tag, and thus help to simplify SUMO spectra, making interpretation of mass spectra and location of the SUMOylation site easier. As described in this report, we demonstrate a method for identifying SUMOylation sites using three commercially available SUMO- modified isoforms, and by employing acid-only and acid/trypsin cleavage strategies. These approaches were carried out using MALDI-TOF and LC/MS instrumentation, along with CID and ETD fragmentation. PMID:22576878

  1. Effects of digestible protein to energy ratios on growth and carcass chemical composition of siamese fighting fish (Betta splendens

    Directory of Open Access Journals (Sweden)

    Marcos Vinicius Antunes de Lemos

    2014-02-01

    Full Text Available The expansion in the market of ornamental fish depends on the intensification of production systems and the generation of appropriate technologies. Therefore, it is necessary to know the nutritional requirements of each species for the formulation of nutritionally balanced diets. A 30-day feeding trial was undertaken to evaluate the effects of different dietary digestible protein to energy ratio in diets for Betta splendens fingerlings. Fifty-six all-male B. splendens with 1.03 ± 0.11 g average weight were randomly stocked into 56 1L-aquaria in a completely randomized design with a 2 x 4 factorial arrangement, comprising eight treatments and seven replication. Each fish was considered an experimental unit. Diets were formulated to contain 28.0, 35.0, 42.0 and 49.0% DP and 3,600 and 3,200 kcal/kg DE. The growth parameters evaluated in this trial were weight gain (WG, feed intake (FI, feed conversion ratio (FCR, protein efficiency ratio (PER and specific growth rate (SGR. No difference (P > 0.05 was observed for WG, FCR and SGR. However, a higher FI was observed for fish fed diets containing 42.0% DP (P < 0.05. A positive effect of DP and DE on PER was observed in this study. Generally, according to the increase of DE levels, a reduction on PER was observed. Based on this experimental condition, we recommend the use of 28.0% DP and 3,200 kcal/kg DE in diets for B. splendens.

  2. In vitro protein digestibility and physicochemical properties of dry red bean (Phaseolus vulgaris) flour: effect of processing and incorporation of soybean and cowpea flour.

    Science.gov (United States)

    Njintang, N Y; Mbofung, C M; Waldron, K W

    2001-05-01

    A study was carried out to determine the effect of germination and drying temperature on the in vitro protein digestibility and physicochemical properties of dry red bean flours. A 2 x 3 factorial experiment with two treatments (germination and nongermination) and three drying temperatures was used for this purpose. The effect of particle size on water absorption capacity of bean flour was investigated. In addition, the effect of incorporating soybean and cowpea into the red bean flour on functional properties was equally investigated. Results reveal that protein digestibility increased with germination and also with drying temperature. Drying at 60 degrees C produced flours of optimum functional characteristics, although the hydrophilic/lipophilic index was high and the solubility index reduced. Germination and particle size as well as drying temperature all affected the water uptake properties of bean flours. Incorporation of soybean and cowpea flour into germinated bean flour at levels of 10 and 30%, respectively, produced a composite with higher functional properties.

  3. Improvement of the relative entropy based protein folding method

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    The "relative entropy" has been used as a minimization function to predict the tertiary structure of a protein backbone, and good results have been obtained. However, in our previous work, the ensemble average of the contact potential was estimated by an approximate calculation. In order to improve the theoretical integrity of the relative-entropy-based method, a new theoretical calculation method of the ensemble average of the contact potential was presented in this work, which is based on the thermodynamic perturbation theory. Tests of the improved algorithm were performed on twelve small proteins. The root mean square deviations of the predicted versus the native structures from Protein Data Bank range from 0.40 to 0.60 nm. Compared with the previous approximate values, the average prediction accuracy is improved by 0.04 nm.

  4. Improvement of the relative entropy based protein folding method

    Institute of Scientific and Technical Information of China (English)

    QI LiSheng; SU JiGuo; CHEN WeiZu; WANG CunXin

    2009-01-01

    The "relative entropy" has been used as a minimization function to predict the tertiary structure of a protein backbone, and good results have been obtained. However, in our previous work, the ensemble average of the contact potential was estimated by an approximate calculation. In order to improve the theoretical integrity of the relative-entropy-based method, a new theoretical calculation method of the ensemble average of the contact potential was presented in this work, which is based on the thermodynamic perturbation theory. Testa of the improved algorithm were performed on twelve small proteins. The root mean square deviations of the predicted versus the native structures from Protein Data Bank range from 0.40 to 0.60 nm. Compared with the previous approximate values, the average prediction accuracy is improved by 0.04 nm.

  5. 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

  6. 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.

  7. Improving accuracy of protein-protein interaction prediction by considering the converse problem for sequence representation

    Directory of Open Access Journals (Sweden)

    Wang Yong

    2011-10-01

    Full Text Available Abstract Background With the development of genome-sequencing technologies, protein sequences are readily obtained by translating the measured mRNAs. Therefore predicting protein-protein interactions from the sequences is of great demand. The reason lies in the fact that identifying protein-protein interactions is becoming a bottleneck for eventually understanding the functions of proteins, especially for those organisms barely characterized. Although a few methods have been proposed, the converse problem, if the features used extract sufficient and unbiased information from protein sequences, is almost untouched. Results In this study, we interrogate this problem theoretically by an optimization scheme. Motivated by the theoretical investigation, we find novel encoding methods for both protein sequences and protein pairs. Our new methods exploit sufficiently the information of protein sequences and reduce artificial bias and computational cost. Thus, it significantly outperforms the available methods regarding sensitivity, specificity, precision, and recall with cross-validation evaluation and reaches ~80% and ~90% accuracy in Escherichia coli and Saccharomyces cerevisiae respectively. Our findings here hold important implication for other sequence-based prediction tasks because representation of biological sequence is always the first step in computational biology. Conclusions By considering the converse problem, we propose new representation methods for both protein sequences and protein pairs. The results show that our method significantly improves the accuracy of protein-protein interaction predictions.

  8. 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.

  9. Improved cell-free RNA and protein synthesis system.

    Directory of Open Access Journals (Sweden)

    Jun Li

    Full Text Available Cell-free RNA and protein synthesis (CFPS is becoming increasingly used for protein production as yields increase and costs decrease. Advances in reconstituted CFPS systems such as the Protein synthesis Using Recombinant Elements (PURE system offer new opportunities to tailor the reactions for specialized applications including in vitro protein evolution, protein microarrays, isotopic labeling, and incorporating unnatural amino acids. In this study, using firefly luciferase synthesis as a reporter system, we improved PURE system productivity up to 5 fold by adding or adjusting a variety of factors that affect transcription and translation, including Elongation factors (EF-Ts, EF-Tu, EF-G, and EF4, ribosome recycling factor (RRF, release factors (RF1, RF2, RF3, chaperones (GroEL/ES, BSA and tRNAs. The work provides a more efficient defined in vitro transcription and translation system and a deeper understanding of the factors that limit the whole system efficiency.

  10. Protein-carbohydrate supplements improve muscle protein balance in muscular dystrophy patients after endurance exercise

    DEFF Research Database (Denmark)

    Andersen, Grete; Ørngreen, Mette C; Preisler, Nicolai

    2015-01-01

    In healthy individuals, postexercise protein supplementation increases muscle protein anabolism. In patients with muscular dystrophies, aerobic exercise improves muscle function, but the effect of exercise on muscle protein balance is unknown. Therefore, we investigated 1) muscle protein balance...... before, during, and after exercise and 2) the effect of postexercise protein-carbohydrate supplementation on muscle protein balance in patients with muscular dystrophies. In 17 patients [7 women and 10 men, aged 33 ± 11 yr (18-52), body mass index: 22 ± 3 kg/m(2) (16-26)] and 8 healthy matched controls...... [3 women and 5 men, age 33 ± 13 years (19-54), body mass index: 23 ± 3 kg/m(2) (19-27)], muscle protein synthesis, breakdown, and fractional synthesis rates (FSR) were measured across the leg using tracer dilution methodology on two occasions, with and without oral postexercise protein...

  11. Crude protein reduction and digestible methionine+cystine and threonine to digestible lysine ratios in diets for Nile tilapia fingerlings Redução de proteína bruta e relações de metionina+cistina e treonina digestíveis com lisina digestível em dietas para alevinos de tilápia-do-nilo

    Directory of Open Access Journals (Sweden)

    Moisés Quadros

    2009-08-01

    Full Text Available The objective of the present study was to investigate crude protein reduction and the methionine+cystine and threonine to lysine ratios in diets for reversed Nile tilapia fingerlings (Oreochromis niloticus, Thai strain. Four hundred and twenty Nile tilapia fingerlings with an average initial weight of 1.83 ± 0.02 g were placed in a completely randomized design, with five treatments, seven replications per treatment and 12 fish per experimental unit, for 37-days. Two crude protein (CP levels were evaluated (32 and 28%. The diet with 32% CP had methionine+cystine:lysine and threonine:lysine ratios of 63 and 74%, respectively. Three diets with 28% CP had methionine+cystine:lysine and threonine:lysine ratios of 63 and 74%, 60 and 74% and 60 and 69%, respectively. The diet with 28% CP and had methionine+cystine:lysine ratio of 63% and threonine:lysine of 74% and was supplemented with 0.3% glutamic acid. The fish were kept in thirty-five 130 L aquariums supplied with single-pass flow-through water, individual aeration and fed ad libitum six times per day. Fish fed the diet with 32% CP had lower protein efficiency ratios and nitrogen efficiency ratios. The diet CP level was 28% and the methionine+cystine and threonine to lysine ratios were 60 and 69%, respectively.Objetivou-se avaliar a redução do nível de proteína bruta e as relações dos aminoácidos metionina+cistina e treonina digestíveis com a lisina digestível em dietas para alevinos revertidos de tilápia-do-nilo (Oreochromis niloticus, da linhagem tailandesa. Um experimento com duração de 37 dias foi realizado utilizando-se 420 alevinos com peso inicial de 1,83 ± 0,02 g, distribuídos em delineamento inteiramente ao acaso, composto de 5 dietas, 7 repetições e 12 peixes por unidade experimental. Foram avaliados dois níveis de PB (32 e 28% em diversas dietas experimentais. Na dieta com 32% de PB, foram mantidas as relações de metionina+cistina e treonina com lisina de 63 e 74

  12. CNNcon: improved protein contact maps prediction using cascaded neural networks.

    Directory of Open Access Journals (Sweden)

    Wang Ding

    Full Text Available BACKGROUNDS: Despite continuing progress in X-ray crystallography and high-field NMR spectroscopy for determination of three-dimensional protein structures, the number of unsolved and newly discovered sequences grows much faster than that of determined structures. Protein modeling methods can possibly bridge this huge sequence-structure gap with the development of computational science. A grand challenging problem is to predict three-dimensional protein structure from its primary structure (residues sequence alone. However, predicting residue contact maps is a crucial and promising intermediate step towards final three-dimensional structure prediction. Better predictions of local and non-local contacts between residues can transform protein sequence alignment to structure alignment, which can finally improve template based three-dimensional protein structure predictors greatly. METHODS: CNNcon, an improved multiple neural networks based contact map predictor using six sub-networks and one final cascade-network, was developed in this paper. Both the sub-networks and the final cascade-network were trained and tested with their corresponding data sets. While for testing, the target protein was first coded and then input to its corresponding sub-networks for prediction. After that, the intermediate results were input to the cascade-network to finish the final prediction. RESULTS: The CNNcon can accurately predict 58.86% in average of contacts at a distance cutoff of 8 Å for proteins with lengths ranging from 51 to 450. The comparison results show that the present method performs better than the compared state-of-the-art predictors. Particularly, the prediction accuracy keeps steady with the increase of protein sequence length. It indicates that the CNNcon overcomes the thin density problem, with which other current predictors have trouble. This advantage makes the method valuable to the prediction of long length proteins. As a result, the effective

  13. Effects of dietary protein and energy levels on digestive enzyme activities and electrolyte composition in the small intestinal fluid of geese.

    Science.gov (United States)

    Yang, Jing; Yang, Lin; Wang, Yongchang; Zhai, Shuangshuang; Wang, Shenshen; Yang, Zhipeng; Wang, Wence

    2017-02-01

    The present study was conducted to evaluate the effects of dietary protein and energy levels on digestive enzymes and electrolyte composition in jejunum of geese. A 3×3 factorial and completely randomized design was adopted with three protein levels and three energy levels. The experiment included four replicates for each treatment, and three geese for each replicate. Isovolumetric supernate from centrifugal jejuna fluid were mixed in each replicate. Activities of digestive enzymes and ions were analyzed. The results showed trypsin and chymotrypsin activities were significantly increased with increasing of dietary protein and energy levels (Penergy levels. However, no significant differences were found for the activities of amylase and cellulase, as well as the concentration of Na(+) among groups with different protein and energy levels. In conclusion, digesta enzymes and electrolytes in the small intestine adapted to the protein and energy levels. The activities of protease, rather than amylase and cellulase were induced with increasing of protein and energy levels. The imbalance of positive and negative ions was possibly adjusted by the fluctuant concentrations of K(+) , Cl(-) and Ca(2+) for maintaining normal physiological function.

  14. Identifying proper agitation interval to prevent floating layers formation of corn stover and improve biogas production in anaerobic digestion.

    Science.gov (United States)

    Tian, Libin; Zou, Dexun; Yuan, Hairong; Wang, Linfeng; Zhang, Xin; Li, Xiujin

    2015-06-01

    Floating tests were conducted in anaerobic digestion with different OLR of corn stover to investigate formation of floating layers and to find proper agitation interval for preventing floating layer formation. Floating layers were formed in the early stage of no-agitation period. The daily biogas production was decreased by 81.87-87.90% in digesters with no agitation and feeding compared with digesters having agitation. Reduction of biogas production was mainly attributed to poor contact of substrate-microorganisms. Agitation intervals of 10 h, 6 h, and 2 h were found to be proper for eliminating floating layer at OLR of 1.44, 1.78 and 2.11 g(TS) L(-1) d(-1), respectively. The proper agitation interval was further validated by anaerobic experiments. It showed that proper agitation interval could not only prevent floating layer formation and achieve high biogas production but also increase energy efficiency of anaerobic digestion. The finding is useful for operating anaerobic digester with corn stover in a cost-effective way.

  15. 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.

  16. Improved Functional Characteristics of Whey Protein Hydrolysates in Food Industry.

    Science.gov (United States)

    Jeewanthi, Renda Kankanamge Chaturika; Lee, Na-Kyoung; Paik, Hyun-Dong

    2015-01-01

    This review focuses on the enhanced functional characteristics of enzymatic hydrolysates of whey proteins (WPHs) in food applications compared to intact whey proteins (WPs). WPs are applied in foods as whey protein concentrates (WPCs), whey protein isolates (WPIs), and WPHs. WPs are byproducts of cheese production, used in a wide range of food applications due to their nutritional validity, functional activities, and cost effectiveness. Enzymatic hydrolysis yields improved functional and nutritional benefits in contrast to heat denaturation or native applications. WPHs improve solubility over a wide range of pH, create viscosity through water binding, and promote cohesion, adhesion, and elasticity. WPHs form stronger but more flexible edible films than WPC or WPI. WPHs enhance emulsification, bind fat, and facilitate whipping, compared to intact WPs. Extensive hydrolyzed WPHs with proper heat applications are the best emulsifiers and addition of polysaccharides improves the emulsification ability of WPHs. Also, WPHs improve the sensorial properties like color, flavor, and texture but impart a bitter taste in case where extensive hydrolysis (degree of hydrolysis greater than 8%). It is important to consider the type of enzyme, hydrolysis conditions, and WPHs production method based on the nature of food application.

  17. Food microstructure and starch digestion.

    Science.gov (United States)

    Singh, Jaspreet; Kaur, Lovedeep; Singh, Harjinder

    2013-01-01

    Microstructural characteristics of starch-based natural foods such as parenchyma or cotyledon cell shape, cell size and composition, and cell wall composition play a key role in influencing the starch digestibility during gastrointestinal digestion. The stability of cell wall components and the arrangement of starch granules in the cells may affect the free access of amylolytic enzymes during digestion. Commonly used food processing techniques such as thermal processing, extrusion cooking, and post-cooking refrigerated storage alter the physical state of starch (gelatinization, retrogradation, etc.) and its digestibility. Rheological characteristics (viscosity) of food affect the water availability during starch hydrolysis and, consequently, the absorption of digested carbohydrates in the gastrointestinal tract. The nonstarch ingredients and other constituents present in food matrix, such as proteins and lipids interact with starch during processing, which leads to an alteration in the overall starch digestibility and physicochemical characteristics of digesta. Starch digestibility can be controlled by critically manipulating the food microstructure, processing techniques, and food composition.

  18. Protein requirements in larvae and adults of Scaptotrigona postica (Hymenoptera: Apidae [correction of Apidia], Meliponinae): midgut proteolytic activity and pollen digestion.

    Science.gov (United States)

    do Carmo Zerbo, A; Silva de Moraes, R L; Brochetto-Braga, M R

    2001-05-01

    The number and degree of digestion of pollen grains in the midgut and rectum, the midgut proteolytic activity and the time of pollen grain passage through the digestive tract in the stingless bee Scaptotrigona postica (Latreille) have been analyzed. The results show similar protein requirements among larvae, nurse bees and queens, as well as between forager bees and old males, but these requirements are higher in individuals from the former groups than in those from the latter. Although protein requirements have been demonstrated to vary according to a bee's activity in the colony, they are similar among bees from different castes or sexes. These changes in feeding behavior are related to the bee's function and to less competition for nourishment among individuals of the colony. It is also noted that pollen grains took between 6 and 28 h to pass through the digestive tract. Pollen grains are irregularly accumulated in the various regions of the midgut, which may reflect functional differentiation throughout the midgut.

  19. The addition of a Buttiauxella sp. phytase to lactating sow diets deficient in phosphorus and calcium reduces weight loss and improves nutrient digestibility.

    Science.gov (United States)

    Wealleans, A L; Bold, R M; Dersjant-Li, Y; Awati, A

    2015-11-01

    Improving the efficiency of P use by pigs is especially important for lactating sows, whose metabolic requirements for P and Ca are high. The effect of a sp. phytase on lactating sow performance and nutrient digestibility was investigated using the combined data set for 6 studies. Treatments included a nutritionally adequate positive control diet (PC), a negative control diet (NC; with an average reduction of 0.16% available phosphorous and 0.15% Ca vs. PC), and NC supplemented with a sp. phytase at 250, 500, 1,000 or 2,000 phytase unit (FTU)/kg, respectively. Phosphorus and Ca deficiency in the NC resulted in significantly higher BW loss compared with the PC. All phytase treatments maintained BW loss at the same level as the PC. Increasing doses of phytase significantly ( phytase supplementation. Digestible P and Ca were significantly improved (linear, phytase supplementation. Significantly lower apparent total tract digestibility of energy, Ca, and P was found in the NC treatment vs. the PC treatment, whereas no significant differences were found between phytase treatment and the PC treatment. In conclusion, phytase supplementation at a level of 250 FTU/kg can replace 0.16% available phosphorous and 0.15% Ca; however, increasing the phytase dose can further reduce BW loss in sows fed P- and Ca- deficient diets.

  20. Linker engineering for fusion protein construction: Improvement and characterization of a GLP-1 fusion protein.

    Science.gov (United States)

    Kong, Yuelin; Tong, Yue; Gao, Mingming; Chen, Chen; Gao, Xiangdong; Yao, Wenbing

    2016-01-01

    Protein engineering has been successfully applied in protein drug discovery. Using this technology, we previously have constructed a fusion protein by linking the globular domain of adiponectin to the C-terminus of a glucagon-like peptide-1 (GLP-1) analog. Herein, to further improve its bioactivity, we reconstructed this fusion protein by introducing linker peptides of different length and flexibility. The reconstructed fusion proteins were overexpressed in Escherichia coli and purified using nickel affinity chromatography. Their agonist activity towards receptors of GLP-1 and adiponectin were assessed in vitro by using luciferase assay and AMP-activated protein kinase (AMPK) immunoblotting, respectively. The effects of the selected fusion protein on glucose and lipid metabolism were evaluated in mice. The fusion protein reconstructed using a linker peptide of AMGPSSGAPGGGGS showed high potency in activating GLP-1 receptor and triggering AMPK phosphorylation via activating the adiponectin receptor. Remarkably, the optimized fusion protein was highly effective in lowering blood glucose and lipids in mice. Collectively, these findings demonstrate that the bioactivity of this GLP-1 fusion protein can be significantly promoted by linker engineering, and indicate that the optimized GLP-1 fusion protein is a promising lead structure for anti-diabetic drug discovery.

  1. 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.

  2. Estrategias para mejorar los resultados academicos para las latinas (Strategies for Improving the Educational Outcomes of Latinas). ERIC Digest.

    Science.gov (United States)

    Schwartz, Wendy

    The educational experiences of Latinas are affected by the interaction of many factors, including poverty, racism, sexual harassment, and lack of English language proficiency. This Spanish-language digest presents a range of strategies that schools can employ to promote the academic achievement of Latinas. Schools should communicate that Hispanic…

  3. Improvement of the Digestibility of Sulfated Hyaluronans by Bovine Testicular Hyaluronidase: A UV Spectroscopic and Mass Spectrometric Study

    Directory of Open Access Journals (Sweden)

    Katharina Lemmnitzer

    2014-01-01

    Full Text Available Glycosaminoglycans (GAGs such as hyaluronan (HA and chondroitin sulfate (CS are important, natural polysaccharides which occur in biological (connective tissues and have various biotechnological and medical applications. Additionally, there is increasing evidence that chemically (oversulfated GAGs possess promising properties and are useful as implant coatings. Unfortunately, a detailed characterization of these GAGs is challenging: although mass spectrometry (MS is one of the most powerful tools to elucidate the structures of (polysaccharides, MS is not applicable to high mass polysaccharides, but characteristic oligosaccharides are needed. These oligosaccharides are normally generated by enzymatic digestion. However, chemically modified (particularly sulfated GAGs are extremely refractive to enzymatic digestion. This study focuses on the investigation of the digestibility of GAGs with different degrees of sulfation by bovine testicular hyaluronidase (BTH. It will be shown by using an adapted spectrophotometric assay that all investigated GAGs can be basically digested if the reaction conditions are carefully adjusted. However, the oligosaccharide yield correlates reciprocally with the number of sulfate residues per polymer repeating unit. Finally, matrix-laser desorption and ionization (MALDI MS will be used to study the released oligosaccharides and their sulfation patterns.

  4. Text mining improves prediction of protein functional sites.

    Science.gov (United States)

    Verspoor, Karin M; Cohn, Judith D; Ravikumar, Komandur E; Wall, Michael E

    2012-01-01

    We present an approach that integrates protein structure analysis and text mining for protein functional site prediction, called LEAP-FS (Literature Enhanced Automated Prediction of Functional Sites). The structure analysis was carried out using Dynamics Perturbation Analysis (DPA), which predicts functional sites at control points where interactions greatly perturb protein vibrations. The text mining extracts mentions of residues in the literature, and predicts that residues mentioned are functionally important. We assessed the significance of each of these methods by analyzing their performance in finding known functional sites (specifically, small-molecule binding sites and catalytic sites) in about 100,000 publicly available protein structures. The DPA predictions recapitulated many of the functional site annotations and preferentially recovered binding sites annotated as biologically relevant vs. those annotated as potentially spurious. The text-based predictions were also substantially supported by the functional site annotations: compared to other residues, residues mentioned in text were roughly six times more likely to be found in a functional site. The overlap of predictions with annotations improved when the text-based and structure-based methods agreed. Our analysis also yielded new high-quality predictions of many functional site residues that were not catalogued in the curated data sources we inspected. We conclude that both DPA and text mining independently provide valuable high-throughput protein functional site predictions, and that integrating the two methods using LEAP-FS further improves the quality of these predictions.

