Combined Mechanical Destruction and Alkaline Pretreatment of Wheat Straw for Enhanced Enzymatic Saccharification

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Qianqian Wang

2014-09-01

Full Text Available Wheat straw was pretreated by combined mechanical destruction and alkaline pretreatments to enhance enzymatic saccharification. Four strategies were employed to evaluate the potential of wheat straw as a feedstock for fermentable sugar production. The effects of the pretreatments on the substrate morphology, size distribution, chemical composition, and cellulose crystallinity, along with the subsequent enzymatic digestibility, were investigated. Optical microscope images showed that mechanical pretreatment alone resulted in poor fiber defibrillation, wherein samples mostly consisted of rigid fiber bundles, while integrated mechanical destruction and alkaline pretreatment led to relatively good fiber defibrillation. Low temperature NaOH/urea pretreatment can fibrillate the rigid fiber bundles into a relatively loose network and alter the structure of the treated substrate to make cellulose more accessible. The glucan conversion rates were 77% and 95% for integrated mechanical destruction and alkaline pretreatments and mechanical destruction followed by low temperature NaOH/urea and ammonium/urea pretreatments, respectively, after 72 h of enzymatic hydrolysis with enzyme loadings of 10 FPU cellulase per g of oven-dry substrate.

Analysing the mechanisms of sludge digestion enhanced by iron.

Science.gov (United States)

Hao, Xiaodi; Wei, Jing; van Loosdrecht, Mark C M; Cao, Daqi

2017-06-15

Carbon-neutral operation of wastewater treatment plants (WWTPs) requires enhancing anaerobic digestion (AD) of excess sludge for a higher energy conversion efficiency. Among others, iron has been identified to function on enhancing methane production in AD. As an industrial residual, waste iron scraps (WISs) have been reported as potentially enhancing CH 4 production in AD. With this study, the mechanisms of AD enhanced by WISs are analysed in a two-phase process: acidogenic phase (AP) and methanogenic phase (MP). Semi-continuous tests substantially excluded ORP reduction and hydrogen-evolution corrosion induced by WISs in enhancing CH 4 production, although WISs (10 g Fe/L) could indeed increase CH 4 production by 10.1% and 21.4% when added in AP and MP respectively. Detection on both FISH and enzymatic activities of involved microorganisms revealed that the stimulating effects of WISs on anaerobes (both catabolism and anabolism) could play an important (96.3%) role in enhancing CH 4 production, which would facilitate hydrolysis of refractory organics and improvement of electron transport rate (ETR). Copyright © 2017 Elsevier Ltd. All rights reserved.

The effects of fermentation and enzymatic treatment of pea on nutrient digestibility and growth performance of broilers.

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Goodarzi Boroojeni, F; Senz, M; Kozłowski, K; Boros, D; Wisniewska, M; Rose, D; Männer, K; Zentek, J

2017-10-01

The present study examined the impacts of native, fermented or enzymatically treated peas (Pisum sativum L.) inclusion in broiler diets, on growth performance and nutrient digestibility. For the fermentation process, Madonna pea was mixed with water (1/1) containing 2.57×108 Bacillus subtilis (GalliPro®) spores/kg pea and then, incubated for 48 h at 30 °C. For the enzymatic treatment process, the used water for dough production contained three enzymes, AlphaGalTM (α-galactosidase), RONOZYME® ProAct and VP (protease and pectinases respectively - DSM, Switzerland) and the pea dough incubated for 24 h at 30°C. Nine corn-wheat-soybean diets were formulated by supplying 10%, 20% and 30% of the required CP with either native, fermented or enzymatically treated peas. Performance was recorded weekly and at the end of the experiment (day 35), apparent ileal digestibility (AID) of CP, amino acids (AA), crude fat, starch, Ca, P and K were determined. Data were subjected to ANOVA using GLM procedure with a 3×3 factorial arrangement of treatments. Both processes reduced α-galactosides, phytate, trypsin inhibitor activity and resistant starch in peas. Increasing levels of pea products up to 300 g/kg diet, reduced BW gain and feed intake (P⩽0.05). Broilers fed diets containing enzymatically treated pea had the best feed conversion ratio at day 35. Different types of pea product and their inclusion levels had no effect on AID of all nutrients. The interaction between type of the pea products and inclusion levels was significant for AID of starch. For native pea diets, 10% group showed similar AID of starch to 20% native pea but it had higher AID than 30% native pea. For fermented and enzymatically treated groups, all three levels displayed similar AID of starch. In conclusion, enzymatic treatment and fermentation could improve the nutritional quality of pea. Inclusion of enzymatically treated pea in broiler diets could improve broiler performance compared with other pea

Release of Antioxidant Capacity from Five Plant Foods during a Multistep Enzymatic Digestion Protocol

NARCIS (Netherlands)

Papillo, V.A.; Vitaglione, P.; Graziani, G.; Gokmen, V.; Fogliano, V.

2014-01-01

This study aimed at elucidating the influence of food matrix on the release of antioxidant activity from five plant foods (apple, spinach, walnut, red bean, and whole wheat). To this purpose a protocol based on sequential enzymatic digestion was adopted. The total antioxidant capacity (TAC) of both

Effects of aqueous ammonia treatment on fiber’s surface morphology and enzymatic digestibility of empty fruit bunch fiber (EFBF)

International Nuclear Information System (INIS)

Ling, Tang Pei; Hassan, Osman

2013-01-01

This study was conducted to investigate the effects of aqueous ammonia reflux and soaked treatment on the fiber’s surface morphology and enzymatic digestibility of empty fruit bunch fiber (EFBF). The surface morphological changes of the fiber after aqueous ammonia treatment was linked to the sugars yield by enzymatic hydrolysis. The effectiveness of 6.25% aqueous ammonia treatment in improving enzymatic digestibility of EFBF was initially studied in reflux system and by soaking. The results showed that soaked treatment was more effective than reflux system. Further study on soaked treatment of EFBF was carried out by increasing the ammonia concentration to 12.50%. Soaking in aqueous ammonia was conducted at 30°C and 50°C for 24 hours. The results of enzymatic hydrolysis showed that sugar yield from EFBF soaked in 12.50% aqueous ammonia at 50°C was the highest. Approximately 242.91±15.50 mg/g EFBF of xylose and 320.49±28.31 mg/g EFBF of glucose were produced by the action of enzyme Cellic Ctec 2. Results of scanning electron microscopic showed that aqueous ammonia treatment by soaking had caused a more severe structural distortion on the fiber’s surface and higher removal of silica bodies that embedded on the fiber than those in reflux system. The changes on the fiber’s surface morphology were believed is the contributing factor that improved the enzymatic digestibility of EFBF after aqueous ammonia treatment

Effects of aqueous ammonia treatment on fiber’s surface morphology and enzymatic digestibility of empty fruit bunch fiber (EFBF)

Energy Technology Data Exchange (ETDEWEB)

Ling, Tang Pei; Hassan, Osman [Department of Food Science, School of Chemical Science and Food Technology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi UKM, Selangor (Malaysia)

2013-11-27

This study was conducted to investigate the effects of aqueous ammonia reflux and soaked treatment on the fiber’s surface morphology and enzymatic digestibility of empty fruit bunch fiber (EFBF). The surface morphological changes of the fiber after aqueous ammonia treatment was linked to the sugars yield by enzymatic hydrolysis. The effectiveness of 6.25% aqueous ammonia treatment in improving enzymatic digestibility of EFBF was initially studied in reflux system and by soaking. The results showed that soaked treatment was more effective than reflux system. Further study on soaked treatment of EFBF was carried out by increasing the ammonia concentration to 12.50%. Soaking in aqueous ammonia was conducted at 30°C and 50°C for 24 hours. The results of enzymatic hydrolysis showed that sugar yield from EFBF soaked in 12.50% aqueous ammonia at 50°C was the highest. Approximately 242.91±15.50 mg/g EFBF of xylose and 320.49±28.31 mg/g EFBF of glucose were produced by the action of enzyme Cellic Ctec 2. Results of scanning electron microscopic showed that aqueous ammonia treatment by soaking had caused a more severe structural distortion on the fiber’s surface and higher removal of silica bodies that embedded on the fiber than those in reflux system. The changes on the fiber’s surface morphology were believed is the contributing factor that improved the enzymatic digestibility of EFBF after aqueous ammonia treatment.

Stability of milk fat globule membrane proteins toward human enzymatic gastrointestinal digestion.

Science.gov (United States)

Le, T T; Van de Wiele, T; Do, T N H; Debyser, G; Struijs, K; Devreese, B; Dewettinck, K; Van Camp, J

2012-05-01

The milk fat globule membrane (MFGM) fraction refers to the thin film of polar lipids and membrane proteins that surrounds fat globules in milk. It is its unique biochemical composition that renders MFGM with some beneficial biological activities, such as anti-adhesive effects toward pathogens. However, a prerequisite for the putative bioactivity of MFGM is its stability during gastrointestinal digestion. We, therefore, subjected MFGM material, isolated from raw milk, to an in vitro enzymatic gastrointestinal digestion. Sodium dodecyl sulfate PAGE, in combination with 2 staining methods, Coomassie Blue and periodic acid Schiff staining, was used to evaluate polypeptide patterns of the digest, whereas mass spectrometry was used to confirm the presence of specific MFGM proteins. Generally, it was observed that glycoproteins showed higher resistance to endogenous proteases compared with non-glycosylated proteins. Mucin 1 displayed the highest resistance to digestion and a considerable part of this protein was still detected at its original molecular weight after gastric and small intestine digestion. Cluster of differentiation 36 was also quite resistant to pepsin. A significant part of periodic acid Schiff 6/7 survived the gastric digestion, provided that the lipid moiety was not removed from the MFGM material. Overall, MFGM glycoproteins are generally more resistant to gastrointestinal digestion than serum milk proteins and the presence of lipids, besides glycosylation, may protect MFGM glycoproteins from gastrointestinal digestion. This gastrointestinal stability makes MFGM glycoproteins amenable to further studies in which their putative health-promoting effects can be explored. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Combined pretreatment with hot compressed water and wet disk milling opened up oil palm biomass structure resulting in enhanced enzymatic digestibility.

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Zakaria, Mohd Rafein; Hirata, Satoshi; Fujimoto, Shinji; Hassan, Mohd Ali

2015-10-01

Combined pretreatment with hot compressed water and wet disk milling was performed with the aim to reduce the natural recalcitrance of oil palm biomass by opening its structure and provide maximal access to cellulase attack. Oil palm empty fruit bunch and oil palm frond fiber were first hydrothermally pretreated at 150-190° C and 10-240 min. Further treatment with wet disk milling resulted in nanofibrillation of fiber which caused the loosening of the tight biomass structure, thus increasing the subsequent enzymatic conversion of cellulose to glucose. The effectiveness of the combined pretreatments was evaluated by chemical composition changes, power consumption, morphological alterations by SEM and the enzymatic digestibility of treated samples. At optimal pretreatment process, approximately 88.5% and 100.0% of total sugar yields were obtained from oil palm empty fruit bunch and oil palm frond fiber samples, which only consumed about 15.1 and 23.5 MJ/kg of biomass, respectively. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enzymatic digestibility of peptides cross-linked by ionizing radiation

International Nuclear Information System (INIS)

Dizdaroglu, M.; Gajewski, E.; Simic, M.G.

1984-01-01

Digestibility by proteolytic enzymes of peptides cross-linked by ionizing radiation was investigated. Small peptides of alanine and phenylalanine were chosen as model compounds and aminopeptidases and carboxypeptidases were used as proteolytic enzymes. Peptides exposed to γ-radiation in aqueous solution were analysed by high-performance liquid chromatography before and after hydrolysis by aminopeptidase M, leucine aminopeptidase carboxypeptidase A and carboxypeptidase Y. The results obtained clearly demonstrate the different actions of these enzymes on cross-linked aliphatic and aromatic peptides. Peptide bonds of cross-linked dipeptides of alanine were completely resistant to enzymatic hydrolysis whereas the enzymes, except for carboxypeptidase Y, cleaved all peptide bonds of cross-linked peptides of phenylalanine. The actions of the enzymes on these particular compounds are discussed in detail. (author)

Efficient sugar release by acetic acid ethanol-based organosolv pretreatment and enzymatic saccharification.

Science.gov (United States)

Zhang, Hongdan; Wu, Shubin

2014-12-03

Acetic acid ethanol-based organosolv pretreatment of sugar cane bagasse was performed to enhance enzymatic hydrolysis. The effect of different parameters (including temperature, reaction time, solvent concentration, and acid catalyst dose) on pretreatment prehydrolyzate and subsequent enzymatic digestibility was determined. During the pretreatment process, 11.83 g of xylose based on 100 g of raw material could be obtained. After the ethanol-based pretreatment, the enzymatic hydrolysis was enhanced and the highest glucose yield of 40.99 g based on 100 g of raw material could be obtained, representing 93.8% of glucose in sugar cane bagasse. The maximum total sugar yields occurred at 190 °C, 45 min, 60:40 ethanol/water, and 5% dosage of acetic acid, reaching 58.36 g (including 17.69 g of xylose and 40.67 g of glucose) based on 100 g of raw material, representing 85.4% of total sugars in raw material. Furthermore, characterization of the pretreated sugar cane bagasse using X-ray diffraction and scanning electron microscopy analyses were also developed. The results suggested that ethanol-based organosolv pretreatment could enhance enzymatic digestibilities because of the delignification and removal of xylan.

Comparison of lab, pilot, and industrial scale low consistency mechanical refining for improvements in enzymatic digestibility of pretreated hardwood.

Science.gov (United States)

Jones, Brandon W; Venditti, Richard; Park, Sunkyu; Jameel, Hasan

2014-09-01

Mechanical refining has been shown to improve biomass enzymatic digestibility. In this study industrial high-yield sodium carbonate hardwood pulp was subjected to lab, pilot and industrial refining to determine if the mechanical refining improves the enzymatic hydrolysis sugar conversion efficiency differently at different refining scales. Lab, pilot and industrial refining increased the biomass digestibility for lignocellulosic biomass relative to the unrefined material. The sugar conversion was increased from 36% to 65% at 5 FPU/g of biomass with industrial refining at 67.0 kWh/t, which was more energy efficient than lab and pilot scale refining. There is a maximum in the sugar conversion with respect to the amount of refining energy. Water retention value is a good predictor of improvements in sugar conversion for a given fiber source and composition. Improvements in biomass digestibility with refining due to lab, pilot plant and industrial refining were similar with respect to water retention value. Published by Elsevier Ltd.

Enzymatic digestive activity and absorption efficiency in Tagelus dombeii upon Alexandrium catenella exposure

Science.gov (United States)

Fernández-Reiriz, M. J.; Navarro, J. M.; Cisternas, B. A.; Babarro, J. M. F.; Labarta, U.

2013-12-01

We analyzed absorption efficiency (AE) and digestive enzyme activity (amylase, cellulase complex, and laminarinase) of the infaunal bivalve Tagelus dombeii originating from two geographic sites, Corral-Valdivia and Melinka-Aysén, which have different long-term paralytic shellfish poisoning (PSP) exposure rates. We report the effects of past feeding history (origin) on T. dombeii exposed to a mixed diet containing the toxic dinoflagellate Alexandrium catenella and another dinoflagellate-free control diet over a 12-day period in the laboratory. Absorption efficiency values of T. dombeii individuals that experienced PSP exposure in their habitat (Melinka-Aysén) remained unchanged during exposure to toxic food in the laboratory. In contrast, T. dombeii from a non-PSP exposure field site (Corral-Valdivia) showed a significant reduction in AE with toxic exposure time. This study established that the amylase and cellulase complexes were the most important enzymes in the digestive glands of Tagelus from both sites. The temporal evolution of enzymatic activity under toxic diet was fitted to exponential (amylase and cellulase) and to a logarithmic (laminarinase) models. In all fits, we found significant effect of origin in the model parameters. At the beginning of the experiment, higher enzymatic activity was observed for clams from Corral-Valdivia. The amylase activity decreased with time exposure for individuals from Corral and increased for individuals from Melinka. Cellulase activity did not vary over time for clams from Corral, but increased for individuals from Melinka and laminarinase activity decreased over time for individuals from Corral and remained unchanged over time for Melinka. A feeding history of exposure to the dinoflagellate A. catenella was reflected in the digestive responses of both T. dombeii populations.

Comparison of various pretreatments for ethanol production enhancement from solid residue after rumen fluid digestion of rice straw.

Science.gov (United States)

Zhang, Haibo; Zhang, Panyue; Ye, Jie; Wu, Yan; Liu, Jianbo; Fang, Wei; Xu, Dong; Wang, Bei; Yan, Li; Zeng, Guangming

2018-01-01

The rumen digested residue of rice straw contains high residual carbohydrates, which makes it a potential cellulosic ethanol feedstock. This study evaluated the feasibility and effectiveness of applying microwave assisted alkali (MAP), ultrasound assisted alkali (UAP), and ball milling pretreatment (BMP) to enhance ethanol production from two digested residues (2.5%-DR and 10%-DR) after rumen fluid digestion of rice straw at 2.5% and 10.0% solid content. Results revealed that 2.5%-DR and 10%-DR had a cellulose content of 36.4% and 41.7%, respectively. MAP and UAP improved enzymatic hydrolysis of digested residue by removing the lignin and hemicellulose, while BMP by decreasing the particle size and crystallinity. BMP was concluded as the suitable pretreatment, resulting in an ethanol yield of 116.65 and 147.42mgg -1 for 2.5%-DR and 10%-DR, respectively. The integrated system including BMP for digested residue at 2.5% solid content achieved a maximum energy output of 7010kJkg -1 . Copyright © 2017 Elsevier Ltd. All rights reserved.

Genetic Engineering of Energy Crops to Reduce Recalcitrance and Enhance Biomass Digestibility

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Monika Yadav

2018-06-01

Full Text Available Bioenergy, biofuels, and a range of valuable chemicals may be extracted from the abundantly available lignocellulosic biomass. To reduce the recalcitrance imposed by the complex cell wall structure, genetic engineering has been proposed over the years as a suitable solution to modify the genes, thereby, controlling the overall phenotypic expression. The present review provides a brief description of the plant cell wall structure and its compositional array i.e., lignin, cellulose, hemicellulose, wall proteins, and pectin, along with their effect on biomass digestibility. Also, this review discusses the potential to increase biomass by gene modification. Furthermore, the review highlights the potential genes associated with the regulation of cell wall structure, which can be targeted for achieving energy crops with desired phenotypes. These genetic approaches provide a robust and assured method to bring about the desired modifications in cell wall structure, composition, and characteristics. Ultimately, these genetic modifications pave the way for achieving enhanced biomass yield and enzymatic digestibility of energy crops, which is crucial for maximizing the outcomes of energy crop breeding and biorefinery applications.

Effect of Enzymatic Digestion of Protein Derivatives Obtained from Mucuna pruriens L. on Production of Proinflammatory Mediators by BALB/c Mouse Macrophages.

Science.gov (United States)

Martínez Leo, Edwin E; Arana Argáez, Victor E; Acevedo Fernández, Juan J; Puc, Rosa Moo; Segura Campos, Maira R

2018-04-25

Inflammation is considered to be a major risk factor for the pathogenesis of chronic non-communicable diseases. Macrophages are important immune cells, which regulate inflammation and host defense by secretion of proinflammatory mediators. Obtaining biopeptides by enzymatic hydrolysis adds value to proteins of vegetative origin, such as Mucuna pruriens L. The present study evaluated the effect of enzymatic digestion of protein derivatives obtained from M. pruriens L. on the production of proinflammatory mediators by BALB/c mouse macrophages. Five different molecular weight peptide fractions were obtained (F > 10, 5-10, 3-5, 1-3, and < 1 kDa, respectively). At 300 μg/mL, F5-10 kDa inhibited 50.26 and 61.00% NO and H 2 O 2 production, respectively. Moreover, F5-10 kDa reduced the IL-6 and TNFα levels to 60.25 and 69.54%, respectively. After enzymatic digestive simulation, F5-10 kDa decreased the inflammatory mediators.

The minor wall-networks between monolignols and interlinked-phenolics predominantly affect biomass enzymatic digestibility in Miscanthus.

Science.gov (United States)

Li, Zhengru; Zhao, Chunqiao; Zha, Yi; Wan, Can; Si, Shengli; Liu, Fei; Zhang, Rui; Li, Fengcheng; Yu, Bin; Yi, Zili; Xu, Ning; Peng, Liangcai; Li, Qing

2014-01-01

Plant lignin is one of the major wall components that greatly contribute to biomass recalcitrance for biofuel production. In this study, total 79 representative Miscanthus germplasms were determined with wide biomass digestibility and diverse monolignol composition. Integrative analyses indicated that three major monolignols (S, G, H) and S/G ratio could account for lignin negative influence on biomass digestibility upon NaOH and H2SO4 pretreatments. Notably, the biomass enzymatic digestions were predominately affected by the non-KOH-extractable lignin and interlinked-phenolics, other than the KOH-extractable ones that cover 80% of total lignin. Furthermore, a positive correlation was found between the monolignols and phenolics at pnetworks against cellulases accessibility. The results indicated that the non-KOH-extractable lignin-complex should be the target either for cost-effective biomass pretreatments or for relatively simply genetic modification of plant cell walls in Miscanthus.