  5. Text mining improves prediction of protein functional sites.

    Directory of Open Access Journals (Sweden)

    Karin M Verspoor

    Full Text Available We present an approach that integrates protein structure analysis and text mining for protein functional site prediction, called LEAP-FS (Literature Enhanced Automated Prediction of Functional Sites. The structure analysis was carried out using Dynamics Perturbation Analysis (DPA, which predicts functional sites at control points where interactions greatly perturb protein vibrations. The text mining extracts mentions of residues in the literature, and predicts that residues mentioned are functionally important. We assessed the significance of each of these methods by analyzing their performance in finding known functional sites (specifically, small-molecule binding sites and catalytic sites in about 100,000 publicly available protein structures. The DPA predictions recapitulated many of the functional site annotations and preferentially recovered binding sites annotated as biologically relevant vs. those annotated as potentially spurious. The text-based predictions were also substantially supported by the functional site annotations: compared to other residues, residues mentioned in text were roughly six times more likely to be found in a functional site. The overlap of predictions with annotations improved when the text-based and structure-based methods agreed. Our analysis also yielded new high-quality predictions of many functional site residues that were not catalogued in the curated data sources we inspected. We conclude that both DPA and text mining independently provide valuable high-throughput protein functional site predictions, and that integrating the two methods using LEAP-FS further improves the quality of these predictions.

  6. 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.

  7. Anaerobic co-digestion of pig manure and crude glycerol at mesophilic conditions: biogas and digestate.

    Science.gov (United States)

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

    2012-04-01

    Crude glycerol derived from biodiesel production is characterized by its high concentration of organic carbon and its solubility in water; properties that make it a suitable co-substrate to improve the efficiency of a manure digester. An increase of about 400% in biogas production was obtained under mesophilic conditions when pig manure was co-digested with 4% of glycerol, on a wet-basis, compared to mono-digestion. The increase in biogas production was mainly a consequence of the increase in organic loading rate. However, the differences could also be related to the synergy between both substrates and the carbon-to-nitrogen ratio. Moreover, the analysis of the macro-compounds, protein, lipids, carbohydrates and fibers, showed lower removal efficiencies in the co-digester as the microorganisms obtained nutrients from the soluble carbohydrates provided by the glycerol. The digestate stability, evaluated through a respirometric assay, showed that co-substrate addition does not exert a negative impact on digestate quality.

  8. Exigência de lisina digestível e planos de nutrição para frangos de corte machos mantendo as relações metionina + cistina e treonina digestível na proteína ideal Digestible lysine requirements for male broilers keeping methionine + cystine and threonine ratios in the ideal protein

    Directory of Open Access Journals (Sweden)

    Douglas Haese

    2012-03-01

    diet. Weight gain and feed conversion improved quadratically by increasing dietary levels of lysine. Digestible lysine requirement of 1-to-7-day-old male broilers is 1.30 and 1.29% for best weight gain and feed conversion, respectively, corresponding to intakes of 2.68 and 2.64g digestible lysine. The nutritional plan PN3 meets the nutritional needs of male broilers from 1 to 21 days of age.

  9. Binding affinity between dietary polyphenols and β-lactoglobulin negatively correlates with the protein susceptibility to digestion and total antioxidant activity of complexes formed.

    Science.gov (United States)

    Stojadinovic, Marija; Radosavljevic, Jelena; Ognjenovic, Jana; Vesic, Jelena; Prodic, Ivana; Stanic-Vucinic, Dragana; Cirkovic Velickovic, Tanja

    2013-02-15

    Non-covalent interactions between β-lactoglobulin (BLG) and polyphenol extracts of teas, coffee and cocoa were studied by fluorescence and CD spectroscopy at pH values of the gastrointestinal tract (GIT). The biological implications of non-covalent binding of polyphenols to BLG were investigated by in vitro pepsin and pancreatin digestibility assay and ABTS radical scavenging activity of complexes formed. The polyphenol-BLG systems were stable at pH values of the GIT. The most profound effect of pH on binding affinity was observed for polyphenol extracts rich in phenolic acids. Stronger non-covalent interactions delayed pepsin and pancreatin digestion of BLG and induced β-sheet to α-helix transition at neutral pH. All polyphenols tested protected protein secondary structure at an extremely acidic pH of 1.2. A positive correlation was found between the strength of protein-polyphenol interactions and (a) half time of protein decay in gastric conditions (R(2)=0.85), (b) masking of total antioxidant capacity of protein-polyphenol complexes (R(2)=0.95).

  10. Characterization of quinoa seed proteome combining different protein precipitation techniques: Improvement of knowledge of nonmodel plant proteomics.

    Science.gov (United States)

    Capriotti, Anna Laura; Cavaliere, Chiara; Piovesana, Susy; Stampachiacchiere, Serena; Ventura, Salvatore; Zenezini Chiozzi, Riccardo; Laganà, Aldo

    2015-03-01

    A shotgun proteomics approach was used to characterize the quinoa seed proteome. To obtain comprehensive proteomic data from quinoa seeds three different precipitation procedures were employed: MeOH/CHCl3 /double-distilled H2 O, acetone either alone or with trichloroacetic acid; the isolated proteins were then in-solution digested and the resulting peptides were analyzed by nano-liquid chromatography coupled to tandem mass spectrometry. However, since quinoa is a nonmodel plant species, only a few protein sequences are included in the most widely known protein sequence databases. To improve the data reliability a UniProt subdatabase, containing only proteins of Caryophillales order, was used. A total of 352 proteins were identified and evaluated both from a qualitative and quantitative point of view. This combined approach is certainly useful to increase the final number of identifications, but no particular class of proteins was extracted and identified in spite of the different chemistries and the different precipitation protocols. However, with respect to the other two procedures, from the relative quantitative analysis, based on the number of spectral counts, the trichloroacetic acid/acetone protocol was the best procedure for sample handling and quantitative protein extraction. This study could pave the way to further high-throughput studies on Chenopodium Quinoa.

  11. Ileal digestibility of amino acids in novel organic protein feedstuffs for pigs: Mussel meal(Mytilus edulis)

    OpenAIRE

    Kortelainen, Tiina; Siljander-Rasi, Hilkka; Tuori, Mikko; Partanen, Kirsi

    2014-01-01

    Abstract The objective of this study was to determine the apparent total tract digestibility (ATTD) of nu-trients and the standardised ileal digestibility (SID) of amino acids in organically produced mus-sel (Mytilus edulis) meal in growing piglets. The use of mussel meal in pig feeding is not allowed for the time being, but feed legislation in the EU concerning the use of mussel meal for pigs is in progress. The experiment was carried out with a total of 24 growing pigs, 13 gilts and ...

  12. Amino acid digestibility and concentration of digestible and metabolizable energy in soybean meal produced from conventional, high-protein, or low-oligosaccharide varieties of soybeans and fed to growing pigs.

    Science.gov (United States)

    Baker, K M; Stein, H H

    2009-07-01

    Two experiments were conducted to determine AA digestibility and the concentration of DE and ME in 5 sources of soybean meal (SBM). The 5 sources included hexane-extracted SBM produced from high-protein soybeans (SBM-HP) and conventional soybeans (SBM-CONV), and mechanically extruded-expelled SBM produced from high-protein soybeans (EE-SBM-HP), low-oligosaccharide soybeans (EE-SBM-LO), and conventional soybeans (EE-SBM-CONV). Five diets that each contained 1 source of SBM and a N-free diet were used in Exp. 1 to determine AA digestibility in each meal. Twelve growing barrows (initial BW: 67.7 +/- 1.34 kg) were allotted to a replicated 6 x 6 Latin square design with 6 periods and 6 diets in each square. Each period lasted 7 d, and ileal digesta were collected on d 6 and 7 of each period. Results of the experiment showed that the standardized ileal digestibility (SID) of all AA except Trp was similar for SBM-HP and SBM-CONV, but EE-SBM-HP and EE-SBM-LO had greater (P < 0.05) SID of His, Ile, Lys, Thr, and Val than EE-SBM-CONV. The SID of all indispensable AA in EE-SBM-HP was greater (P < 0.05) than in SBM-HP. The SID of Arg, Ile, Leu, and Phe in EE-SBM-CONV was greater (P < 0.05) than in SBM-CONV, but the SID of Trp was also greater (P < 0.05) in SBM-CONV than in EE-SBM-CONV. Experiment 2 was conducted to measure DE and ME in the same 5 sources of SBM as used in Exp. 1. Forty-eight growing barrows (initial BW: 38.6 +/- 3.46 kg) were placed in metabolism cages and randomly allotted to 6 diets with 8 replicates per diet. A corn-based diet and 5 diets based on a mixture of corn and each source of SBM were formulated. Urine and feces were collected during a 5-d collection period, and values for DE and ME in each source of SBM were calculated using the difference procedure. Results showed that the ME in SBM-HP tended to be greater (P = 0.10) than in SBM-CONV (4,074 vs. 3,672 kcal/kg of DM). The ME in EE-SBM-HP also tended to be greater (P = 0.10) than in EE-SBM-CONV and

  13. 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.

  14. The digestive system of 1-week-old Jersey calves is well suited to digest, absorb, and incorporate protein and energy into tissue growth even when calves are fed a high plane of milk replacer.

    Science.gov (United States)

    Liang, Yu; Carroll, Jeffery A; Ballou, Michael A

    2016-03-01

    The objectives of the current study were to determine the apparent digestibilities of nitrogen, organic matter, ash, and energy as well as investigate the nitrogen retention of calves fed different planes of milk replacer nutrition during the first week of life. Twelve Jersey calves were blocked by body weight at birth and randomly assigned to either a high plane of nutrition (HPN) or low plane of nutrition (LPN) treatment. The HPN calves were offered 19.2g of dry matter/kg of body weight of a 28% all-milk crude protein and 20% fat milk replacer. The LPN calves were fed 11.6g of dry matter/kg of body weight of a 20% all-milk crude protein and 20% fat milk replacer. All calves were given 3 L of pooled colostrum within 1h of birth after which they were assigned to treatments; no starter was offered during the study. Calves were given 1 d to adapt to their treatments and environment, so calves were 30 to 36 h old at the start of data collection. The study was divided into two 72-h periods. Total collection of feces occurred over each 72 h period, and total urine was collected for the last 24h of each period. Peripheral blood samples were collected at the beginning and end of each period and analyzed for plasma glucose and urea nitrogen concentrations. Data are reported as HPN vs. LPN, respectively. Fecal scores were greater for HPN calves during both periods; however, no difference was found in the dry matter percentage of feces (30.9 vs. 31.9 ± 0.06). No differences were found between treatments in either digestible or metabolizable energy efficiencies, which averaged 93.3 and 83.7%, respectively. A treatment × period interaction was found on the percentage of intake nitrogen retained, in which calves fed the HPN had a greater percentage of intake nitrogen retained during period 1 (87.9 vs. 78.4 ± 1.79%), but was not different from calves fed the LPN during period 2 (85.4 vs. 84.9 ± 1.79%). From these data therefore, we conclude that healthy neonatal calves have

  15. 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.

  16. Metabolic engineering of Escherichia coli to improve recombinant protein production.

    Science.gov (United States)

    Liu, Min; Feng, Xinjun; Ding, Yamei; Zhao, Guang; Liu, Huizhou; Xian, Mo

    2015-12-01

    Escherichia coli is one of the most widely used strains for recombinant protein production. However, obstacles also exist in both academic researches and industrial applications, such as the metabolic burden, the carbon source waste, and the cells' physiological deterioration. This article reviews recent approaches for improving recombinant protein production in metabolic engineering, including workhorse selection, stress factor application, and carbon flux regulation. Selecting a suitable host is the first key point for recombinant protein production. In general, it all depends on characteristics of the strains and the target proteins. It will be triggered cells physiological deterioration when the medium is significantly different from the cell's natural environment. Coexpression of stress factors can help proteins to fold into their native conformation. Carbon flux regulation is a direct approach for redirecting more carbon flux toward the desirable pathways and products. However, some undesirable consequences are usually found in metabolic engineering, such as glucose transport inhibition, cell growth retardation, and useless metabolite accumulation. More efficient regulators and platform cell factories should be explored to meet a variety of production demands.

  17. 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.

  18. Effects of extruding wheat dried distillers grains with solubles with peas or canola meal on ruminal fermentation, microbial protein synthesis, nutrient digestion, and milk production in dairy cows.

    Science.gov (United States)

    Claassen, R M; Christensen, D A; Mutsvangwa, T

    2016-09-01

    Our objective was to examine the effects of feeding coextruded and nonextruded supplements consisting of wheat dried distillers grains with solubles with peas (WDDGS-peas) or canola meal (WDDGS-CM) on ruminal fermentation, omasal flow, and production performance in Holstein cows. Eight cows (4 ruminally cannulated) were used in a replicated 4×4 Latin square with 28-d periods and a 2×2 factorial arrangement of dietary treatments. Dietary treatments were coextruded or nonextruded mixtures of WDDGS-peas and WDDGS-CM that were included in total mixed rations at 15.1% [dry matter (DM) basis]. Diet had no effect on DM intake. Milk yield was greater in cows fed coextruded diets compared with those fed nonextruded diets. Milk fat content was greater in cows fed nonextruded diets compared with those fed coextruded diets, but milk fat yield was greater in cows fed coextruded diets compared with those fed nonextruded diets. Milk yield tended to be greater and milk protein yield was greater in cows fed WDDGS-peas compared with those fed WDDGS-CM. Cows fed nonextruded diets had a greater milk urea-N concentration compared with those fed coextruded diets. Cows fed coextruded diets had greater ruminal digestion of DM and tended to have greater ruminal digestion of organic matter compared with those fed nonextruded diets. Total-tract digestibilities of organic matter, crude protein, ether extract, and starch were greater, whereas that of acid detergent fiber and neutral detergent fiber tended to be greater in cows fed coextruded compared with those fed nonextruded diets. Total-tract digestibility of ether extract was lower whereas that of starch was greater and that of crude protein tended to be greater in cows fed WDDGS-peas compared with those fed WDDGS-CM. Total N excretion and milk N efficiency were unaffected by diet. Ruminal NH3-N concentration tended to be greater in cows fed WDDGS-CM compared with those fed WDDGS-peas. Ruminal propionate concentration was greater whereas

  19. Comparison of HgO and CuSO4 as digestion catalysts in manual Kjeldahl determination of crude protein in animal feeds: collaborative study.

    Science.gov (United States)

    Kane, P F

    1984-01-01

    The official AOAC manual Kjeldahl method for determining crude protein in animal feeds, 7.015, uses HgO as a catalyst in the digestion step. Because of environmental considerations, there is considerable interest in alternative catalysts. A collaborative study compares the official HgO-catalyzed method and an alternative using CuSO4. Fifty-four samples consisting of blind duplicates of closely matched pairs, representing a range of animal feed materials and 2 standard materials, were analyzed once by each method. Results were returned by 22 laboratories. Means and standard deviations between methods were comparable. The CuSO4-catalyzed method has been adopted official first action.

  20. Recovery and characterization of Balanites aegyptiaca Del. kernel proteins. Effect of defatting, air classification, wet sieving and aqueous ethanol treatment on solubility, digestibility, amino acid composition and sapogenin content.

    Science.gov (United States)

    Mohamed, A M; Wolf, W; Spiess, W E

    2000-02-01

    In order to find alternative protein sources in African regions where protein deficiency in nutrition is prevailing, solubility, in-vitro digestibility, amino acid composition and chemical score of Balanites aegyptiaca Del. kernel proteins were investigated as a function of different processing steps including defatting, air classification, wet sieving and aqueous ethanol treatment. Air classification delivered a fine fraction of 58.1% of the total protein. Applying a wet sieving process, a protein concentrate of 72.9% protein content was achieved but the recovery was very low (35.6%). However, in case of isoelectric precipitation followed by aqueous ethanol treatment both protein content (78.2%) and recovery (53.7%) were high. Data concerning the chemical score revealed, that lysine content of the defatted kernel flour amounted to 74.2% of the recommended FAO/WHO standard level. In-vitro protein digestibility was found to be higher than of legume proteins. The digestible protein of the full fat flour, defatted flour, air classified and wet sieved fine fractions and protein concentrate were 91.9, 93.7, 82.0, 86.4 and 94.2%, respectively. The sapogenin content per 100 g protein of the investigated protein preparations was significantly lower (46% to 62%) than of the initial material (oilcake).

  1. 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.

    NARCIS (Netherlands)

    Oosting, S.J.; Viets, T.C.; Lammers-Wienhoven, S.C.W.; Bruchem, van 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).

  2. An early-life hypoxia event has a long-term impact on protein digestion and growth in European sea bass juvenile.

    Science.gov (United States)

    Zambonino-Infante, José L; Mazurais, David; Servili, A; Cahu, C; Vanderplancke, G; Le Bayon, N; Huelvan, C; Claireaux, Guy

    2017-03-16

    Ocean warming, eutrophication and consequent decrease in oxygen lead to smaller average fish size. Although such responses are well-known in an evolutionary context, involving multiple generations, it appears to be incompatible with current rapid environmental change. Rather, phenotypic plasticity could provide a means for marine fish to cope with rapid environmental changes. However, little is known about the mechanisms underlying plastic responses to environmental conditions that favour small phenotypes.Our aim was to investigate how and why European sea bass that had experienced a short episode of moderate hypoxia during their larval stage subsequently exhibited a growth depression at the juvenile stage compared to the control group.We examined whether energy was used to cover higher costs for maintenance, digestion or activity metabolisms, as a result of differing metabolic rate. The lower growth was not a consequence of lower feed intake.We measured several respirometry parameters and we only found a higher SDA (Specific Dynamic Action) duration and lower SDA amplitude in a fish phenotype with lower growth; this phenotype was also associated with a lower protein digestive capacity in the intestine.Our results contribute to the understanding of the observed decrease in growth in response to climate change. They demonstrate that the reduced growth of juvenile fishes as a consequence of an early-life hypoxia event was not due to a change of fish aerobic scope, but to a specific change in the efficiency of protein digestive functions. The question remains of whether this effect is epigenetic and could be reversible in the offspring.

  3. Cry1Ie蛋白的模拟胃肠液消化稳定性及热稳定性分析%Analysis of Digestive Stability in Simulative Digestive Tract Fluid and Heat Stability of Cry1Ie Protein

    Institute of Scientific and Technical Information of China (English)

    李欣竹; 耿丽丽; 高继国; 张杰

    2015-01-01

    Cry1Ie蛋白对亚洲玉米螟(Ostrinia furnacalis)具有高毒力,cry1Ie基因已经应用于转基因抗虫玉米的种质创制。为评价Cry1Ie蛋白的食用安全性,开展了Cry1Ie蛋白的消化及热稳定性研究。利用实验室已经构建的表达载体,在大肠杆菌中表达了分子量为81 kD的可溶性Cry1Ie蛋白,经过Ni-NTA亲和层析和Superdex-75分子筛层析获得纯度达91%蛋白。模拟消化液实验结果表明,Cry1Ie蛋白在模拟胃肠液中15 s内即被消化,SDS-PAGE未检测到蛋白残留。热稳定性实验中,Cry1Ie蛋白在玉米粉提取液中不稳定,100℃30 min内蛋白基本降解。对样品进行了生物活性测定发现,经模拟胃肠液消化处理和热处理后的Cry1Ie蛋白对亚洲玉米螟无杀虫活性。Cry1Ie蛋白在胃肠液系统中和热处理条件下均不稳定,并丧失了对亚洲玉米螟的杀虫活性。%Due to the high virulence to Asian corn borer(Ostrinia furnacalis)of Cry1Ie, this protein had applied into the germplasm creating of transgenic insect-resistant maize. To evaluate the food safety of Cry1Ie protein, this study carried out the digestion and heat stability assays. Using the expressing vector which the laboratory has constructed, the Cry1Ie protein(81 kD)was expressed inEscherichia coli, and the high purity protein was obtained using Ni-NTA chromatography and Superdex-75 size-exclusion chromatography. The results of artificial digestion assay showed that Cry1Ie was degraded rapidly within 15 s in simulated gastric fluid and intestinal fluid, without any peptide left in SDS-PAGE. In the heat stability assay, Cry1Ie was not stable in the extracting solution of maize meal, and most was degraded within 30 min under 100℃. In the bioassay of the Asian corn borer(Ostrinia furnacalis), it suggested that Cry1Ie lost the bio-activity after the treatment of digestion and heat. In conclusion, Cry1Ie was not stable in the gastrointestinal fliud and the

  4. Production of Lupinus angustifolius protein hydrolysates with improved functional properties

    Directory of Open Access Journals (Sweden)

    Millán, Francisco

    2005-06-01

    Full Text Available Protein hydrolysates wer e obtained from lupin flour and from the purified globulin α -conglutin, and their functional properties were studied. Hydrolysis with alcalase for 60 minutes yielded degrees of hydrolysis ranging from 4 % to 11 % for lupin flour, and from 4 % to 13% for α -conglutin. Protein solubility, oil absorption, foam capacity and stability, emulsifying activity, and emulsion stability of hydrolysates with 6% degree of hydrolysis were determined and compared with the properties of the original flour. The protein hydrolysates showed better functional properties than the original proteins. Most importantly, the solubility of the α -conglutin and L. angustifolius flour hydrolysates was increased by 43 % and 52 %, respectively. Thus, lupin seed protein hydrolysates have improved functional properties and could be used in the elaboration of a variety of products such as breads, cakes, and salad dressings.Se obtuvieron hidrolizados proteicos de la harina del altramuz y de la globulina α - conglutina purificada y se estudiaron sus propiedades funcionales. La hidrólisis con alcalasa durante 60 minutos produjo hidrolizados con grados de hidrólisis entre el 4 % y el 11 % para la harina y entre el 4 % y el 13 % para la α - conglutina. Se estudió en un hidrolizado con un 6 % de grado de hidrólisis la solubilidad proteica, absorción de aceite, capacidad y estabilidad espumante y actividad y estabilidad emulsificante. Los hidrolizados proteicos mostraron mejores propiedades funcionales que las proteínas originales. Más aún, la solubilidad de los hidrolizados de α - conglutina y la harina se incrementó en un 43 % y 52 % respectivamente. Así pues, hidrolizados de proteínas de semilla de lupino presentan mejores propiedades funcionales y podrían usarse en la elaboración de productos como pan, dulces, salsas o cremas.

  5. Ileal digestibility of amino acids in novel organic protein feedstuffs for pigs: Black soldier fly larvae meal(Hermetia illucens)

    OpenAIRE

    Kortelainen, Tiina; Siljander-Rasi, Hilkka; Tuori, Mikko; Partanen, Kirsi

    2014-01-01

    Abstract The objective of this study was to determine the standardised ileal digestibility (SID) of amino acids in organically produced black soldier fly larvae (Hermeti illucens) meal in growing piglets. The use of Hermetia meal in pig feeding is not allowed for the time being, but feed legislation in the EU concerning the use of Hermetia meal for pigs is in progress. Two batches of Hermetia meal arrived from Switzerland (FiBL Research Institute of Organic Agriculture). In batch 1, fa...

  6. Digestive Diseases

    Science.gov (United States)

    ... Celiac Disease Bowel Control Problems (Fecal Incontinence) Gas Lactose Intolerance Diarrhea Diverticulosis & Diverticulitis Acid Reflux (GER & GERD) More Digestive Disease Topics Children and Teens Acid Reflux (GER & GERD) in Infants Acid Reflux (GER & GERD) in Children & Teens Chronic ...

  7. Effect of protein source and protease addition on performance, blood metabolites and nutrient digestibility of turkeys fed on low-protein diets from 28 to 55 d post hatch.

    Science.gov (United States)

    Shahir, M H; Rahimi, R; Taheri, H R; Heidariniya, A; Baradaran, N; Asadi Kermani, Z

    2016-06-01

    The objective of this study was to investigate the effect of a monocomponent protease and dietary inclusion of canola meal (CM) and poultry by-product meal (PBM) on growth performance, carcass characteristics and blood metabolites of turkeys fed on low crude protein (CP) diets from 28 to 55 d post hatch. Experimental treatments included control, maize-soybean meal diet including 258.3 g/kg CP; negative control 1 (NC1), maize-soybean meal diet with reduced CP (232.4 g/kg); NC2, control diet (CP, 258.3 g/kg) including CM (80 g/kg) and PBM (80 g/kg); NC3, maize-soybean meal diet with reduced CP (232.4 g/kg) including CM (80 g/kg) and PBM (80 g/kg). Also, the NC1 + P and NC3 + P diets were created by addition of protease enzyme (30 000 units/kg of diet) to the NC1 and NC3 diets, respectively. The NC3 group had lower body weight gain (BWG) compared to those fed on the control diet, and no improvement with enzyme addition (NC3 + P) was achieved. The protease addition to the NC1 diet (NC1 + P) improved BWG to the level of the control diet. The NC1 group had higher feed conversion ratio (FCR) compared to the control and NC3 + P, but protease addition to the NC1 diet improved FCR. Protease addition to the low CP diets resulted in higher nitrogen (N) retention than in the control and NC2 groups. Also, the NC1 + P and NC3 + P diets increased apparent ileal digestibility (AID) of CP compared to the control group. It was concluded that addition of CM (up to 80 g/kg) and PBM (up to 80 g/kg) to turkey diets had no negative effect on growth performance from 28 to 55 d of age. The NC1 + P group achieved the BWG of the control group which was partially due to increases in N retention and AID of CP, but the NC3 + P group failed to recover the growth losses. This difference implies that the efficacy of the protease may depend upon the protein source in the ration.

  8. A meta-analysis of feed digestion in dairy cows. 1. The effects of forage and concentrate factors on total diet digestibility.

    Science.gov (United States)

    Nousiainen, J; Rinne, M; Huhtanen, P

    2009-10-01

    A meta-analysis based on published experiments with lactating dairy cows was conducted to study the effects of dietary forage and concentrate factors on apparent total diet digestibility. A data set was collected that included a total of 497 dietary treatment means from 92 studies. The diets were based on grass silage or on legume or whole-crop cereal silages partly or completely substituted for grass silage. The silages were supplemented with concentrates given at a flat rate within a dietary comparison. For the statistical evaluation, the data were divided into 5 subsets to quantify silage (digestibility, 42 diets in 17 studies; fermentation characteristics, 108 diets in 39 studies) and concentrate (amount of supplementation, 142 diets in 59 studies; concentration of crude protein, 215 diets in 82 studies; carbohydrate composition, 66 diets in 23 studies) factors on total diet digestibility. The diet digestibility of dairy cows was determined by total fecal collection or by using acid-insoluble ash as an internal marker. Diet organic matter digestibility (OMD) at a maintenance level of feeding (OMD(m)) was estimated using sheep in vivo or corresponding in vitro digestibility values for the forage and reported ingredient and chemical composition values, with tabulated digestibility coefficients for the concentrate components of the diet. A mixed model regression analysis was used to detect the responses of different dietary factors on apparent total diet digestibility. Improved silage OMD(m) resulting from earlier harvest was translated into improved production-level OMD in cows (OMD(p)). The effects of silage fermentation characteristics on OMD(p) were quantitatively small, although sometimes significant. Concentrate supplementation improved total diet OMD(m), but this was not realized in lactating dairy cows because of linearly decreased neutral detergent fiber (NDF) digestibility as concentrate intake increased. Increasing the concentrate crude protein amount

  9. Improving Methane Production through Co-Digestion of Canola Straw and Buffalo Dung by H2O2 Pretreatment

    Directory of Open Access Journals (Sweden)

    ALTAF ALAM NOONARI

    2017-01-01

    Full Text Available In this study an effect of acidic pre-treatment on the CS (Canola Straw and BD (Buffalo Dung by anaerobic co-digestion was investigated. H2O2 (Hydrogen Peroxide is a mainly accustomed reagent, used as a bleaching agent in the different industries such as paper and wood. In the present study, it was used as a pre-treatment chemical at varying concentrations in batch reactors. The co-digestion of CS and BD was carried out in SAMPTS (Semi-Automatic Methane Potential Test System at mesophilic (37±1oC conditions. The CS was pretreated in glass bottles with different concentrations of the H2O2 for seven days. The inoculum used in the present study was an effluent of the CSTR (Continuous Stirred Tank Reactor, which was treating BD at mesophilic conditions. The specific methane production from the codigestion of canola straw and BD, by the pre-treatment of H2O2 at concentrations of 0.5, 1.0, and 1.5% were 530.8, 544.5, and 510.3 NmL CH4 g/VS, respectively. The significant reduction in the volatile solids of CS was observed at the optimum pre-treatment of 1.0% H2O2.

  10. Biogas production improvement and C/N control by natural clinoptilolite addition into anaerobic co-digestion of Phragmites australis, feces and kitchen waste.

    Science.gov (United States)

    Wang, Xiaowei; Zhang, Lieyu; Xi, Beidou; Sun, Wenjun; Xia, Xunfeng; Zhu, Chaowei; He, Xiaosong; Li, Mingxiao; Yang, Tianxue; Wang, Pengfei; Zhang, Zhonglei

    2015-03-01

    Anaerobic co-digestion (A co-D) performance of Phragmites australis, feces and kitchen waste with addition of clinoptilolite (one main kind of zeolite) was investigated to evaluate the improvement of biogas/methane production and internal mechanism of nitrogen and organics control. A better biogas/methane production was observed by 10% clinoptilolite (v/v) than bentonite and diatomite, with the shortest lag phase of 0.070d(-1), the max rate of 15.89L/(kgVSday) and ultimate biogas production of 308.2L/kgVS as the modified Gompertz equation predicted. Accordingly, the content of methane in the biogas was increased from 44.10% to 65.30%. Furthermore, the clinoptilolite inhibited the acidification of digestion liquid (optimum pH 7.0-7.5) and enhanced the VFAs (acetic acid, propionic acid and butyric acid) destruction. Moreover, 10% of clinoptilolite optimally enhanced the microbial utilization of Ca(2+)/Mg(2+), controlled the C/N ratio, and improved the biogas production as well as NH3-N/NO3-N inhibition efficiency.

  11. Common Prairie feeds with different soluble and insoluble fractions used for CPM diet formulation in dairy cattle: Impact of carbohydrate-protein matrix structure on protein and other primary nutrient digestion

    Science.gov (United States)

    Peng, Quanhui; Wang, Zhisheng; Zhang, Xuewei; Yu, Peiqiang

    2014-03-01

    An experiment was conducted to investigate the relationship of carbohydrates molecular spectral characteristics to rumen degradability of primary nutrients in Prairie feeds in dairy cattle. In total, 12 different types of feeds were selected, each type of feed was from three different source with total 37 samples. Six types of them were energy-sourced feeds and the others were protein-sourced feeds. The carbohydrates molecular spectral intensity of various functional groups were collected using Fourier transform infrared attenuated total reflectance (ATR-FT/IR) spectroscopy. In the in situ study, the results showed that the rumen digestibility and digestible fractions of primary nutrients (DM, OM, NCP, and CP) were significantly different (P carbohydrates as a whole have an effect on protein rumen degradation, rather than cellulose alone, indicating carbohydrate-protein matrix structure impact protein utilization in dairy cattle. The non-invasive molecular spectral technique (ATR-FT/IR) could be used as a rapid potential tool to predict rumen protein degradation of feedstuffs by using molecular spectral bands intensities in carbohydrate fingerprint region.

  12. ffects of different levels of digestible arginine and protein in starter diets containing ideal amino acids ratio on Eperformance, carcass traits and serum parameters in broiler chickens

    Directory of Open Access Journals (Sweden)

    Mostafa Gholami

    2015-12-01

    Full Text Available Introduction: Nutrition and health during the first days of life has critical effect on broiler chickens performance. It is well known that diet formulation based on digestible nutrients is superior to formulation based on total nutrients. The suitable supply of essential amino acids in broilers’ diets requires proper knowledge on their metabolic effects in the body. The excessive or unbalanced intake of essential and non-essential amino acids can be harmful to broilers’ metabolism, due to amino acid antagonisms. Arginine is an essential amino acid for broilers since the urea cycle is not functional in birds. Arginine involves in the synthesis of ornithine, a precursor of polyamines that have a key role in cell division, DNA synthesis, nitric oxide (NO synthesis, and cell cycle regulation. Also, arginine increases the release of insulin, growth hormone, and IGF-A and luteinizing hormone (LH in the blood stream. On the other hand, in corn- soybean meal based diets arginine is the fifth limiting amino acid after methionine, lysine, threonine, and valine. Thus, this study was carried out to investigate the effects of different digestible arginine (DA and digestible protein (DP levels of starter diets (1-10 d based on ideal amino acids ratio on performance, carcass traits and serum parameters in broiler chickens. Materials and Methods: Four handed day-old male broiler chickes (Ross 308 were distributed in 10 treatments of 4 replicates (floor pens each. The experiment was designed as a 2×5 factorial arrangement in a completely randomized design. Experimental diets were formulated with five levels of digestible arginine (1.05, 1.18, 1.31, 1.44 and 1.57% and 2 levels of digestible protein (18 and 20%. Chicken were fed with experimental diets during 1 to 10 days of age, and then received similar diets formulated according to Ross 308 (2009 recommendations. All birds had free access to feed and water during the whole rearing period. Temperature was

  13. Improved "optical highlighter" probes derived from discosoma red fluorescent protein.

    Science.gov (United States)

    Robinson, Lisbeth C; Marchant, Jonathan S

    2005-02-01

    The tetrameric red fluorescent protein, DsRed, undergoes a rapid red to green color change evoked by short wavelength (lambda highlighter" probe for tracking live cells, organelles, and fusion proteins. This color change results from selective bleaching of the "mature" red-emitting species of DsRed and an enhancement of emission from the "immature" green species, likely caused by dequenching of fluorescence resonance energy transfer occurring within the protein tetramer. Here, we have examined the role of residues known to influence the rate and completeness of chromophore maturation on the cellular and biophysical properties of DsRed mutants. Surprisingly, a single amino acid mutation (N42Q) with increased basal green emission yet rapid chromophore maturation displayed a multiphoton-evoked color change that was brighter, more consistent, more vivid, and easier to evoke than DsRed, despite the larger proportion of green chromophores. Rapidly maturing mutants with more complete chromophore maturation, exhibited little color change and increased resistance to multiphoton bleaching. We describe improved optical and cell biological properties for two DsRed-derived variants which we showcase in photolabeling studies, and discuss these data in terms of implications for fluorescence resonance energy transfer-based probes.

  14. Effects of varying ruminally undegradable protein supplementation on forage digestion, nitrogen metabolism, and urea kinetics in Nellore cattle fed low-quality tropical forage.

    Science.gov (United States)

    Batista, E D; Detmann, E; Titgemeyer, E C; Valadares Filho, S C; Valadares, R F D; Prates, L L; Rennó, L N; Paulino, M F

    2016-01-01

    Effects of supplemental RDP and RUP on nutrient digestion, N metabolism, urea kinetics, and muscle protein degradation were evaluated in Nellore heifers () consuming low-quality signal grass hay (5% CP and 80% NDF, DM basis). Five ruminally and abomasally cannulated Nellore heifers (248 ± 9 kg) were used in a 5 × 5 Latin square. Treatments were the control (no supplement) and RDP supplementation to meet 100% of the RDP requirement plus RUP provision to supply 0, 50, 100, or 150% of the RUP requirement. Supplemental RDP (casein plus NPN) was ruminally dosed twice daily, and RUP supply (casein) was continuously infused abomasally. Jugular infusion of [NN]-urea with measurement of enrichment in urine was used to evaluate urea kinetics. The ratio of urinary 3-methylhistidine to creatinine was used to estimate skeletal muscle protein degradation. Forage NDF intake (2.48 kg/d) was not affected ( ≥ 0.37) by supplementation, but supplementation did increase ruminal NDF digestion ( Urea entry rate and gastrointestinal entry rate of urea were increased by supplementation ( urea entry rate and tended ( = 0.07) to linearly increase gastrointestinal entry rate of urea. Urea use for anabolic purposes tended ( = 0.07) to be increased by supplementation, and RUP provision also tended ( = 0.08) to linearly increase the amount of urea used for anabolism. The fraction of recycled urea N incorporated into microbial N was greater ( urea from the renal tubule than did supplemented heifers. Overall, unsupplemented heifers had greater mobilization of AA from myofibrillar protein, which provided N for urea synthesis and subsequent recycling. Supplemental RUP, when RDP was supplied, not only increased N retention but also supported increased urea N recycling and increased ruminal microbial protein synthesis.

  15. Effect of pectin extracted from citrus pulp on digesta characteristics and nutrient digestibility in broilers chickens

    Directory of Open Access Journals (Sweden)

    Vanessa Karla Silva

    2013-08-01

    Full Text Available 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.

  16. Improving protein coreference resolution by simple semantic classification

    Directory of Open Access Journals (Sweden)

    Nguyen Ngan

    2012-11-01

    Full Text Available Abstract Background Current research has shown that major difficulties in event extraction for the biomedical domain are traceable to coreference. Therefore, coreference resolution is believed to be useful for improving event extraction. To address coreference resolution in molecular biology literature, the Protein Coreference (COREF task was arranged in the BioNLP Shared Task (BioNLP-ST, hereafter 2011, as a supporting task. However, the shared task results indicated that transferring coreference resolution methods developed for other domains to the biological domain was not a straight-forward task, due to the domain differences in the coreference phenomena. Results We analyzed the contribution of domain-specific information, including the information that indicates the protein type, in a rule-based protein coreference resolution system. In particular, the domain-specific information is encoded into semantic classification modules for which the output is used in different components of the coreference resolution. We compared our system with the top four systems in the BioNLP-ST 2011; surprisingly, we found that the minimal configuration had outperformed the best system in the BioNLP-ST 2011. Analysis of the experimental results revealed that semantic classification, using protein information, has contributed to an increase in performance by 2.3% on the test data, and 4.0% on the development data, in F-score. Conclusions The use of domain-specific information in semantic classification is important for effective coreference resolution. Since it is difficult to transfer domain-specific information across different domains, we need to continue seek for methods to utilize such information in coreference resolution.

  17. Methane production improvement by modulation of solid phase immersion in dry batch anaerobic digestion process: Dynamic of methanogen populations.

    Science.gov (United States)

    André, L; Ndiaye, M; Pernier, M; Lespinard, O; Pauss, A; Lamy, E; Ribeiro, T

    2016-05-01

    Several 60L dry batch anaerobic digestion (AD) reactors were implemented with or without liquid reserve on cattle manure. The immersed part modulation of cattle manure increased the methane flow of about 13%. The quantitative real time PCR and the optimized DNA extraction were implemented and validated to characterize and quantify the methanogen dynamic in dry batch AD process. Final quantities of methanogens converged toward the same level in several inocula at the end of AD. Methanogen dynamic was shown by dominance of Methanosarcinaceae for acetotrophic methanogens and Methanobacteriales for the hydrogenotrophic methanogens. Overall, methanogens populations were stabilized in liquid phase, except Methanosaetaceae. Solid phase was colonized by Methanomicrobiales and Methanosarcinaceae populations giving a support to biofilm development. The methane increase could be explained by a raise of Methanosarcinaceae population in presence of a total contact between solid and liquid phases. Methanosarcinaceae was a bio-indicator of the methane production.

  18. Kinetics of digestion of low-quality forage grazed by beef cattle fed supplements containing increasing levels of rumen undegradable protein

    Directory of Open Access Journals (Sweden)

    Raphael dos Santos Gomes

    Full Text Available ABSTRACT This experiment was conducted to evaluate the degradation kinetics and microbial efficiency of beef cattle grazing on low-quality forage and receiving supplements with different levels of rumen undegradable protein (RUP. The animals grazed on palisade grass pasture solely or this pasture and supplement containing 40 or 60 g of RUP per 100 g of crude protein (CP. The degradation profiles of neutral detergent fiber, fiber carbohydrates, and neutral detergent insoluble protein were interpreted kinetically by using a decreasing logistic model. Treatments (no supplement, or RUP at 40 or 60 g−1 100 g CP did not affect rumen fill; however, the increase in the indigestible fiber carbohydrate fraction that occurred at the expense of the digestible fiber carbohydrate fraction resulted in a greater rumen fill effect. The palisade grass showed a significant proportion of its nitrogen in the form of slowly degradable protein as neutral detergent insoluble protein, which amounted to 26 g per 100 g CP. Supplementation with 40 g of RUP per 100 g CP decresead the indigestible fraction of the low-quality forage. However, the absence of a rumen-fill effect demonstrates that the additional supply of nutrients contributes greatly to increasing growth efficiency and use of the available energy from the forage by the ruminal microorganisms.

  19. Molecular basis of processing-induced changes in protein structure in relation to intestinal digestion in yellow and green type pea (Pisum sativum L.): A molecular spectroscopic analysis.

    Science.gov (United States)

    Yu, Gloria Qingyu; Warkentin, Tom; Niu, Zhiyuan; Khan, Nazir A; Yu, Peiqiang

    2015-12-01

    The objectives of this study were (1) to quantify the protein inherent molecular structural features of green cotyledon (CDC Striker) and yellow cotyledon (CDC Meadow) pea (Pisum sativum L.) seeds using molecular spectroscopic technique (FT/IR-ATR); (2) measure the denaturation of protein molecular makeup in the two types of pea during dry roasting (120°C for 60 min), autoclaving (120°C for 60 min) or microwaving (for 5 min); and (3) correlate the heat-induced changes in protein molecular makeup to the corresponding changes in protein digestibility determined using modified three-step in vitro procedure. Compared with yellow-type, the green-type peas had higher (Ppeas had lower (Ppea-types. However, across the pea types the correlation was not significant. Principal component and hierarchical cluster analyses on the entire spectral data from the amide region (ca. 1727-1480 cm(-1)) were able to visualize and discriminate the structural difference between pea varieties and processing treatments. This study shows that the molecular spectroscopy can be used as a rapid tool to screen the protein value of raw and heat-treated peas.

  20. Evolutionary optimization of kernel weights improves protein complex comembership prediction.

    Science.gov (United States)

    Hulsman, Marc; Reinders, Marcel J T; de Ridder, Dick

    2009-01-01

    In recent years, more and more high-throughput data sources useful for protein complex prediction have become available (e.g., gene sequence, mRNA expression, and interactions). The integration of these different data sources can be challenging. Recently, it has been recognized that kernel-based classifiers are well suited for this task. However, the different kernels (data sources) are often combined using equal weights. Although several methods have been developed to optimize kernel weights, no large-scale example of an improvement in classifier performance has been shown yet. In this work, we employ an evolutionary algorithm to determine weights for a larger set of kernels by optimizing a criterion based on the area under the ROC curve. We show that setting the right kernel weights can indeed improve performance. We compare this to the existing kernel weight optimization methods (i.e., (regularized) optimization of the SVM criterion or aligning the kernel with an ideal kernel) and find that these do not result in a significant performance improvement and can even cause a decrease in performance. Results also show that an expert approach of assigning high weights to features with high individual performance is not necessarily the best strategy.

  1. 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; Young, Clifford

    2013-01-01

    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....

  2. Improvement of anaerobic digestion performance by continuous nitrogen removal with a membrane contactor treating a substrate rich in ammonia and sulfide.

    Science.gov (United States)

    Lauterböck, B; Nikolausz, M; Lv, Z; Baumgartner, M; Liebhard, G; Fuchs, W

    2014-04-01

    The effect of reduced ammonia levels on anaerobic digestion was investigated. Two reactors were fed with slaughterhouse waste, one with a hollow fiber membrane contractor for ammonia removal and one without. Different organic loading rates (OLR) and free ammonia and sulfide concentrations were investigated. In the reactor with the membrane contactor, the NH4-N concentration was reduced threefold. At a moderate OLR (3.1 kg chemical oxygen demand - COD/m(3)/d), this reactor performed significantly better than the reference reactor. At high OLR (4.2 kg COD/m(3)/d), the reference reactor almost stopped producing methane (0.01 Nl/gCOD). The membrane reactor also showed a stable process with a methane yield of 0.23 Nl/g COD was achieved. Both reactors had predominantly a hydrogenotrophic microbial consortium, however in the membrane reactor the genus Methanosaeta (acetoclastic) was also detected. In general, all relevant parameters and the methanogenic consortium indicated improved anaerobic digestion of the reactor with the membrane.

  3. Validação das relações dos aminoácidos metionina, treonina, triptofano e valina com a lisina digestível na proteína ideal em rações para porcas em lactação Validation of the relationships of methionine, threonine, tryptophan and valine amino acids with the digestible lysine in the ideal protein in feedings for lactating sows

    Directory of Open Access Journals (Sweden)

    Douglas Haese

    2010-07-01

    Full Text Available Com o objetivo de validar as relações de aminoácidos digestíveis com a lisina digestível propostas com base no conceito da proteína ideal, foi realizado um experimento com 120 matrizes suínas em lactação, distribuídas em delineamento em blocos, composto de seis dietas, 20 blocos e uma matriz por unidade experimental. As dietas foram formuladas a partir de uma dieta-referência - contendo 19,5% de proteína bruta (PB e 0,95% de lisina digestível - suplementada com metionina, treonina, triptofano e/ou valina digestíveis para atingir as relações com a lisina digestível determinadas com base na proteína ideal. As dietas não influenciaram a perda e a variação de peso, a espessura e a variação da espessura de toucinho, a proteína e a variação de proteína corporal, a gordura e a variação de gordura corporal nem alteraram o intervalo desmame-estro ou a produção de leite das porcas. As relações dos aminoácidos metionina + cistina, treonina, triptofano e valina digestíveis com a lisina digestível propostas com base na proteína ideal atendem às exigências de porcas em lactação para melhores desempenhos produtivo e reprodutivo.This experiment aimed at validating the relationship between digestible amino acids with the digestible lysine proposed on the basis of the ideal protein concept. This experiment used 120 lactating sows distributed in a complete randomized block design with six treatments and 20 repetitions and one sow per experimental unity. The diets were formulated based on a reference diet - which contained 19.5% of crude protein (CP and 0.95% of digestible lysine supplemented with digestible methionine, threonine, tryptophan and/or valine to meet the ideal protein based determined relationships with digestible lysine. The diets did not change weight loss and variation, thickness and thickness variation of backfat, protein and body protein variation, fat and body fat variation, neither they changed the

  4. [Effects of the size of magnetic particles of immobilized enzyme reactors on the digestion performance].