Lignosulfonate and elevated pH can enhance enzymatic saccharification of lignocelluloses

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Wang ZJ

2013-01-01

Full Text Available Abstract Background Nonspecific (nonproductive binding (adsorption of cellulase by lignin has been identified as a key barrier to reduce cellulase loading for economical sugar and biofuel production from lignocellulosic biomass. Sulfite Pretreatment to Overcome Recalcitrance of Lignocelluloses (SPORL is a relatively new process, but demonstrated robust performance for sugar and biofuel production from woody biomass especially softwoods in terms of yields and energy efficiencies. This study demonstrated the role of lignin sulfonation in enhancing enzymatic saccharification of lignocelluloses – lignosulfonate from SPORL can improve enzymatic hydrolysis of lignocelluloses, contrary to the conventional belief that lignin inhibits enzymatic hydrolysis due to nonspecific binding of cellulase. Results The study found that lignosulfonate from SPORL pretreatment and from a commercial source inhibits enzymatic hydrolysis of pure cellulosic substrates at low concentrations due to nonspecific binding of cellulase. Surprisingly, the reduction in enzymatic saccharification efficiency of a lignocellulosic substrate was fully recovered as the concentrations of these two lignosulfonates increased. We hypothesize that lignosulfonate serves as a surfactant to enhance enzymatic hydrolysis at higher concentrations and that this enhancement offsets its inhibitive effect from nonspecific binding of cellulase, when lignosulfonate is applied to lignocellulosic solid substrates. Lignosulfonate can block nonspecific binding of cellulase by bound lignin on the solid substrates, in the same manner as a nonionic surfactant, to significantly enhance enzymatic saccharification. This enhancement is linearly proportional to the amount of lignosulfonate applied which is very important to practical applications. For a SPORL-pretreated lodgepole pine solid, 90% cellulose saccharification was achieved at cellulase loading of 13 FPU/g glucan with the application of its

Extractability and digestibility of plant cell wall polysaccharides during hydrothermal and enzymatic degradation of wheat straw (Triticum aestivum L.)

DEFF Research Database (Denmark)

Hansen, Mads A.T.; Ahl, Louise I.; Pedersen, Henriette L.

2014-01-01

to about 20, but mostly around 3-8, and notably more acetylated in stems. Arabinoxylan (AX) and mixed-linkage glucan (MLG) became water-extractable while xylan, xyloglucan (XG), mannan and glucan remained only alkali-extractable. All polysaccharides became partly digestible after pretreatment however......, regardless their extractability in water or only alkali. Based on the results, AX and MLG appear to be loosely bound in the cell wall matrix while the other polysaccharides are bound more tightly and shielded from enzymatic attack by AX and MLG until pretreatment. The gradual solubilisation and digestion...

The minor wall-networks between monolignols and interlinked-phenolics predominantly affect biomass enzymatic digestibility in Miscanthus.

Directory of Open Access Journals (Sweden)

Zhengru Li

Full Text Available Plant lignin is one of the major wall components that greatly contribute to biomass recalcitrance for biofuel production. In this study, total 79 representative Miscanthus germplasms were determined with wide biomass digestibility and diverse monolignol composition. Integrative analyses indicated that three major monolignols (S, G, H and S/G ratio could account for lignin negative influence on biomass digestibility upon NaOH and H2SO4 pretreatments. Notably, the biomass enzymatic digestions were predominately affected by the non-KOH-extractable lignin and interlinked-phenolics, other than the KOH-extractable ones that cover 80% of total lignin. Furthermore, a positive correlation was found between the monolignols and phenolics at p<0.05 level in the non-KOH-extractable only, suggesting their tight association to form the minor wall-networks against cellulases accessibility. The results indicated that the non-KOH-extractable lignin-complex should be the target either for cost-effective biomass pretreatments or for relatively simply genetic modification of plant cell walls in Miscanthus.

Steam explosion distinctively enhances biomass enzymatic saccharification of cotton stalks by largely reducing cellulose polymerization degree in G. barbadense and G. hirsutum.

Science.gov (United States)

Huang, Yu; Wei, Xiaoyang; Zhou, Shiguang; Liu, Mingyong; Tu, Yuanyuan; Li, Ao; Chen, Peng; Wang, Yanting; Zhang, Xuewen; Tai, Hongzhong; Peng, Liangcai; Xia, Tao

2015-04-01

In this study, steam explosion pretreatment was performed in cotton stalks, leading to 5-6 folds enhancements on biomass enzymatic saccharification distinctive in Gossypium barbadense and Gossypium hirsutum species. Sequential 1% H2SO4 pretreatment could further increase biomass digestibility of the steam-exploded stalks, and also cause the highest sugar-ethanol conversion rates probably by releasing less inhibitor to yeast fermentation. By comparison, extremely high concentration alkali (16% NaOH) pretreatment with raw stalks resulted in the highest hexoses yields, but it had the lowest sugar-ethanol conversion rates. Characterization of wall polymer features indicated that biomass saccharification was enhanced with steam explosion by largely reducing cellulose DP and extracting hemicelluloses. It also showed that cellulose crystallinity and arabinose substitution degree of xylans were the major factors on biomass digestibility in cotton stalks. Hence, this study has provided the insights into cell wall modification and biomass process technology in cotton stalks and beyond. Copyright © 2015 Elsevier Ltd. All rights reserved.

Size effects on acid bisulfite pretreatment efficiency: multiple product yields in spent liquor and enzymatic digestibility of pretreated solids

Science.gov (United States)

Yalan Liu; Jinwu Wang; Michael P. Wolcott

2017-01-01

Currently, feedstock size effects on chemical pretreatment performance were not clear due to the complexity of the pretreatment process and multiple evaluation standards such as the sugar recovery in spent liquor or enzymatic digestibility. In this study, we evaluated the size effects by various ways: the sugar recovery and coproduct yields in spent liquor, the...

Using temperature-responsive zwitterionic surfactant to enhance the enzymatic hydrolysis of lignocelluloses and recover cellulase by cooling.

Science.gov (United States)

Cai, Cheng; Pang, Yuxia; Zhan, Xuejuan; Zeng, Meijun; Lou, Hongming; Qian, Yong; Yang, Dongjie; Qiu, Xueqing

2017-11-01

Some zwitterionic surfactants exhibit upper critical solution temperature (UCST) in aqueous solutions. For the zwitterionic surfactant solution mixed with cellulase, when its temperature is below UCST, the cellulase can be recovered by coprecipitation with zwitterionic surfactant. In this work, 3-(Hexadecyldimethylammonio) propanesulfonate (SB3-16) was selected to enhance the enzymatic hydrolysis of lignocelluloses and recover the cellulase. After adding 2mmol/L of SB3-16, the enzymatic digestibility of eucalyptus pretreated by dilute acid (Eu-DA) and by sulfite (Eu-SPORL) increased from 27.9% and 35.1% to 72.6% and 89.7%, respectively. The results showed that SB3-16 could reduce the non-productive adsorption of cellulase on hydrophobic interface, while it did not significantly inhibit the activity of cellulase. For the solution contained 1wt% SB3-16 and 200mg protein/L CTec2 cellulase, 55.2% of protein could be recovered by cooling. The filter paper activity of the recovered cellulase was 1.93FPU/mg protein, which was 95.8% of its initial activity. Copyright © 2017. Published by Elsevier Ltd.

Chemical composition and enzymatic digestibility of sugarcane clones selected for varied lignin content

Directory of Open Access Journals (Sweden)

Masarin Fernando

2011-12-01

Full Text Available Abstract Background The recalcitrance of lignocellulosic materials is a major limitation for their conversion into fermentable sugars. Lignin depletion in new cultivars or transgenic plants has been identified as a way to diminish this recalcitrance. In this study, we assessed the success of a sugarcane breeding program in selecting sugarcane plants with low lignin content, and report the chemical composition and agronomic characteristics of eleven experimental hybrids and two reference samples. The enzymatic digestion of untreated and chemically delignified samples was evaluated to advance the performance of the sugarcane residue (bagasse in cellulosic-ethanol production processes. Results The ranges for the percentages of glucan, hemicellulose, lignin, and extractive (based on oven-dry biomass of the experimental hybrids and reference samples were 38% to 43%, 25% to 32%, 17% to 24%, and 1.6% to 7.5%, respectively. The samples with the smallest amounts of lignin did not produce the largest amounts of total polysaccharides. Instead, a variable increase in the mass of a number of components, including extractives, seemed to compensate for the reduction in lignin content. Hydroxycinnamic acids accounted for a significant part of the aromatic compounds in the samples, with p-coumaric acid predominating, whereas ferulic acid was present only in low amounts. Hydroxycinnamic acids with ester linkage to the hemicelluloses varied from 2.3% to 3.6%. The percentage of total hydroxycinnamic acids (including the fraction linked to lignin through ether linkages varied from 5.0% to 9.2%, and correlated to some extent with the lignin content. These clones released up to 31% of glucose after 72 hours of digestion with commercial cellulases, whereas chemically delignified samples led to cellulose conversion values of more than 80%. However, plants with lower lignin content required less delignification to reach higher efficiencies of cellulose conversion during

Multi-Scale Computational Enzymology: Enhancing Our Understanding of Enzymatic Catalysis

OpenAIRE

Rami Gherib; Hisham M. Dokainish; James W. Gauld

2013-01-01

Elucidating the origin of enzymatic catalysis stands as one the great challenges of contemporary biochemistry and biophysics. The recent emergence of computational enzymology has enhanced our atomistic-level description of biocatalysis as well the kinetic and thermodynamic properties of their mechanisms. There exists a diversity of computational methods allowing the investigation of specific enzymatic properties. Small or large density functional theory models allow the comparison of a pleth...

Correlation between anatomical characteristics of ethanol organosolv pretreated Buddleja davidii and its enzymatic conversion to glucose.

Science.gov (United States)

Hallac, Bassem B; Ray, Michael; Murphy, Richard J; Ragauskas, Arthur J

2010-12-01

Buddleja davidii is a unique biomass that has many attractive agroenergy features, especially its wide range of growth habitat. The anatomical characteristics of B. davidii were investigated before and after ethanol organosolv pretreatment (one of the leading pretreatment technologies) in order to further understand the alterations that occur to the cellular structure of the biomass which can then be correlated with its enzymatic digestibility. Results showed that the ethanol organosolv pretreatment of B. davidii selectively removes lignin from the middle lamella (ML), which does not significantly disrupt the crystalline structure of cellulose. The removal of ML lignin is a major factor in enhancing enzymatic cellulose-to-glucose hydrolysis. The pretreatment also causes cell deformation, resulting in cracks and breaks in the cell wall. These observations, together with characterization analysis of the cell wall polymer material, lend support to the hypothesis that the physical distribution of lignin in the biomass matrix is an important structural feature affecting biomass enzymatic digestibility. © 2010 Wiley Periodicals, Inc.

Mechano-Enzymatic Deconstruction with a New Enzymatic Cocktail to Enhance Enzymatic Hydrolysis and Bioethanol Fermentation of Two Macroalgae Species

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Sameh Amamou

2018-01-01

Full Text Available The aim of this study was to explore the efficiency of a mechano-enzymatic deconstruction of two macroalgae species for sugars and bioethanol production, by using a new enzymatic cocktail (Haliatase and two types of milling modes (vibro-ball: VBM and centrifugal milling: CM. By increasing the enzymatic concentration from 3.4 to 30 g/L, the total sugars released after 72 h of hydrolysis increased (from 6.7 to 13.1 g/100 g TS and from 7.95 to 10.8 g/100 g TS for the green algae U. lactuca and the red algae G. sesquipedale, respectively. Conversely, total sugars released from G. sesquipedale increased (up to 126% and 129% after VBM and CM, respectively. The best bioethanol yield (6 geth/100 g TS was reached after 72 h of fermentation of U. lactuca and no increase was obtained after centrifugal milling. The latter led to an enhancement of the ethanol yield of G. sesquipedale (from 2 to 4 g/100 g TS.

Investigation of bi-enzymatic reactor based on hybrid monolith with nanoparticles embedded and its proteolytic characteristics.

Science.gov (United States)

Shangguan, Lulu; Zhang, Lingyi; Xiong, Zhichao; Ren, Jun; Zhang, Runsheng; Gao, Fangyuan; Zhang, Weibing

2015-04-03

The bottom-up strategy of proteomic profiling study based on mass spectrometer (MS) has drawn high attention. However, conventional solution-based digestion could not satisfy the demands of highly efficient and complete high throughput proteolysis of complex samples. We proposed a novel bi-enzymatic reactor by immobilizing two different enzymes (trypsin/chymotrypsin) onto a mixed support of hybrid organic-inorganic monolith with SBA-15 nanoparticles embedded. Typsin and chymotrypsin were crossly immobilized onto the mixed support by covalent bonding onto the monolith with glutaraldehyde as bridge reagent and chelation via copper ion onto the nanoparticles, respectively. Compared with single enzymatic reactors, the bi-enzymatic reactor improved the overall functional analysis of membrane proteins of rat liver by doubling the number of identified peptides (from 1184/1010 with trypsin/chymotrypsin enzymatic reactors to 2891 with bi-enzymatic reactor), which led to more proteins identified with deep coverage (from 452/336 to 620); the efficiency of the bi-enzymatic reactor is also better than that of solution-based tandem digestion, greatly shorting the digestion time from 24h to 50s. Moreover, more transmembrane proteins were identified by bi-enzymatic reactor (106) compared with solution-based tandem digestion (95) with the same two enzymes and enzymatic reactors with single enzyme immobilized (75 with trypsin and 66 with chymotrypsin). The proteolytic characteristics of the bi-enzymatic reactors were evaluated by applying them to digestion of rat liver proteins. The reactors showed good digestion capability for proteins with different hydrophobicity and molecular weight. Copyright © 2015 Elsevier B.V. All rights reserved.

ENHANCED ANAEROBIC DIGESTION OF FOOD WASTE BY SUPPLEMENTING TRACE ELEMENTS: ROLE OF SELENIUM (VI AND IRON (II

Directory of Open Access Journals (Sweden)

Javkhlan eAriunbaatar

2016-02-01

Full Text Available This paper discusses the potential to enhance the anaerobic digestion of food waste FW by supplementing trace elements (Fe, Co, Ni, Zn, Mn, Cu, Se, and Mo individually as well as in cocktails. A series of batch experiments on the biomethane potential of synthetic food waste were performed with low (FW-A and high (FW-B trace element background concentrations prepared in, respectively, Delft (The Netherlands and Tampa (Florida, USA. The most effective trace elements for FW-A were Fe with an increase of 39.2 (± 0.6 % of biomethane production, followed by Se (34.1 ± 5.6 % increase, Ni (26.4 ± 0.2 % increase and Co (23.8 ± 0.2 % increase. For FW-B supplementing these trace elements did not result in enhancement of the biomethane production, except for Se. FW-B had a Se concentration of 1.3 (± 0. 5 µg/gTS, while it was below the detection limit for FW-A. Regardless of the FW source, Se resulted in 30 – 35% increase of biomethane production at a concentration range of 25-50 µg/L (0.32 – 0.63 µM. Volatile fatty acids analysis revealed that TE supplementation enhances their consumption, thus yielding a higher biomethane production. Moreover, additional experiments on sulfide inhibition showed the enhancing effects of trace elements on the anaerobic digestion of food waste were not related with sulfide toxicity, but with the enzymatic reactions and/or microbial biomass aggregation.

Two-Step Hot-Compressed Water Treatment of Douglas Fir for Efficient Total Sugar Recovery by Enzymatic Hydrolysis

Directory of Open Access Journals (Sweden)

Hiroyuki Inoue

2016-04-01

Full Text Available The non-catalytic hydrothermal pretreatment of softwood is generally less effective for subsequent enzymatic hydrolysis. In this study, the efficacy of hot-compressed water (HCW treatment of Douglas fir was investigated between 180 °C and 260 °C, allowing solubilization of the cellulose components. The enzymatic digestibility of cellulosic residues increased significantly under HCW conditions > 250 °C, and the enhanced glucan digestibility was closely related to the decomposition of the cellulose component. Combination of the first-stage HCW treatment (220 °C, 5 min to recover hemicellulosic sugars with the second-stage HCW treatment (260 °C, 5 min to improve cellulose digestibility gave a total sugar recovery of 56.2% based on the dried raw materials. This yield was 1.4 times higher than that from the one-step HCW-treated sample (260 °C, 5 min. Additionally, an enzymatic hydrolysate from the two-step HCW-treated sample exceeded 90% of the ethanol fermentation yield based on the total sugars present in the hydrolysates. These results suggest the potential of the two-step HCW treatment of softwood as a pretreatment technology for efficient total sugar recovery and ethanol production.

Effects of enzymatic hydrolysis and ultrasounds pretreatments on corn cob and vine trimming shoots for biogas production.

Science.gov (United States)

Pérez-Rodríguez, N; García-Bernet, D; Domínguez, J M

2016-12-01

Due to their lignocellulosic nature, corn cob and vine trimming shoots (VTS) could be valorized by anaerobic digestion for biogas production. To enhance the digestibility of substrates, pretreatments of lignocellulosic materials are recommended. The effect of enzymatic hydrolysis, ultrasounds pretreatments (US) and the combination of both was assayed in lignocellulosic composition, methane, and biogas yields. The pretreatments leaded to a reduction in lignin and an increase in neutral detergent soluble compounds making corn cob and VTS more amendable for biogas conversion. The US were negative for biogas production from both substrates and in particular strongly detrimental for VTS. On the opposite side, the enzymatic hydrolysis was certainly beneficial increasing 59.8% and 14.6% the methane production from VTS and corn cob, respectively. The prior application of US did not potentiate (or not sufficiently) the improvement in the methane production reflected by the enzymatic hydrolysis pretreatment of VTS and corn cob. Copyright © 2016 Elsevier Ltd. All rights reserved.

Aged refuse enhances anaerobic digestion of waste activated sludge.

Science.gov (United States)

Zhao, Jianwei; Gui, Lin; Wang, Qilin; Liu, Yiwen; Wang, Dongbo; Ni, Bing-Jie; Li, Xiaoming; Xu, Rui; Zeng, Guangming; Yang, Qi

2017-10-15

In this work, a low-cost alternative approach (i.e., adding aged refuse (AR) into waste activated sludge) to significantly enhance anaerobic digestion of sludge was reported. Experimental results showed that with the addition dosage of AR increasing from 0 to 400 mg/g dry sludge soluble chemical oxygen demand (COD) increased from 1150 to 5240 mg/L at the digestion time of 5 d, while the maximal production of volatile fatty acids (VFA) increased from 82.6 to 183.9 mg COD/g volatile suspended solids. Although further increase of AR addition decreased the concentrations of both soluble COD and VFA, their contents in these systems with AR addition at any concentration investigated were still higher than those in the blank, which resulted in higher methane yields in these systems. Mechanism studies revealed that pertinent addition of AR promoted solubilization, hydrolysis, and acidogenesis processes and did not affect methanogenesis significantly. It was found that varieties of enzymes and anaerobes in AR were primary reason for the enhancement of anaerobic digestion. Humic substances in AR benefited hydrolysis and acidogenesis but inhibited methanogenesis. The effect of heavy metals in AR on sludge anaerobic digestion was dosage dependent. Sludge anaerobic digestion was enhanced by appropriate amounts of heavy metals but inhibited by excessive amounts of heavy metals. The relative abundances of microorganisms responsible for sludge hydrolysis and acidogenesis were also observed to be improved in the system with AR addition, which was consistent with the performance of anaerobic digestion. Copyright © 2017 Elsevier Ltd. All rights reserved.