    Science.gov (United States)

    Zhang, Jiao; Zhou, Lianqi; Tian, Fang; Zhang, Yangjun; Qian, Xiaohong

    2013-02-01

    We applied immobilized enzyme reactors prepared with different sizes of magnetic particles into protein and proteome digestion. In addition, the influences of different sizes of the magnetic particles were studied on the reunion, enzyme efficiency and leakage sites. The experimental results showed that in comparison with the submicron magnetic particles, the amount of trypsin immobilized on the magnetic nanoparticles was 3. 5 times more than that of the submicron magnetic particles. However, the enzymatic efficiency was at the same level when the same amount of trypsin was used, and the reunion phenomenon was obviously improved when the size of the magnetic nanoparticles increased. Taking the immobilized enzyme reactor of 20 nm magnetic nanoparticles as an example, the digestion performance was further examined. The experimental results showed that rapid digestion could be achieved within 1 mm when the mass ratio of the trypsin and bovine serum albumin was 1:1. The peptide number of 0 missed cleavage site and the sequence coverage changed little after the protein was digested for 10 mm. It was concluded that the digestion efficiency of the immobilized enzyme reactor was much better than that of the in-solution digestion. When the immobilized enzyme reactors and the free trypsin were used for digestion, little differences of the leakage sites were found. Therefore, the immobilized enzyme reactors prepared with different sizes of magnetic particles can be applied in proteomic research for quick and efficient digestion.

  5. Improved protein quality in transgenic soybean expressing a de novo synthetic protein, MB-16.

    Science.gov (United States)

    Zhang, Yunfang; Schernthaner, Johann; Labbé, Natalie; Hefford, Mary A; Zhao, Jiping; Simmonds, Daina H

    2014-06-01

    To improve soybean [Glycine max (L.) Merrill] seed nutritional quality, a synthetic gene, MB-16 was introduced into the soybean genome to boost seed methionine content. MB-16, an 11 kDa de novo protein enriched in the essential amino acids (EAAs) methionine, threonine, lysine and leucine, was originally developed for expression in rumen bacteria. For efficient seed expression, constructs were designed using the soybean codon bias, with and without the KDEL ER retention sequence, and β-conglycinin or cruciferin seed specific protein storage promoters. Homozygous lines, with single locus integrations, were identified for several transgenic events. Transgene transmission and MB-16 protein expression were confirmed to the T5 and T7 generations, respectively. Quantitative RT-PCR analysis of developing seed showed that the transcript peaked in growing seed, 5-6 mm long, remained at this peak level to the full-sized green seed and then was significantly reduced in maturing yellow seed. Transformed events carrying constructs with the rumen bacteria codon preference showed the same transcription pattern as those with the soybean codon preference, but the transcript levels were lower at each developmental stage. MB-16 protein levels, as determined by immunoblots, were highest in full-sized green seed but the protein virtually disappeared in mature seed. However, amino acid analysis of mature seed, in the best transgenic line, showed a significant increase of 16.2 and 65.9 % in methionine and cysteine, respectively, as compared to the parent. This indicates that MB-16 elevated the sulfur amino acids, improved the EAA seed profile and confirms that a de novo synthetic gene can enhance the nutritional quality of soybean.

  6. Effects of concentrate crude protein content on nutrient digestibility, energy utilization, and methane emissions in lactating dairy cows fed fresh-cut perennial grass.

    Science.gov (United States)

    Hynes, D N; Stergiadis, S; Gordon, A; Yan, T

    2016-11-01

    Although many studies have investigated mitigation strategies for methane (CH4) output from dairy cows fed a wide variety of diets, research on the effects of concentrate crude protein (CP) content on CH4 emissions from dairy cows offered fresh grass is limited. The present study was designed to evaluate the effects of cow genotype and concentrate CP level on nutrient digestibility, energy utilization, and CH4 emissions in dairy cows offered fresh-grass diets. Twelve multiparous lactating dairy cows (6 Holstein and 6 Holstein × Swedish Red) were blocked into 3 groups for each breed and assigned to a low-, medium-, or high-CP concentrate diet [14.1, 16.1, and 18.1% CP on a dry matter (DM) basis, respectively], in a 3-period changeover study (25d per period). Total diets contained (DM basis) 32.8% concentrates and 67.2% perennial ryegrass, which was harvested daily. All measurements were undertaken during the final 6d of each period: digestibility measurements for 6d and calorimetric measurements in respiration chambers for 3d. Feed intake and milk production data were reported in a previous paper. We observed no significant interaction between concentrate CP level and cow genotype on any parameter. Concentrate CP level had no significant effect on any energy utilization parameter, except for urinary energy output, which was positively related to concentrate CP level. Similarly, concentrate CP content had no effect on CH4 emission (g/d), CH4 per kg feed intake, or nutrient digestibility. Cross breeding of Holstein cows significantly reduced gross energy, digestible energy, and metabolizable energy intake, heat production, and milk energy output. However, cow genotype had no significant effect on energy utilization efficiency or CH4 parameters. Furthermore, the present study yielded a value for gross energy lost as CH4 (5.6%) on fresh grass-based diets that was lower than the widely accepted value of 6.5%. The present findings indicate that reducing concentrate CP

  7. 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 α-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.

  8. Post-polymerization photografting on methacrylate-based monoliths for separation of intact proteins and protein digests with comprehensive two-dimensional liquid chromatography hyphenated with high-resolution mass spectrometry.

    Science.gov (United States)

    Vonk, Rudy J; Wouters, Sam; Barcaru, Andrei; Vivó-Truyols, Gabriel; Eeltink, Sebastiaan; de Koning, Leo J; Schoenmakers, Peter J

    2015-05-01

    Post-polymerization photografting is a versatile tool to alter the surface chemistry of organic-based monoliths so as to obtain desired stationary phase properties. In this study, 2-acrylamido-2-methyl-1-propanesulfonic acid was grafted to a hydrophobic poly(butyl methacrylate-co-ethylene glycol dimethacrylate) monolith to create a strong cation exchange stationary phase. Both single-step and two-step photografting were addressed, and the effects of grafting conditions were assessed. An experimental design has been applied in an attempt to optimize three of the key parameters of the two-step photografting chemistry, i.e. the grafting time of the initiator, the monomer concentration and the monomer irradiation time. The photografted columns were implemented in a comprehensive two-dimensional column liquid chromatography ( (t) LC ×  (t) LC) workflow and applied for the separation of intact proteins and peptides. A baseline separation of 11 intact proteins was obtained within 20 min by implementing a gradient across a limited RP composition window in the second dimension. (t) LC ×  (t) LC with UV detection was used for the separation of cytochrome c digest, bovine serum insulin digest and a digest of a complex protein mixture. A semi-quantitative estimation of the occupation of separation space, the orthogonality, of the (t) LC ×  (t) LC system yielded 75%. The (t) LC ×  (t) LC setup was hyphenated to a high-resolution Fourier transform ion cyclotron resonance mass spectrometer instrument to identify the bovine serum insulin tryptic peptides and to demonstrate the compatibility with MS analysis.

  9. 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 6.427 mg/L of TN, NH4-N and PO4-P respectively after 40 d of fed-batch cultivation, while in batch cultivation the quantities of TN, NH4-N and PO4-P removed were 33.331, 37.227 and 1.323 mg/L. Biomass production of Desmodesmus sp. was also enhanced in fed-batch cultivation, when ADW loading was carried out every 2 days; the biomass concentration peaked at 1.039 g/L, which was three times higher than that obtained in batch cultivation (0.385 g/L). The highest lipid production (261.8 mg/L) was also recorded in fed-batch cultivation as compared to batch cultivation (83.3 mg/L). Fed-batch cultivation of Desmodesmus sp. could provide effective control of nutrients limitation and/or ammonia inhibition on microalgae cultivation.

  10. 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.

  11. 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 μg/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.

  12. 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.

  13. Utilizing enzymatic digestion procedures in the bioanalytical laboratory.

    Science.gov (United States)

    Szapacs, Matthew; Mencken, Thomas; Williams, Jon; Li, Yi

    2016-01-01

    In recent years, the use of LC-MS technologies in the bioanlytical laboratory for quantitation of peptide/protein biomarkers and biotherapeutics has increased dramatically. The increased interest is due to the improvement in sensitivity of MS instruments over the last 5-10 years, as well as its proven ability to overcome some common issues associated with immunoassay, namely selectivity and reagent availability. However, large proteins (>10 kDa) chromatograph and ionize poorly. To overcome this challenge, LC-MS/MS workflows for proteins larger than 10 kDa utilize enzymatic digestion procedures with subsequent quantitation of one or more of these enzymatically derived peptides to act as a surrogate for the intact protein. Here, recommendations of digestion technique and potential internal standards are summarized.

  14. Improvement of wheat protein quality and quantity by breeding.

    Science.gov (United States)

    Johnson, V A; Mattern, P J

    1978-01-01

    Substantial genetic, variability for grain protein content in wheat has been identified. In appropriate combinations known genes can increase protein content of wheat grain by 5 percentage points. Productive high protein experimental lines with good agronomic traits and satisfactory processing attributes have been identified. A high protein hard red winter variety developed in Nebraska was released for commercial production in 1975 under the name "Lancota". The high protein of Lancota resides entirely in the starchy endosperm portion of the kernel and is fully transmissible to white milled flour. The high protein of Lancota results from elevated NO3 reductase activity, increased N-absorption by the roots, and more complete translocation of N to the grain. Despite strong environmental influence on wheat protein level, genes for high protein have been demonstrated to effectively increase protein content in many different production environments. Lysine % of protein decreases but lysine % of grain increases as protein is increased. Genetic variability for lysine of sufficient magnitude to overcome the normal depression of lysine % of protein as protein is increased has been uncovered. Experimental lines have been developed in the ARS-Nebraska program in which genes for high protein and high lysine were combined. The lines have been widely distributed for use in other breeding programs.

  15. Improved protein structure selection using decoy-dependent discriminatory functions

    Directory of Open Access Journals (Sweden)

    Levitt Michael

    2004-06-01

    -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.

  16. Lipid digestion of protein stabilized emulsions investigated in a dynamic in vitro gastro-intestinal model system

    NARCIS (Netherlands)

    Helbig, A.; Silletti, E.; Aken, G.A. van; Oosterveld, A.; Minekus, M.; Hamer, R.J.; Gruppen, H.

    2013-01-01

    This study investigated the effect of gastric passage of protein stabilized emulsions, i.e., whey protein isolate (WPI) and lysozyme, under dynamic in vitro conditions on both the gastric and intestinal lipolysis. Emulsions were prepared at neutral pH to enable an opposite surface charge. Experiment

  17. Lipid Digestion of Protein Stabilized Emulsions Investigated in a Dynamic In Vitro Gastro-Intestinal Model System

    NARCIS (Netherlands)

    Helbig, A.; Silletti, E.; Aken, van G.A.; Oosterveld, A.; Minekus, M.; Hamer, R.J.; Gruppen, H.

    2013-01-01

    This study investigated the effect of gastric passage of protein stabilized emulsions, i.e., whey protein isolate (WPI) and lysozyme, under dynamic in vitro conditions on both the gastric and intestinal lipolysis. Emulsions were prepared at neutral pH to enable an opposite surface charge. Experiment

  18. Local e extensão da digestão em terneiros desmamados alimentados "Ad Libitum" com dietas ricas em concentrado contendo diferentes fontes de proteína suplementar Site and extent of digestion by weaned calves fed "Ad Libitum" on high concentrate-rich diets with different supplemental protein sources

    Directory of Open Access Journals (Sweden)

    Gilberto Vilmar Kozloski

    1999-06-01

    Full Text Available Foram utilizados nove terneiros da raça Holandesa (139 ± 2,4kg, fistulados no duodeno, em um delineamento experimental completamente casualizado para estudar o efeito da suplementação com diferentes fontes protéicas (2% de uréia ou 10% de farinha de carne na dieta total de dietas ricas em concentrado (milho e farelo de soja sobre o local e extensão da digestão do alimento, em condições de consumo ad libitum. A relação volumoso:concentrado das dietas foi de 30:70. A digestibilidade ruminal e total da matéria seca, matéria orgânica e nitrogênio, assim como o fluxo duodenal de nitrogênio microbiano e residual do alimento, e a eficiência da síntese protéica microbiana foram semelhantes nos dois tratamentos (P>0,10. A digestibilidade ruminal do amido também foi semelhante, mas a digestibilidade total foi menor no tratamento com uréia (PNine weaned Holstein calves (139 ± 2.4kg fitted with duodenal cannulas were used in a completly ramdomized design to study the effects of supplemental protein sources (2% of urea or 10% of meat meal of high concentrate diets (corn and soybean meal on site and extent of digestion of feed, in ad libitum feeding conditions. The forage:concentrate ratio was 30:70. The ruminal and total digestibility of dry matter, organic matter and nitrogen, as well as microbial and feed residual nitrogen duodenal flow, and the microbial protein synthesis efficiency were similar for both treatments (P>0.10. The ruminal digestibility of starch was also similar, but total digestibility was lower for urea treatment (P<0.10. However, this is probably due more to the limited capacity of ruminants to digest starch in the intestines than to an effect of the protein supplement. The use of urea or meat meal, as supplemental protein sources for corn and soybean meal based diets, formulated to supply the growth requirements of calves fed ad libitum, did not affect the digestion process.Thus, the choise of the protein

  19. Phytic acid reduction in soy protein improves zinc bioavailability

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, J.R.; Wong, M.S.; Burns, R.A.; Erdman, J.W. Jr. (Univ. of Illinois, Urbana (United States) Mead Johnson Research Center, Evansville, IN (United States))

    1991-03-15

    The objective of this study was to confirm previous studies that have suggested that reduction of phytic acid in soy improved zinc bioavailability (BV). Two commercially-produced soybean isolates containing either a normal phytic acid level or a reduced level were formulated into diets so as to provide 6 or 9 ppm zinc. Control diets were egg white protein-based and contained 3, 6 or 9 ppm zinc from zinc carbonate. Weanling male rats were fed these diets for 21 days and food intake and weight gain monitored. Slope ratio analysis of total tibia zinc content compared to total zinc intake revealed that zinc BV from reduced phytic acid soy isolate-containing diets was indistinguishable from control egg white diets. In contrast, zinc BV from normal soy isolate diets was significantly reduced compared to reduced phytic acid and control diets. These results coupled with other results indicate that phytic acid is the inhibitory factor in soybean products that results in reduced zinc BV.

  20. Directed evolution methods for improving polypeptide folding and solubility and superfolder fluorescent proteins generated thereby

    Science.gov (United States)

    Waldo, Geoffrey S.

    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.

  1. Study of Raphanus sativus L. on Improving Digestion Function%白萝卜促消化功能的研究

    Institute of Scientific and Technical Information of China (English)

    彭章蒙; 孟晓伟; 刘朋朋; 张宏桂

    2013-01-01

    To study the Effects of Raphanus sativus L. on improving digestive function. The distance of ink moving in small intestine in KM mice was used to evaluate the effect. The body weight,food intakes,food efficiency ratio and the secretion amount of SD rats'gastric juice and pepsin activity were also used to evaluate the effect. Raphanus sativus L. could increase the rate of ink promoting,which was superior to that in the model control group,it also could promote the secretion amount of gastric juice and pepsin,which was superior to that in the control group. Raphanus sativus L. has the function of promoting the digestion and absorption function.%研究白萝卜的促消化作用。根据卫生部《保健食品检验与评价技术规范》2003年版规定的方法,取雄性昆明小鼠60只,进行小鼠小肠墨汁推进试验;选用清洁级雄性SD大鼠40只,进行大鼠体重、体重增重、摄食量、食物利用率和大鼠胃蛋白酶测定试验。白萝卜能显著提高小肠推进率;显著增强大鼠胃蛋白酶排出量。白萝卜具有助消化作用。

  2. 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 positive

  3. Influence of ruminal or duodenal soybean oil infusion on intake, ruminal fermentation, site and extent of digestion, and microbial protein synthesis in beef heifers consuming grass hay.

    Science.gov (United States)

    Krysl, L J; Judkins, M B; Bohman, V R

    1991-06-01

    Six heifers (two Hereford X Jersey, four Hereford X Longhorn; average BW 278 kg) cannulated at the rumen and duodenum and fed a grass hay (fescue/orchardgrass) diet were used in a replicated 3 X 3 Latin square. Treatments were either no infusion (C), 150 ml of duodenally infused soybean oil (DI), or 150 ml of ruminally infused soybean oil (RI)/heifer twice daily for a total daily infusion of 300 ml of soybean oil. Periods of the Latin square included 18 d for adaptation and 5 d for collection. Forage OM, ADF, NDF, and N intakes were not affected (P greater than .10) by soybean oil infusion. Ruminal (P = .11) and total tract (P less than .10) OM digestibilities were decreased by RI compared with C or DI, but ADF and NDF digestibilities were not affected by treatment. Duodenal N (P less than .05) and microbial N flows were increased (P less than .10) for C and RI compared with DI. Microbial efficiency (g of N/kg of OM truly fermented) was improved (P less than .10) by RI compared with DI but did not differ (P greater than .10) from C. Ruminal pH was lower (P less than .05) with RI than with either C or DI. Ruminal NH3 N, total VFA, and acetate were not affected (P greater than .10) by treatment. Propionate (mol/100 mol) was greater (P less than .05) with RI than with DI and C, but the proportion of butyrate did not differ among treatments. These data indicate minimal direct benefits for improving forage usage as a result of soybean oil infusion with a 100% grass diet; however, animals should realize benefits from additional dietary energy provided by infused lipid.

  4. Improving protein array performance: focus on washing and storage conditions.

    Science.gov (United States)

    Nath, Nidhi; Hurst, Robin; Hook, Brad; Meisenheimer, Poncho; Zhao, Kate Q; Nassif, Nadine; Bulleit, Robert F; Storts, Douglas R

    2008-10-01

    For protein microarrays, maintaining protein stability during the slide processing steps of washing, drying, and storage is of major concern. Although several studies have focused on the stability of immobilized antibodies in antibody microarrays, studies on protein-protein interaction arrays and enzyme arrays are lacking. In this paper we used five bait-prey protein interaction pairs and three enzymes to optimize the washing, drying, and storage conditions for protein arrays. The protein arrays for the study were fabricated by combining HaloTag technology and cell-free protein expression. The HaloTag technology, in combination with cell-free expression, allowed rapid expression and immobilization of fusion proteins on hydrogel-coated glass slides directly from cell extracts without any prior purification. Experimental results indicate enzyme captured on glass slides undergoes significant loss of activity when washed and spin-dried using only phosphate buffer, as is typically done with antibody arrays. The impact of washing and spin-drying in phosphate buffer on protein-protein interaction arrays was minimal. However, addition of 5% glycerol to the wash buffer helps retain enzyme activity during washing and drying. We observed significant loss of enzyme activity when slides were stored dry at 4 degrees C, however immobilized enzymes remained active for 30 days when stored at -20 degrees C in 50% glycerol. We also found that cell-free extract containing HaloTag-fused enzymes could undergo multiple freeze/thaw cycles without any adverse impact on enzyme activity. The findings indicate that for large ongoing studies, proteins of interest expressed in cell-free extract can be stored at -70 degrees C and repeatedly used to print small batches of protein array slides to be used over a few weeks.

  5. Small intestinal digestion of raw cornstarch in cattle consuming a soybean hull-based diet is improved by duodenal casein infusion.

    Science.gov (United States)

    Brake, D W; Titgemeyer, E C; Bailey, E A; Anderson, D E

    2014-09-01

    Six duodenally and ileally cannulated steers were used in 3 sequential studies to measure 1) basal nutrient flows from a soybean hull-based diet, 2) small intestinal digestibility of raw cornstarch continuously infused into the duodenum, and 3) responses of small intestinal starch digestion to duodenal infusion of 200 or 400 g/d casein. Our objective was to evaluate responses in small intestinal starch digestion in cattle over time and to measure responses in small intestinal starch digestion to increasing amounts of MP. On average, cattle consumed 3.7 kg/d DM, 68 g/d dietary N, and 70 g/d dietary starch. Starch flow to the duodenum was small (38 g/d), and N flow was 91 g/d. Small intestinal digestibility of duodenal N was 57%, and small intestinal digestion of duodenal starch flow was extensive (92%). Small intestinal starch digestibility was 34% when 1.5 kg/d raw cornstarch was continuously infused into the duodenum. Subsequently, cattle were placed in 1 of 2 replicated Latin squares that were balanced for carryover effects to determine response to casein infusions and time required for adaptation. Duodenal infusion of casein linearly increased (P ≤ 0.05) small intestinal starch digestibility, and small intestinal starch digestion adapted to infusion of casein in 6 d. Ethanol-soluble starch and unpolymerized glucose flowing to the ileum increased linearly (P ≤ 0.05) with increasing infusion of casein. Plasma cholecystokinin was not affected by casein infusion, but circulating levels of glucose were increased by casein supplementation (P ≤ 0.05). Responses in small intestinal starch digestion in cattle adapted to casein within 6 d, and increases in duodenal supply of casein up to 400 g/d increased small intestinal starch digestion in cattle.

  6. 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...

  7. Rapid and simultaneous determination of essential minerals and trace elements in human milk by improved flame atomic absorption spectroscopy (FAAS) with microwave digestion.

    Science.gov (United States)

    Luo, Yang; Zhang, Bo; Chen, Ming; Wang, Jue; Zhang, Xue; Gao, Wei-Yin; Huang, Jun-Fu; Fu, Wei-Ling

    2010-09-08

    A method for the simultaneous and economical determination of many trace elements in human milk is developed. Two multi-element hollow cathode lamps (HCLs) were used instead of single-element HCLs to improve the sample throughput of flame atomic absorption spectroscopy (FAAS). The microwave digestion of milk is optimized prior to detection, and the performance characteristics of the improved analysis method are identified. Clinical samples are detected by both FAAS and inductively coupled plasma-optical emission spectroscopy (ICP-OES) for methodology evaluation. Results reveal that the proposed FAAS with multi-element HCLs could determine six essential minerals and trace elements within 15 min. This method provides a linear analytical range of 0.01-10 mg L(-1). For Ca, Cu, Fe, Mg, Mn, and Zn, the limits of determination are 1.5, 3, 1.8, 2.2, 2.1, and 1.3 microg L(-1), respectively. The mean relative standard deviations (RSDs) of intra- and interassays are lower than 7%. Excellent operational characteristics of rapidity, simplicity, and economy make the proposed method a promising one for the quantification of trace elements in human milk in clinics of underdeveloped areas.

  8. Pretreatment of oil palm empty fruit bunch (OPEFB) by N-methylmorpholine-N-oxide (NMMO) for biogas production: structural changes and digestion improvement.

    Science.gov (United States)

    Purwandari, Fiametta Ayu; Sanjaya, Adhitya Pitara; Millati, Ria; Cahyanto, Muhammad Nur; Horváth, Ilona Sárvári; Niklasson, Claes; Taherzadeh, Mohammad J

    2013-01-01

    Pretreatment of OPEFB (oil palm empty fruit bunch) by NMMO (N-methylmorpholine-N-oxide) on its subsequent digestions was investigated. The pretreatments were carried out at 90 and 120 °C for 1, 3, and 5h in three different modes of dissolution (by 85% NMMO solution), ballooning (79% NMMO solution), and swelling (73% NMMO solution). The total solid recovery after the pretreatment was 89-94%. The pretreatment process did not have a major impact on the composition of OPEFB, other than a reduction of ash from 5.4% up to 1.3%. The best improvement in biogas production was achieved by a dissolution mode pretreatment of OPEFB, using conditions of 85% NMMO, 3h, and 120 °C. It resulted in 0.408 Nm(3)/kg VS methane yield and 0.032 Nm(3)CH(4)/kg VS/day initial methane production rate, which correspond in improving by 48% and 167% compared to the untreated OPEFB, respectively.

  9. Improving protein function prediction methods with integrated literature data

    Directory of Open Access Journals (Sweden)

    Gabow Aaron P

    2008-04-01

    Full Text Available Abstract Background Determining the function of uncharacterized proteins is a major challenge in the post-genomic era due to the problem's complexity and scale. Identifying a protein's function contributes to an understanding of its role in the involved pathways, its suitability as a drug target, and its potential for protein modifications. Several graph-theoretic approaches predict unidentified functions of proteins by using the functional annotations of better-characterized proteins in protein-protein interaction networks. We systematically consider the use of literature co-occurrence data, introduce a new method for quantifying the reliability of co-occurrence and test how performance differs across species. We also quantify changes in performance as the prediction algorithms annotate with increased specificity. Results We find that including information on the co-occurrence of proteins within an abstract greatly boosts performance in the Functional Flow graph-theoretic function prediction algorithm in yeast, fly and worm. This increase in performance is not simply due to the presence of additional edges since supplementing protein-protein interactions with co-occurrence data outperforms supplementing with a comparably-sized genetic interaction dataset. Through the combination of protein-protein interactions and co-occurrence data, the neighborhood around unknown proteins is quickly connected to well-characterized nodes which global prediction algorithms can exploit. Our method for quantifying co-occurrence reliability shows superior performance to the other methods, particularly at threshold values around 10% which yield the best trade off between coverage and accuracy. In contrast, the traditional way of asserting co-occurrence when at least one abstract mentions both proteins proves to be the worst method for generating co-occurrence data, introducing too many false positives. Annotating the functions with greater specificity is harder

  10. Improving the quality of rice straw by urea and calcium hydroxide on rumen ecology, microbial protein synthesis in beef cattle.

    Science.gov (United States)

    Polyorach, S; Wanapat, M

    2015-06-01

    Four rumen-fistulated beef cattle were randomly assigned to four treatments according to a 4 × 4 Latin square design to study the influence of urea and calcium hydroxide [Ca(OH)2 ] treatment of rice straw to improve the nutritive value of rice straw. Four dietary treatments were as follows: untreated rice straw, 50 g/kg urea-treated rice straw, 20 g/kg urea + 20 g/kg calcium hydroxide-treated rice straw and 30 g/kg urea + 20 g/kg calcium hydroxide-treated rice straw. All animals were kept in individual pens and fed with concentrate at 0.5 g/kg of BW (DM), rice straw was fed ad libitum. The experiment was conducted for four periods, and each period lasted for 21 days. During the first 14 days, DM feed intake measurements were made while during the last 7 days, all cattle were moved to metabolism crates for total faeces and urine collections. The results revealed that 20 g/kg urea + 20 g/kg calcium hydroxide-treated rice straw improved the nutritive value of rice straw, in terms of dry matter intake, digestibility, ruminal volatile fatty acids, population of bacteria and fungi, nitrogen retention and microbial protein synthesis. Based on this study, it could be concluded that using urea plus calcium hydroxide was one alternative method to improve the nutritive value of rice straw, rumen ecology and fermentation and thus a reduction of treatment cost.

  11. Bovine milk proteome in the first 9 days: protein interactions in maturation of the immune and digestive system of the newborn.

    Directory of Open Access Journals (Sweden)

    Lina Zhang

    Full Text Available In order to better understand the milk proteome and its changes from colostrum to mature milk, samples taken at seven time points in the first 9 days from 4 individual cows were analyzed using proteomic techniques. Both the similarity in changes from day 0 to day 9 in the quantitative milk proteome, and the differences in specific protein abundance, were observed among four cows. One third of the quantified proteins showed a significant decrease in concentration over the first 9 days after calving, especially in the immune proteins (as much as 40 fold. Three relative high abundant enzymes (XDH, LPL, and RNASE1 and cell division and proliferation protein (CREG1 may be involved in the maturation of the gastro-intestinal tract. In addition, high correlations between proteins involved in complement and blood coagulation cascades illustrates the complex nature of biological interrelationships between milk proteins. The linear decrease of protease inhibitors and proteins involved in innate and adaptive immune system implies a protective role for protease inhibitor against degradation. In conclusion, the results found in this study not only improve our understanding of the role of colostrum in both host defense and development of the newborn calf but also provides guidance for the improvement of infant formula through better understanding of the complex interactions between milk proteins.

  12. Ruminal degradability and intestinal digestibility of individual amino acids in mixed diets with different crude protein levels measured by the modified in vitro three-step and mobile nylon bag technique.

    Science.gov (United States)

    Gao, Wei; Zhang, Bowen; Lv, Bo; Liu, Chenli; Chen, Daofu

    2016-04-01

    The ruminal degradability and intestinal digestibility of dry matter (DM), crude protein (CP) and amino acids (AA) in three total mixed rations with different CP levels were estimated using the modified in vitro three-step procedure (TSP) and mobile nylon bag (MNB) technique on growing lambs. The ruminal effective degradability of DM and CP did not respond with increasing dietary CP level. However, the intestinal digestibility of DM was significantly increased with increasing dietary CP level estimated by TSP (P < 0.05) or MNB method (P < 0.01). Intestinal digestibility coefficients of CP determined by TSP were lower than those of the MNB method. Histidine was extensively degraded by rumen micro-organisms, while tyrosine was the most anti-degradable AA among the samples. The ruminal AA degradability exhibited no significant differences except for threonine, tryptophan, alanine, aspartic acid and proline for the three diets. Similarly, only a few AAs (i.e. histidine, methionine, tryptophan, aspartic acid and cysteine in TSP; histidine, tryptophan, aspartic acid and serine in MNB) had significant differences in their intestinal digestibility; in addition, values of MNB were lower than that of the TSP method, indicating that intestinal digestibility of DM seems to be overestimated in TSP, while that of CP might be overestimated in the MNB method.

  13. Protein sequence classification with improved extreme learning machine algorithms.

    Science.gov (United States)

    Cao, Jiuwen; Xiong, Lianglin

    2014-01-01

    Precisely classifying a protein sequence from a large biological protein sequences database plays an important role for developing competitive pharmacological products. Comparing the unseen sequence with all the identified protein sequences and returning the category index with the highest similarity scored protein, conventional methods are usually time-consuming. Therefore, it is urgent and necessary to build an efficient protein sequence classification system. In this paper, we study the performance of protein sequence classification using SLFNs. The recent efficient extreme learning machine (ELM) and its invariants are utilized as the training algorithms. The optimal pruned ELM is first employed for protein sequence classification in this paper. To further enhance the performance, the ensemble based SLFNs structure is constructed where multiple SLFNs with the same number of hidden nodes and the same activation function are used as ensembles. For each ensemble, the same training algorithm is adopted. The final category index is derived using the majority voting method. Two approaches, namely, the basic ELM and the OP-ELM, are adopted for the ensemble based SLFNs. The performance is analyzed and compared with several existing methods using datasets obtained from the Protein Information Resource center. The experimental results show the priority of the proposed algorithms.

  14. Heat shock response improves heterologous protein secretion in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Hou, Jin; Österlund, Tobias; Liu, Zihe

    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...

  15. Strain engineering for improved expression of recombinant proteins in bacteria.

    Science.gov (United States)

    Makino, Tomohiro; Skretas, Georgios; Georgiou, George

    2011-05-14

    Protein expression in Escherichia coli represents the most facile approach for the preparation of non-glycosylated proteins for analytical and preparative purposes. So far, the optimization of recombinant expression has largely remained a matter of trial and error and has relied upon varying parameters, such as expression vector, media composition, growth temperature and chaperone co-expression. Recently several new approaches for the genome-scale engineering of E. coli to enhance recombinant protein expression have been developed. These methodologies now enable the generation of optimized E. coli expression strains in a manner analogous to metabolic engineering for the synthesis of low-molecular-weight compounds. In this review, we provide an overview of strain engineering approaches useful for enhancing the expression of hard-to-produce proteins, including heterologous membrane proteins.

  16. Super High Dosing with a Novel Buttiauxella Phytase Continuously Improves Growth Performance, Nutrient Digestibility, and Mineral Status of Weaned Pigs.

    Science.gov (United States)

    Zeng, Zhikai; Li, Qingyun; Tian, Qiyu; Zhao, Panfeng; Xu, Xiao; Yu, Shukun; Piao, Xiangshu

    2015-11-01

    This study was conducted to evaluate the efficacy of a novel Buttiauxella phytase to pigs fed P-deficient, corn-soybean meal diets. One hundred and twenty crossbred piglets (9.53 ± 0.84 kg) were allocated to one of five treatments which consisted of four low P diets (0.61 % Ca and 0.46 % total P) supplemented with 0, 500, 1,000, or 20,000 FTU/kg phytase as well as a positive control diet (0.77 % Ca and 0.62 % total P). Each treatment had six replicated pens with four pigs per pen. Pigs were fed the experimental diets for 28 days. Phytase supplementation linearly improved (P phytase (20,000 FTU/kg) further increased (P phytase inclusion group, as well as ATTD of Ca and P. Metacarpal bone characteristics and several trace mineral concentration in bone, plasma, or organ tissues were linearly (P phytase. Super high dosing with phytase (20,000 FTU/kg) supplementation improved (P phytase supplementation (500 or 1,000 FTU/kg). In conclusion, supplementation of 500 FTU of Buttiauxella phytase/kg and above effectively hydrolyzed phytate in a low-P corn-soybean diet for pigs. In addition, a super high dosing with phytase (20,000 FTU/kg) improved macro- or micro mineral availability and growth performance.

  17. Changes over lactation in breast milk serum proteins involved in the maturation of immune and digestive system of the infant

    NARCIS (Netherlands)

    Zhang, Lina; Waard, de Marita; Verheijen, Hester; Boeren, Sjef; Hageman, Jos A.; Hooijdonk, van Toon; Vervoort, Jacques; Goudoever, van Johannes B.; Hettinga, Kasper

    2016-01-01

    To objective of this study was to better understand the biological functions of breast milk proteins in relation to the growth and development of infants over the first six months of life. Breast milk samples from four individual women collected at seven time points in the first six months after

  18. Lipid transfer protein : a pan-allergen in plant-derived foods that is highly resistant to pepsin digestion

    NARCIS (Netherlands)

    Asero, R.; Mistrello, G.; Roncarolo, D.; Vries, de S.C.; Gautier, M.F.; Ciurana, C.L.; Verbeek, E.; Mohammadi, T.; Knul-Brettlova, V.; Akkerdaas, J.H.; Bulder, I.; Aalberse, R.C.; Ree, van R.

    2000-01-01

    Lipid transfer proteins (LTPs) are small molecules of approximately 10 kD that demonstrate high stability. They have recently been identified as allergens in the Rosaceae subfamilies of the Prunoideae (peach, apricot, plum) and of the Pomoideae (apple). They belong to a family of structurally highly

  19. Identifying Gel-Separated Proteins Using In-Gel Digestion, Mass Spectrometry, and Database Searching: Consider the Chemistry

    Science.gov (United States)

    Albright, Jessica C.; Dassenko, David J.; Mohamed, Essa A.; Beussman, Douglas J.

    2009-01-01

    Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry is an important bioanalytical technique in drug discovery, proteomics, and research at the biology-chemistry interface. This is an especially powerful tool when combined with gel separation of proteins and database mining using the mass spectral data. Currently, few hands-on…

  20. Performance of piglets in response to the standardized ileal digestible phenylalanine and tyrosine supply in low-protein diets.

    Science.gov (United States)

    Gloaguen, M; Le Floc'h, N; Primot, Y; Corrent, E; van Milgen, J

    2014-09-01

    Reducing the CP level of the diet allows for a reduction in N excretion without limiting performance as long as the amino acid (AA) requirements are covered. The availability of crystalline AA has permitted for a considerable reduction in the CP level of diets, practically used in pig nutrition. The adoption of low CP diets and the extent to which the CP content can be reduced further depends on the knowledge of the minimum levels of indispensable AA that maximize growth. The standardized ileal digestible (SID) Phe : Lys and Tyr : Lys requirements and the possibility to substitute Tyr by Phe have never been studied in piglets. The objectives of this study were to estimate these requirements in 10 to 20 kg pig as well as to determine the extent to which Phe can be used to cover the Tyr requirement. In three dose-response studies, six pigs within each of 14 blocks were assigned to six low CP diets (14.5% CP) sub-limiting in Lys at 1.00% SID. In experiment 1, the SID Phe : Lys requirement estimate was assessed by supplementing a Phe-deficient diet with different levels of l-Phe to attain 33%, 39%, 46%, 52%, 58%, and 65% SID Phe : Lys. Because Phe can be used for Tyr synthesis, the diets provided a sufficient Tyr supply. A similar approach was used in experiment 2 with six levels of l-Tyr supplementation to attain 21%, 27%, 33%, 39%, 45% and 52% SID Tyr : Lys. Phenylalanine was supplied at a level sufficient to sustain maximum growth (estimated in experiment 1). The SID Phe : Lys and SID Tyr : Lys requirements for maximizing daily gain were 54% and 40% using a curvilinear-plateau model, respectively. A 10% deficiency in Phe and Tyr reduced daily gain by 3.0% and 0.7%, respectively. In experiment 3, the effect of the equimolar substitution of dietary SID Tyr by Phe to obtain 50%, 57%, and 64% SID Phe : (Phe+Tyr) was studied at two limiting levels of Phe+Tyr. From 57% to 64% SID Phe : (Phe+Tyr), performance was slightly reduced. In conclusion, it is recommended not to

  1. Improving Protein Fold Recognition by Deep Learning Networks

    Science.gov (United States)

    Jo, Taeho; Hou, Jie; Eickholt, Jesse; Cheng, Jianlin

    2015-12-01

    For accurate recognition of protein folds, a deep learning network method (DN-Fold) was developed to predict if a given query-template protein pair belongs to the same structural fold. The input used stemmed from the protein sequence and structural features extracted from the protein pair. We evaluated the performance of DN-Fold along with 18 different methods on Lindahl’s benchmark dataset and on a large benchmark set extracted from SCOP 1.75 consisting of about one million protein pairs, at three different levels of fold recognition (i.e., protein family, superfamily, and fold) depending on the evolutionary distance between protein sequences. The correct recognition rate of ensembled DN-Fold for Top 1 predictions is 84.5%, 61.5%, and 33.6% and for Top 5 is 91.2%, 76.5%, and 60.7% at family, superfamily, and fold levels, respectively. We also evaluated the performance of single DN-Fold (DN-FoldS), which showed the comparable results at the level of family and superfamily, compared to ensemble DN-Fold. Finally, we extended the binary classification problem of fold recognition to real-value regression task, which also show a promising performance. DN-Fold is freely available through a web server at http://iris.rnet.missouri.edu/dnfold.

  2. 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 diet, the determined AID were greater (P diet, the determined AID were greater (P diet were lower (P 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.

  3. An improved strategy for easy process monitoring and advanced purification of recombinant proteins.

    Science.gov (United States)

    Miladi, Baligh; Dridi, Cyrine; El Marjou, Ahmed; Boeuf, Guilhem; Bouallagui, Hassib; Dufour, Florence; Di Martino, Patrick; Elm'selmi, Abdellatif

    2013-11-01

    In this work, a multifunctional expression cassette, termed Multitags, combining different and complementary functionalities, was designed and used to monitor the expression and the purification of two model proteins (Pfu DNA polymerase and Myosin-VIIa- and Rab-Interracting protein : MyRIP). Multitags contains two affinity purification tags, a polyhistidine sequence (10× His) and the streptavidin-binding peptide (SBP) and as a marker tag the heme-binding domain of rat cytochrome b5 followed by the TEV cleavage site. Using the Multitags as fusion partner, more than 90 % of both fusion proteins were produced in soluble form when expressed in Escherichia coli KRX. In addition, high purity (99 %) of recombinant proteins was achieved after two consecutive affinity purification steps. The expression cassette also demonstrated an accurate monitoring capability comparable to that of a dual recognition-based method. The choice of the SBP tag was considered as an integral process that included a method for tag removal. Thus, an immobilized TEV protease fixed on streptavidin-agarose matrix was used for the cleavage of fusion proteins. After digestion, both unprocessed fusion proteins and Multitags were retained on the proteolytic column via their SBP sequence, allowing cleavage and recovery of target proteins on one step. This combined approach may accelerate the development of optimized production processes, while insuring high product quality and a low production cost.

  4. A never ending race for new and improved fluorescent proteins

    Directory of Open Access Journals (Sweden)

    Jones Alexander M

    2012-05-01

    Full Text Available Abstract Bioluminescent and fluorescent proteins are now used as tools for research in all organisms. There has been massive progress over the past 15 years in creating a palette of fluorescent proteins with a wide spectrum of specific properties. One of the big challenges is to decide which variant may be best for a certain application. A recent article by Mann et al. in BMC Biotechnology describes a new orange fluorescent protein in plants. See research article http://www.biomedcentral.com/1472-6750/12/17

  5. A never ending race for new and improved fluorescent proteins

    Science.gov (United States)

    2012-01-01

    Bioluminescent and fluorescent proteins are now used as tools for research in all organisms. There has been massive progress over the past 15 years in creating a palette of fluorescent proteins with a wide spectrum of specific properties. One of the big challenges is to decide which variant may be best for a certain application. A recent article by Mann et al. in BMC Biotechnology describes a new orange fluorescent protein in plants. See research article http://www.biomedcentral.com/1472-6750/12/17 PMID:22554191

  6. Proteína bruta e energia digestível em dietas para alevinos de curimbatá (Prochilodus affins Crude protein and digestible energy in the diets for curimbatá fingerlings (Prochilodus affins

    Directory of Open Access Journals (Sweden)

    Marcos Antonio Delmondes Bomfim

    2005-12-01

    Full Text Available Objetivando-se determinar as exigências de proteína bruta (PB em função do nível de energia digestível (ED da dieta para alevinos de curimbatá (Prochilodus affins, foram utilizados 256 peixes com peso inicial de 2,72 ± 0,11 g, mantidos em 32 aquários de 100 L, dotados de abastecimento de água, temperatura controlada e aeração individuais. O experimento foi realizado seguindo-se esquema fatorial 4 x 2 (quatro níveis de PB: 18,0; 22,0; 26,0; e 30,0%, combinados com dois níveis de ED: 2.700 e 3.000 kcal/kg, em um delineamento inteiramente ao acaso, com quatro repetições e oito peixes por unidade experimental. Os peixes foram alimentados ad libitum em duas refeições diárias durante 78 dias. Avaliaram-se o ganho de peso, o consumo aparente de racão, a conversão alimentar aparente, a taxa de crescimento específico, a taxa de eficiência protéica, a eficiência de retenção de nitrogênio, a umidade, a proteína e a gordura corporais e as porcentagens de nitrogênio e de gordura no ganho de peso. Verificou-se que não houve interação dos níveis de PB ´ ED, à exceção da porcentagem de gordura no ganho de peso. Com o nível de 3.000 kcal de ED/kg, obtiveram-se peixes com maiores níveis de matéria seca, gordura corporal e porcentagem de gordura no ganho de peso. Concluiu-se que a exigência de PB e ED para alevinos de curimbatá é de 26,05% e 2.700 kcal/kg, respectivamente, que corresponde a uma relação ED:PB de 10,36 kcal de ED/g de PB, por proporcionar as melhores respostas em ganho de peso e composição de carcaça.Two hundred and fifty six curimbatá (Prochilodus affins fingerlings averaging initial weight of 2.72 ± 0.11 g, stocked in 32 aquariums (100 L with water renewal, controlled temperature and individual aeration, were used to determine the dietary crude protein (CP requirements, according to the digestible energy level (DE. The experiment was carried out in to a 4 x 2 factorial scheme (four CP levels: 18

  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

    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

  8. Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein

    NARCIS (Netherlands)

    Shaner, Nathan C; Campbell, Robert E; Steinbach, Paul A; Giepmans, Ben N G; Palmer, Amy E; Tsien, Roger Y

    2004-01-01

    Fluorescent proteins are genetically encoded, easily imaged reporters crucial in biology and biotechnology. When a protein is tagged by fusion to a fluorescent protein, interactions between fluorescent proteins can undesirably disturb targeting or function. Unfortunately, all wild-type yellow-to-red

  9. Lisina digestível em dietas de baixa proteína para frangos de corte tipo caipira de um aos 28 dias Digestible lysine in diets containing low protein concentrations for broiler type redneck from 1 to 28 days

    Directory of Open Access Journals (Sweden)

    H.G. Oliveira

    2013-04-01

    Full Text Available Avaliou-se a inclusão de diferentes porcentagens de lisina digestível em dietas de baixa proteína para frangos de corte tipo caipira, machos e fêmeas, com idade entre um e 28 dias. Utilizaram-se 792 machos e 792 fêmeas de linhagem comercial, de um dia de idade, distribuídos em delineamento inteiramente ao acaso, em esquema fatorial 2x6 (sexo x porcentagem de lisina digestível: 0,85; 0,90; 0,95; 1,00; 1,05 e 1,10% com seis repetições de 22 aves. Não foi constatada interação (P>0,05 de sexo versus inclusão de lisina. Observou-se superioridade (P0,05 para conversão alimentar. Da composição centesimal da carcaça, apenas o percentual de matéria mineral foi influenciada (PThe inclusion of different percentages of digestible lysine in low-protein diets was evaluated for male and female free-range broiler chickens, from 1 to 28 days. We used 792 female and 792 male naked neck chickens a day in a completely randomized 2x6 factorial scheme (sex x percentages of digestible lysine: 0.85, 0.90, 0.95, 1.00, 1.05 and1.10% with six replicates of 22 broilers. No interaction of sex versus lysine was found. Superiority was observed (P0.05 in food conversion.In the chemical composition of the carcass, only thepercentage of mineral matter was influenced (P<0.05 bydigestible lysine levels.It was concluded that the level of 0.85% for type redneck broiler meets the nutritional requirement of digestible lysine in diets containing lower protein concentrations.