A new approach to protein enzymatic digestion for fast protein identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Czech Academy of Sciences Publication Activity Database

Dyčka, Filip; Laštovičková, Markéta; Bobálová, Janette

2008-01-01

Roč. 102, č. 15 (2008), s967-s968 ISSN 1803-2389. [Meeting on Chemistry and Life /4./. Brno, 09.09.2008-11.09.2008] R&D Projects: GA AV ČR IAA600040701 Institutional research plan: CEZ:AV0Z40310501 Keywords : enzymatic digest ion * fast approach Subject RIV: CB - Analytical Chemistry, Separation

Enzymatic saccharification of dilute acid pretreated saline crops for fermentable sugar production

Energy Technology Data Exchange (ETDEWEB)

Zheng, Yi; Zhang, Ruihong [Biological and Agricultural Engineering Department, University of California, Davis One Shields Avenue, Davis, CA 95616 (United States); Pan, Zhongli [Biological and Agricultural Engineering Department, University of California, Davis One Shields Avenue, Davis, CA 95616 (United States); Processed Foods Research Unit, USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710 (United States); Wang, Donghai [Biological and Agricultural Engineering Department, Kansas State University, Manhattan, KS 66506 (United States)

2009-11-15

Four saline crops [athel (Tamarix aphylla L), eucalyptus (Eucalyptus camaldulensis), Jose Tall Wheatgrass (Agropyron elongatum), and Creeping Wild Ryegrass (Leymus triticoides)] that are used in farms for salt uptake from soil and drainage irrigation water have the potential for fuel ethanol production because they don't take a large number of arable lands. Dilute sulfuric acid pretreatment and enzymatic hydrolysis were conducted to select the optimum pretreatment conditions and the best saline crop for further enzymatic hydrolysis research. The optimum dilute acid pretreatment conditions included T = 165 C, t = 8 min, and sulfuric acid concentration 1.4% (w/w). Creeping Wild Ryegrass was decided to be the best saline crop. Solid loading, cellulase and {beta}-glucosidase concentrations had significant effects on the enzymatic hydrolysis of dilute acid pretreated Creeping Wild Ryegrass. Glucose concentration increased by 36 mg/mL and enzymatic digestibility decreased by 20% when the solid loading increased from 4 to 12%. With 8% solid loading, enzymatic digestibility increased by over 30% with the increase of cellulase concentration from 5 to 15 FPU/g-cellulose. Under given cellulase concentration of 15 FPU/g-cellulose, 60% increase of enzymatic digestibility of pretreated Creeping Wild Ryegrass was obtained with the increase of {beta}-glucosidase concentration up to 15 CBU/g-cellulose. With a high solid loading of 10%, fed-batch operation generated 12% and 18% higher enzymatic digestibility and glucose concentration, respectively, than batch process. (author)

Preliminary experimental results of Sewage Sludge (SS) Co-digestion with Palm Oil Mill Effluent (POME) for Enhanced Biogas Production in Laboratory Scale Anaerobic Digester

International Nuclear Information System (INIS)

Sivasankari, R; Kumaran, P; Normanbhay, Saifuddin; Shamsuddin, Abd Halim

2013-01-01

An investigation on the feasibility of co-digesting Sewage Sludge with Palm Oil Mill Effluent for enhancing the biogas production and the corresponding effect of the co-digestion substrate ratio on the biogas production has been evaluated. Anaerobic co-digestion of POME with SS was performed at ratios of 100:0, 70:30, 60:40 and 0:100 to find the optimum blend required for enhanced waste digestion and biogas production. Single stage batch digestion was carried out for 12 days in a laboratory scale anaerobic digester. Co-digestion of sludge's at the 70:30 proportion resulted in optimal COD and C: N ratio which subsequently recorded the highest performance with regards to biogas production at 28.1 L's compared to the 1.98 L's of biogas produced from digestion of SS alone. From the results obtained, it is evident that co-digestion of POME and SS is an attractive option to be explored for enhancement of biogas production in anaerobic digesters.

Impact of enzymatic digestion on bacterial community composition in CF airway samples.

Science.gov (United States)

Williamson, Kayla M; Wagner, Brandie D; Robertson, Charles E; Johnson, Emily J; Zemanick, Edith T; Harris, J Kirk

2017-01-01

Previous studies have demonstrated the importance of DNA extraction methods for molecular detection of Staphylococcus, an important bacterial group in cystic fibrosis (CF). We sought to evaluate the effect of enzymatic digestion (EnzD) prior to DNA extraction on bacterial communities identified in sputum and oropharyngeal swab (OP) samples from patients with CF. DNA from 81 samples (39 sputum and 42 OP) collected from 63 patients with CF was extracted in duplicate with and without EnzD. Bacterial communities were determined by rRNA gene sequencing, and measures of alpha and beta diversity were calculated. Principal Coordinate Analysis (PCoA) was used to assess differences at the community level and Wilcoxon Signed Rank tests were used to compare relative abundance (RA) of individual genera for paired samples with and without EnzD. Shannon Diversity Index (alpha-diversity) decreased in sputum and OP samples with the use of EnzD. Larger shifts in community composition were observed for OP samples (beta-diversity, measured by Morisita-Horn), whereas less change in communities was observed for sputum samples. The use of EnzD with OP swabs resulted in significant increase in RA for the genera Gemella ( p  microbial community composition. We show that the application of EnzD to CF airway samples, particularly OP swabs, results in differences in microbial communities detected by sequencing. Use of EnzD can result in large changes in bacterial community composition, and is particularly useful for detection of Staphylococcus in CF OP samples. The enhanced identification of Staphylococcus aureus is a strong indication to utilize EnzD in studies that use OP swabs to monitor CF airway communities.

Recombinant EXLX1 from Bacillus subtilis for enhancing enzymatic ...

African Journals Online (AJOL)

AJL

2012-06-21

Jun 21, 2012 ... enhancing enzymatic hydrolysis of corn stover with low cellulase loadings. Zhang Yan1, He Ming-Xiong2,3*, Wu Bo1, ... University, Chengdu 610064, China. 2Biogas Institute of Ministry of Agriculture, Biomass Energy Technology Research Centre, Section 4-13, Renming Nanlu,. Chengdu 610041, China.

Structural and Digestion Properties of Soluble-, Slowly Digestible and Resistant Maltodextrin from Cassava Starch by Enzymatic Modification

DEFF Research Database (Denmark)

Sorndech, Waraporn

The combination of branching enzyme (BE) and amylomaltase (AM) were selected to modify cassava starch. AM were used to elongate the glucan chains in order to enhance BE activity to create branching linkages. Cassava starch were gelatinized and incubated with BE or AMBE or BEAMBE or simultaneous...... AM and BE. The molecular analysis of the products including amylopectin chain length distribution, content of α-1,6 glucosidic linkages, absolute molecular weight distribution and digestibility were examined. Only BE catalysis showed 7.8% of branching linkages. The sequential AMBE-treated starch...... showed 9.9%-10.0% branching linkages, while the sequential BEAMBE-treated starch gained 10.9%-13.1% of branching linkages. Moreover, the sequential AMBE and BEAMBE-treated starch retarded the digestion rate of α-amylase and glucoamylase. Overall, sequential BEAMBE catalysis resulted in more...

Enhancement of enzymatic saccharification of Eucalyptus globulus: steam explosion versus steam treatment.

Science.gov (United States)

Martin-Sampedro, Raquel; Revilla, Esteban; Villar, Juan C; Eugenio, Maria E

2014-09-01

Steam explosion and steam pre-treatment have proved capable of enhancing enzymatic saccharification of lignocellulosic materials. However, until now, these methods had not been compared under the same operational conditions and using the same raw material. Both pre-treatments lead to increased yields in the saccharification of Eucalyptus globulus; but results have been better with steam pre-treatments, despite the more accessible surface of exploded samples. The reason for this finding could be enzymatic inhibition: steam explosion causes a more extensive extraction of hemicelluloses and releases a greater amount of degradation products which can inhibit enzymatic action. Enzymatic inhibition is also dependent on the amount and chemical structure of lignin, which was also a contributing factor to the lower enzymatic yields obtained with the most severe pre-treatment. Thus, the highest yields (46.7% glucose and 73.4% xylose yields) were obtained after two cycle of steam treatment, of 5 and 3 min, at 183°C. Copyright © 2014 Elsevier Ltd. All rights reserved.

Influence of Feeding Enzymatically Hydrolyzed Yeast Cell Wall on Growth Performance and Digestive Function of Feedlot Cattle during Periods of Elevated Ambient Temperature

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J. Salinas-Chavira

2015-09-01

Full Text Available In experiment 1, eighty crossbred steers (239±15 kg were used in a 229-d experiment to evaluate the effects of increasing levels of enzymatically hydrolyzed yeast (EHY cell wall in diets on growth performance feedlot cattle during periods of elevated ambient temperature. Treatments consisted of steam-flaked corn-based diets supplemented to provide 0, 1, 2, or 3 g EHY/hd/d. There were no effects on growth performance during the initial 139-d period. However, from d 139 to harvest, when 24-h temperature humidity index averaged 80, EHY increased dry matter intake (DMI (linear effect, p0.10 on carcass characteristics. In experiment 2, four Holstein steers (292±5 kg with cannulas in the rumen and proximal duodenum were used in a 4×4 Latin Square design experiment to evaluate treatments effects on characteristics of ruminal and total tract digestion in steers. There were no treatment effects (p>0.10 on ruminal pH, total volatile fatty acid, molar proportions of acetate, butyrate, or estimated methane production. Supplemental EHY decreased ruminal molar proportion of acetate (p = 0.08, increased molar proportion of propionate (p = 0.09, and decreased acetate:propionate molar ratio (p = 0.07 and estimated ruminal methane production (p = 0.09. It is concluded that supplemental EHY may enhance DMI and ADG of feedlot steers during periods of high ambient temperature. Supplemental EHY may also enhance ruminal fiber digestion and decrease ruminal acetate:propionate molar ratios in feedlot steers fed steam-flaked corn-based finishing diets.

Actinidin enhances protein digestion in the small intestine as assessed using an in vitro digestion model.

Science.gov (United States)

Kaur, Lovedeep; Rutherfurd, Shane M; Moughan, Paul J; Drummond, Lynley; Boland, Mike J

2010-04-28

This paper describes an in vitro study that tests the proposition that actinidin from green kiwifruit influences the digestion of proteins in the small intestine. Different food proteins, from sources including soy, meat, milk, and cereals, were incubated in the presence or absence of green kiwifruit extract (containing actinidin) using a two-stage in vitro digestion system consisting of an incubation with pepsin at stomach pH (simulating gastric digestion) and then with added pancreatin at small intestinal pH, simulating upper tract digestion in humans. The digests from the small intestinal stage (following the gastric digestion phase) were subjected to gel electrophoresis (SDS-PAGE) to assess loss of intact protein and development of large peptides during the in vitro simulated digestion. Kiwifruit extract influenced the digestion patterns of all of the proteins to various extents. For some proteins, actinidin had little impact on digestion. However, for other proteins, the presence of kiwifruit extract resulted in a substantially greater loss of intact protein and different peptide patterns from those seen after digestion with pepsin and pancreatin alone. In particular, enhanced digestion of whey protein isolate, zein, gluten, and gliadin was observed. In addition, reverse-phase HPLC (RP-HPLC) analysis showed that a 2.5 h incubation of sodium caseinate with kiwifruit extract alone resulted in approximately 45% loss of intact protein.

Microwave enhanced digestion of aerobic SBR sludge | Kennedy ...

African Journals Online (AJOL)

MWs) for improving characteristics of aerobic sequencing batch reactor (SBR) sludge to enhance mesophilic anaerobic digestion. Effects of pretreatment temperature, MW irradiation intensity and solids concentration on sludge characterisation ...

Multi-Scale Computational Enzymology: Enhancing Our Understanding of Enzymatic Catalysis

Science.gov (United States)

Gherib, Rami; Dokainish, Hisham M.; Gauld, James W.

2014-01-01

Elucidating the origin of enzymatic catalysis stands as one the great challenges of contemporary biochemistry and biophysics. The recent emergence of computational enzymology has enhanced our atomistic-level description of biocatalysis as well the kinetic and thermodynamic properties of their mechanisms. There exists a diversity of computational methods allowing the investigation of specific enzymatic properties. Small or large density functional theory models allow the comparison of a plethora of mechanistic reactive species and divergent catalytic pathways. Molecular docking can model different substrate conformations embedded within enzyme active sites and determine those with optimal binding affinities. Molecular dynamics simulations provide insights into the dynamics and roles of active site components as well as the interactions between substrate and enzymes. Hybrid quantum mechanical/molecular mechanical (QM/MM) can model reactions in active sites while considering steric and electrostatic contributions provided by the surrounding environment. Using previous studies done within our group, on OvoA, EgtB, ThrRS, LuxS and MsrA enzymatic systems, we will review how these methods can be used either independently or cooperatively to get insights into enzymatic catalysis. PMID:24384841

Multi-Scale Computational Enzymology: Enhancing Our Understanding of Enzymatic Catalysis

Directory of Open Access Journals (Sweden)

Rami Gherib

2013-12-01

Full Text Available Elucidating the origin of enzymatic catalysis stands as one the great challenges of contemporary biochemistry and biophysics. The recent emergence of computational enzymology has enhanced our atomistic-level description of biocatalysis as well the kinetic and thermodynamic properties of their mechanisms. There exists a diversity of computational methods allowing the investigation of specific enzymatic properties. Small or large density functional theory models allow the comparison of a plethora of mechanistic reactive species and divergent catalytic pathways. Molecular docking can model different substrate conformations embedded within enzyme active sites and determine those with optimal binding affinities. Molecular dynamics simulations provide insights into the dynamics and roles of active site components as well as the interactions between substrate and enzymes. Hybrid quantum mechanical/molecular mechanical (QM/MM can model reactions in active sites while considering steric and electrostatic contributions provided by the surrounding environment. Using previous studies done within our group, on OvoA, EgtB, ThrRS, LuxS and MsrA enzymatic systems, we will review how these methods can be used either independently or cooperatively to get insights into enzymatic catalysis.

The In Vitro Effects of Enzymatic Digested Gliadin on the Functionality of the Autophagy Process

Directory of Open Access Journals (Sweden)

Federico Manai

2018-02-01

Full Text Available Gliadin, the alcohol-soluble protein fraction of wheat, contains the factor toxic for celiac disease (CD, and its toxicity is not reduced by digestion with gastro-pancreatic enzymes. Importantly, it is proved that an innate immunity to gliadin plays a key role in the development of CD. The immune response induces epithelial stress and reprograms intraepithelial lymphocytes into natural killer (NK-like cells, leading to enterocyte apoptosis and an increase in epithelium permeability. In this contribution, we have reported that in Caco-2 cells the administration of enzymatically digested gliadin (PT-gliadin reduced significantly the expression of the autophagy-related marker LC3-II. Furthermore, electron and fluorescent microscope analysis suggested a compromised functionality of the autophagosome apparatus. The rescue of the dysregulated autophagy process, along with a reduction of PT-gliadin toxicity, was obtained with a starvation induction protocol and by 3-methyladenine administration, while rapamycin, a well-known autophagy inducer, did not produce a significant improvement in the clearance of extra- and intra-cellular fluorescent PT-gliadin amount. Altogether, our results highlighted the possible contribution of the autophagy process in the degradation and in the reduction of extra-cellular release of gliadin peptides and suggest novel molecular targets to counteract gliadin-induced toxicity in CD.

Sustainable coatings from bio-based, enzymatically synthesized polyesters with enhanced functionalities

NARCIS (Netherlands)

Gustini, L.; Lavilla, C.; Finzel, L.; Noordover, B.A.J.; Hendrix, M.M.R.M.; Koning, C.E.

2016-01-01

Bio-based sorbitol-containing polyester polyols were synthesized via enzymatic polycondensation. The selectivity of the biocatalyst for primary vs. secondary hydroxyl groups allowed for the preparation of close to linear renewable polyester polyols with enhanced hydroxyl functionalities, both as

Wet explosion pretreatment of sugarcane bagasse for enhanced enzymatic hydrolysis

DEFF Research Database (Denmark)

Biswas, Rajib; Uellendahl, Hinrich; Ahring, Birgitte Kiær

2014-01-01

.7% of the theoretical maximum value. Pretreatment at 200 C with oxygen exhibited enhanced enzymatic efficiency but lower xylose recovery and formation of the degradation products such as acetate, furfural and HMF of 7.6, 3.3 and 1.0 g/L, respectively. In the hydrolysis, the total sugars (glucose + xylose) yielded...

Effects of sodium carbonate pretreatment on the chemical compositions and enzymatic saccharification of rice straw.

Science.gov (United States)

Yang, Linfeng; Cao, Jie; Jin, Yongcan; Chang, Hou-min; Jameel, Hasan; Phillips, Richard; Li, Zhongzheng

2012-11-01

The effects of sodium carbonate (Na(2)CO(3)) pretreatment on the chemical compositions and enzymatic saccharification of rice straw were investigated. The enzymatic digestibility of rice straw is enhanced after pretreatment since pretreated solids show significant delignification with high sugar availability. During pretreatment, an increasing temperature and Na(2)CO(3) charge leads to enhanced delignification, whereas an increased degradation of polysaccharides as well, of which xylan acts more susceptible than glucan. The sugar recovery of enzymatic hydrolysis goes up rapidly with the total titratable alkali (TTA) increasing from 0% to 8%, and then it reaches a plateau. The highest sugar recovery of rice straw after pretreatment, 71.7%, 73.2%, and 76.1% for total sugar, glucan, and xylan, respectively, is obtained at 140°C, TTA 8% and cellulase loading of 20 FPU/g-cellulose. In this condition, the corresponding delignification ratio of pretreated solid is 41.8%, while 95% of glucan and 76% of xylan are conserved. Copyright © 2012 Elsevier Ltd. All rights reserved.

Enhanced enzymatic hydrolysis and acetone-butanol-ethanol fermentation of sugarcane bagasse by combined diluted acid with oxidate ammonolysis pretreatment.

Science.gov (United States)

Li, Hailong; Xiong, Lian; Chen, Xuefang; Wang, Can; Qi, Gaoxiang; Huang, Chao; Luo, Mutan; Chen, Xinde

2017-03-01

This study aims to propose a biorefinery pretreatment technology for the bioconversion of sugarcane bagasse (SB) into biofuels and N-fertilizers. Performance of diluted acid (DA), aqueous ammonia (AA), oxidate ammonolysis (OA) and the combined DA with AA or OA were compared in SB pretreatment by enzymatic hydrolysis, structural characterization and acetone-butanol-ethanol (ABE) fermentation. Results indicated that DA-OA pretreatment improves the digestibility of SB by sufficiently hydrolyzing hemicellulose into fermentable monosaccharides and oxidating lignin into soluble N-fertilizer with high nitrogen content (11.25%) and low C/N ratio (3.39). The enzymatic hydrolysates from DA-OA pretreated SB mainly composed of glucose was more suitable for the production of ABE solvents than the enzymatic hydrolysates from OA pretreated SB containing high ratio of xylose. The fermentation of enzymatic hydrolysates from DA-OA pretreated SB produced 12.12g/L ABE in 120h. These results suggested that SB could be utilized efficient, economic, and environmental by DA-OA pretreatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

Enhancement of anaerobic sludge digestion by high-pressure homogenization.

Science.gov (United States)

Zhang, Sheng; Zhang, Panyue; Zhang, Guangming; Fan, Jie; Zhang, Yuxuan

2012-08-01

To improve anaerobic sludge digestion efficiency, the effects of high-pressure homogenization (HPH) conditions on the anaerobic sludge digestion were investigated. The VS and TCOD were significantly removed with the anaerobic digestion, and the VS removal and TCOD removal increased with increasing the homogenization pressure and homogenization cycle number; correspondingly, the accumulative biogas production also increased with increasing the homogenization pressure and homogenization cycle number. The optimal homogenization pressure was 50 MPa for one homogenization cycle and 40 MPa for two homogenization cycles. The SCOD of the sludge supernatant significantly increased with increasing the homogenization pressure and homogenization cycle number due to the sludge disintegration. The relationship between the biogas production and the sludge disintegration showed that the accumulative biogas and methane production were mainly enhanced by the sludge disintegration, which accelerated the anaerobic digestion process and improved the methane content in the biogas. Copyright © 2012 Elsevier Ltd. All rights reserved.

Lignin-based polyoxyethylene ether enhanced enzymatic hydrolysis of lignocelluloses by dispersing cellulase aggregates.

Science.gov (United States)

Lin, Xuliang; Qiu, Xueqing; Yuan, Long; Li, Zihao; Lou, Hongming; Zhou, Mingsong; Yang, Dongjie

2015-06-01

Water-soluble lignin-based polyoxyethylene ether (EHL-PEG), prepared from enzymatic hydrolysis lignin (EHL) and polyethylene glycol (PEG1000), was used to improve enzymatic hydrolysis efficiency of corn stover. The glucose yield of corn stover at 72h was increased from 16.7% to 70.1% by EHL-PEG, while increase in yield with PEG4600 alone was 52.3%. With the increase of lignin content, EHL-PEG improved enzymatic hydrolysis of microcrystalline cellulose more obvious than PEG4600. EHL-PEG could reduce at least 88% of the adsorption of cellulase on the lignin film measured by quartz crystal microbalance with dissipation monitoring (QCM-D), while reduction with PEG4600 was 43%. Cellulase aggregated at 1220nm in acetate buffer analyzed by dynamic light scattering. EHL-PEG dispersed cellulase aggregates and formed smaller aggregates with cellulase, thereby, reduced significantly nonproductive adsorption of cellulase on lignin and enhanced enzymatic hydrolysis of lignocelluloses. Copyright © 2015 Elsevier Ltd. All rights reserved.

Selective enhancement and verification of woody biomass digestibility as a denitrification carbon source.

Science.gov (United States)

Hu, Rongting; Zheng, Xilai; Xin, Jia; Sun, Zhaoyue; Zheng, Tianyuan

2017-11-01

The denitrification efficiency of woody biomass as carbon source is low because of its poor carbon availability. In this study, representative poplar sawdust was pretreated with lime and peracetic acid to enhance the biomass digestibility to different degrees; sawdust was then mixed with soil to investigate its denitrification efficiency. Under controllable conditions (25-95°C, 12-24h, varying dosages), sawdust digestibility (characterized by reducing sugar yield) was selectively enhanced 1.0-21.8 times over that of the raw sawdust (28.8mgeq.glucoseg -1 dry biomass). This increase was mainly attributed to the removal of lignin from the biomass. As a carbon source, the sawdust (digestibility enhanced by 5.4 times) increased the nitrate removal rate by 4.7 times, without N 2 O emission. However, the sawdust with high digestibility (12.6 or 18.0 times), despite releasing more dissolved organic carbon (DOC), did not exhibit further increase in denitrification efficiency, and emitted N 2 O. Copyright © 2017 Elsevier Ltd. All rights reserved.