  10. Engineering new strategies to improve pharmacokinetics of therapeutic proteins

    OpenAIRE

    Cantante, Cátia Sofia de Carvalho, 1982-

    2014-01-01

    Tese de doutoramento, Farmácia (Biotecnologia Farmacêutica), Universidade de Lisboa, Faculdade de Farmácia, 2014 Recombinant antibody formats and therapeutic proteins have found increasing applications as therapeutics, e.g. for the treatment of cancer or inflammatory diseases. While whole antibodies have an exceptionally long half-life, small antibody formats and small molecular weight therapeutic proteins often suffer from rapid elimination from circulation. Usually, this involves frequen...

  11. Strain engineering for improved expression of recombinant proteins in bacteria

    OpenAIRE

    Skretas Georgios; Makino Tomohiro; Georgiou George

    2011-01-01

    Abstract Protein expression in Escherichia coli represents the most facile approach for the preparation of non-glycosylated proteins for analytical and preparative purposes. So far, the optimization of recombinant expression has largely remained a matter of trial and error and has relied upon varying parameters, such as expression vector, media composition, growth temperature and chaperone co-expression. Recently several new approaches for the genome-scale engineering of E. coli to enhance re...

  12. Digestion of Nucleic Acids Starts in the Stomach.

    Science.gov (United States)

    Liu, Yu; Zhang, Yanfang; Dong, Ping; An, Ran; Xue, Changhu; Ge, Yinlin; Wei, Liangzhou; Liang, Xingguo

    2015-07-14

    The ingestion of nucleic acids (NAs) as a nutritional supplement or in genetically modified food has attracted the attention of researchers in recent years. Discussions over the fate of NAs led us to study their digestion in the stomach. Interestingly, we found that NAs are digested efficiently by human gastric juice. By performing digests with commercial, recombinant and mutant pepsin, a protein-specific enzyme, we learned that the digestion of NAs could be attributed to pepsin rather than to the acidity of the stomach. Further study showed that pepsin cleaved NAs in a moderately site-specific manner to yield 3'-phosphorylated fragments and the active site to digest NAs is probably the same as that used to digest protein. Our results rectify the misunderstandings that the digestion of NAs in the gastric tract begins in the intestine and that pepsin can only digest protein, shedding new light on NA metabolism and pepsin enzymology.

  13. Improving protein protein interaction prediction based on phylogenetic information using a least-squares support vector machine.

    Science.gov (United States)

    Craig, Roger A; Liao, Li

    2007-12-01

    Predicting protein-protein interactions has become a key step of reverse-engineering biological networks to better understand cellular functions. The experimental methods in determining protein-protein interactions are time-consuming and costly, which has motivated vigorous development of computational approaches for predicting protein-protein interactions. A set of recently developed bioinformatics methods utilizes coevolutionary information of the interacting partners (e.g., as exhibited in the form of correlations between distance matrices, where, for each protein, a matrix stores the pairwise distances between the protein and its orthologs in a group of reference genomes). We proposed a novel method to account for the intra-matrix correlations in improving predictive accuracy. The distance matrices for a pair of proteins are transformed and concatenated into a phylogenetic vector. A least-squares support vector machine is trained and tested on pairs of proteins, represented as phylogenetic vectors, whose interactions are known. The intra-matrix correlations are accounted for by introducing a weighted linear kernel, which determines the dot product of two phylogenetic vectors. The performance, measured as receiver operator characteristic (ROC) score in cross-validation experiments, shows significant improvement of our method (ROC score 0.928) over that obtained by Pearson correlations (0.659).

  14. Interaction of sorghum tannins with wheat proteins and effects on in vitro starch digestibility in wheat flour tortillas

    Science.gov (United States)

    Excess calorie intake and associated disease risks are a major problem in many parts of the world. Carbohydrates contribute the most calories in the diet, making starch-based foods a logical target for improvement. This study investigated the potential interaction of sorghum bran proanthocyanidins (...

  15. Growth and survival of juvenile queen conch, Strombus gigas fed artificial diets containing varying levels of digestible protein and energy

    Science.gov (United States)

    Aquaculture methods for queen conch, Strombus gigas, have been established for several decades. However, there is a need to improve husbandry techniques for the growout of juveniles. The purpose of this study was to determine growth and survival of juvenile queen conch fed artificial diets with incr...

  16. Improving microbial protein synthesis in the rumen of sheep fed fresh tofu waste by crude tannin extract of Acacia mangium

    Directory of Open Access Journals (Sweden)

    Elizabeth Wina

    2012-10-01

    Full Text Available Tannin can be found in many tropical plants and the presence of tannin may reduce the nutritional value of feed. However, it can give beneficial effect as it protects the protein from its excessive degradation in the rumen. Tofu waste is abundant in a country, including Indonesia, where tofu is produced widely. A feeding trial on sheep to study the effect of tannin on microbial protein synthesis and condition of rumen fermentation, digestibility and N retention of feed containing tofu waste was conducted. Fifteen sheep were allocated to one of 3 treatments, i.e: 1 control treatment, feed without tannin, 2 feed + tannin treatment and 3 feed + tannin + polyethylene glycol (PEG treatment. The sheep was placed in the individual metabolism cage. They were fed with treatment feed for 14 days and then, the following 7 days, faeces and urine were collected. At the end of the treatment, rumen fluid was taken from each sheep before and after feeding. Rumen parameter (pH, ammonia, SCFA, protozoa and microbial protein synthesis, digestibility, nitrogen retention were observed. Results showed that inclusion of 4% tannin in the feed depressed ammonia production in the rumen of sheep before and 3 hours after morning feeding. Total SCFA and individual SCFA were not different among treatments. Tannin did not affect daily feed intake. Tannin lowered the dry matter, organic matter and protein digestibilities but it significantly reduced ADF digestibility (P < 0.05 and tended to reduce NDF digestibility (P < 0.1. However, tannin increased the microbial protein synthesis. PEG addition was expected to bind tannin and alleviate the negative effect of tannin.

  17. Analysis of digester design concepts

    Energy Technology Data Exchange (ETDEWEB)

    Ashare, E.; Wilson, E. H.

    1979-01-29

    Engineering economic analyses were performed on various digester design concepts to determine the relative performance for various biomass feedstocks. A comprehensive literature survey describing the state-of-the-art of the various digestion designs is included. The digester designs included in the analyses are CSTR, plug flow, batch, CSTR in series, multi-stage digestion and biomethanation. Other process options investigated included pretreatment processes such as shredding, degritting, and chemical pretreatment, and post-digestion processes, such as dewatering and gas purification. The biomass sources considered include feedlot manure, rice straw, and bagasse. The results of the analysis indicate that the most economical (on a unit gas cost basis) digester design concept is the plug flow reactor. This conclusion results from this system providing a high gas production rate combined with a low capital hole-in-the-ground digester design concept. The costs determined in this analysis do not include any credits or penalties for feedstock or by-products, but present the costs only for conversion of biomass to methane. The batch land-fill type digester design was shown to have a unit gas cost comparable to that for a conventional stirred tank digester, with the potential of reducing the cost if a land-fill site were available for a lower cost per unit volume. The use of chemical pretreatment resulted in a higher unit gas cost, primarily due to the cost of pretreatment chemical. A sensitivity analysis indicated that the use of chemical pretreatment could improve the economics provided a process could be developed which utilized either less pretreatment chemical or a less costly chemical. The use of other process options resulted in higher unit gas costs. These options should only be used when necessary for proper process performance, or to result in production of a valuable by-product.

  18. 基于质谱检测转基因生物外源蛋白质的消化稳定性%Mass Spectrometry-Based Analysis of Digestive Stability of Target Protein in Genetically Modified Organism

    Institute of Scientific and Technical Information of China (English)

    毛劼; 孙兴; 程娟献; 王心正; 赵永强; 王红霞; 何昆; 夏晴

    2016-01-01

    基于无标记定量质谱检测技术建立了一种新的转基因生物外源蛋白质消化稳定性评价方法。以牛β-乳球蛋白、牛血清白蛋白和大豆胰蛋白酶抑制剂为标准蛋白质,经模拟人体胃/肠消化液消化后进行凝胶电泳分离,切取目标蛋白质条带进行酶切,提取肽段进行纳升液相色谱-电喷雾串联质谱分析。基于Mascot数据库检索鉴定目标蛋白质。计算各消化时间点蛋白质匹配肽段数与消化前蛋白质匹配肽段数的比值,当比值≤0.50时判断为目标蛋白质在该时间段内已消化。将此方法应用于转基因抗虫水稻“华恢1号”外源蛋白Cry1Ab/1Ac的消化稳定性分析,结果显示在模拟胃液中消化2 min时,比值下降到0.50以下,在模拟肠液中消化15 s后比值下降到0.50以下,表明该蛋白在胃/肠消化液中具有消化不稳定性。%Digestive stability analysis of exogenous protein is one of the important indexes of geneticaly modified organisms (GMO) safety assessment. In the present study, we established a novel assessment assay for protein digestive stability based on label-free quantification of mass spectrometry using bovineβ-lacto globulin (BLG), bovine serum albumin (BSA) and soybean trypsin inhibitor (STI) as standard proteins. Protein samples were treated with simulated gastric/intestinal fluids (SGF/SIF), and then separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The target bands were then cut out and trypsinized. The extracted peptides were analyzed by nano liquid chromatography-electronic spray ion-mass spectrum/mass spectrum and identified by Mascot software. By calculating the ratio between the matched peptide numbers of the target protein before and after SGF/SIF treatment, the digestibility of the target protein was estimated. When the ratio was lower than 0.50, the target protein was considered digestible. This newly developed assay was applied to Cry1Ab/1

  19. Influence of low protein diets on gene expression of digestive enzymes and hormone secretion in the gastrointestinal tract of young weaned piglets

    Institute of Scientific and Technical Information of China (English)

    Zhi-mei TIAN; Xian-yong MA; Xue-fen YANG; Qiu-li FAN; Yun-xia XIONG; Yue-qin QIU; Li WANG; Xiao-lu WEN; Zong-yong JIANG‡

    2016-01-01

    中文概要题目:低蛋白日粮对断奶仔猪胃肠道消化酶表达及激素分泌的影响目的:研究蛋白日粮水平对断奶仔猪消化的影响机制。创新点:从营养物质消化及激素变化方面研究低蛋白日粮对仔猪胃肠道消化吸收的影响,探究低蛋白日粮饲养的可行性。方法:18头28日龄断奶仔猪经过7天适应期后平均体重为(9.57±0.64) kg。仔猪随机分为三组,每组6头,分别饲喂20%、17%和14%粗蛋白日粮并自由采食45天。根据国家研究委员会(NRC)2012年的标准,通过添加赖氨酸、蛋氨酸、半胱氨酸、苏氨酸及色氨酸等理想蛋白模型满足仔猪生长需求。实验结束后,进行颈静脉采血并屠宰取样。结论:结果表明基于蛋白日粮水平对内分泌及消化酶影响,与14%及20%粗蛋白组相比,17%蛋白日粮更有利于胃肠道和胰腺等组织中三种主要营养物质的消化。因此,17%粗蛋白水平更适合断奶仔猪的营养需求,证实了低蛋白日粮对断奶仔猪饲养的可行性。%To investigate dietary protein level effects on digestive mechanisms, weaned piglets were fed for 45 d with diets containing 20%, 17%, or 14% crude protein (CP) supplemented to meet requirements for essential amino acids. This article describes the influence of dietary protein on gastrointestinal hormones and expression of an array of digestive enzymes in the gastrointestinal tract and pancreas. Results indicated that there were no significant differ-ences in expression of enzymes involved in carbohydrate digestion, except for maltase in the duodenum. In the je-junum, amylase expression in pigs fed 20% CP was much higher than that in pigs fed other diets (P0.05), there was a trend towards higher expression of various proteases in pigs fed 17% CP. The duodenal expression of en-teropeptidase in diets with 14% and 17% CP was significantly higher than that with 20% CP (P0.05). The expression of GPR93 as

  20. Latest improvements in CIEF: from proteins to microorganisms.

    Science.gov (United States)

    Šalplachta, Jiří; Kubesová, Anna; Horká, Marie

    2012-10-01

    In recent years, characterization and identification of microorganisms has become very important in different fields of human activity. Conventional laboratory methods are time consuming, laborious, and they may provide both false positive or negative results, especially for closely related microorganisms. On that account, new methods for fast and reliable microbial characterization are of great interest. In particular, capillary electrophoretic techniques have a great potential for characterization of microorganisms due to their unique surface properties. Cell surface proteins play a key role in this respect. Since CIEF represents one of the most efficient techniques for protein separation, it was consequently applied to the analysis of microbial cells. This review describes, after a brief introduction to CIEF of proteins, recent developments in CIEF of diverse microorganisms (viruses, bacteria, yeasts, and fungi). Possible application schemes in human and veterinary medicine as well as in plant protection and in biosecurity are outlined.

  1. DNA Nanoparticles for Improved Protein Synthesis In Vitro.

    Science.gov (United States)

    Galinis, Robertas; Stonyte, Greta; Kiseliovas, Vaidotas; Zilionis, Rapolas; Studer, Sabine; Hilvert, Donald; Janulaitis, Arvydas; Mazutis, Linas

    2016-02-24

    The amplification and digital quantification of single DNA molecules are important in biomedicine and diagnostics. Beyond quantifying DNA molecules in a sample, the ability to express proteins from the amplified DNA would open even broader applications in synthetic biology, directed evolution, and proteomics. Herein, a microfluidic approach is reported for the production of condensed DNA nanoparticles that can serve as efficient templates for in vitro protein synthesis. Using phi29 DNA polymerase and a multiple displacement amplification reaction, single DNA molecules were converted into DNA nanoparticles containing up to about 10(4)  clonal gene copies of the starting template. DNA nanoparticle formation was triggered by accumulation of inorganic pyrophosphate (produced during DNA synthesis) and magnesium ions from the buffer. Transcription-translation reactions performed in vitro showed that individual DNA nanoparticles can serve as efficient templates for protein synthesis in vitro.

  2. Starch digestion capacity of poultry.

    Science.gov (United States)

    Svihus, B

    2014-09-01

    Starch is quantitatively the most important nutrient in poultry diets and will to a large extent be present as intact starch granules due to very limited extent of gelatinization during pelleting. Although native starch is difficult to digest due to a semi-crystalline structure, even fast-growing broiler chickens appears to be able to digest this starch more or less completely during passage through the jejunum. However, reduced starch digestibility has been observed, particularly in pelleted diets containing large quantities of wheat. Although properties of the starch granule such as size and components on the granule surface may affect digestibility, the entrapment of starch granules in cell walls and a protein matrix may be even more important factors impeding starch digestion. In that case, this and the fact that amylase secretion is normally very high in poultry may explain the lack of convincing effects of exogenous α-amylase added to the diet. However, few well-designed experiments assessing mechanisms of starch digestion and the effect of α-amylase supplementation have been carried out, and thus more research is needed in this important area.

  3. Evolutionary Optimization of Kernel Weights Improves Protein Complex Comembership Prediction

    NARCIS (Netherlands)

    Hulsman, M.; Reinders, M.J.T.; De Ridder, D.

    2008-01-01

    In recent years, more and more high-throughput data sources useful for protein complex prediction have become available (e.g., gene sequence, mRNA expression, and interactions). The integration of these different data sources can be challenging. Recently, it has been recognized that kernel-based cla

  4. New promoters to improve heterologous protein production in Aspergillus vadensis

    NARCIS (Netherlands)

    Culleton, H.; Bouzid, O.; McKie, V.; de Vries, Ronald P.

    2014-01-01

    Aspergillus is a widely used host organism for the industrial production of homologous and heterologous proteins. Although Aspergillus niger is most commonly used, a close relative of this species, Aspergillus vadensis, has been suggested as a suitable and more favourable alternative due in part, to

  5. Improvements in the Protein Identifier Cross-Reference service.

    Science.gov (United States)

    Wein, Samuel P; Côté, Richard G; Dumousseau, Marine; Reisinger, Florian; Hermjakob, Henning; Vizcaíno, Juan A

    2012-07-01

    The Protein Identifier Cross-Reference (PICR) service is a tool that allows users to map protein identifiers, protein sequences and gene identifiers across over 100 different source databases. PICR takes input through an interactive website as well as Representational State Transfer (REST) and Simple Object Access Protocol (SOAP) services. It returns the results as HTML pages, XLS and CSV files. It has been in production since 2007 and has been recently enhanced to add new functionality and increase the number of databases it covers. Protein subsequences can be Basic Local Alignment Search Tool (BLAST) against the UniProt Knowledgebase (UniProtKB) to provide an entry point to the standard PICR mapping algorithm. In addition, gene identifiers from UniProtKB and Ensembl can now be submitted as input or mapped to as output from PICR. We have also implemented a 'best-guess' mapping algorithm for UniProt. In this article, we describe the usefulness of PICR, how these changes have been implemented, and the corresponding additions to the web services. Finally, we explain that the number of source databases covered by PICR has increased from the initial 73 to the current 102. New resources include several new species-specific Ensembl databases as well as the Ensembl Genome ones. PICR can be accessed at http://www.ebi.ac.uk/Tools/picr/.

  6. An improved Coomassie Brilliant Blue (CBB R-250) staining to proteins in gels

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    An improved CBB staining with higher sensitivity than that of the typical CBB staining was reported. The main improvement was using a fixing step of 25% trichloroacetic acid (TCA) before CBB staining. For most proteins studied, the sensitivity of the improved CBB staining was about twice as high as that of the typical method. For basic and low molecular weight proteins such as ribosomal proteins, the sensitivity of this improved staining method was about 3.5-28 times that of the typical method. It was speculated that the improved procedure would be suitable for exact quantitative analysis of proteins fractionated by SDS-PAGE, especially for basic and low molecular weight proteins. On the other hand, this new modified method might be also applied to multidisciplinary studies, such as biological researches and nuclear sciences.

  7. Assessment of two-phase olive mill solid waste and microalgae co-digestion to improve methane production and process kinetics.

    Science.gov (United States)

    Fernández-Rodríguez, M J; Rincón, B; Fermoso, F G; Jiménez, A M; Borja, R

    2014-04-01

    Olive mill solid waste (OMSW) is a pollutant waste coming from olive oil elaboration by the two-phase centrifugation system. OMSW has a high organic matter content and unbalanced carbon to nitrogen (C/N) ratio, 31/1, which avoids obtaining high methane yields in the anaerobic digestion of this waste. In the present study a microalgae, Dunaliella salina, was employed as co-substrate for the OMSW anaerobic digestion in order to decrease the C/N ratio and increase its biodegradability. Different co-digestion mixtures (C/N ratios) were studied. The increase of D. salina from 25% to 50% in the co-digestion mixture clearly increased the biodegradability of the sole substrates. The highest biodegradability was found for the co-digestion mixture 50% OMSW-50% D. salina. Nevertheless, the maximum methane production, 330mLCH4/gVSadded, and the highest methane production rate were obtained for the co-digestion mixture 75% OMSW-25% D. salina, keeping a C/N ratio near to 26.7/1.

  8. Molecular cloning and characterization of novel Morus alba germin-like protein gene which encodes for a silkworm gut digestion-resistant antimicrobial protein.

    Directory of Open Access Journals (Sweden)

    Bharat Bhusan Patnaik

    Full Text Available BACKGROUND: Silkworm fecal matter is considered one of the richest sources of antimicrobial and antiviral protein (substances and such economically feasible and eco-friendly proteins acting as secondary metabolites from the insect system can be explored for their practical utility in conferring broad spectrum disease resistance against pathogenic microbial specimens. METHODOLOGY/PRINCIPAL FINDINGS: Silkworm fecal matter extracts prepared in 0.02 M phosphate buffer saline (pH 7.4, at a temperature of 60°C was subjected to 40% saturated ammonium sulphate precipitation and purified by gel-filtration chromatography (GFC. SDS-PAGE under denaturing conditions showed a single band at about 21.5 kDa. The peak fraction, thus obtained by GFC wastested for homogeneityusing C18reverse-phase high performance liquid chromatography (HPLC. The activity of the purified protein was tested against selected Gram +/- bacteria and phytopathogenic Fusarium species with concentration-dependent inhibitionrelationship. The purified bioactive protein was subjected to matrix-assisted laser desorption and ionization-time of flight mass spectrometry (MALDI-TOF-MS and N-terminal sequencing by Edman degradation towards its identification. The N-terminal first 18 amino acid sequence following the predicted signal peptide showed homology to plant germin-like proteins (Glp. In order to characterize the full-length gene sequence in detail, the partial cDNA was cloned and sequenced using degenerate primers, followed by 5'- and 3'-rapid amplification of cDNA ends (RACE-PCR. The full-length cDNA sequence composed of 630 bp encoding 209 amino acids and corresponded to germin-like proteins (Glps involved in plant development and defense. CONCLUSIONS/SIGNIFICANCE: The study reports, characterization of novel Glpbelonging to subfamily 3 from M. alba by the purification of mature active protein from silkworm fecal matter. The N-terminal amino acid sequence of the purified protein was

  9. Analysis of the role of the gene coding the Amyloid-Precursor Protein Binding Protein 1 (APP-BP1) in the radio-sensitivity of epidermoid carcinomas of the upper aero-digestive tract infected by the human papillomavirus; Analyse du role du gene codant l'Amyloid-Precursor Protein Binding Protein 1 (APP-BP1) dans la radiosensibilite des carcinomes epidermoides des voies aero-digestives superieures infectees par le papillomavirus humain

    Energy Technology Data Exchange (ETDEWEB)

    Guihard, S.; Altmeyer, A.; Ramolu, L.; Macabre, C.; Abecassis, J.; Noel, G.; Jung, A.C. [Centre de lutte contre le cancer Paul-Strauss, 67 - Strasbourg (France)

    2010-10-15

    As the human papillomavirus (HPV) is at the origin of 25% of upper aero-digestive tract cancers, and as these tumours present an increased radio-sensitivity compared to other tumours, probably due to a greater transcriptional activity of p53, the authors report the study on the influence of a decrease of the expression of the APP-BP1 in these tumours which could favour a radio-induced apoptosis. By using a reverse transcriptase polymerase chain reaction (RT-PCR), they assessed the APP-BP1 expression levels as well as expression levels of transcriptions coding onco-proteins known to be over-expressed in HPV+ tumours. They compared the radio-sensitivities of HPV+ and HPV- cells, the first one appearing to be greater than the second one. Short communication

  10. Heat-treatment reduces anti-nutritional phytochemicals and maintains protein quality in genetically improved hulled soybean flour

    Directory of Open Access Journals (Sweden)

    Ariela Werneck de Carvalho

    2013-06-01

    Full Text Available The soybean is a protein source of high biological value. However, the presence of anti-nutritional factors affects its protein quality and limits the bioavailability of other nutrients. The effect of heat-treatment, 150 ºC for 30 minutes, on hulled and hull-less soybean flour from the cultivar UFVTN 105AP on urease, trypsin inhibitor activity, protein solubility, amino acid profile, and in vivo protein quality was investigated. The treatment reduced the trypsin inhibitor activity and urease, but it did not affect protein solubility. Protein Efficiency Coefficient (PER values of the flours were similar, and the PER of the hull-less soybean flour did not differ from casein. The Net Protein Ratio (NPR did not differ between the experimental groups. The True Digestibility (TD of the flours did not differ, but both were lower in casein and the Protein Digestibility Corrected Amino Acid Score (PDCCAS was lower than the TD, due to limited valine determined by the chemical score. Therefore, the flours showed reduced anti-nutritional phytochemicals and similar protein quality, and therefore the whole flours can be used as a source of high quality protein.