Pretreatment of forest residues of Douglas fir by wet explosion for enhanced enzymatic saccharification.

Science.gov (United States)

Biswas, Rajib; Teller, Philip J; Ahring, Birgitte K

2015-09-01

The logging and lumbering industry in the Pacific Northwest region generates huge amount of forest residues, offering an inexpensive raw material for biorefineries. Wet explosion (WEx) pretreatment was applied to the recalcitrant biomass to optimize process conditions including temperature (170-190 °C), time (10-30 min), and oxygen loading (0.5-7.5% of DM) through an experimental design. Optimal pH for enzymatic hydrolysis of the optimized samples and a complete mass balance have been evaluated. Results indicated that cellulose digestibility improved in all conditions tested with maximum digestibility achieved at 190 °C, time 30 min, and oxygen loading of 7.5%. Glucose yield at optimal pH of 5.5 was 63.3% with an excellent recovery of cellulose and lignin of 99.9% and 96.3%, respectively. Hemicellulose sugars recovery for xylose and mannose was found to be 69.2% and 76.0%, respectively, indicating that WEx is capable of producing relative high sugar yield even from the recalcitrant forest residues. Copyright © 2015 Elsevier Ltd. All rights reserved.

Radiation-induced enhancement of enzymatic cell lysis of Micrococcus radiodurans

International Nuclear Information System (INIS)

Watanabe, H.; Takehisa, M.; Iizuka, H.

1981-01-01

The intact cells of M. radiodurans were rendered sensitive to the action of lytic enzyme (P2-2 enzyme) by irradiation. The radiation-induced enhancement of cell lysis with P2-2 enzyme was completely prevented by the addition of t-butanol and irradiation at liquid nitrogen temperature. These results indicate that the enhancement is due to indirect action resulting from OH radicals. Cell lysis by lysozyme was enhanced only when the cells were irradiated under N 2 O. The enhancement of cell lysis with lysozyme was also prevented by adding alcohols. On the other hand, when lipid components in cells were removed by extraction with n-butanol, the radiation-induced enhancement of cell lysis with P2-2 enzyme and lysozyme was not observed. From these results it is concluded that the enhancement of enzymatic cell lysis by irradiation is attributable to alteration in the lipid-rich layer of the cell wall caused by OH radicals

Radiation-induced enhancement of enzymatic cell lysis of Micrococcus radiodurans

Energy Technology Data Exchange (ETDEWEB)

Watanabe, H.; Takehisa, M. [Japan Atomic Energy Research Inst., Takasaki, Gunma, Takasaki Radiation Chemistry Research Establishment (Japan); Iizuka, H.

1981-10-15

The intact cells of M. radiodurans were rendered sensitive to the action of lytic enzyme (P2-2 enzyme) by irradiation. The radiation-induced enhancement of cell lysis with P2-2 enzyme was completely prevented by the addition of t-butanol and irradiation at liquid nitrogen temperature. These results indicate that the enhancement is due to indirect action resulting from OH radicals. Cell lysis by lysozyme was enhanced only when the cells were irradiated under N{sub 2}O. The enhancement of cell lysis with lysozyme was also prevented by adding alcohols. On the other hand, when lipid components in cells were removed by extraction with n-butanol, the radiation-induced enhancement of cell lysis with P2-2 enzyme and lysozyme was not observed. From these results it is concluded that the enhancement of enzymatic cell lysis by irradiation is attributable to alteration in the lipid-rich layer of the cell wall caused by OH radicals.

Steam-exploded biomass saccharification is predominately affected by lignocellulose porosity and largely enhanced by Tween-80 in Miscanthus.

Science.gov (United States)

Sun, Dan; Alam, Aftab; Tu, Yuanyuan; Zhou, Shiguang; Wang, Yanting; Xia, Tao; Huang, Jiangfeng; Li, Ying; Zahoor; Wei, Xiaoyang; Hao, Bo; Peng, Liangcai

2017-09-01

In this study, total ten Miscanthus accessions exhibited diverse cell wall compositions, leading to largely varied hexoses yields at 17%-40% (% cellulose) released from direct enzymatic hydrolysis of steam-exploded (SE) residues. Further supplied with 2% Tween-80 into the enzymatic digestion, the Mis7 accession showed the higher hexose yield by 14.8-fold than that of raw material, whereas the Mis10 had the highest hexoses yield at 77% among ten Miscanthus accessions. Significantly, this study identified four wall polymer features that negatively affect biomass saccharification as pbiomass enzymatic digestion. Hence, this study provides the potential strategy to enhance biomass saccharification using optimal biomass process technology and related genetic breeding in Miscanthus and beyond. Copyright © 2017 Elsevier Ltd. All rights reserved.

Enhancement of biogas production from olive mill effluent (OME) by co-digestion

Energy Technology Data Exchange (ETDEWEB)

Azbar, Nuri; Keskin, Tugba; Yuruyen, Aysegul [Bioengineering Department, Faculty of Engineering, Ege University, 35100 Bornova, Izmir (Turkey)

2008-12-15

The olive oil has a healthy image during its consumption due to its oleic acid content, which may prevent some human diseases. Ironically, by-products of olive mill production such as olive mill effluent (OME) and olive cake pose a serious environmental risk where it is produced. In this study, feasibility of using some agro-industrial residue streams such as cheese whey (CW) and laying hen litter (LHL) in order to enhance the methane production of OME was investigated. For this purpose, biochemical methane potential (BMP) assay was carried out for both raw OME alone and OME mixed with varying amount of other substrates such as LHL and CW in the serum bottles, respectively. Corresponding methane production values for various mixtures of the organic residue streams used in this study were determined. It was demonstrated that co-digestion of OME with LHL significantly enhanced the biodegradability of OME which was too low if it was digested alone. Over 90% increase in biogas production was obtained when digesting OME with LHL. The biogas production increased only 22%, when CW was used for the same purpose. It was demonstrated that the biodegradability of OME could be significantly enhanced by co-digestion and thereby integrated management of OME using anaerobic degradation could be proposed as an economically viable and ecologically acceptable solution for the safe disposal of OME. (author)

Impact of enzymatic digestion on bacterial community composition in CF airway samples

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Kayla M. Williamson

2017-05-01

Full Text Available Background Previous studies have demonstrated the importance of DNA extraction methods for molecular detection of Staphylococcus, an important bacterial group in cystic fibrosis (CF. We sought to evaluate the effect of enzymatic digestion (EnzD prior to DNA extraction on bacterial communities identified in sputum and oropharyngeal swab (OP samples from patients with CF. Methods DNA from 81 samples (39 sputum and 42 OP collected from 63 patients with CF was extracted in duplicate with and without EnzD. Bacterial communities were determined by rRNA gene sequencing, and measures of alpha and beta diversity were calculated. Principal Coordinate Analysis (PCoA was used to assess differences at the community level and Wilcoxon Signed Rank tests were used to compare relative abundance (RA of individual genera for paired samples with and without EnzD. Results Shannon Diversity Index (alpha-diversity decreased in sputum and OP samples with the use of EnzD. Larger shifts in community composition were observed for OP samples (beta-diversity, measured by Morisita-Horn, whereas less change in communities was observed for sputum samples. The use of EnzD with OP swabs resulted in significant increase in RA for the genera Gemella (p < 0.01, Streptococcus (p < 0.01, and Rothia (p < 0.01. Staphylococcus (p < 0.01 was the only genus with a significant increase in RA from sputum, whereas the following genera decreased in RA with EnzD: Veillonella (p < 0.01, Granulicatella (p < 0.01, Prevotella (p < 0.01, and Gemella (p = 0.02. In OP samples, higher RA of Gram-positive taxa was associated with larger changes in microbial community composition. Discussion We show that the application of EnzD to CF airway samples, particularly OP swabs, results in differences in microbial communities detected by sequencing. Use of EnzD can result in large changes in bacterial community composition, and is particularly useful for detection of Staphylococcus in CF OP

Enhancing anaerobic digestion of poultry blood using activated carbon

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Maria José Cuetos

2017-05-01

Full Text Available The potential of using anaerobic digestion for the treatment of poultry blood has been evaluated in batch assays at the laboratory scale and in a mesophilic semi-continuous reactor. The biodegradability test performed on residual poultry blood was carried out in spite of high inhibitory levels of acid intermediaries. The use of activated carbon as a way to prevent inhibitory conditions demonstrated the feasibility of attaining anaerobic digestion under extreme ammonium and acid conditions. Batch assays with higher carbon content presented higher methane production rates, although the difference in the final cumulative biogas production was not as sharp. The digestion of residual blood was also studied under semi-continuous operation using granular and powdered activated carbon. The average specific methane production was 216 ± 12 mL CH4/g VS. This result was obtained in spite of a strong volatile fatty acid (VFA accumulation, reaching values around 6 g/L, along with high ammonium concentrations (in the range of 6–8 g/L. The use of powdered activated carbon resulted in a better assimilation of C3-C5 acid forms, indicating that an enhancement in syntrophic metabolism may have taken place. Thermal analysis and scanning electron microscopy (SEM were applied as analytical tools for measuring the presence of organic material in the final digestate and evidencing modifications on the carbon surface. The addition of activated carbon for the digestion of residual blood highly improved the digestion process. The adsorption capacity of ammonium, the protection this carrier may offer by limiting mass transfer of toxic compounds, and its capacity to act as a conductive material may explain the successful digestion of residual blood as the sole substrate.

pH-Induced Lignin Surface Modification to Reduce Nonspecific Cellulase Binding and Enhance Enzymatic Saccharification of Lignocelluloses

Science.gov (United States)

Hongming Lou; J.Y. Zhu; Tian Qing Lan; Huranran Lai; Xueqing Qiu

2013-01-01

We studied the mechanism of the significant enhancement in the enzymatic saccharification of lignocelluloses at an elevated pH of 5.5–6.0. Four lignin residues with different sulfonic acid contents were isolated from enzymatic hydrolysis of lodgepole pine pretreated by either dilute acid (DA) or sulfite pretreatment to overcome recalcitrance of lignocelluloses (SPORL...

Comparative Analysis of End Point Enzymatic Digests of Arabino-Xylan Isolated from Switchgrass (Panicum virgatum L of Varying Maturities using LC-MSn

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Michael J. Bowman

2012-11-01

Full Text Available Switchgrass (Panicum virgatum L., SG is a perennial grass presently used for forage and being developed as a bioenergy crop for conversion of cell wall carbohydrates to biofuels. Up to 50% of the cell wall associated carbohydrates are xylan. SG was analyzed for xylan structural features at variable harvest maturities. Xylan from each of three maturities was isolated using classical alkaline extraction to yield fractions (Xyl A and B with varying compositional ratios. The Xyl B fraction was observed to decrease with plant age. Xylan samples were subsequently prepared for structure analysis by digesting with pure endo-xylanase, which preserved side-groups, or a commercial carbohydrase preparation favored for biomass conversion work. Enzymatic digestion products were successfully permethylated and analyzed by reverse-phase liquid chromatography with mass spectrometric detection (RP-HPLC-MSn. This method is advantageous compared to prior work on plant biomass because it avoids isolation of individual arabinoxylan oligomers. The use of RP-HPLC- MSn differentiated 14 structural oligosaccharides (d.p. 3–9 from the monocomponent enzyme digestion and nine oligosaccharide structures (d.p. 3–9 from hydrolysis with a cellulase enzyme cocktail. The distribution of arabinoxylan oligomers varied depending upon the enzyme(s applied but did not vary with harvest maturity.

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.

Malting process optimization for protein digestibility enhancement in finger millet grain.

Science.gov (United States)

Hejazi, Sara Najdi; Orsat, Valérie

2016-04-01

Finger millet (Eleusine coracana) is a nutritious, gluten-free, and drought resistant cereal containing high amounts of protein, carbohydrate, and minerals. However, bio-availability of these nutrients is restricted due to the presence of an excessive level of anti-nutrient components, mainly phytic acid, tannin, and oxalate. It has been shown that a well-designed malting/germination process can significantly reduce these anti-nutrients and consequently enhance the nutrient availability. In the present study, the effects of two important germination factors, duration and temperature, on the enhancement of in-vitro protein digestibility of finger millet were thoroughly investigated and optimized. Based on a central composite design, the grains were germinated for 24, 36, and 48 h at 22, 26, and 30 °C. For all factor combinations, protein, peptide, phytic acid, tannin, and oxalate contents were evaluated and digestibility was assessed. It was shown that during the malting/germinating process, both temperature and duration factors significantly influenced the investigated quantities. Germination of finger millet for 48 h at 30 °C increased protein digestibility from 74 % (for native grain) up to 91 %. Besides, it notably decreased phytic acid, tannin, and oxalate contents by 45 %, 46 %, and 29 %, respectively. Linear correlations between protein digestibility and these anti-nutrients were observed.

Digestibility of gluten proteins is reduced by baking and enhanced by starch digestion.

Science.gov (United States)

Smith, Frances; Pan, Xiaoyan; Bellido, Vincent; Toole, Geraldine A; Gates, Fred K; Wickham, Martin S J; Shewry, Peter R; Bakalis, Serafim; Padfield, Philip; Mills, E N Clare

2015-10-01

Resistance of proteins to gastrointestinal digestion may play a role in determining immune-mediated adverse reactions to foods. However, digestion studies have largely been restricted to purified proteins and the impact of food processing and food matrices on protein digestibility is poorly understood. Digestibility of a total gliadin fraction (TGF), flour (cv Hereward), and bread was assessed using in vitro batch digestion with simulated oral, gastric, and duodenal phases. Protein digestion was monitored by SDS-PAGE and immunoblotting using monoclonal antibodies specific for celiac-toxic sequences (QQSF, QPFP) and starch digestion by measuring undigested starch. Whereas the TGF was rapidly digested during the gastric phase the gluten proteins in bread were virtually undigested and digested rapidly during the duodenal phase only if amylase was included. Duodenal starch digestion was also slower in the absence of duodenal proteases. The baking process reduces the digestibility of wheat gluten proteins, including those containing sequences active in celiac disease. Starch digestion affects the extent of protein digestion, probably because of gluten-starch complex formation during baking. Digestion studies using purified protein fractions alone are therefore not predictive of digestion in complex food matrices. © 2015 The Authors. Molecular Nutrition & Food Research published by Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Enhanced high-solids anaerobic digestion of waste activated sludge by the addition of scrap iron.

Science.gov (United States)

Zhang, Yaobin; Feng, Yinghong; Yu, Qilin; Xu, Zibin; Quan, Xie

2014-05-01

Anaerobic digestion of waste activated sludge usually requires pretreatment procedure to improve the bioavailability of sludge, which involves considerable energy and high expenditures. This study proposes a cost-effective method for enhanced anaerobic digestion of sludge without a pretreatment by directly adding iron into the digester. The results showed that addition of Fe(0) powder could enhance 14.46% methane yield, and Fe scrap (clean scrap) could further enhance methane yield (improving rate 21.28%) because the scrap has better mass transfer efficiency with sludge and liquid than Fe(0) powder. The scrap of Fe with rust (rusty scrap) could induce microbial Fe(III) reduction, which resulted in achieving the highest methane yield (improving rate 29.51%), and the reduction rate of volatile suspended solids (VSS) was also highest (48.27%) among Fe powder, clean scrap and rusty scrap. PCR-DGGE proved that the addition of rusty scrap could enhance diversity of acetobacteria and enrich iron-reducing bacteria to enhance degradation of complex substrates. Copyright © 2014 Elsevier Ltd. All rights reserved.

[Anaerobic digestion of lignocellulosic biomass with animal digestion mechanisms].

Science.gov (United States)

Wu, Hao; Zhang, Pan-Yue; Guo, Jian-Bin; Wu, Yong-Jie

2013-02-01

Lignocellulosic material is the most abundant renewable resource in the earth. Herbivores and wood-eating insects are highly effective in the digestion of plant cellulose, while anaerobic digestion process simulating animal alimentary tract still remains inefficient. The digestion mechanisms of herbivores and wood-eating insects and the development of anaerobic digestion processes of lignocellulose were reviewed for better understanding of animal digestion mechanisms and their application in design and operation of the anaerobic digestion reactor. Highly effective digestion of lignocellulosic materials in animal digestive system results from the synergistic effect of various digestive enzymes and a series of physical and biochemical reactions. Microbial fermentation system is strongly supported by powerful pretreatment, such as rumination of ruminants, cellulase catalysis and alkali treatment in digestive tract of wood-eating insects. Oxygen concentration gradient along the digestive tract may stimulate the hydrolytic activity of some microorganisms. In addition, the excellent arrangement of solid retention time, digesta flow and end product discharge enhance the animal digestion of wood cellulose. Although anaerobic digestion processes inoculated with rumen microorganisms based rumen digestion mechanisms were developed to treat lignocellulose, the fermentation was more greatly limited by the environmental conditions in the anaerobic digestion reactors than that in rumen or hindgut. Therefore, the anaerobic digestion processes simulating animal digestion mechanisms can effectively enhance the degradation of wood cellulose and other organic solid wastes.

Three-stage enzymatic digestive system for a gut-on-a-chip

NARCIS (Netherlands)

de Haan, Pim; Ianovska, Margaryta A.; Mathwig, Klaus; Bouwmeester, Hans; Verpoorte, Elisabeth

2017-01-01

In this work, we present the development of a three-stage microfluidic system as a cell-free model for digestion in the human gastrointestinal (GI) tract. Larger-scale digestion models are currently being used for pharmacological, toxicological and nutritional studies to determine the possible

Hydroxypropylation of pigeon pea (cajanus cajan) starch: Preparation, functional characterizations and enzymatic digestibility

International Nuclear Information System (INIS)

Lawal, O.S.

2008-05-01

Hydroxypropyl starch derivatives were prepared from pigeon pea starch (NPPS) which is an unconventional starch source. Functional parameters and characterization of both native and modified starches were carried out. The starch granules appeared oval or elliptical in shape with sizes ranging from 7 - 40 μm in width and 10 . 30 μm in length. Hydroxypropylation did not alter the shape of the starch granules in a pronounced way. Generally, the x-ray diffractograms of both native and hydroxypropyl derivatives showed the 'C' pattern. However, slight reductions were observed in the intensity of starches after modification. At all temperatures studied (30 - 90 deg. C), swelling and solubility of hydroxypropylated starches were higher than the NPPS. Progressive increases in swelling capacity and solubility were observed as the MS increased among the hydroxypropylated starches. Hydroxypropylation reduced starch paste turbidity on storage. Also, studies showed that syneresis reduced after hydroxypropylation. In addition, syneresis reduced as the MS of the hydroxypropyl starches increased. The results indicate that pasting temperature and peak temperature reduced after modification but peak viscosity increased in hydroxypropylated starch derivatives compared with the native starch. Setback reduced in hydroxypropylated starches compared with the native starch. Enthalpy of gelatinization and percentage retrogradation reduced after hydroxypropylation and progressive reductions were observed as the MS increased among the starch derivatives. Hydroxypropylation increased enzymatic digestibility. (author)

Enzymatic formulation capable of degrading scrapie prion under mild digestion conditions.

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Emeka A Okoroma

Full Text Available The prion agent is notoriously resistant to common proteases and conventional sterilisation procedures. The current methods known to destroy prion infectivity such as incineration, alkaline and thermal hydrolysis are harsh, destructive, environmentally polluting and potentially hazardous, thus limit their applications for decontamination of delicate medical and laboratory devices, remediation of prion contaminated environment and for processing animal by-products including specified risk materials and carcases. Therefore, an environmentally friendly, non-destructive enzymatic degradation approach is highly desirable. A feather-degrading Bacillus licheniformis N22 keratinase has been isolated which degraded scrapie prion to undetectable level of PrP(Sc signals as determined by Western Blot analysis. Prion infectivity was verified by ex vivo cell-based assay. An enzymatic formulation combining N22 keratinase and biosurfactant derived from Pseudomonas aeruginosa degraded PrP(Sc at 65 °C in 10 min to undetectable level -. A time-course degradation analysis carried out at 50 °C over 2 h revealed the progressive attenuation of PrP(Sc intensity. Test of residual infectivity by standard cell culture assay confirmed that the enzymatic formulation reduced PrP(Sc infectivity to undetectable levels as compared to cells challenged with untreated standard scrapie sheep prion (SSBP/1 (p-value = 0.008 at 95% confidence interval. This novel enzymatic formulation has significant potential application for prion decontamination in various environmentally friendly systems under mild treatment conditions.

Preliminary characterization of digestive enzymes in freshwater mussels

Science.gov (United States)

Sauey, Blake W.; Amberg, Jon J.; Cooper, Scott T.; Grunwald, Sandra K.; Newton, Teresa J.; Haro, Roger J.

2015-01-01

Resource managers lack an effective chemical tool to control the invasive zebra mussel Dreissena polymorpha. Zebra mussels clog water intakes for hydroelectric companies, harm unionid mussel species, and are believed to be a reservoir of avian botulism. Little is known about the digestive physiology of zebra mussels and unionid mussels. The enzymatic profile of the digestive glands of zebra mussels and native threeridge (Amblema plicata) and plain pocketbook mussels (Lampsilis cardium) are characterized using a commercial enzyme kit, api ZYM, and validated the kit with reagent-grade enzymes. A linear correlation was shown for only one of nineteen enzymes, tested between the api ZYM kit and a specific enzyme kit. Thus, the api ZYM kit should only be used to make general comparisons of enzyme presence and to observe trends in enzyme activities. Enzymatic trends were seen in the unionid mussel species, but not in zebra mussels sampled 32 days apart from the same location. Enzymatic classes, based on substrate, showed different trends, with proteolytic and phospholytic enzymes having the most change in relative enzyme activity.

Does the commonly used pH-stat method with back titration really quantify the enzymatic digestibility of lipid drug delivery systems? A case study on solid lipid nanoparticles (SLN).

Science.gov (United States)

Heider, Martha; Hause, Gerd; Mäder, Karsten

2016-12-01

Enzymatic digestion of lipid drug carriers is very important. Commonly, pancreatin induced formation of fatty acids is monitored by the pH-stat method, which provides a fast, but unspecific readout. However, according to the literature, the pKa values of long chain fatty acids are strongly dependent on the local environment and might vary between 4.2 and 10.15. The high pKa values would lead to an incomplete detection of the lipid digestion and false results. In order to investigate these issues in more detail, we produced cetyl palmitate solid lipid nanoparticles (CP-SLN) stabilized with poloxamer 188 or polysorbate 80. The digestion of CP-SLN was investigated by two different and independent readouts. A HPTLC assay was used in addition to the pH-stat method (with or without back titration). An incomplete digestion of CP-SLN was observed with all methods. Partial digestion of polysorbate 80 contributed to the formation of fatty acids. Depending on the investigated system and the experimental conditions (FaSSIF or FeSSIF) the results of both readout methods were comparable or not. For example, in FeSSIF conditions, the values detected by HPTLC were roughly twice as high as the pH-stat results. Our findings on solid lipids agree with data from Helbig et al. on lipid emulsions, where a gas chromatography method detected much higher values than the pH-stat assay (Food Hydrocoll. 28 (2012) 10-19). The results of our pH-stat experiments with back titration at different pH values showed increased values for fatty acids from pH 7.5 to pH 10. The values obtained by back titration at high pH values (pH 9 or higher) did exceed the digestion values measured by HPTLC. Therefore, we conclude that the pH-stat method might give the same results as more specific reference methods, but it might also both under- (without back titration) or overestimate (with back titration) the enzymatic digestion of lipid drug delivery systems. A further outcome of our study was the proof that

In vitro digestion with bile acids enhances the bioaccessibility of kale polyphenols.

Science.gov (United States)

Yang, Isabelle; Jayaprakasha, Guddarangavvanahally K; Patil, Bhimanagouda

2018-02-21

Kale (Brassica oleracea) is a leafy green vegetable belonging to the Brassicaceae family, and kale leaves have large amounts of dietary fiber and polyphenolics. Dietary fiber can bind bile acids, thus potentially decreasing cholesterol levels; however, whether the polyphenols from kale contribute to in vitro bile acid binding capacity remains unclear. In the present study, kale was extracted with hexane, acetone, and MeOH : water and the dried extracts, as well as the fiber-rich residue, were tested for their bile acid binding capacity. The fiber-rich residue bound total bile acids in amounts equivalent to that bound by raw kale. The lyophilized acetone extract bound significantly more glycochenodeoxycholate and glycodeoxycholate and less of other bile acids. To test whether bile acid binding enhanced the bioaccessibility of polyphenolic compounds from kale, we used ultra-performance liquid chromatography coupled with electrospray ionization/quadrupole-time-of-flight mass spectrometry to identify chemical constituents and measure their bioaccessibility in an in vitro digestion reaction. This identified 36 phenolic compounds in kale, including 18 kaempferol derivatives, 13 quercetin derivatives, 4 sinapoyl derivatives, and one caffeoylquinic acid. The bioaccessibility of these phenolics was significantly higher (69.4%) in digestions with bile acids. Moreover, bile acids enhanced the bioaccessibility of quercetin by 25 times: only 2.7% of quercetin derivatives were bioaccessible in the digestion without bile acids, but with bile acids, their accessibility increased to 69.5%. Bile acids increased the bioaccessibility of kaempferol from 37.7% to 69.2%. The extractability and biostability of total phenolics in the digested residue increased 1.8 fold in the digestions with bile acids. These results demonstrated the potential use of kale to improve human health.

Facilitating the enzymatic saccharification of pulped bamboo residues by degrading the remained xylan and lignin-carbohydrates complexes.

Science.gov (United States)

Huang, Caoxing; He, Juan; Li, Xin; Min, Douyong; Yong, Qiang

2015-09-01

Kraft pulping was performed on bamboo residues and its impact on the chemical compositions and the enzymatic digestibility of the samples were investigated. To improve the digestibility of sample by degrading the xylan and lignin-carbohydrates complexes (LCCs), xylanase and α-L-arabinofuranosidase (AF) were supplemented with cellulase. The results showed more carbohydrates were remained in the samples pulped with low effective alkali (EA) charge, compared to conventional kraft pulping. When 120 IU/g xylanase and 15 IU/g AF were supplemented with 20 FPU/g cellulase, the xylan degradation yield of the sample pulped with 12% EA charge increased from 68.20% to 88.35%, resulting in an increased enzymatic saccharification efficiency from 58.98% to 83.23%. The amount of LCCs in this sample decreased from 8.63/100C9 to 2.99/100C9 after saccharification with these enzymes. The results indicated that degrading the remained xylan and LCCs in the pulp could improve its enzymatic digestibility. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enzymatic cell wall degradation of high-pressure-homogenized tomato puree and its effect on lycopene bioaccessibility.

Science.gov (United States)

Palmero, Paola; Colle, Ines; Lemmens, Lien; Panozzo, Agnese; Nguyen, Tuyen Thi My; Hendrickx, Marc; Van Loey, Ann

2016-01-15

High-pressure homogenization disrupts cell structures, assisting carotenoid release from the matrix and subsequent micellarization. However, lycopene bioaccessibility of tomato puree upon high-pressure homogenization is limited by the formation of a process-induced barrier. In this context, cell wall-degrading enzymes were applied to hydrolyze the formed barrier and enhance lycopene bioaccessibility. The effectiveness of the enzymes in degrading their corresponding substrates was evaluated (consistency, amount of reducing sugars, molar mass distribution and immunolabeling). An in vitro digestion procedure was applied to evaluate the effect of the enzymatic treatments on lycopene bioaccessibility. Enzymatic treatments with pectinases and cellulase were proved to effectively degrade their corresponding cell wall polymers; however, no further significant increase in lycopene bioaccessibility was obtained. A process-induced barrier consisting of cell wall material is not the only factor governing lycopene bioaccessibility upon high-pressure homogenization. © 2015 Society of Chemical Industry.

Enhancement mechanisms of short-time aerobic digestion for waste activated sludge in the presence of cocoamidopropyl betaine.

Science.gov (United States)

Xia, Siqing; Zhou, Yun; Eustance, Everett; Zhang, Zhiqiang

2017-10-18

Cocoamidopropyl betaine (CAPB), which is a biodegradable ampholytic surfactant, has recently been found to dramatically enhance the aerobic digestion of waste activated sludge (WAS) in short-time aerobic digestion (STAD) systems. Therefore, it is important to understand the mechanisms in which CAPB enhances WAS aerobic digestion performance. Results showed that CAPB could dramatically enhance the solubilization of soluble proteins (PN), polysaccharides (PS), nucleic acids (NA) and humic-like substances (HS) in the STAD system within the initial 2 h. Then PN, PS and NA gradually decreased, while HS showed only minor decease. In addition, CAPB increased the proportion of low MW fractions (biodegradable. Specific oxygen uptake rates and dehydrogenase enzyme activity results indicated that CAPB markedly improved the aerobic microorganism activities. Microbial community analyses and principle coordinate analyses (PCoA) revealed that CAPB increased the proportion of some functional microorganisms, including Proteobacteria, Planctomycetales, Acinetobacter, Pseudomonas and Aeromonas. The changes driven by CAPB could explain the enhanced performance of the STAD system for WAS aerobic treatment.

Chemical characterization and hydrothermal pretreatment of Salicornia bigelovii straw for enhanced enzymatic hydrolysis and bioethanol potential

DEFF Research Database (Denmark)

Cybulska, Iwona; Chaturvedi, Tanmay; Brudecki, Grzegorz P.

2014-01-01

equipment and avoid inhibition of enzymes and yeast. Composition of the washed biomass was comparable to traditional lignocellulosic biomasses with relatively high glucan and xylan content (26 and 22. g/100. gDM, respectively) but with lower lignin content (7. g/100. gDM). The washed feedstock was subjected...... to hydrothermal pretreatment, producing highly digestible (up to 92% glucan-to-glucose conversion) and fermentable (up to 100% glucose-to-ethanol conversion) fiber fractions. Liquid fractions obtained in the pretreatment did not show inhibition towards Saccharomyces cerevisiae. No significant differences among...... the enzymatic convertibility and microbial fermentability of the fibers as well as low xylose recoveries suggest that lower severity pretreatment conditions could be exploited for S. bigelovii. © 2013 Elsevier Ltd....

The development of orally administrable gemcitabine prodrugs with D-enantiomer amino acids: enhanced membrane permeability and enzymatic stability.

Science.gov (United States)

Tsume, Yasuhiro; Incecayir, Tuba; Song, Xueqin; Hilfinger, John M; Amidon, Gordon L

2014-04-01

Gemcitabine prodrugs with D- and L-configuration amino acids were synthesized and their chemical stability in buffers, resistance to glycosidic bond metabolism, enzymatic activation, permeability in Caco-2 cells and mouse intestinal membrane, anti-proliferation activity in cancer cell were determined and compared to that of parent drug, gemcitabine. Prodrugs containing D-configuration amino acids were enzymatically more stable than ones with L-configuration amino acids. The activation of all gemcitabine prodrugs was 1.3-17.6-fold faster in cancer cell homogenate than their hydrolysis in buffer, suggesting enzymatic action. The enzymatic activation of amino acid monoester prodrugs containing D-configuration amino acids in cell homogenates was 2.2-10.9-fold slower than one of amino acid monoester prodrugs with L-configuration amino acids. All prodrugs exhibited enhanced resistance to glycosidic bond metabolism by thymidine phosphorylase compared to parent gemcitabine. Gemcitabine prodrugs showed superior the effective permeability in mouse jejunum to gemcitabine. More importantly, the high plasma concentration of d-amino acid gemcitabine prodrugs was observed more than one of L-amino acid gemcitabine prodrugs. In general, the 5'-mono-amino acid monoester gemcitabine prodrugs exhibited higher permeability and uptake than their parent drug, gemcitabine. Cell proliferation assays in AsPC-1 pancreatic ductal cell line indicated that gemcitabine prodrugs were more potent than their parent drug, gemcitabine. The transport and enzymatic profiles of 5'-D-valyl-gemcitabine and 5'-D-phenylalanyl-gemcitabine suggest their potential for increased oral uptake and delayed enzymatic bioconversion as well as enhanced uptake and cytotoxic activity in cancer cells, would facilitate the development of oral dosage form for anti-cancer agents and, hence, improve the quality of life for the cancer patients. Copyright © 2014. Published by Elsevier B.V.

Influence of season, environment and feeding habits on the enzymatic activity of peptidase and β-glucosidase in the gastrointestinal tract of two Siluriformes fishes (Teleostei

Directory of Open Access Journals (Sweden)

Silvana Duarte

2013-06-01

Full Text Available The enzymatic activities involved in the digestion of proteins and carbohydrates were compared among three organs of the digestive track of two Siluriformes fish species, and between two areas: a reservoir, and an area downriver of it. Our aim was to test the hypothesis that the digestive organs of species with varied feeding habits have different enzymatic activities, and that the enzymatic activity differs among seasons and environmental conditions. The iliophagous/herbivorous species Hypostomus auroguttatus Kner, 1854 had higher trypsin-like, chymotrypsin-like peptidase and β-glucosidase activity in the intestine when compared with the omnivorous species Pimelodus maculatus Lacepède, 1803, whereas the latter had more hepatic trypsin-like activity than the former. The peak of activity of the three enzymes in H. auroguttatus was recorded in the winter and spring. On the other hand, P. maculatus tended to have the more prominent peptidase and β-glucosidase activity in the summer, and the smallest in the winter. The intestine of H. auroguttatus had higher enzymatic (trypsin, chymotrypsin and β-glucosidase activity than the stomach and the liver. For P. maculatus, the highest β-glucosidase activity was found in the liver. The enzymatic activity of H. aurogutattus did not differ between lotic and lentic systems, whereas P. maculatus had comparatively higher stomach and hepatic trypsin levels and hepatic chymotrypsin-like activities in the reservoir than down in the river. These findings indicate that, in H. auroguttatus, most digestive activity occurs in the intestine, which is long and adapted for the digestion of bottom-river vegetable matter and detritus. The seasons and the type of the system (lentic vs. lotic seem to affect the enzymatic activity for these two species differently, a likely consequence of their different lifestyles.

Enzymatic Saccharification and Ethanol Fermentation of Reed Pretreated with Liquid Hot Water

Directory of Open Access Journals (Sweden)

Jie Lu

2012-01-01

Full Text Available Reed is a widespread-growing, inexpensive, and readily available lignocellulosic material source in northeast China. The objective of this study is to evaluate the liquid hot water (LHW pretreatment efficiency of reed based on the enzymatic digestibility and ethanol fermentability of water-insoluble solids (WISs from reed after the LHW pretreatment. Several variables in the LHW pretreatment and enzymatic hydrolysis process were optimized. The conversion of glucan to glucose and glucose concentrations are considered as response variables in different conditions. The optimum conditions for the LHW pretreatment of reed area temperature of 180°C for 20min and a solid-to-liquid ratio of 1 : 10. These optimum conditions for the LHW pretreatment of reed resulted in a cellulose conversion rate of 82.59% in the subsequent enzymatic hydrolysis at 50°C for 72 h with a cellulase loading of 30 filter paper unit per gram of oven-dried WIS. Increasing the pretreatment temperature resulted in a higher enzymatic digestibility of the WIS from reed. Separate hydrolysis and fermentation of WIS showed that the conversion of glucan to ethanol reached 99.5% of the theoretical yield. The LHW pretreatment of reed is a suitable method to acquire a high recovery of fermentable sugars and high ethanol conversion yield.

Influence of In vitro Digestion on Antioxidative Activity of Coconut ...

African Journals Online (AJOL)

Conclusion: CMPHs are high value-added antioxidants and possess a potential ..... side chain groups were expected to be exposed, ..... enzymatic digestion on antioxidant activity of coffee ... yam bean seed (Sphenostylis stenocarpa) protein.

Review of enhanced processes for anaerobic digestion treatment of sewage sludge

Science.gov (United States)

Liu, Xinyuan; Han, Zeyu; Yang, Jie; Ye, Tianyi; Yang, Fang; Wu, Nan; Bao, Zhenbo

2018-02-01

Great amount of sewage sludge had been produced each year, which led to serious environmental pollution. Many new technologies had been developed recently, but they were hard to be applied in large scales. As one of the traditional technologies, anaerobic fermentation process was capable of obtaining bioenergy by biogas production under the functions of microbes. However, the anaerobic process is facing new challenges due to the low fermentation efficiency caused by the characteristics of sewage sludge itself. In order to improve the energy yield, the enhancement technologies including sewage sludge pretreatment process, co-digestion process, high-solid digestion process and two-stage fermentation process were widely studied in the literatures, which were introduced in this article.

Microbial community structure in a thermophilic aerobic digester used as a sludge pretreatment process for the mesophilic anaerobic digestion and the enhancement of methane production.

Science.gov (United States)

Jang, Hyun Min; Park, Sang Kyu; Ha, Jeong Hyub; Park, Jong Moon

2013-10-01

An effective two-stage sewage sludge digestion process, consisting of thermophilic aerobic digestion (TAD) followed by mesophilic anaerobic digestion (MAD), was developed for efficient sludge reduction and methane production. Using TAD as a biological pretreatment, the total volatile suspended solid reduction (VSSR) and methane production rate (MPR) in the MAD reactor were significantly improved. According to denaturing gradient gel electrophoresis (DGGE) analysis, the results indicated that the dominant bacteria species such as Ureibacillus thermophiles and Bacterium thermus in TAD were major routes for enhancing soluble organic matter. TAD pretreatment using a relatively short SRT of 1 day showed highly increased soluble organic products and positively affected an increment of bacteria populations which performed interrelated microbial metabolisms with methanogenic species in the MAD; consequently, a quantitative real-time PCR indicated greatly increased Methanosarcinales (acetate-utilizing methanogens) in the MAD, resulting in enhanced methane production. Copyright © 2013 Elsevier Ltd. All rights reserved.

Towards a sustainable paradigm of waste-to-energy process: Enhanced anaerobic digestion of sludge with woody biochar

Energy Technology Data Exchange (ETDEWEB)

Shen, Yanwen; Linville, Jessica L.; Ignacio-de Leon, Patricia Anne A.; Schoene, Robin P.; Urgun-Demirtas, Meltem

2016-11-01

This study presents an integrated waste-to-energy process, using two waste streams, sludge generated from the municipal wastewater treatment plants (WWTPs) and biochar generated from the biomass gasification systems, to produce fungible biomethane and nutrient-rich digestate with fertilizer value. Two woody biochar, namely pinewood (PBC) and white oak biochar (WOBC) were used as additives during anaerobic digestion (AD) of WWTP sludge to enhance methane production at mesophilic and thermophilic temperatures. The PBC and WOBC have porous structure, large surface area and desirable chemical properties to be used as AD amendment material to sequester CO2 from biogas in the digester. The biochar-amended digesters achieved average methane content in biogas of up to 92.3% and 79.0%, corresponding to CO2 sequestration by up to 66.2% and 32.4% during mesophilic and thermophilic AD, respectively. Biochar addition enhanced process stability by increasing the alkalinity, but inhibitory effects were observed at high dosage. It also alleviated free ammonia inhibition by up to 10.5%. The biochar-amended digesters generated digestate rich in macro- and micronutrients including K (up to 300 m/L), Ca (up to 750 mg/L), Mg (up to 1800 mg/L) and Fe (up to 390 mg/L), making biochar-amended digestate a potential alternative used as agricultural lime fertilizer.

Microbial Insight into a Pilot-Scale Enhanced Two-Stage High-Solid Anaerobic Digestion System Treating Waste Activated Sludge.

Science.gov (United States)

Wu, Jing; Cao, Zhiping; Hu, Yuying; Wang, Xiaolu; Wang, Guangqi; Zuo, Jiane; Wang, Kaijun; Qian, Yi

2017-11-30

High solid anaerobic digestion (HSAD) is a rapidly developed anaerobic digestion technique for treating municipal sludge, and has been widely used in Europe and Asia. Recently, the enhanced HSAD process with thermal treatment showed its advantages in both methane production and VS reduction. However, the understanding of the microbial community is still poor. This study investigated microbial communities in a pilot enhanced two-stage HSAD system that degraded waste activated sludge at 9% solid content. The system employed process "thermal pre-treatment (TPT) at 70 °C, thermophilic anaerobic digestion (TAD), and mesophilic anaerobic digestion (MAD)". Hydrogenotrophic methanogens Methanothermobacter spp. dominated the system with relative abundance up to about 100% in both TAD and MAD. Syntrophic acetate oxidation (SAO) bacteria were discovered in TAD, and they converted acetate into H₂ and CO₂ to support hydrogenotrophic methanogenesis. The microbial composition and conversion route of this system are derived from the high solid content and protein content in raw sludge, as well as the operational conditions. This study could facilitate the understanding of the enhanced HSAD process, and is of academic and industrial importance.

Nanosilver: A Catalyst in Enzymatic Hydrolysis of Starch

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Falkowska Marta

2014-09-01

Full Text Available Silver nanoparticles are widely used, because of their antimicrobial properties. In this paper, the rate of starch digestion in the presence of nanocatalyst was compared with the rate of reaction without nanosilver. The rate of enzymatic degradation of starch was found to be increased in the presence of silver nanoparticles. It is considered that α-amylase was immobilized onto the surface of nanoparticles.

[Enhancement of anaerobic digestion of excess sludge by acid-alkali pretreatment].