  11. Gold nanoparticle-protein arrays improve resolution for cryo-electron microscopy

    OpenAIRE

    Hu, Minghui; Qian, Luping; Briñas, Raymond P.; Lymar, Elena S.; Kuznetsova, Larisa; Hainfeld, James F.

    2007-01-01

    Cryo-electron microscopy single particle analysis shows limited resolution due to poor alignment precision of noisy images taken under low electron exposure. Certain advantages can be obtained by assembling proteins into two-dimensional (2D) arrays since protein particles are locked into repetitive orientations, thus improving alignment precision. We present a labeling method to prepare protein 2D arrays using gold nanoparticles (NPs) interconnecting genetic tag sites on proteins. As an examp...

  12. Robust protein microarray image segmentation using improved seeded region growing algorithm

    Institute of Scientific and Technical Information of China (English)

    Liqiang Wang(王立强); Xuxiang Ni(倪旭翔); Zukang Lu(陆祖康)

    2003-01-01

    Protein microarray technology has recently emerged as a powerful tool for biomedical research. Before automatic analysis the protein microarray images, protein spots in the images must be determined appropriately by spot segmentation algorithm. In this paper, an improved seeded region growing (ISRG)algorithm for protein microarray segmentation is presented, the seeds are obtained by finding the positions of the printed spots, and the protein spot regions are grown through these seeds. The experiment results show that the presented algorithm is accurate for adaptive shape segmentation and robust for protein microarray images contaminated by noise.

  13. Two-stage anaerobic and post-aerobic mesophilic digestion of sewage sludge: Analysis of process performance and hygienization potential.

    Science.gov (United States)

    Tomei, M Concetta; Mosca Angelucci, Domenica; Levantesi, Caterina

    2016-03-01

    Sequential anaerobic-aerobic digestion has been demonstrated to be effective for enhanced sludge stabilization, in terms of increased solid reduction and improvement of sludge dewaterability. In this study, we propose a modified version of the sequential anaerobic-aerobic digestion process by operating the aerobic step under mesophilic conditions (T=37 °C), in order to improve the aerobic degradation kinetics of soluble and particulate chemical oxygen demand (COD). Process performance has been assessed in terms of "classical parameters" such as volatile solids (VS) removal, biogas production, COD removal, nitrogen species, and polysaccharide and protein fate. The aerobic step was operated under intermittent aeration to achieve nitrogen removal. Aerobic mesophilic conditions consistently increased VS removal, providing 32% additional removal vs. 20% at 20 °C. Similar results were obtained for nitrogen removal, increasing from 64% up to 99% at the higher temperature. Improved sludge dewaterability was also observed with a capillary suction time decrease of ~50% during the mesophilic aerobic step. This finding may be attributable to the decreased protein content in the aerobic digested sludge. The post-aerobic digestion exerted a positive effect on the reduction of microbial indicators while no consistent improvement of hygienization related to the increased temperature was observed. The techno-economic analysis of the proposed digestion layout showed a net cost saving for sludge disposal estimated in the range of 28-35% in comparison to the single-phase anaerobic digestion.

  14. Digestion proteomics: tracking lactoferrin truncation and peptide release during simulated gastric digestion.

    Science.gov (United States)

    Grosvenor, Anita J; Haigh, Brendan J; Dyer, Jolon M

    2014-11-01

    The extent to which nutritional and functional benefit is derived from proteins in food is related to its breakdown and digestion in the body after consumption. Further, detailed information about food protein truncation during digestion is critical to understanding and optimising the availability of bioactives, in controlling and limiting allergen release, and in minimising or monitoring the effects of processing and food preparation. However, tracking the complex array of products formed during the digestion of proteins is not easily accomplished using classical proteomics. We here present and develop a novel proteomic approach using isobaric labelling to mapping and tracking protein truncation and peptide release during simulated gastric digestion, using bovine lactoferrin as a model food protein. The relative abundance of related peptides was tracked throughout a digestion time course, and the effect of pasteurisation on peptide release assessed. The new approach to food digestion proteomics developed here therefore appears to be highly suitable not only for tracking the truncation and relative abundance of released peptides during gastric digestion, but also for determining the effects of protein modification on digestibility and potential bioavailability.

  15. ON-COLUMN ENRICHMENT OF HYDROPHOBIC CYP450 PROTEINS IN HPLC FRACTIONATION OF MOUSE MICROSOMES PRIOR TO PROTEIN DIGESTION AND NANOSPRAY-LC/MSMS ANALYSIS

    Science.gov (United States)

    IntroductionMembrane proteins play crucial role in many cellular processes and are promising candidates for biomarker discovery but are under-represented in the field of proteomics due to their hydrophobic nature. Although standard reversed-phase LC methods often exhibit ...

  16. Effects of supplementary urea-minerals lick block on the kinetics of fibre digestion, nutrient digestibility and nitrogen utilization of low quality roughages

    Institute of Scientific and Technical Information of China (English)

    WU Yue-ming; HU Wei-lian; LIU Jian-xin

    2005-01-01

    Three yearling lambs with a rumen cannula were used to investigate the effects of supplementation with an urea-minerals lick block (ULB) on the kinetics ofruminal fibre digestion, nutrient digestibility and nitrogen (N) utilization of rice straw (RS), ammonia bicarbonate (AB)-treated RS (ABRS) and hay prepared from natural pasture. The digestibility of dry matter and organic matter of RS increased by 13.1% and 12.7% (P<0.05) when the diet was supplemented with ULB, and approached to that of ABRS, indicating that the effect of ULB on digestibility of RS is similar to that of AB treatment. The digestibility of ABRS was slightly improved by the ULB feeding. Nitrogen retention was highest in lambs fed on ABRS alone, followed by hay with ULB, and was lowest in animals fed on RS with ULB. However, both the amount and proportion of N retention to N intake were enhanced by ULB supplementation to lambs fed on hay. The proportion of N retained to N digested decreased due to ULB supplementation to lambs fed on RS or ABRS. Supplementing ULB did not greatly influence the rumen degradation of either dry matter or crude protein in each of the three diets. RS and hay had similar values in the potential extent of digestion (PED) and digestion rate of PED (kd) of fibrous materials, but the discrete lag time for RS was lower than that for hay. The AB treatment significantly increased the PED (P<0.05) and kd (P<0.05) of RS. Neither the PED nor kd for RS and ABRS was influenced by ULB supplementation, but the kd for hay significantly increased due to ULB. The lag time for hay was also shortened by the ULB feeding. The ULB improved the digestion of fibre in the rumen of lambs fed on low quality roughage. It is inferred that while ULB is effective in increasing nutrient digestibility of low quality roughages by improving ruminal fibre digestion. A synchronized supply of N and energy to rumen microbes should be considered to improve the efficiency of N utilization when the basal

  17. Lipid digestion, absorption and uptake in Solea senegalensis.

    Science.gov (United States)

    Borges, Pedro; Medale, Françoise; Veron, Vincent; Pires, Maria Dos Anjos; Dias, Jorge; Valente, Luísa M P

    2013-09-01

    Dietary lipids are the major energy source for metabolic purposes in most fish species, and improve dietary protein utilization for growth. In a previous study we have reported a low tolerance of Senegalese sole juveniles to dietary lipid levels and suggested a maximal dietary inclusion level of 8% lipids for both optimal growth and nutrient utilization. The mechanisms behind this apparent poor utilization of the dietary lipids are still to be elucidated. The primary aim of the present study was to investigate the overall process of digestion and lipid absorption in relation to dietary lipid levels. Triplicate groups of twenty fish (mean initial mass 29g) were fed two isonitrogenous diets (54% of protein dry matter basis) with different lipid levels (L4 and L17, 4 and 17% lipids dry matter basis), for 88days. Protein and lipid apparent digestibility coefficients as well as lipase activity were similar in both groups suggesting that Solea senegalensis has the ability to digest equally well a low fat or a high fat diet. Plasma triglyceride concentrations were significantly higher 5 and 16h after feeding in fish fed the L17 compared to those fed L4, following dietary lipid supply, demonstrating effective lipid absorption. Expression of proteins related to lipid transport (microsomal triglyceride transfer protein), trafficking (Fatty acid binding protein 11) and fatty acid uptake (VLDL-r) was significantly higher in liver of fish fed the high fat diet 16h after the meal, but remained unchanged in muscle. In conclusion, it seems that high fat diets do not impair lipid digestion and absorption.

  18. Screening of protease producing fungi for microbial digestion of seed proteins and synthesis of amino acids-metalnutrient chelates.

    Science.gov (United States)

    Deore, G B; Limaye, A S; Dushing, Y A; Dhobale, S B; Kale, S; Laware, S L

    2013-01-15

    The problem of metalnutrient deficiency is becoming more serious with the introduction of modern agricultural practices. As a result, metalnutrient deficiency is recognized as one of the critical yield limiting factors. Metalnutrients are generally offered in their sulphate or oxide forms. However, it is reported that organically bound minerals generally have a higher bioavailability than inorganic minerals. Chelation makes otherwise unavailable metalnutrients plant available. Amino acids are well known among various chelating agents. In present investigation the fungus Paecilomyces variotii PR-4 was isolated from soil and was used for production of protease and determination of its activity. Proteins from germinating seeds of chick pea, mung bean, soybean and cowpea were hydrolyzed for the production of amino acids. Amino acids were recovered, estimated and utilized for chelation of metalnutrients viz., Zn, Cu, Fe, Mn, Mg, B and Mo. The resultant chelates were employed to detect with Fourier Transform Infra-Red Spectrophotometer (FTIR) analysis. The peaks of most intensive bands in the IR spectra of ligands recorded were present in the intervals of the wave numbers 3500-3300 and 1720-1700 cm(-1). Chelation of metalnutrients led to the broadening of peak and changes of the peak position of hydroxyl groups, which indicated the binding of the carboxylic groups and primary amine groups of amino acids to the metalnutrients. The resultant amino acids-metalnutrient chelates can be utilized as organic fertilizer.

  19. ENERGIA DIGESTÍVEL E DIGESTIBILIDADE APARENTE DA PROTEÍNA BRUTA E MATÉRIA SECA DO FENO DE PLANTAS AQUÁTICAS PARA SUÍNOS EM FASE DE CRESCIMENTO DIGESTIBLE ENERGY AND APPARENT DIGESTIBILITY OF THE CRUDE PROTEIN AND DRY MATTER OF THE AQUATIC PLANTS HAY FOR GROWTH PHASE SWINE

    Directory of Open Access Journals (Sweden)

    Fábio Enrique Lemos Budiño

    2009-09-01

    Full Text Available Foi realizado um experimento com o intuito de avaliar o feno de plantas aquáticas como ingrediente de dietas para suínos em crescimento. Utilizaram-se dez suínos de peso inicial de 28,85 ± 2,95kg, distribuídos entre os tratamentos – ração-referência (RR e ração-teste (RT – sendo que a RT correspondeu a 80% da RR acrescida de 20% de feno de plantas aquáticas. Desenvolveu-se ensaio de digestibilidade utilizando-se como marcador 1,0% de óxido férrico adicionado às dietas, e no sétimo dia deu início à coleta das fezes. O feno de plantas aquáticas apresentou os seguintes valores nutricionais de energia digestível: 812,26 kcal/kg; de proteína digestível: -2,38% e de matéria seca digestível: 12,90%.

    PALAVRAS-CHAVES: Fibra, leitão, metabolismo.
    An experiment was carried out to evaluate the use of the aquatic plants hay as ingredient of diets for growth swine. Ten swine with initial weight of 28.85 ± 2.95kg were used, distributed among the treatments: ration reference (RR and ration tests (RT, where RT corresponded to 80% of added RR of 20% of aquatic plants hay. The digestibility assay was accomplished, being used as marker 1.0% of ferric oxide added to the diets and in the seventh day the collection of the feces was begun. The aquatic plants hay presented the following nutritional values of digestible energy: 812.26 kcal/kg; of digestible protein: -2.38% and of dry matter digestible: 12.90%.

    KEY WORDS: Fiber, metabolism, pig.

  20. Site and extent of amino acid digestion in dairy cattle fed with corn and its byproducts

    Directory of Open Access Journals (Sweden)

    Reginaldo Nassar Ferreira

    2015-02-01

    Full Text Available The study was conducted to evaluated the site and extent of dry matter (DM, crude protein (CP, methionine (Met, lysine (Lys, and threonine (Thr digestion of corn and byproducts obtained from corn germ mixed with different amounts of extruded or non-extruded ether extract (EE in dairy cattle. Treatments consisted in eight types of feed and two processing in a 4 × 2 factorial design. There were four feeds: corn grain cracked (Corn, corn germ meal with 1% EE (CG1, corn germ meal with 7% EE (CG7, and corn germ meal with 10% EE (CG10. The feeds were processed in one of two ways: extruded (Ex and not extruded. In situ techniques were used to determine DM, CP, Met, Lys, and Thr partial and total tract digestion. A basic diet was compounded of corn germ meal, soybean meal and coastcross hay in a 70:30 roughage to concentrate ratio. There was no interaction (P>0.05 between feeds and processing method. Extrusion improved (P0.05 for corn and corn germ meal mixed with 7 and 10% EE, regardless of EE processing method. The CP total tract digestibility of corn germ meal with 1% nonextruded EE was 16.62% higher (P<0.05 than that of the extruded form. The best total CP digestibility was obtained for corn germ meal with 7% EE, independently of the processing method. The effects of EE processing method on partial and total digestibility differed between amino acid. Corn and corn byproduct extrusion may improve dry matter digestibility, but do not necessarily influence crude protein digestion. Ruminal and intestinal digestibility of Met, Lys, and Thr depends on both feed type and processing method. Therefore, amino acid availability should be considered individually.

  1. potentially pathogenic gastrointestinal microorganisms (ID 2972), improved lactose digestion (ID 2972) and increasing IL-10 production (ID 2973) (further assessment) pursuant to Article 13(1) of Regulation (EC) No 1924/2006

    DEFF Research Database (Denmark)

    Tetens, Inge

    of Lactobacillus paracasei CNCM I-1688 and Lactobacillus salivarius CNCM I-1794 and reduction of gastro-intestinal discomfort, decreasing potentially pathogenic gastro-intestinal microorganisms, improved lactose digestion and increasing IL-10 production. The food constituent that is the subject of the health......-intestinal discomfort, is a beneficial physiological effect for the general population. The claimed effect, decreasing potentially pathogenic gastro-intestinal microorganisms, might be a beneficial physiological effect for the general population. The claimed effect, improved lactose digestion, is a beneficial...... physiological effect for individuals with lactose maldigestion. No human intervention studies were provided from which conclusions could be drawn for the scientific substantiation of the above-mentioned claims. On the basis of the data provided, the Panel concludes that a cause and effect relationship has...

  2. Engineering Digestion: Multiscale Processes of Food Digestion.

    Science.gov (United States)

    Bornhorst, Gail M; Gouseti, Ourania; Wickham, Martin S J; Bakalis, Serafim

    2016-03-01

    Food digestion is a complex, multiscale process that has recently become of interest to the food industry due to the developing links between food and health or disease. Food digestion can be studied by using either in vitro or in vivo models, each having certain advantages or disadvantages. The recent interest in food digestion has resulted in a large number of studies in this area, yet few have provided an in-depth, quantitative description of digestion processes. To provide a framework to develop these quantitative comparisons, a summary is given here between digestion processes and parallel unit operations in the food and chemical industry. Characterization parameters and phenomena are suggested for each step of digestion. In addition to the quantitative characterization of digestion processes, the multiscale aspect of digestion must also be considered. In both food systems and the gastrointestinal tract, multiple length scales are involved in food breakdown, mixing, absorption. These different length scales influence digestion processes independently as well as through interrelated mechanisms. To facilitate optimized development of functional food products, a multiscale, engineering approach may be taken to describe food digestion processes. A framework for this approach is described in this review, as well as examples that demonstrate the importance of process characterization as well as the multiple, interrelated length scales in the digestion process.

  3. Amaranth peptides from simulated gastrointestinal digestion: antioxidant activity against reactive species.

    Science.gov (United States)

    Delgado, María C Orsini; Galleano, Mónica; Añón, María C; Tironi, Valeria A

    2015-03-01

    We evaluated the capacity of simulated gastrointestinal digests or alcalase hydrolysates of protein isolates from amaranth to scavenge diverse physiologically relevant reactive species. The more active hydrolysate was obtained with the former method. Moreover, a prior alcalase treatment of the isolate followed by the same simulated gastrointestinal digestion did not improve the antioxidant capacity in any of the assays performed and even produced a negative effect under some conditions. Gastrointestinal digestion produced a strong increment in the scavenging capacity against peroxyl radicals (ORAC assay), hydroxyl radicals (ESR-OH assay), and peroxynitrites; thus decreasing the IC50 values to approximately 20, 25, and 20%, respectively, of the levels attained with the nonhydrolyzed proteins. Metal chelation (HORAC assay) also enhanced respect to isolate levels, but to a lesser extent (decreasing IC50 values to only 50%). The nitric-oxide- and superoxide-scavenging capacities of the digests were not relevant with respect to the methodologies used. The gastrointestinal digests from amaranth proteins acted against reactive species by different mechanisms, thus indicating the protein isolate to be a potential polyfunctional antioxidant ingredient.

  4. MALDI MS peptide mapping performance by in-gel digestion on a probe with prestructured sample supports

    DEFF Research Database (Denmark)

    Klenø, Tina Guldberg; Andreasen, Christian Maaløv; Kjeldal, Helle Ørsted;

    2004-01-01

    Matrix-assisted laser desorption/ionization (tandem) mass spectrometry (MALDI MS) is widely used in protein chemistry and proteomics research for the identification and characterization of proteins isolated by polyacrylamide gel electrophoresis. In an effort to minimize sample handling and increase...... sample throughput, we have developed a novel in-gel digestion protocol where sample preparation is performed directly on a MALDI probe with prestructured sample support. The protocol consists of few sample-handling steps and has minimal consumption of reagents, making the protocol sensitive, timesaving...... and MALDI peptide mass mapping was 89%. The on-probe in-gel digestion procedure provided superior sensitivity and peptide mass mapping performance as compared to our standard in-gel digestion protocol. The on-probe digestion technique resulted in significantly improved amino acid sequence coverage...

  5. Improved functional overview of protein complexes using inferred epistatic relationships

    LENUS (Irish Health Repository)

    Ryan, Colm

    2011-05-23

    Abstract Background Epistatic Miniarray Profiling(E-MAP) quantifies the net effect on growth rate of disrupting pairs of genes, often producing phenotypes that may be more (negative epistasis) or less (positive epistasis) severe than the phenotype predicted based on single gene disruptions. Epistatic interactions are important for understanding cell biology because they define relationships between individual genes, and between sets of genes involved in biochemical pathways and protein complexes. Each E-MAP screen quantifies the interactions between a logically selected subset of genes (e.g. genes whose products share a common function). Interactions that occur between genes involved in different cellular processes are not as frequently measured, yet these interactions are important for providing an overview of cellular organization. Results We introduce a method for combining overlapping E-MAP screens and inferring new interactions between them. We use this method to infer with high confidence 2,240 new strongly epistatic interactions and 34,469 weakly epistatic or neutral interactions. We show that accuracy of the predicted interactions approaches that of replicate experiments and that, like measured interactions, they are enriched for features such as shared biochemical pathways and knockout phenotypes. We constructed an expanded epistasis map for yeast cell protein complexes and show that our new interactions increase the evidence for previously proposed inter-complex connections, and predict many new links. We validated a number of these in the laboratory, including new interactions linking the SWR-C chromatin modifying complex and the nuclear transport apparatus. Conclusion Overall, our data support a modular model of yeast cell protein network organization and show how prediction methods can considerably extend the information that can be extracted from overlapping E-MAP screens.

  6. Anaerobic Digestion: Process

    DEFF Research Database (Denmark)

    Angelidaki, Irini; Batstone, Damien J.

    2011-01-01

    with very little dry matter may also be called a digest. The digest should not be termed compost unless it specifically has been composted in an aerated step. This chapter describes the basic processes of anaerobic digestion. Chapter 9.5 describes the anaerobic treatment technologies, and Chapter 9.......6 addresses the mass balances and environmental aspects of anaerobic digestion....

  7. Functional improvement of antibody fragments using a novel phage coat protein III fusion system

    DEFF Research Database (Denmark)

    Jensen, Kim Bak; Larsen, Martin; Pedersen, Jesper Søndergaard;

    2002-01-01

    Functional expressions of proteins often depend on the presence of host specific factors. Frequently recombinant expression strategies of proteins in foreign hosts, such as bacteria, have been associated with poor yields or significant loss of functionality. Improvements in the performance...... of heterologous expression systems will benefit present-day quests in structural and functional genomics where high amounts of active protein are required. One example, which has been the subject of considerable interest, is recombinant antibodies or fragments thereof as expressions of these in bacteria......(s) of the filamentous phage coat protein III. Furthermore, it will be shown that the observed effect is neither due to improved stability nor increased avidity....

  8. Contribution of protein, starch, and fat to the apparent ileal digestible energy of corn- and wheat-based broiler diets in response to exogenous xylanase and amylase without or with protease.

    Science.gov (United States)

    Romero, L F; Sands, J S; Indrakumar, S E; Plumstead, P W; Dalsgaard, S; Ravindran, V

    2014-10-01

    The ileal energy contribution of protein, starch, and fat in response to 2 exogenous enzyme combinations was studied in 2 digestibility assays with 21- (experiment 1; 432 birds) and 42-d-old (experiment 2; 288 birds) Ross 308 broiler chickens. A 2 × 2 × 3 factorial arrangement of treatments with 2 base grains (corn or wheat), without or with high fiber ingredients (corn distillers dried grains with solubles and canola meal), and 3 enzyme treatments was implemented. Enzyme treatments, fed from 12 to 21 d or 32 to 42 d, were 1) without enzymes, 2) with xylanase from Trichoderma ressei (2,000 U/kg) and amylase from Bacillus licheniformis (200 U/kg; XA), or 3) with XA plus protease from Bacillus subtilis (4,000 U/kg; XAP). All diets contained Escherichia coli phytase (500 FTU/kg). Apparent ileal digestibility (AID) of protein, starch, and fat, as well as the apparent ileal digestible energy, were determined using titanium dioxide as inert marker. A generalized mixed model was used to test main effects and 2-way interactions at P starch at 21 and 42 d, and AID of fat at 21 d, with greater effects of enzymes in wheat-based compared with corn-based diets, but significant increments due to enzymes compared with controls in both diet types. Apparent ileal digestibility of fat at 42 d increased with enzyme supplementation compared with the control treatments. The XA and XAP treatments gradually (P starch, fat, and protein were affected differentially by base grain and the presence of fibrous ingredients at 21 and 42 d of age.