Science.gov (United States)

Yuan, Guang-Huan; Zhou, Xing-Qiu; Wu, Jian-Dong

2012-06-01

In order to enhance the efficiency of anaerobic digestion of excess sludge, acid-alkali pretreatment method was studied. Three different pretreatment methods (alkali alone,acid-alkali, alkali-acid) were compared to investigate their impacts on hydrolysis and acidification of activated sludge. In addition, their influences on methane-producing in subsequent anaerobic digestion process were also studied. The results showed that the soluble chemical oxygen demand (SCOD) of alkaline treatment alone was about 16% higher than the combining of acid and alkali treatment, SCOD concentration increased to 5406.1 mg x L(-1) after 8 d pretreatment. After treated by acid (pH 4.0, 4 d) and alkali (pH 10.0, 4 d), the acetic acid production and its content in short-chain fatty acids (SCFAs) were higher than other pretreatment methods. And the acetic acid production (as COD/VSS) could reach 74.4 mg x g(-1), accounting for 60.5% of SCFAs. After acid-alkali pretreatment, the C: N ratio of the sludge mixed liquor was about 25, and the C: P ratio was between 35-40, which was more favorable than C: N and C: P ratio of alkali alone and alkali-acid to subsequent anaerobic digestion. The control experiments showed that, after acid-alkali pretreatment, anaerobic digestion cumulative methane yield (CH4/VSS(in)) reached to 136.1 mL x g(-1) at 15 d, which was about 2.5-, 1.6-, and 1.7-fold of the blank (unpretreated), alkali alone pretreatment and alkali-acid pretreatment, respectively. After acid-alkali pretreatment for 8 d and anaerobic digestion for 15 d, the removal efficiency of VSS was about 60.9%, and the sludge reduction effect was better than other pretreatments. It is obvious that the acid-alkali pretreatment method was more favorable to anaerobic digestion and sludge reduction.

Microwave irradiation of lignocellulosic materials, 4: Enhancement of enzymatic susceptibility of microwave-irradiated softwoods

International Nuclear Information System (INIS)

Azuma, J.; Higashino, J.; Isaka, M.; Koshijima, T.

1985-01-01

Effect of microwave irradiation on the enzymatic susceptibility of various softwoods was investigated. The pH values of the reaction liquor dropped with increasing temperature to 2.9-3.3 at 230°C, consistent with increase in acidity (0.5-0.85 meq at 230-239° C). Above approximately 180°C, hemicellulose underwent acid-mediated autohydrolysis and became water-soluble yielding a mixture of oligosaccharides and monosaccharides. The composition of water-soluble portion was similar for all wood species tested. The maximum extents of saccharification below 240°C ranged between 36-62% for softwoods, while those for hardwoods were between 88-93%. The present investigation confirmed that microwave pretreatment enhanced the enzymatic susceptibility of various softwoods. However, further attempt should be needed to give higher values equal to those for hardwoods. (author)

Optimization of municipal sludge and grease co-digestion using disintegration technologies.

Science.gov (United States)

Bouchy, L; Pérez, A; Camacho, P; Rubio, P; Silvestre, G; Fernández, B; Cano, R; Polanco, M; Díaz, N

2012-01-01

Many drivers tend to foster the development of renewable energy production in wastewater treatment plants as many expectations rely upon energy recovery from sewage sludge, for example through biogas use. This paper is focused on the assessment of grease waste (GW) as an adequate substrate for co-digestion with municipal sludge, as it has a methane potential of 479-710 LCH(4)/kg VS, as well as the evaluation of disintegration technologies as a method to optimize the co-digestion process. With this objective three different pre-treatments have been selected for evaluation: thermal hydrolysis, ultrasound and enzymatic treatment. Results have shown that co-digestion processes without pre-treatment had a maximum increment of 128% of the volumetric methane productivity when GW addition was 23% inlet (at 20 days of HRT and with an OLR of 3.0 kg COD/m(3)d), compared with conventional digestion of sewage sludge alone. Concerning the application of the selected disintegration technologies, all pre-treatments showed improvements in terms of methane yield (51.8, 89.5 and 57.6% more for thermal hydrolysis, ultrasound and enzymatic treatment, respectively, compared with non-pretreated wastes), thermal hydrolysis of GW and secondary sludge being the best configuration as it improved the solubilization of the organic matter and the hydrodynamic characteristics of digestates.

Isohexide and Sorbitol-Derived, Enzymatically Synthesized Renewable Polyesters with Enhanced Tg.

Science.gov (United States)

Gustini, Liliana; Lavilla, Cristina; de Ilarduya, Antxon Martínez; Muñoz-Guerra, Sebastián; Koning, Cor E

2016-10-10

Sugar-based polyesters derived from sorbitol and isohexides were obtained via solvent-free enzymatic catalysis. Pendant hydroxyl groups, coming from the sorbitol units, were present along the polyester backbone, whereas the two isohexides, namely, isomannide and isoidide dimethyl ester monomers, were selected to introduce rigidity into the polyester chains. The feasibility of incorporating isomannide as a diol compared to the isoidide dimethyl ester as acyl-donor via lipase-catalyzed polycondensation was investigated. The presence of bicyclic units resulted in enhanced T g with respect to the parent sorbitol-containing polyester lacking isohexides. The different capability of the two isohexides to boost the thermal properties confirmed the more flexible character provided by the isoidide diester derivative. Solvent-borne coatings were prepared by cross-linking the sugar-based polyester polyols with polyisocyanates. The increased rigidity of the obtained sugar-based polyester polyols led to an enhancement in hardness of the resulting coatings.

Rapid and efficient protein enzymatic digestion: An experimental comparison

Czech Academy of Sciences Publication Activity Database

Dyčka, Filip; Bobáľ, P.; Mazanec, Karel; Bobálová, Janette

2012-01-01

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

In vitro gastrointestinal digestion study of a novel bio-tofu with special emphasis on the impact of microbial transglutaminase

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Guangliang Xing

2016-12-01

Full Text Available We have developed a novel bio-tofu, made from mixed soy and cow milk (MSCM, using Lactobacillus helveticus MB2-1 and Lactobacillus plantarum B1-6 incorporated with microbial transglutaminase (MTGase as coagulant. MTGase was added to improve the textural properties and suit for cooking. However, the effect of MTGase on the digestion of mixed-protein fermented by lactic acid bacteria was unclear. This study aimed at evaluating the effect of MTGase on protein digestion of bio-tofu under simulated gastrointestinal digestion condition. The results showed that addition of MTGase could affect the particle size distribution, degree of hydrolysis, the content of soluble proteins and free amino acids. Based on the electrophoresis data, MTGase addition enhanced protein polymerization. During gastric and intestinal digestion process, proteins from bio-tofu were degraded into low molecular mass peptides. Our results suggested that incorporation of MTGase could lead to enzymatic modification of proteins of bio-tofu which may help in controlling energy intake and decrease the chance of food allergy.

In vitro gastrointestinal digestion study of a novel bio-tofu with special emphasis on the impact of microbial transglutaminase

Science.gov (United States)

Xing, Guangliang; Rui, Xin; Jiang, Mei; Xiao, Yu; Guan, Ying; Wang, Dan

2016-01-01

We have developed a novel bio-tofu, made from mixed soy and cow milk (MSCM), using Lactobacillus helveticus MB2-1 and Lactobacillus plantarum B1-6 incorporated with microbial transglutaminase (MTGase) as coagulant. MTGase was added to improve the textural properties and suit for cooking. However, the effect of MTGase on the digestion of mixed-protein fermented by lactic acid bacteria was unclear. This study aimed at evaluating the effect of MTGase on protein digestion of bio-tofu under simulated gastrointestinal digestion condition. The results showed that addition of MTGase could affect the particle size distribution, degree of hydrolysis, the content of soluble proteins and free amino acids. Based on the electrophoresis data, MTGase addition enhanced protein polymerization. During gastric and intestinal digestion process, proteins from bio-tofu were degraded into low molecular mass peptides. Our results suggested that incorporation of MTGase could lead to enzymatic modification of proteins of bio-tofu which may help in controlling energy intake and decrease the chance of food allergy. PMID:27994970

Novel anti-oxidative peptides from enzymatic digestion of human milk

DEFF Research Database (Denmark)

Tsopmo, Apollinaire; Romanowski, Andrea; Banda, Lyness

2011-01-01

Humanmilk pepsin and pancreatin digests were separated using molecular membrane and reverse phase chromatography. Chemical screening of the resulting fractions using the ORAC antioxidant assay yielded a peptide fraction (PF-23) with high antioxidant activity (5207 μM Trolox Equivalents (TE...

Study of bioleaching under different hydraulic retention time for enhancing the dewaterability of digestate.

Science.gov (United States)

Li, Linshuai; Gao, Jingqing; Zhu, Songfeng; Li, Yonghong; Zhang, Ruiqin

2015-12-01

Dewatering of kitchen waste digestate is a key problem to solve so as to increase the application of kitchen waste after anaerobic digestion. In this study, the effects of bioleaching under different hydraulic retention time (HRT = 2, 2.5, and 3 days) on dewaterability of kitchen waste digestate were evaluated. A 12-stage plug flow bioreactor with 180 L working volume was used for digestate bioleaching. The bioleached digestate under different HRTs were collected and dewatered by plate-and-frame filter press. The results showed that the moisture contents of digestate cakes were 67.87 % at 2 days of HRT, 58.06 % at 2.5 days of HRT, and 54.45 % at 3 days of HRT, respectively, indicating the longer the HRT, the lower the moisture content of filter cake. Balanced between the cost and practical need, 2.5 days can be used as the HRT in engineering application. Under the condition of HRT of 2.5 days, the pH, specific resistance to filtration (SRF), capillary suction time (CST), and sedimentation rate of digestate changed from the initial values of 8.08, 210.6 s, 23.4 × 10(12) m kg(-1) and 10 % to 3.21, 32.7 s, 2.44 × 10(12) m kg(-1) and 76.8 %, respectively. Based on the observations above, the authors conclude that bioleaching technology is an effective method to enhance digestate dewaterability and reduce the cost of subsequent reutilization.

Thermal and enzymatic pretreatment of sludge containing phthalate esters prior to mesophilic anaerobic digestion

DEFF Research Database (Denmark)

Gavala, Hariklia N.; Yenal, U.; Ahring, Birgitte Kiær

2004-01-01

The present study aimed at investigating the effect of thermal pretreatment of sludge at 70degreesC on the anaerobic degradation of three commonly found phthalic acid esters (PAE): di-ethyl phthalate (DEP), di-butyl phthalate (DBP), and di-ethylhexyl phthalate (DEHP). Also, the enzymatic treatment...... at 28degreesC with a commercial lipase was studied as a way to enhance PAE removal. Pretreatment at 70degreesC of the sludge containing PAE negatively influenced the anaerobic biodegradability of phthalate esters at 37degreesC. The observed reduction of PAE biodegradation rates after the thermal...... pretreatment was found to be proportional to the PAE solubility in water: the higher the solubility, the higher the percentage of the reduction (DEP > DBP > DEHP). PAE were slowly degraded during the pretreatment at 70degreesC, yet this was probably due to physicochemical reactions than to microbial...

Changes on antioxidant activity of microwave-treated protein hydrolysates after simulated gastrointestinal digestion: Purification and identification.

Science.gov (United States)

Ketnawa, Sunantha; Wickramathilaka, Malithi; Liceaga, Andrea M

2018-07-15

Two samples of trout frame protein hydrolysates were prepared by Microwave Pretreatment followed by Conventional Enzymatic hydrolysis (MPCE) and Non-Pretreated followed by Microwave-assisted Enzymatic hydrolysis (NPME), respectively, were subjected to simulated gastrointestinal digestion. Changes on degree of hydrolysis, antioxidant activity, molecular weight, and amino acid composition between undigested and after gastrointestinal digestion of peptides were investigated. Comparing to undigested peptides, a breakdown of MPCE and NPME into smaller molecules was observed. Degree of hydrolysis, ABTS + radical scavenging activity and reducing power increased (P digestion. A purified peptide from GI-MPCE had two possible sequences, NGRLGYSEGVM or GNRLGYSWDD (1182.65 Da). Whereas GI-NPME had two peptides IRGPEEHMHR or RVAPEEHMHR (1261.77 Da) and SAGVPRHK or SARPRHK (962.63 Da). These results indicate that digested hydrolysates can be a rich source of antioxidants. Isolated peptides extracted from trout frame by-products could be new food ingredients used as natural antioxidants. Copyright © 2018 Elsevier Ltd. All rights reserved.

The human digestive tract has proteases capable of gluten hydrolysis

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Sergio Gutiérrez

2017-07-01

Conclusion: The digestive tracts of patients with CD and healthy subjects have enzymatic machinery needed for gluten degradation. Patients with CD showed more gluten hydrolysis than did healthy individuals, although, in both cases, a fraction of 33-mer peptide remained intact. Gliadin peptides derived from gastrointestinal digestion, especially the 33-mer, can potentially be used by commensal microbiota from both CD-positive and CD-negative individuals, and differences in bacterial hydrolysis can modify its immunogenic capacity.

The Effect of Irradiation Treatment on the Non-Enzymatic Browning Reaction in Legume Seeds

International Nuclear Information System (INIS)

El-Niely, H.F.G.

2013-01-01

The present study was conducted to evaluate the effects of gamma irradiation treatment, at room temperature, on the non-enzymatic browning reaction (Millerd reaction products, MRPs) generated in soybeans, broad beans and dried peas seeds at dose levels of 10, 30 and 60 kGy and their effects on the chemical constituents, soluble protein, available lysine and in vitro protein digestibility. The formation of MRPs in the studied legumes was assayed by monitoring the formation of brown pigments (browning intensity) by spectrophotometric method. The results revealed that the chemical composition of irradiated legumes showed non-significant differences relative to the raw one. A dose dependent decrease in soluble proteins and available lysine in the three legumes were observed. The non-enzymatic browning reaction was significantly increased with increasing the radiation dose, which was proved by changes in browning index tests. At the same time, the in vitro protein digestibility was increased after irradiation up to 60 kGy. Irradiation of dried peas with 60 kGy produced higher browning index than the other legumes. A positive correlation was observed between the radiation dose and the browning index for soybeans (R2= 0.96), broad beans (R2 = 0.81) and dried peas (R2 = 0.97) which means that 96%, 81% and 97 of the variation in the incidence of non-enzymatic browning reaction in soybean, broad bean and dried peas, respectively, are due to the effect of irradiation treatments. The present study suggests that the formation of non-enzymatic browning reaction did not impair the nutritional quality of legumes, therefore, the process of irradiation was helpful in increasing the in vitro protein digestibility of studied legumes. These results clearly indicated that gamma irradiation processing at the studied doses can add valuable effects to the studied legumes

Extrusion enhances metabolizable energy and ileal amino acids digestibility of canola meal for broiler chickens

Directory of Open Access Journals (Sweden)

Aljuobori Ahmed

2014-01-01

Full Text Available The aim of the current study was to determine the effect of extrusion process on apparent metabolizable energy (AME, crude protein (CP and amino acid (AA digestibility of canola meal (CM in broiler chickens. A total of 36, 42-day-old broilers were randomly assigned into adaptation diets (no CM or 30% CM with six replicates. After 4 days of adaptation period, on day 47, birds were allowed to consume the assay diets that contain CM or extruded canola meal (ECM as the sole source of energy and protein. Following 4 h after feeding, the birds were killed and ileal contents were collected. The results showed that ECM had greater (P<0.001 AME (10.87 vs 9.39 MJ/kg compared to CM. The extrusion also significantly enhanced apparent ileal digestibility of CP and some of AA such as Asp, Glu, Ser, Thr and Trp. In conclusion, the extrusion treatment appeared to be a practical and effective approach in enhancing the digestibility of AME, CP and some AA of CM in broiler chickens.

Bioelectrochemical enhancement of methane production in low temperature anaerobic digestion at 10 °C

NARCIS (Netherlands)

Liu, Dandan; Zhang, Lei; Chen, Si; Buisman, Cees; Heijne, ter Annemiek

2016-01-01

Anaerobic digestion at low temperature is an attractive technology especially in moderate climates, however, low temperature results in low microbial activity and low rates of methane formation. This study investigated if bioelectrochemical systems (BESs) can enhance methane production from

Low melting point pyridinium ionic liquid pretreatment for enhancing enzymatic saccharification of cellulosic biomass.

Science.gov (United States)

Uju; Nakamoto, Aya; Shoda, Yasuhiro; Goto, Masahiro; Tokuhara, Wataru; Noritake, Yoshiyuki; Katahira, Satoshi; Ishida, Nobuhiro; Ogino, Chiaki; Kamiya, Noriho

2013-05-01

The potential of 1-hexylpyridinium chloride ([Hpy][Cl]), to pretreat cellulosic feedstocks was investigated using microcrystalline cellulose (Avicel) and Bagasse at 80 °C or 100 °C. Short [Hpy][Cl] pretreatments, conversion of pretreated Avicel to glucose was attained after 24h enzymatic saccharification under optimal conditions, whereas regenerated Bagasse showed 1-3-fold higher conversion than untreated biomass. FT-IR analysis of both Avicel and Bagasse samples pretreated with [Hpy][Cl] or 1-ethyl-3-methyimidazolium acetate ([Emim][OAc]) revealed that these ionic liquids behaved differently during pretreatment. [Hpy][Cl] pretreatment for an extended duration (180 min) released mono- and disaccharides without using cellulase enzymes, suggesting [Hpy][Cl] has capability for direct saccharification of cellulosic feedstocks. On the basis of the results obtained, [Hpy][Cl] pretreatment enhanced initial reaction rates in enzymatic saccharification by either crystalline polymorphic alteration of cellulose or partial degradation of the crystalline cellulosic fraction in biomass. Copyright © 2012 Elsevier Ltd. All rights reserved.

A qualitative and quantitative evaluation of the peptide characteristics of microwave- and ultrasound-assisted digestion in discovery and targeted proteomic analyses.

Science.gov (United States)

Guo, Zhengguang; Cheng, Jie; Sun, Haidan; Sun, Wei

2017-08-30

Fast digestion methods can dramatically accelerate enzyme digestion and increase the throughput of proteomic analysis. However, the peptide characteristics of fast digestion methods and their performance in discovery and targeted proteomic analysis must be systematically evaluated. Three digestion methods, including overnight digestion, microwave-assisted protein enzymatic digestion (MAPED), and high-intensity focused ultrasonic-assisted enzymatic digestion (HIFUSAED), in trypsin or in trypsin/Lys-C were comprehensively compared in both discovery and targeted proteomics analysis using the HeLa cell proteome. In discovery proteomic analysis, the highest numbers of peptides and proteins were identified when the sample was digested via the MAPED method with trypsin/Lys-C. The fast digestion methods showed a higher mis-cleavage rate and a lower semi-tryptic rate than the overnight digestion method. In both label-free quantitative analysis and targeted proteomic analysis, both fully cleaved peptides (FCPs) and mis-cleaved peptides (MCPs) from the fast digestion methods and the overnight digestion method showed good reproducibility if they showed good abundance. When both the FCPs and MCPs were included in the analysis, the MAPED with trypsin/Lys-C method showed the best results for both discovery proteomic analysis and relative quantitative targeted proteomic analysis. These results will be beneficial for the application of fast digestion methods to proteomics. Copyright © 2017 John Wiley & Sons, Ltd.

Enhanced cellulase hydrolysis of eucalyptus waste fibers from pulp mill by Tween80-assisted ferric chloride pretreatment.

Science.gov (United States)

Chen, Liheng; Fu, Shiyu

2013-04-03

Pretreatment combining FeCl3 and Tween80 was performed for cellulose-to-ethanol conversion of eucalyptus alkaline peroxide mechanical pulping waste fibers (EAWFs). The FeCl3 pretreatment alone showed a good effect on the enzymatic hydrolysis of EAWFs, but inhibited enzyme activity to some extent. A surfactant, Tween80, added during FeCl3 pretreatment was shown to significantly enhance enzyme reaction by eluting enzymatic inhibitors such as iron(III) that are present at the surface of the pretreated biomass. Treatment temperature, liquid-solid ratio, treatment time, FeCl3 concentration, and Tween80 dosage for pretreatment were optimized as follows: 180 °C, 8:1, 30 min, 0.15 mol/L, and 1% (w/v). Pretreated EAWFs under such optimal conditions provided enzymatic glucose (based on 100 g of oven-dried feedstock) and substrate enzymatic digestibility of EAWFs of 34.8 g and 91.3% after 72 h of enzymatic hydrolysis, respectively, with an initial cellulase loading of 20 FPU/g substrate.

Methods to enhance hydrolysis during one and two-stage anaerobic digestion of energy crops and crop residues

Energy Technology Data Exchange (ETDEWEB)

Jagadabhi, P. S.

2011-07-01

The objective of this thesis was to evaluate methods to enhance hydrolysis (measured as specific SCOD production, g SCOD g-1 VS) during one and two-stage anaerobic digestion (AD) of energy crops and crop residues. Addition of macro (NH{sub 4}Cl), micro nutrients (Fe, Ni, Co and Mo) and leachate replacement during mono-digestion of grass silage in one-stage leach bed reactors (LBRs) enhanced hydrolysis by 18 % (0.56 g SCOD g-1 VS), 7 % (0.45 g SCOD g-1 VS) and 34 % (0.51 g SCOD g-1 VS) respectively compared to respective controls. On the other hand, creating micro-aerobic conditions (at 1 l min-1, 2.5 l of air) did not improve hydrolysis but enhanced VFA production by 4 fold (from 2.2 g l-1 to 9 g l-1). Application of rumen cultures improved hydrolysis by 10 % (0.33 g SCOD g-1 VS) more than control (0.30 g SCOD g-1 VS). Similarly, during two-stage AD in LBR-UASB reactor configuration leachate replacement enhanced hydrolysis in cucumber and grass silage (0.5 g SCOD g-1 VS) than in tomato and common reed (0.35 and 0.15 g SCOD g-1 VS respectively). During co-digestion of grass silage and cow manure at a ratio of 30:70 (VS) in CSTR, re-circulation of alkali treated solid fraction of digestate did not improve the anaerobic biodegradation rates or methane yields. Results from batch experiments showed that methane potential of grass silage varied from 0.28-0.39 m3 CH{sub 4} kg-1 VS{sub added} in all the experiments. On the other hand, methane potentials of the studied crop residues were 0.32 m3 CH{sub 4} kg-1 VS{sub added} for tomato and 0.26 m3 CH{sub 4} kg-1 VS{sub added} for cucumber and common reed. Alkali pretreatment of solids, obtained from digestate (during co-digestion of grass silage and cow manure in one-stage CSTRs), at a low concentration of 20 g NaOH kg-1 VS resulted in higher methane yield (0.34 m3 CH{sub 4} kg-1 VS{sub added}) than the other tested dosages (40 and 60 g NaOH kg-1 VS). Addition of macro nutrient (NH{sub 4}Cl) enhanced methane potential of

Enhanced waste activated sludge digestion using a submerged anaerobic dynamic membrane bioreactor: performance, sludge characteristics and microbial community

Science.gov (United States)

Yu, Hongguang; Wang, Zhiwei; Wu, Zhichao; Zhu, Chaowei

2016-02-01

Anaerobic digestion (AD) plays an important role in waste activated sludge (WAS) treatment; however, conventional AD (CAD) process needs substantial improvements, especially for the treatment of WAS with low solids content and poor anaerobic biodegradability. Herein, we propose a submerged anaerobic dynamic membrane bioreactor (AnDMBR) for simultaneous WAS thickening and digestion without any pretreatment. During the long-term operation, the AnDMBR exhibited an enhanced sludge reduction and improved methane production over CAD process. Moreover, the biogas generated in the AnDMBR contained higher methane content than CAD process. Stable carbon isotopic signatures elucidated the occurrence of combined methanogenic pathways in the AnDMBR process, in which hydrogenotrophic methanogenic pathway made a larger contribution to the total methane production. It was also found that organic matter degradation was enhanced in the AnDMBR, thus providing more favorable substrates for microorganisms. Pyrosequencing revealed that Proteobacteria and Bacteroidetes were abundant in bacterial communities and Methanosarcina and Methanosaeta in archaeal communities, which played an important role in the AnDMBR system. This study shed light on the enhanced digestion of WAS using AnDMBR technology.

Digestibility of nutrients and aspects of the digestive physiology of ...

African Journals Online (AJOL)

The greater cane rat, Thryonomys swinderianus, utilizes high fibrous plant material and is an important meat source in West Africa. An insight in its digestive physiology will enhance our understanding of its feeding habits. Digestibility coefficients of the food were determined during two seasons before the animals were ...

Processed Meat Protein and Heat-Stable Peptide Marker Identification Using Microwave-Assisted Tryptic Digestion

Directory of Open Access Journals (Sweden)

Magdalena Montowska

2016-01-01

Full Text Available New approaches to rapid examination of proteins and peptides in complex food matrices are of great interest to the community of food scientists. The aim of the study is to examine the influence of microwave irradiation on the acceleration of enzymatic cleavage and enzymatic digestion of denatured proteins in cooked meat of five species (cattle, horse, pig, chicken and turkey and processed meat products (coarsely minced, smoked, cooked and semi-dried sausages. Severe protein aggregation occurred not only in heated meat under harsh treatment at 190 °C but also in processed meat products. All the protein aggregates were thoroughly hydrolyzed aft er 1 h of trypsin treatment with short exposure times of 40 and 20 s to microwave irradiation at 138 and 303 W. There were much more missed cleavage sites observed in all microwave-assisted digestions. Despite the incompleteness of microwave-assisted digestion, six unique peptide markers were detected, which allowed unambiguous identification of processed meat derived from the examined species. Although the microwave-assisted tryptic digestion can serve as a tool for rapid and high-throughput protein identification, great caution and pre-evaluation of individual samples is recommended in protein quantitation.

Understanding of alkaline pretreatment parameters for corn stover enzymatic saccharification

Directory of Open Access Journals (Sweden)

Chen Ye

2013-01-01

Full Text Available Abstract Background Previous research on alkaline pretreatment has mainly focused on optimization of the process parameters to improve substrate digestibility. To achieve satisfactory sugar yield, extremely high chemical loading and enzyme dosages were typically used. Relatively little attention has been paid to reduction of chemical consumption and process waste management, which has proven to be an indispensable component of the bio-refineries. To indicate alkali strength, both alkali concentration in pretreatment solution (g alkali/g pretreatment liquor or g alkali/L pretreatment liquor and alkali loading based on biomass solids (g alkali/g dry biomass have been widely used. The dual approaches make it difficult to compare the chemical consumption in different process scenarios while evaluating the cost effectiveness of this pretreatment technology. The current work addresses these issues through pretreatment of corn stover at various combinations of pretreatment conditions. Enzymatic hydrolysis with different enzyme blends was subsequently performed to identify the effects of pretreatment parameters on substrate digestibility as well as process operational and capital costs. Results The results showed that sodium hydroxide loading is the most dominant variable for enzymatic digestibility. To reach 70% glucan conversion while avoiding extensive degradation of hemicellulose, approximately 0.08 g NaOH/g corn stover was required. It was also concluded that alkali loading based on total solids (g NaOH/g dry biomass governs the pretreatment efficiency. Supplementing cellulase with accessory enzymes such as α-arabinofuranosidase and β-xylosidase significantly improved the conversion of the hemicellulose by 6–17%. Conclusions The current work presents the impact of alkaline pretreatment parameters on the enzymatic hydrolysis of corn stover as well as the process operational and capital investment costs. The high chemical consumption for alkaline

Evaluation of soluble fraction and enzymatic residual fraction of dilute dry acid, ethylenediamine, and steam explosion pretreated corn stover on the enzymatic hydrolysis of cellulose.

Science.gov (United States)

Qin, Lei; Liu, Li; Li, Wen-Chao; Zhu, Jia-Qing; Li, Bing-Zhi; Yuan, Ying-Jin

2016-06-01

This study is aimed to examine the inhibition of soluble fraction (SF) and enzymatic residual fraction (ERF) in dry dilute acid (DDA), ethylenediamine (EDA) and steam explosion (SE) pretreated corn stover (CS) on the enzymatic digestibility of cellulose. SF of DDA, EDA and SE pretreated CS has high xylose, soluble lignin and xylo-oligomer content, respectively. SF of EDA pretreated CS leads to the highest inhibition, followed by SE and DDA pretreated CS. Inhibition of ERF of DDA and SE pretreated CS is higher than that of EDA pretreated CS. The inhibition degree (A0/A) of SF is 1.76 and 1.21 times to that of ERF for EDA and SE pretreated CS, respectively. The inhibition degree of ERF is 1.05 times to that of SF in DDA pretreated CS. The quantitative analysis shows that SF of EDA pretreated CS, SF and ERF of SE pretreated CS cause significant inhibition during enzymatic hydrolysis. Copyright © 2016 Elsevier Ltd. All rights reserved.

The effects of gastric digestion on codfish allergenicity

DEFF Research Database (Denmark)

Untersmayr, Eva; Poulsen, Lars K.; Platzer, Michael H

2005-01-01

In a recent murine study, we showed that impaired gastric digestion supports the induction of fish allergy by protecting the digestion-sensitive major allergen parvalbumin and thus enhancing its sensitizing properties.......In a recent murine study, we showed that impaired gastric digestion supports the induction of fish allergy by protecting the digestion-sensitive major allergen parvalbumin and thus enhancing its sensitizing properties....

pH catalyzed pretreatment of corn bran for enhanced enzymatic arabinoxylan degradation

DEFF Research Database (Denmark)

Agger, Jane; Johansen, Katja Salomon; Meyer, Anne S.

2011-01-01

Corn bran is mainly made up of the pericarp of corn kernels and is a byproduct stream resulting from the wet milling step in corn starch processing. Through statistic modeling this study examined the optimization of pretreatment of corn bran for enzymatic hydrolysis. A low pH pretreatment (pH 2......, 150°C, 65min) boosted the enzymatic release of xylose and glucose and maximized biomass solubilization. With more acidic pretreatment followed by enzymatic hydrolysis the total xylose release was maximized (at pH 1.3) reaching ∼50% by weight of the original amount present in destarched corn bran......, but the enzyme catalyzed xylose release was maximal after pretreatment at approx. pH 2. The total glucose release peaked after pretreatment of approx. pH 1.5 with an enzymatic release of approx. 68% by weight of the original amounts present in destarched corn bran. For arabinose the enzymatic release...

Influence of enzymatic hydrolysis on the allergenic reactivity of processed cashew and pistachio.

Science.gov (United States)

Cuadrado, Carmen; Cheng, Hsiaopo; Sanchiz, Africa; Ballesteros, Isabel; Easson, Michael; Grimm, Casey C; Dieguez, M Carmen; Linacero, Rosario; Burbano, Carmen; Maleki, Soheila J

2018-02-15

Cashew and pistachio allergies are considered a serious health problem. Previous studies have shown that thermal processing, pressurization and enzymatic hydrolysis may reduce the allergenic properties of food by changing the protein structure. This study assesses the allergenic properties of cashew and pistachio after thermal treatment (boiling and autoclaving), with or without pressure (autoclaving), and multiple enzymatic treatments under sonication, by SDS-PAGE, western blot and ELISA, with serum IgE of allergic individuals, and mass spectroscopy. Autoclaving and enzymatic hydrolysis under sonication separately induced a measurable reduction in the IgE binding properties of pastes made from treated cashew and pistachio nuts. These treatments were more effective with pistachio allergens. However, heat combined with enzymatic digestion was necessary to markedly lower IgE binding to cashew allergens. The findings identify highly effective simultaneous processing conditions to reduce or even abolish the allergenic potency of cashew and pistachio. Copyright © 2017 Elsevier Ltd. All rights reserved.

Radiation pretreatments of cellulose materials for the enhancement of enzymatic hydrolysis

International Nuclear Information System (INIS)

Ardica, S.; Calderaro, E.; Cappadona, C.

1985-01-01

The effect of γ-ray pre-irradiation of cellulose materials such as wood chips, paper, grain straw, hay and kapok on glucose production on enzymatic hydrolysis by cellulase has been investigated. These materials have been irradiated in air, water and acetate buffer solution over the dose range 10 3 to 4 x 10 6 Gy. In the relatively low dose range, up to about 5 x 10 5 Gy, the glucose yields after enzymatic hydrolysis are practically insensitive to radiation. At higher dose levels, up to 1.7 to 2 x 10 6 Gy, the pre-irradiation becomes very effective on enzymatic cellulose conversion. It has been found that the radiation-induced degradation of cellulose into low molecular weight polysaccharides is dependent on the nature and chemical composition of the cellulose materials and on the radiation environmental conditions. Further increases of dose causes radiation-induced structural modifications in polysaccharides previously produced, which can lead to a decrease in glucose production by enzymatic hydrolysis. (author)

Enhanced non-enzymatic glucose sensing based on copper nanoparticles decorated nitrogen-doped graphene.

Science.gov (United States)

Jiang, Ding; Liu, Qian; Wang, Kun; Qian, Jing; Dong, Xiaoya; Yang, Zhenting; Du, Xiaojiao; Qiu, Baijing

2014-04-15

Copper nanoparticles (NPs) decorated nitrogen-doped graphene (Cu-N-G) was prepared by a facile thermal treatment, and further employed as a novel sensing material for fabricating the sensitive non-enzymatic glucose sensor. Compared with pure Cu NPs, the Cu-N-G showed enhanced electrocatalytic activity to glucose oxidation due to the integration of N-G, which exhibited the oxidation peak current of glucose ca. 23-fold higher than that of pure Cu NPs. The presented sensor showed excellent performances for glucose detection including wide linear range of 0.004-4.5 mM, low detection limit (1.3 μM, S/N=3), high sensitivity (48.13 μA mM(-1)), fast response time (doped graphene as enhanced materials in fabricating sensors for chemical and biochemical analysis. © 2013 Published by Elsevier B.V.

Surface properties correlate to the digestibility of hydrothermally pretreated lignocellulosic Poaceae biomass feedstocks

DEFF Research Database (Denmark)

Tristan Djajadi, Demi; Hansen, Aleksander R.; Jensen, Anders

2017-01-01

physical and chemical features of the biomass surfaces, specifically contact angle measurements (wettability) and attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy (surfacebiopolymer composition) produced data correlating pretreatment severity and enzymatic digestibility......, and they also revealed differences that correlated to enzymatic glucose yield responses among the three different biomass types. Conclusion: The study revealed that to a large extent, factors related to physico-chemical surface properties, namely surface wettability as assessed by contact angle measurements...

Evaluation of lime and hydrothermal pretreatments for efficient enzymatic hydrolysis of raw sugarcane bagasse.

Science.gov (United States)

Grimaldi, Maira Prearo; Marques, Marina Paganini; Laluce, Cecília; Cilli, Eduardo Maffud; Sponchiado, Sandra Regina Pombeiro

2015-01-01

Ethanol production from sugarcane bagasse requires a pretreatment step to disrupt the cellulose-hemicellulose-lignin complex and to increase biomass digestibility, thus allowing the obtaining of high yields of fermentable sugars for the subsequent fermentation. Hydrothermal and lime pretreatments have emerged as effective methods in preparing the lignocellulosic biomass for bioconversion. These pretreatments are advantageous because they can be performed under mild temperature and pressure conditions, resulting in less sugar degradation compared with other pretreatments, and also are cost-effective and environmentally sustainable. In this study, we evaluated the effect of these pretreatments on the efficiency of enzymatic hydrolysis of raw sugarcane bagasse obtained directly from mill without prior screening. In addition, we evaluated the structure and composition modifications of this bagasse after lime and hydrothermal pretreatments. The highest cellulose hydrolysis rate (70 % digestion) was obtained for raw sugarcane bagasse pretreated with lime [0.1 g Ca(OH)2/g raw] for 60 min at 120 °C compared with hydrothermally pretreated bagasse (21 % digestion) under the same time and temperature conditions. Chemical composition analyses showed that the lime pretreatment of bagasse promoted high solubilization of lignin (30 %) and hemicellulose (5 %) accompanied by a cellulose accumulation (11 %). Analysis of pretreated bagasse structure revealed that lime pretreatment caused considerable damage to the bagasse fibers, including rupture of the cell wall, exposing the cellulose-rich areas to enzymatic action. We showed that lime pretreatment is effective in improving enzymatic digestibility of raw sugarcane bagasse, even at low lime loading and over a short pretreatment period. It was also demonstrated that this pretreatment caused alterations in the structure and composition of raw bagasse, which had a pronounced effect on the enzymes accessibility to the

Rhythmicity and plasticity of digestive physiology in a euryhaline teleost fish, permit (Trachinotus falcatus)

DEFF Research Database (Denmark)

Lazado, Carlo Cabacang; Pedersen, Per Bovbjerg; Nguyen, Huy Quang

2017-01-01

Digestive physiology is considered to be under circadian control, but there is little evidence in teleost fish. The present study explored the rhythmicity and plasticity to feeding schedules of enzymatic digestion in a candidate aquaculture fish, the permit (Trachinotus falcatus). The first...... experiment identified the rhythms of digestive factors throughout the light-dark (LD) cycle. Gastric luminal pH and pepsin activity showed significant daily variation albeit not rhythmic. These dynamic changes were likewise observed in several digestive enzymes, in which the activities of intestinal protease......, chymotrypsin and lipase exhibited significant daily rhythms. In the second experiment, the existence of feed anticipatory activity in the digestive factors was investigated by subjecting the fish to either periodic or random feeding. Anticipatory gastric acidification prior to feeding was identified...

Waste management and enzymatic treatment of Municipal Solid Waste

DEFF Research Database (Denmark)

Jensen, Jacob Wagner

generation for subsequent biogas production. Municipal solid waste (MSW) is produced in large amounts every year in the developed part of the world. The household waste composition varies between geographical areas and between seasons. However the overall content of organic and degradable material is rather......The work carried out during the Ph.D. project is part of the Danish Energy Authority funded research project called PSO REnescience and is focussed on studying the enzymatic hydrolysis and liquefaction of waste biomass. The purpose of studying the liquefaction of waste biomass is uniform slurry...... constant between 50 - 60 % wet weight and therefore holds a potential for bioenergy production. The degradable fraction has positive effects for anaerobic digestion when evaluated to desired parameters of anaerobic digestion plants. Wanted parameters are: 1) high organic content (high volatile solid...

Enhanced enzymatic cellulose degradation by cellobiohydrolases via product removal

DEFF Research Database (Denmark)

Ahmadi Gavlighi, Hassan; Meyer, Anne S.; Mikkelsen, Jørn Dalgaard

2013-01-01

Product inhibition by cellobiose decreases the rate of enzymatic cellulose degradation. The optimal reaction conditions for two Emericella (Aspergillus) nidulans-derived cellobiohydrolases I and II produced in Pichia pastoris were identified as CBHI: 52 °C, pH 4.5–6.5, and CBHII: 46 °C, pH 4.......8. The optimum in a mixture of the two was 50 °C, pH 4.9. An almost fourfold increase in enzymatic hydrolysis yield was achieved with intermittent product removal of cellobiose with membrane filtration (2 kDa cut-off): The conversion of cotton cellulose after 72 h was ~19 % by weight, whereas the conversion...

Enzymatic characterization of lipid-based drug delivery systems

DEFF Research Database (Denmark)

Ljusberg-Wahren, Helena; Seier Nielsen, Flemming; Brogård, Mattias

2005-01-01

The present work introduces a simple and robust in vitro method for enzymatic characterisation of surface properties of lipid dispersions in aqueous media. The initial lipolysis rate in biorelevant media, using pancreatic lipase and a self-microemulsifying formulation (SMEDDS) containing digestible...... lipids as substrate, was determined. The impact of incorporating two sparingly water soluble model drugs, probucol and halofantrine, into the SMEDDS was studied. It was found that both model drugs reduced the initial rate of lipolysis compared with the vehicle, probucol having a larger effect than...

Impact of food processing and simulated gastrointestinal digestion on gliadin immunoreactivity in rolls.

Science.gov (United States)

Brzozowski, Bartosz

2018-07-01

The enzymatic modification of wheat proteins during dough fermentation and its digestion as supported by peptidases of microbiological origin can result in the degradation of important peptides in the pathogenesis of coeliac disease. However, baking bread and the high temperature associated with this could change the physicochemical and immunological properties of proteins. Thermal changes in the spatial structure of proteins and their hydrolysis can lead to a masking or degrading of immunoreactive peptides. The addition of prolyl endopeptidase (PEP), comprising peptidases isolated from Lactobacillus acidophilus 5e2 (LA) or transglutaminase (TG) in the course of fermentation, decreases its immunoreactivity by 83.9%, 51.9% and 18.5%, respectively. An analysis of the fractional composition of gliadins revealed that γ- and ω-gliadins are the proteins most susceptible to enzymatic modification. Hydrolysis of wheat storage proteins with PEP and LA reduces the content of αβ-, γ- and ω-gliadins by 13.7%, 60.2% and 41.9% for PEP and by 22.1%, 43.5% and 36.9% for LA, respectively. Cross-linking of proteins with TG or their hydrolysis by PEP and LA peptidases during the process of forming wheat dough, followed by digesting bread samples with PEP and LA peptidases, decreases the immunoreactivity of bread hydrolysates from 2.4% to 0.02%. The content of peptide detected in polypeptide sequences is 263.4 ± 3.3, 30.9 ± 1.5 and 7.9 ± 0.4 mg kg -1 in samples of hydrolysates of bread digested with PEP, as produced from dough modified by TG, PEP and LA, respectively. Enzymatic pre-modification of proteins during the process of dough fermentation decreases their immunoreactive potential, such that fewer peptides recognised by R5 antibodies are released during the digestion process from the bread matrix. Immunoreactive peptides are degraded more effectively when digestive enzymes are supported by the addition of PEP. © 2017 Society of Chemical Industry. © 2017

Relationship between fiber porosity and cellulose digestibility in steam-exploded Pinus radiata

Energy Technology Data Exchange (ETDEWEB)

Wong, K.K.Y.; Deverell, K.F.; Mackie, K.L.; Clark, T.A.; Donaldson, L.A.

1988-04-05

The use of lignocellulosic materials in bioconversion processes may be improved if the critical factors limiting conversion are better understood. Steam explosion after sulfur dioxide impregnation of wood chips is an effective method for improving the enzymatic digestibility of cellulose in the softwood Pinus radiata. Digestibility of pretreated fiber was progressively increased by altering the conditions of steam explosion. With increasing digestibility, there was an observed increase in fiber porosity as measured by the solute exclusion technique. Accessible pore volume and accessible surface area to a 5-nm dextran probe positively correlated with both 2- and 24-h digestion yields from pretreated fiber. The increase in accessibility was probably the result of hemicellulose extraction and lignin redistribution. A subsequent loss in accessibility, brought about by structural collapse or further lignin redistribution, resulted in a corresponding loss in digestibility. It appears that steam explosion increases cellulose digestibility in P. radiata by increasing fiber porosity.