  9. 不同热处理对牛奶乳清蛋白消化的影响%Effects of Various Heat Treatments on Digestibility of Milk Whey Proteins

    Institute of Scientific and Technical Information of China (English)

    李倩; 李艳莉; 田晓宇; 霍贵成

    2012-01-01

    Demineralised whey powder is the major ingredient of formula for sale in market which was obtained by repeated heating and spray drying. In order to study the protein denaturation after repeat heating as well as the impact on the digestibility, HPLC (high performance liquid chromatography) was applied to determine the α-lactalbumin and β-1actoglobulin denaturation rate in different heat treatment of whey protein, and then pepsin-trypsin (two step method) was applied to determine the in vitro digestibility of different denatured whey protein. The results show that there is a positive correlation between heating and the whey protein denaturation rate, the higher heating temperature, the longer time, the greater the rate of whey protein denaturation, the lower digestibility. The digestibility of demineralised whey powder is only 65.25%, which is not conducive to the baby's digestion and absorption.%反复加热喷雾干燥所得的脱盐乳清粉是目前市售的婴儿配方奶粉中主要的配料,为了了解蛋白质在反复加热后的变性情况以及对消化率的影响,采用高效液相色谱法测定不同热处理的乳清蛋白中α-乳白蛋白和β-乳球蛋白的变性率,然后通过胃蛋白酶-胰蛋白酶两步法测定了不同变性乳清蛋白的体外消化率。结果表明加热对乳清蛋白变性率是正相关,加热温度越高,时间越长,乳清蛋白变性率越大,消化率也越低。脱盐乳清粉的消化率仅为65.25%,不利于婴儿的消化吸收。

  10. Digestive System

    Science.gov (United States)

    ... absorbable atoms and molecules. During the process of absorption, nutrients that come from the food (including carbohydrates, proteins, fats, vitamins, and minerals) pass through channels in the intestinal wall and into the bloodstream. The blood works ...

  11. Coupling Protein Side-Chain and Backbone Flexibility Improves the Re-design of Protein-Ligand Specificity.

    Directory of Open Access Journals (Sweden)

    Noah Ollikainen

    Full Text Available Interactions between small molecules and proteins play critical roles in regulating and facilitating diverse biological functions, yet our ability to accurately re-engineer the specificity of these interactions using computational approaches has been limited. One main difficulty, in addition to inaccuracies in energy functions, is the exquisite sensitivity of protein-ligand interactions to subtle conformational changes, coupled with the computational problem of sampling the large conformational search space of degrees of freedom of ligands, amino acid side chains, and the protein backbone. Here, we describe two benchmarks for evaluating the accuracy of computational approaches for re-engineering protein-ligand interactions: (i prediction of enzyme specificity altering mutations and (ii prediction of sequence tolerance in ligand binding sites. After finding that current state-of-the-art "fixed backbone" design methods perform poorly on these tests, we develop a new "coupled moves" design method in the program Rosetta that couples changes to protein sequence with alterations in both protein side-chain and protein backbone conformations, and allows for changes in ligand rigid-body and torsion degrees of freedom. We show significantly increased accuracy in both predicting ligand specificity altering mutations and binding site sequences. These methodological improvements should be useful for many applications of protein-ligand design. The approach also provides insights into the role of subtle conformational adjustments that enable functional changes not only in engineering applications but also in natural protein evolution.

  12. Effect of dietary protein content on ileal amino acid digestibility, growth performance, and formation of microbial metabolites in ileal and cecal digesta of early-weaned pigs.

    Science.gov (United States)

    Htoo, J K; Araiza, B A; Sauer, W C; Rademacher, M; Zhang, Y; Cervantes, M; Zijlstra, R T

    2007-12-01

    Diarrhea incidence in weaned pigs may be associated with the concentration of intestinal microbial metabolites (ammonia, amines, and VFA) that are influenced by dietary CP content. Three experiments were conducted to determine effects of a low-protein, AA-supplemented diet on ileal AA digestibility, growth performance, diarrhea incidence, and concentration of microbial metabolites in ileal and cecal digesta of pigs weaned at 14 d of age. In Exp. 1, 8 pigs fitted with a simple T-cannula at the distal ileum were assigned in a crossover design to 2 diets containing 24 or 20% CP using wheat, corn, full-fat soybeans, whey powder, fish meal, and blood plasma as the main ingredients. Supplemental AA were added to the diets to meet the AA standards according to the 1998 NRC recommendations. Chromic oxide was used as an indigestible marker. Diets were fed at 2.5 times the ME requirement for maintenance. The reduction of dietary CP decreased (P acid, isobutyric acid, isovaleric acid, total VFA, and putrescine concentrations by 28 to 39%. In Exp. 3, 32 pigs were assigned to 2 diets, similar to Exp. 1, according to a randomized complete block design. Pigs had free access to feed and water. Dietary CP content did not affect growth performance or fecal consistency scores during the 3-wk study, and diarrhea was not observed. The results of these experiments indicate that lowering the dietary CP content combined with supplementation of AA markedly reduced the production of potentially harmful microbial metabolites in cecal digesta of early-weaned pigs without affecting growth performance.

  13. Improved proteomic analysis following trichloroacetic acid extraction of Bacillus anthracis spore proteins.

    Science.gov (United States)

    Deatherage Kaiser, Brooke L; Wunschel, David S; Sydor, Michael A; Warner, Marvin G; Wahl, Karen L; Hutchison, Janine R

    2015-11-01

    Proteomic analysis of bacterial samples provides valuable information about cellular responses and functions under different environmental pressures. Analysis of cellular proteins is dependent upon efficient extraction from bacterial samples, which can be challenging with increasing complexity and refractory characteristics. While no single method can recover 100% of the bacterial proteins, selected protocols can improve overall protein isolation, peptide recovery, or enrichment for certain classes of proteins. The method presented here is technically simple, does not require specialized equipment such as a mechanical disrupter, and is effective for protein extraction of the particularly challenging sample type of Bacillus anthracis Sterne spores. The ability of Trichloroacetic acid (TCA) extraction to isolate proteins from spores and enrich for spore-specific proteins was compared to the traditional mechanical disruption method of bead beating. TCA extraction improved the total average number of proteins identified within a sample as compared to bead beating (547 vs 495, respectively). Further, TCA extraction enriched for 270 spore proteins, including those typically identified by first isolating the spore coat and exosporium layers. Bead beating enriched for 156 spore proteins more typically identified from whole spore proteome analyses. The total average number of proteins identified was equal using TCA or bead beating for easily lysed samples, such as B. anthracis vegetative cells. As with all assays, supplemental methods such as implementation of an alternative preparation method may simplify sample preparation and provide additional insight to the protein biology of the organism being studied.

  14. An Improvement on the Protein Extraction Procedure from Eggshell Membrane

    Institute of Scientific and Technical Information of China (English)

    Peng Ruo-ting; Guo Yulong

    2010-01-01

    @@ 1 Introduction Collagen is a frequently occurring protein found in poultry,livestock and human body,esp.in the hair,skin,bone cartilage,tendon and so on.It forms the strongly insoluble fibers and serves as connective tissue supporting materials.Nowadays it became one of the most important raw materials of bio-engineering materials which are widely used in various fields including cosmetics,biomedical and foods industries.With the outburst of multiple diseases from cows and pigs such as bovine spongiform encephalopathy(BSE),related authorities in different countries have estabhshed many restrictions on trading collagen from mammalians.Due to this ground scientists began to identify new resources of collagens.

  15. Níveis de lisina digestível em rações, utilizando-se o conceito de proteína ideal, para suínos machos castrados de alto potencial genético dos 15 aos 30 kg Dietary digestible lysine levels, using the ideal protein concept, for barrows with high genetic potential from 15 to 30 kg

    Directory of Open Access Journals (Sweden)

    Márvio Lobão Teixeira de Abreu

    2006-06-01

    , were used to evaluate the effect of digestible lysine levels, using the ideal protein concept, on the performance and carcass composition. The treatments were composed of a basal diet containing 19.46% crude protein and 3,280 kcal of ME/kg, that were supplemented with different levels of HCl-L-Lysine, resulting in diets with 0.90, 1.00, 1,10, and 1.20% of digestible lysine. The diets were supplemented with increasing levels of synthetic amino acids, resulting in diets with ratios of methionine + cystine, threonine, tryptophan and valine:lysine constant in 60, 60, 19 e 69%, respectively, based on the true digestibility. No treatment effects on feed intake, concentration of plasma urea and percentages of water, protein and fat in the carcass were observed in this trial. However, significant quadratic effects on weight gain and feed:gain ratio with quadractic maximum and minimum of 1.10 and 1.12% of lysine, respectively, were detected. Intake of digestible lysine increased linearly among treatments. The depositions of protein and fat in the carcass answered quadractily with quadratic maxima at 1.12 and 1.08%, respectively. The level of digestible lysine that provided the best results of performance and carcass characteristics of barrows with high genetic potential from 15 to 30 kg was of 1.12% of true digestible lysine, corresponding to a digestible lysine intake of 12.03 g/day (3.42 g of Lys/Mcal of ME.

  16. Improved Proteomic Analysis Following Trichloroacetic Acid Extraction of Bacillus anthracis Spore Proteins

    Energy Technology Data Exchange (ETDEWEB)

    Kaiser, Brooke LD; Wunschel, David S.; Sydor, Michael A.; Warner, Marvin G.; Wahl, Karen L.; Hutchison, Janine R.

    2015-08-07

    Proteomic analysis of bacterial samples provides valuable information about cellular responses and functions under different environmental pressures. Proteomic analysis is dependent upon efficient extraction of proteins from bacterial samples without introducing bias toward extraction of particular protein classes. While no single method can recover 100% of the bacterial proteins, selected protocols can improve overall protein isolation, peptide recovery, or enrich for certain classes of proteins. The method presented here is technically simple and does not require specialized equipment such as a mechanical disrupter. Our data reveal that for particularly challenging samples, such as B. anthracis Sterne spores, trichloroacetic acid extraction improved the number of proteins identified within a sample compared to bead beating (714 vs 660, respectively). Further, TCA extraction enriched for 103 known spore specific proteins whereas bead beating resulted in 49 unique proteins. Analysis of C. botulinum samples grown to 5 days, composed of vegetative biomass and spores, showed a similar trend with improved protein yields and identification using our method compared to bead beating. Interestingly, easily lysed samples, such as B. anthracis vegetative cells, were equally as effectively processed via TCA and bead beating, but TCA extraction remains the easiest and most cost effective option. As with all assays, supplemental methods such as implementation of an alternative preparation method may provide additional insight to the protein biology of the bacteria being studied.

  17. 蛋白质测定方法改进%Improvement of Determination Method for Protein

    Institute of Scientific and Technical Information of China (English)

    曹坎涛

    2011-01-01

    应用消化-紫外分光光度法定氮测定蛋白质含量。该法较常规凯氏定氮法具有快速、简便、准确、灵敏度高等优点,且所用消化设备简便、常用,易于推广。%To detemine the content of protein by Digestion-UV spectrophotometric.Compared with kjeldahl determination,this method was rapid,simple,convenient,accurate and high sensitivity,and so on.The assimilation equipments are simple and commonly used.The method is easy to popularize.

  18. Comparison of HgO and CuSO4/TiO2 as catalysts in manual Kjeldahl digestion for determination of crude protein in animal feed: collaborative study.

    Science.gov (United States)

    Kane, P F

    1987-01-01

    Because of environmental concerns about HgO, and because of lengthy digestion requirements for HgO and CuSO4, interest in alternative catalysts for the Kjeldahl determination of animal feeds remains high. A digestion system using a mixed CuSO4/TiO2 catalyst has been found to reduce digestion times to 40 min. A collaborative study was carried out to compare this system to the official AOAC HgO method, 7.015. Thirty-eight samples, consisting of blind duplicates of closely matched pairs and 2 standard materials, were analyzed once by each method. Results were received from 13 laboratories. Means and standard deviations of individual samples were comparable, with an overall difference of grand means of 0.005% protein. With only one exception, analyses of variance showed no significant method difference at the 95% confidence level. The CuSO4/TiO2 method has been approved interim official first action as an alternative method for determination of crude protein in animal feed.

  19. Improving analytical methods for protein-protein interaction through implementation of chemically inducible dimerization

    DEFF Research Database (Denmark)

    Andersen, Tonni Grube; Nintemann, Sebastian; Marek, Magdalena;

    2016-01-01

    into the widely used split ubiquitin-, bimolecular fluorescence complementation (BiFC)- and Forster resonance energy transfer (FRET)-based methods and investigated different protein-protein interactions in yeast and plants. We demonstrate the functionality of this concept by the analysis of weakly interacting...... proteins from specialized metabolism in the model plant Arabidopsis thaliana. Our results illustrate that chemically induced dimerization can function as a built-in control for split-based systems that is easily implemented and allows for direct evaluation of functionality....

  20. Effects of replacing rapeseed meal with fava bean at 2 concentrate crude protein levels on feed intake, nutrient digestion, and milk production in cows fed grass silage-based diets.

    Science.gov (United States)

    Puhakka, L; Jaakkola, S; Simpura, I; Kokkonen, T; Vanhatalo, A

    2016-10-01

    The objective of this study was to evaluate the production and physiological responses of dairy cows to the substitution of fava bean for rapeseed meal at 2 protein supplementation levels in grass silage-based diets. We used 6 primiparous and 6 multiparous Finnish Ayrshire cows in a cyclic changeover trial with a 2×3 factorial arrangement of treatments. The experimental diets consisted of formic acid-treated timothy-meadow fescue silage and 3 isonitrogenous concentrates containing either rapeseed meal, fava bean, or a 1:1 mixture of rapeseed meal and fava bean at low and high inclusion rates, resulting in concentrate crude protein (CP) levels of 15.4 and 19.0% in dry matter. Silage dry matter intake decreased linearly when rapeseed meal was replaced with fava bean, the negative effect being more distinct at the high CP level than the low (-2.3 vs. -0.9kg/d, respectively). Similarly, milk and milk protein yields decreased linearly with fava bean, the change tending to be greater at the high CP level than the low. Yield of milk fat was lower for fava bean compared with rapeseed meal, the difference showing no interaction with CP level. Especially at the high CP level, milk urea concentration was higher with fava bean compared with rapeseed meal indicating better utilization of protein from the rapeseed meal. The apparent total-tract organic matter digestibility did not differ between treatments at the low CP level, but digestibility was higher for fava bean than for rapeseed meal at the high CP level. Plasma concentrations of essential amino acids, including methionine and lysine, were lower for fava bean than for rapeseed meal. Compared with rapeseed meal, the use of fava bean in dairy cow diets as the sole protein supplement decreased silage intake and milk production in highly digestible formic acid-treated grass silage-based diets.

  1. 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

  2. Coupling Protein Side-Chain and Backbone Flexibility Improves the Re-design of Protein-Ligand Specificity

    Science.gov (United States)

    Ollikainen, Noah; de Jong, René M.; Kortemme, Tanja

    2015-01-01

    Interactions between small molecules and proteins play critical roles in regulating and facilitating diverse biological functions, yet our ability to accurately re-engineer the specificity of these interactions using computational approaches has been limited. One main difficulty, in addition to inaccuracies in energy functions, is the exquisite sensitivity of protein–ligand interactions to subtle conformational changes, coupled with the computational problem of sampling the large conformational search space of degrees of freedom of ligands, amino acid side chains, and the protein backbone. Here, we describe two benchmarks for evaluating the accuracy of computational approaches for re-engineering protein-ligand interactions: (i) prediction of enzyme specificity altering mutations and (ii) prediction of sequence tolerance in ligand binding sites. After finding that current state-of-the-art “fixed backbone” design methods perform poorly on these tests, we develop a new “coupled moves” design method in the program Rosetta that couples changes to protein sequence with alterations in both protein side-chain and protein backbone conformations, and allows for changes in ligand rigid-body and torsion degrees of freedom. We show significantly increased accuracy in both predicting ligand specificity altering mutations and binding site sequences. These methodological improvements should be useful for many applications of protein – ligand design. The approach also provides insights into the role of subtle conformational adjustments that enable functional changes not only in engineering applications but also in natural protein evolution. PMID:26397464

  3. Microalgal Cultivation in Treating Liquid Digestate from Biogas Systems.

    Science.gov (United States)

    Xia, Ao; Murphy, Jerry D

    2016-04-01

    Biogas production via anaerobic digestion (AD) has rapidly developed in recent years. In addition to biogas, digestate is an important byproduct. Liquid digestate is the major fraction of digestate and may contain high levels of ammonia nitrogen. Traditional processing technologies (such as land application) require significant energy inputs and raise environmental risks (such as eutrophication). Alternatively, microalgae can efficiently remove the nutrients from digestate while producing high-value biomass that can be used for the production of biochemicals and biofuels. Both inorganic and organic carbon sources derived from biogas production can significantly improve microalgal production. Land requirement for microalgal cultivation is estimated as 3% of traditional direct land application of digestate.

  4. Effects of protein conformation in docking: improved pose prediction through protein pocket adaptation.

    Science.gov (United States)

    Jain, Ajay N

    2009-06-01

    Computational methods for docking ligands have been shown to be remarkably dependent on precise protein conformation, where acceptable results in pose prediction have been generally possible only in the artificial case of re-docking a ligand into a protein binding site whose conformation was determined in the presence of the same ligand (the "cognate" docking problem). In such cases, on well curated protein/ligand complexes, accurate dockings can be returned as top-scoring over 75% of the time using tools such as Surflex-Dock. A critical application of docking in modeling for lead optimization requires accurate pose prediction for novel ligands, ranging from simple synthetic analogs to very different molecular scaffolds. Typical results for widely used programs in the "cross-docking case" (making use of a single fixed protein conformation) have rates closer to 20% success. By making use of protein conformations from multiple complexes, Surflex-Dock yields an average success rate of 61% across eight pharmaceutically relevant targets. Following docking, protein pocket adaptation and rescoring identifies single pose families that are correct an average of 67% of the time. Consideration of the best of two pose families (from alternate scoring regimes) yields a 75% mean success rate.

  5. Modelling anaerobic co-digestion in Benchmark Simulation Model No. 2: Parameter estimation, substrate characterisation and plant-wide integration.

    Science.gov (United States)

    Arnell, Magnus; Astals, Sergi; Åmand, Linda; Batstone, Damien J; Jensen, Paul D; Jeppsson, Ulf

    2016-07-01

    Anaerobic co-digestion is an emerging practice at wastewater treatment plants (WWTPs) to improve the energy balance and integrate waste management. Modelling of co-digestion in a plant-wide WWTP model is a powerful tool to assess the impact of co-substrate selection and dose strategy on digester performance and plant-wide effects. A feasible procedure to characterise and fractionate co-substrates COD for the Benchmark Simulation Model No. 2 (BSM2) was developed. This procedure is also applicable for the Anaerobic Digestion Model No. 1 (ADM1). Long chain fatty acid inhibition was included in the ADM1 model to allow for realistic modelling of lipid rich co-substrates. Sensitivity analysis revealed that, apart from the biodegradable fraction of COD, protein and lipid fractions are the most important fractions for methane production and digester stability, with at least two major failure modes identified through principal component analysis (PCA). The model and procedure were tested on bio-methane potential (BMP) tests on three substrates, each rich on carbohydrates, proteins or lipids with good predictive capability in all three cases. This model was then applied to a plant-wide simulation study which confirmed the positive effects of co-digestion on methane production and total operational cost. Simulations also revealed the importance of limiting the protein load to the anaerobic digester to avoid ammonia inhibition in the digester and overloading of the nitrogen removal processes in the water train. In contrast, the digester can treat relatively high loads of lipid rich substrates without prolonged disturbances.

  6. Improved anaerobic digestion of a thermally pretreated mixture of physicochemical sludge; broiler excreta and sugar cane wastes (SCW): Effect on organic matter solubilization, biodegradability and bioenergy production.

    Science.gov (United States)

    Nava-Valente, Noemí; Alvarado-Lassman, Alejandro; Nativitas-Sandoval, Liliana S; Mendez-Contreras, Juan M

    2016-01-01

    Thermal pretreatment effect of a mixture of organic wastes (physicochemical sludge, excreta of broiler chickens and sugarcane wastes (SCW)) in the solubilization and biodegradability organic matter as well as bioenergy production by anaerobic digestion was evaluated. Two different mixtures of physicochemical sludge, excreta of broiler chickens and SCW (70%, 15%, 15% and 60%, 20%, 20% of VS, respectively) were treated at different temperatures (80 °C, 85 °C and 90 °C) and contact time (30, 60 and 90 min). Results indicate that, organic matter solubilization degree increased from 1.14 to 6.56%; subsequently, in the anaerobic digestion process, an increase of 50% in the volatile solids removal and 10% in biogas production was observed, while, retention time decreased from 23 up to 9 days. The results obtained were similar to pilot-scale. In both experimental scales it showed that the synergy produced by the simultaneous anaerobic digestion of different substrates could increase bioenergy production up to 1.3 L bio g(-1) VS removed and 0.82 L CH4 g(-1) VS removed. The treatment conditions presented in this study allow for large residue quantities to be treated and large bioenergy quantities to be produced (10% higher than during conventional treatment) without increasing the anaerobic digester volume.

  7. Digestive Tract

    Institute of Scientific and Technical Information of China (English)

    2004-01-01

    8.1 Stomach and duodenum2004380 Increased expression of heparanase mRNA is correlated with metastasis of gastric carcinoma and is probably correlated with c-met protein. CAIYongguo (蔡永国),et al. 3rd Milk Univ.Dept Gas-troenterol, South-West Hosp, Chongqing 400038. 2004;84(12):974-978.

  8. Reduced β-lactoglobulin IgE binding upon in vitro digestion as a result of the interaction of the protein with casein glycomacropeptide.

    Science.gov (United States)

    Martinez, María J; Martos, Gustavo; Molina, Elena; Pilosof, Ana M R

    2016-02-01

    The aim of this work was to evaluate the effect of the presence of casein glycomacropeptide (CMP) on the in vitro digestibility and potential allergenicity of β-lactoglobulin (β-lg)-CMP mixtures. The digestion products were analyzed by RP-HPLC and RP-HPLC-ESI-MS/MS. The potential allergenicity of the digestion products was studied by human IgE binding by inhibition ELISA with serum samples from children with clinical allergic symptoms to β-lg. No differences were observed by HPLC in the mixtures hydrolysates due to CMP-β-lg interactions. RP-HPLC-ESI-MS/MS results showed different peptides occurring in the mixtures hydrolysates. Additionally, it was observed a significant reduction of β-lg IgE binding in the presence of CMP. The disappearance of epitopes in the digested mixtures could explain the lower IgE binding observed in these systems compared to β-lg. It can be concluded that the presence of CMP in products containing β-lg may modify the digestion products that may reduce the potential allergenicity of β-lg.

  9. Two-phase anaerobic digestion of vegetable market waste fraction of municipal solid waste and development of improved technology for phase separation in two-phase reactor.

    Science.gov (United States)

    Majhi, Bijoy Kumar; Jash, Tushar

    2016-12-01

    Biogas production from vegetable market waste (VMW) fraction of municipal solid waste (MSW) by two-phase anaerobic digestion system should be preferred over the single-stage reactors. This is because VMW undergoes rapid acidification leading to accumulation of volatile fatty acids and consequent low pH resulting in frequent failure of digesters. The weakest part in the two-phase anaerobic reactors was the techniques applied for