Enhancement of biogas production at the municipal wastewater treatment plant by co-digestion with poultry industry waste

International Nuclear Information System (INIS)

Budych-Gorzna, Magdalena; Smoczynski, Marcin; Oleskowicz-Popiel, Piotr

2016-01-01

Highlights: • Laboratory and full-scale trials on co-digestion of sludge and poultry waste were performed. • Successful scaling-up of the results from laboratory to full-scale was accomplished. • Incremental addition of poultry waste to the full-scale anaerobic digesters did not cause any inhibition of the process. • WWTP energy dependency can be reduced significantly by co-digestion of sludge and external source of waste. - Abstract: Municipal wastewater treatment plants (WWTPs) are energy-intensive and thus cost-intensive facilities; therefore, it is crucial to increase energy production directly at the WWTP. Enhancement of biogas production by addition of external substrates is one of the solutions to increase energy self-sufficiency of the WWTPs with an additional benefit of cutting down the greenhouse gas emission. The main aim of the work was to investigate full utilization of the capacity of full-scale digesters at the municipal WWTP by addition of poultry industry waste. At first, laboratory trials were conducted in order to identify the most suitable dose for co-digestion with primary and waste activated sludge and finally, based on the achieved laboratory results, full-scale trials were carried out directly at the municipal WWTP. Poultry industrial waste yielded between 0.39 and 0.88 m 3 of methane per kg of volatile solids during laboratory trials, depending on the added concentration. During full-scale investigation yield of 0.81 m 3 /kg VS was achieved. Enhanced biogas production improved WWTP energy self-sufficiency bringing closer to the aim of increasing the share of self-produced energy up to 80%.

Three-stage enzymatic digestive system for a gut-on-a-chip

NARCIS (Netherlands)

de Haan, Pim; Ianovska, Margaryta A.; Bouwmeester, Hans; Verpoorte, Elisabeth

2017-01-01

Several different devices to model the human gastrointestinal (GI) tract have been developed, which may find applications as pharmacological or toxicological model systems.1 To create a fully functional gut-on-a-chip, it is essential to incorporate the digestive functions of the GI tract into such a

Antioxidant activity of camel milk casein before and after in vitro simulated enzymatic digestion

Directory of Open Access Journals (Sweden)

Zeineb Jrad

2014-11-01

Full Text Available The effect of a successive in vitro hydrolysis by pepsin and pancreatin on the free radical scavenging activity of camel milk casein was investigated in order to assess the effect of gastro-intestinal digestion. Hydrolysis of camel casein was controlled by reversed-phase high performance liquid chromatography. Anti-oxidant activity was measured by the 2,2’-azino-bis-(3-ethylbensothiazoline-6- sulfonic acid (ABTS method. The Trolox equivalent antioxidant capacity (TEAC values of camel casein and its hydrolysate were 1.6±0.12 μmol TE/mg protein and 0.25 μmol TE/μmol eq. NH2, respectively. After digestion, the scavenging activity of the casein peptides was more efficient than those reported in the literature regarding digestive hydrolysates of camel milk, colostrum and whey proteins.

Development of a new bioethanol feedstock - Anaerobically digested fiber from confined dairy operations using different digestion configurations

International Nuclear Information System (INIS)

Yue, Zhengbo; Teater, Charles; MacLellan, James; Liu, Yan; Liao, Wei

2011-01-01

Two types of digesters, continuous stirring-tank reactor (CSTR) and plug flow reactor (PFR), were integrated into a biorefining concept to generate a new cellulosic ethanol feedstock -anaerobically digested fiber (AD fiber) from dairy cow feces. Cellulose content in AD fibers was significantly increased during the anaerobic digestion. CSTR and PFR AD fibers had cellulose contents of 357 and 322 g kg -1 dried AD fiber. The AD fibers were enzymatically hydrolyzed after being pretreated by dilute sulfuric acid or dilute sodium hydroxide, and the hydrolysates were used to produce ethanol. Alkali pretreatment was concluded as a suitable pretreatment method for AD fibers. Under the optimal conditions the AD fibers processed by CSTR and PFR produced ethanol of 26 g kg -1 and 23 g kg -1 dry feces, respectively. Energy balance analysis further indicated that CSTR was a preferred digestion method to prepare AD fiber for ethanol production. -- Highlights: → Anaerobic digestion process has been discovered as a process that is not only a downstream process, but also a pretreatment method to prepare cellulosic feedstock for biorefining. → In this study the effects of two different AD reactor configurations (CSTR and PFR) on AD fiber quality and bioethanol conversion of the AD fiber have been explored. → Mass and energy balance analysis elucidated that compared to PFR, CSTR is better AD treatment to prepare AD fiber for bioethanol production.

Free nitrous acid pre-treatment of waste activated sludge enhances volatile solids destruction and improves sludge dewaterability in continuous anaerobic digestion.

Science.gov (United States)

Wei, Wei; Wang, Qilin; Zhang, Liguo; Laloo, Andrew; Duan, Haoran; Batstone, Damien J; Yuan, Zhiguo

2018-03-01

Previous work has demonstrated that pre-treatment of waste activated sludge (WAS) with free nitrous acid (FNA i.e. HNO 2 ) enhances the biodegradability of WAS, identified by a 20-50% increase in specific methane production in biochemical methane potential (BMP) tests. This suggests that FNA pre-treatment would enhance the destruction of volatile solids (VS) in an anaerobic sludge digester, and reduce overall sludge disposal costs, provided that the dewaterability of the digested sludge is not negatively affected. This study experimentally evaluates the impact of FNA pre-treatment on the VS destruction in anaerobic sludge digestion and on the dewaterability of digested sludge, using continuously operated bench-scale anaerobic digesters. Pre-treatment of full-scale WAS for 24 h at an FNA concentration of 1.8 mg NN/L enhanced VS destruction by 17 ± 1% (from 29.2 ± 0.9% to 34.2 ± 1.1%) and increased dewaterability (centrifuge test) from 12.4 ± 0.4% to 14.1 ± 0.4%. Supporting the VS destruction data, methane production increased by 16 ± 1%. Biochemical methane potential tests indicated that the final digestate stability was also improved with a lower potential from FNA treated digestate. Further, a 2.1 ± 0.2 log improvement in pathogen reduction was also achieved. With inorganic solids representing 15-22% of the full-scale WAS used, FNA pre-treatment resulted in a 16-17% reduction in the volume of dewatered sludge for final disposal. This results in significantly reduced costs as assessed by economic analysis. Copyright © 2017 Elsevier Ltd. All rights reserved.

Quantification of peptides released during in vitro digestion of cooked meat.

Science.gov (United States)

Sayd, T; Chambon, C; Santé-Lhoutellier, V

2016-04-15

We aimed to identify and quantify the peptides generated during in vitro digestion of cooked meat by liquid chromatography coupled with high resolution mass spectrometer. A total of 940 non-redundant peptides in the gastric compartment and 989 non-redundant peptides in the intestinal compartment were quantified and identified. Among the 71 different proteins identified, 43 meat proteins were found in the two digestive compartments, 20 proteins were specific to the gastric compartment and 8 proteins to the intestinal compartment. In terms of estimation, the proteins involved in muscle contraction and structure were preferentially enzymatically hydrolyzed in the small intestine. The effect of cooking provided different but less clear patterns of digestion. To the best of our knowledge, this constitutes the highest number of peptides identified in beef meat digests and provides a comprehensive database for meat protein digestion associated with cooking conditions. Such quantitative and qualitative differences may have important nutritional consequences. Copyright © 2015 Elsevier Ltd. All rights reserved.

Curcumin Encapsulated in Milk Exosomes Resists Human Digestion and Possesses Enhanced Intestinal Permeability in Vitro.

Science.gov (United States)

Vashisht, Monika; Rani, Payal; Onteru, Suneel Kumar; Singh, Dheer

2017-11-01

Exosomes, the extracellular secretary nano-vesicles, act as carriers of biomolecules to the target cells. They exhibit several attributes of an efficient drug delivery system. Curcumin, despite having numerous bioactive and therapeutic properties, has limited pharmaceutical use due to its poor water solubility, stability, and low systemic bioavailability. Hence, this study aims to enhance the therapeutic potential of curcumin, a model hydrophobic drug, by its encapsulation into milk exosomes. In the present study, we investigated the stability of free curcumin and exosomal curcumin in PBS and in vitro digestive processes. Additionally, their uptake and trans-epithelial transport were studied on Caco-2 cells. Curcumin in milk exosomes had higher stability in PBS, sustained harsh digestive processes, and crossed the intestinal barrier than free curcumin. In conclusion, the encapsulation of curcumin into the exosomes enhances its stability, solubility, and bioavailability. Therefore, the present study demonstrated that milk exosomes act as stable oral drug delivery vehicles.

Zero-valent iron enhanced methanogenic activity in anaerobic digestion of waste activated sludge after heat and alkali pretreatment.

Science.gov (United States)

Zhang, Yaobin; Feng, Yinghong; Quan, Xie

2015-04-01

Heat or alkali pretreatment is the effective method to improve hydrolysis of waste sludge and then enhance anaerobic sludge digestion. However the pretreatment may inactivate the methanogens in the sludge. In the present work, zero-valent iron (ZVI) was used to enhance the methanogenic activity in anaerobic sludge digester under two methanogens-suppressing conditions, i.e. heat-pretreatment and alkali condition respectively. With the addition of ZVI, the lag time of methane production was shortened, and the methane yield increased by 91.5% compared to the control group. The consumption of VFA was accelerated by ZVI, especially for acetate, indicating that the acetoclastic methanogenesis was enhanced. In the alkali-condition experiment, the hydrogen produced decreased from 27.6 to 18.8 mL when increasing the ZVI dosage from 0 to 10 g/L. Correspondingly, the methane yield increased from 1.9 to 32.2 mL, which meant that the H2-utilizing methanogenes was enriched. These results suggested that the addition of ZVI into anaerobic digestion of sludge after pretreated by the heat or alkali process could efficiently recover the methanogenic activity and increase the methane production and sludge reduction. Copyright © 2015 Elsevier Ltd. All rights reserved.

Particle size distribution of rice flour affecting the starch enzymatic hydrolysis and hydration properties.

Science.gov (United States)

de la Hera, Esther; Gomez, Manuel; Rosell, Cristina M

2013-10-15

Rice flour is becoming very attractive as raw material, but there is lack of information about the influence of particle size on its functional properties and starch digestibility. This study evaluates the degree of dependence of the rice flour functional properties, mainly derived from starch behavior, with the particle size distribution. Hydration properties of flours and gels and starch enzymatic hydrolysis of individual fractions were assessed. Particle size heterogeneity on rice flour significantly affected functional properties and starch features, at room temperature and also after gelatinization; and the extent of that effect was grain type dependent. Particle size heterogeneity on rice flour induces different pattern in starch enzymatic hydrolysis, with the long grain having slower hydrolysis as indicated the rate constant (k). No correlation between starch digestibility and hydration properties or the protein content was observed. It seems that in intact granules interactions with other grain components must be taken into account. Overall, particle size fractionation of rice flour might be advisable for selecting specific physico-chemical properties. Copyright © 2013. Published by Elsevier Ltd.

Enhancing biogas production from recalcitrant lignocellulosic residue

DEFF Research Database (Denmark)

Tsapekos, Panagiotis

Lignocellulosic substrates are abundant in agricultural areas around the world and lately, are utilized for biogas production in full-scale anaerobic digesters. However, the anaerobic digestion (AD) of these substrates is associated with specific difficulties due to their recalcitrant nature which...... protects them from enzymatic attack. Hence, the main purpose of this work was to define diverse ways to improve the performance of AD systems using these unconventional biomasses. Thus, mechanical and thermal alkaline pretreatments, microaeration and bioaugmentation with hydrolytic microbes were examined...... conductivity, soluble chemical oxygen demand and enzymatic hydrolysis) as a rapid way to predict the methane production. However, the precision of methane yield prediction was not high (R2

Use of alkaline or enzymatic sample pretreatment prior to characterization of gold nanoparticles in animal tissue by single-particle ICPMS

DEFF Research Database (Denmark)

Löschner, Katrin; Brabrand, Myung Suk Jung; Sloth, Jens Jørgen

2014-01-01

, not much is known about the applicability of spICPMS for determination of NPs in complex matrices such as biological tissues. In the present study, alkaline and enzymatic treatments were applied to solubilize spleen samples from rats, which had been administered 60-nm gold nanoparticles (Au......NPs) intravenously. The results showed that similar size distributions of AuNPs were obtained independent of the sample preparation method used. Furthermore, the quantitative results for AuNP mass concentration obtained with spICPMS following alkaline sample pretreatment coincided with results for total gold...... concentration obtained by conventional ICPMS analysis of acid-digested tissue. The recovery of AuNPs from enzymatically digested tissue, however, was approximately four times lower. Spiking experiments of blank spleen samples with AuNPs showed that the lower recovery was caused by an inferior transport...

Carbon balances for in vitro digestion an fermentation of potential roughages for pregnant sows

NARCIS (Netherlands)

Becker, P.M.; Gelder, van A.H.; Wikselaar, van P.G.; Jongbloed, A.W.; Cone, J.W.

2003-01-01

Ad libitum feeding of pregnant sows requires satiating, intake-restricting feed components to prevent sows from getting excessively fat. Because hindgut fermentation starts only after and proceeds much slower than enzymatic digestion in the small intestine, fermentation products might, as nutrients,

Comparative Digestive Physiology

Science.gov (United States)

Karasov, William H.; Douglas, Angela E.

2015-01-01

In vertebrates and invertebrates, morphological and functional features of gastrointestinal (GI) tracts generally reflect food chemistry, such as content of carbohydrates, proteins, fats, and material(s) refractory to rapid digestion (e.g., cellulose). The expression of digestive enzymes and nutrient transporters approximately matches the dietary load of their respective substrates, with relatively modest excess capacity. Mechanisms explaining differences in hydrolase activity between populations and species include gene copy number variations and single-nucleotide polymorphisms. Transcriptional and posttranscriptional adjustments mediate phenotypic changes in the expression of hydrolases and transporters in response to dietary signals. Many species respond to higher food intake by flexibly increasing digestive compartment size. Fermentative processes by symbiotic microorganisms are important for cellulose degradation but are relatively slow, so animals that rely on those processes typically possess special enlarged compartment(s) to maintain a microbiota and other GI structures that slow digesta flow. The taxon richness of the gut microbiota, usually identified by 16S rRNA gene sequencing, is typically an order of magnitude greater in vertebrates than invertebrates, and the interspecific variation in microbial composition is strongly influenced by diet. Many of the nutrient transporters are orthologous across different animal phyla, though functional details may vary (e.g., glucose and amino acid transport with K+ rather than Na+ as a counter ion). Paracellular absorption is important in many birds. Natural toxins are ubiquitous in foods and may influence key features such as digesta transit, enzymatic breakdown, microbial fermentation, and absorption PMID:23720328

High Hydrostatic Pressure-Assisted Enzymatic Treatment Improves Antioxidant and Anti-inflammatory Properties of Phosvitin.

Science.gov (United States)

Yoo, Heejoo; Bamdad, Fatemeh; Gujral, Naiyana; Suh, Joo-Won; Sunwoo, Hoon

2017-01-01

Phosvitin (PV) is a highly-phosphorylated metal-binding protein in egg yolk. Phosphoserine clusters make PV resistant to enzymatic digestion, which might be nutritionally undesirable. This study was designed to determine the effects of high hydrostatic pressure and enzymatic hydrolysis (HHP-EH) on the antioxidant and anti-inflammatory properties of PV hydrolysates (PVHs). PV was hydrolyzed by alcalase, elastase, savinase, thermolysin, and trypsin at 0.1, 50, and 100 MPa pressure levels. PVHs were evaluated for degree of hydrolysis, molecular weight distribution patterns, antioxidant and anti-inflammatory properties in chemical and cellular models. The effect of PVH on gene expression of pro-inflammatory cytokines (TNF-α and IL-1β) was also evaluated using real time-PCR. The hydrolysate with most potent antioxidant and anti-inflammatory properties was subjected to LC-MS/MS analysis to identify the peptide sequence. Hydrolysates produced at 100 MPa exhibited higher degree of hydrolysis and greater reducing power and free radical scavenging activity compared to those obtained at atmospheric pressure. After adjusting the phosphate content, alcalase- and trypsin-digested PVHs showed superior iron chelation capacity (69-73%), regardless of pressure. Both alcalase- and trypsin-digested PVHs significantly inhibited nitric oxide production by RAW264.7 macrophage cells. LPS-stimulated up-regulation of proinflammatory cytokines was also suppressed by alcalase-digested PVH. The HHP-EH method could play a promising role in the production of bioactive peptides from hydrolysis-resistant proteins. HHP-assisted PVH may be useful in preparing a potential pharmaceutical with antioxidant and anti-inflammatory properties. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

BSA treatment to enhance enzymatic hydrolysis of cellulose in lignin containing substrates.

Science.gov (United States)

Yang, Bin; Wyman, Charles E

2006-07-05

Cellulase and bovine serum albumin (BSA) were added to Avicel cellulose and solids containing 56% cellulose and 28% lignin from dilute sulfuric acid pretreatment of corn stover. Little BSA was adsorbed on Avicel cellulose, while pretreated corn stover solids adsorbed considerable amounts of this protein. On the other hand, cellulase was highly adsorbed on both substrates. Adding a 1% concentration of BSA to dilute acid pretreated corn stover prior to enzyme addition at 15 FPU/g cellulose enhanced filter paper activity in solution by about a factor of 2 and beta-glucosidase activity in solution by about a factor of 14. Overall, these results suggested that BSA treatment reduced adsorption of cellulase and particularly beta-glucosidase on lignin. Of particular note, BSA treatment of pretreated corn stover solids prior to enzymatic hydrolysis increased 72 h glucose yields from about 82% to about 92% at a cellulase loading of 15 FPU/g cellulose or achieved about the same yield at a loading of 7.5 FPU/g cellulose. Similar improvements were also observed for enzymatic hydrolysis of ammonia fiber explosion (AFEX) pretreated corn stover and Douglas fir treated by SO(2) steam explosion and for simultaneous saccharification and fermentation (SSF) of BSA pretreated corn stover. In addition, BSA treatment prior to hydrolysis reduced the need for beta-glucosidase supplementation of SSF. The results are consistent with non-specific competitive, irreversible adsorption of BSA on lignin and identify promising strategies to reduce enzyme requirements for cellulose hydrolysis. (c) 2006 Wiley Periodicals, Inc.

Pretreatment of wheat straw by nonionic surfactant-assisted dilute acid for enhancing enzymatic hydrolysis and ethanol production.

Science.gov (United States)

Qi, Benkun; Chen, Xiangrong; Wan, Yinhua

2010-07-01

Pretreating wheat straw (WS) with combined use of varied sulfuric acid concentration (0-3%, w/v) and Tween 20 concentration (0-1%) was investigated in an attempt to enhance the hydrolysis and fermentability of pretreated WS. Enzymatic hydrolysis yield of glucan and xylan and ethanol production by simultaneous saccharification and fermentation (SSF) of water-insoluble solids (WIS) were significantly affected by the amount of Tween 20 added during acid pretreatment. Any further addition of Tween 20 in either hydrolysis stage or fermentation stage only led to small increase in glucan conversion and ethanol production. Determination of adsorption of cellulases during hydrolysis showed that Tween 20-assisted acid treated straw solution contained more free cellulases than individual acid treated straw solution, indicating that modification of lignin surface by Tween 20 added during pretreatment likely occurred. In addition, the effects of pretreatment conditions on overall recovery of glucose and xylose after pretreatment and enzymatic hydrolysis were also investigated. Copyright 2010 Elsevier Ltd. All rights reserved.

Enhanced sugar production from pretreated barley straw by additive xylanase and surfactants in enzymatic hydrolysis for acetone-butanol-ethanol fermentation.

Science.gov (United States)

Yang, Ming; Zhang, Junhua; Kuittinen, Suvi; Vepsäläinen, Jouko; Soininen, Pasi; Keinänen, Markku; Pappinen, Ari

2015-01-01

This study aims to improve enzymatic sugar production from dilute sulfuric acid-pretreated barley straw for acetone-butanol-ethanol (ABE) fermentation. The effects of additive xylanase and surfactants (polyethylene glycol [PEG] and Tween) in an enzymatic reaction system on straw hydrolysis yields were investigated. By combined application of 2g/100g dry-matter (DM) xylanase and PEG 4000, the glucose yield was increased from 53.2% to 86.9% and the xylose yield was increased from 36.2% to 70.2%, which were considerably higher than results obtained with xylanase or surfactant alone. The ABE fermentation of enzymatic hydrolysate produced 10.8 g/L ABE, in which 7.9 g/L was butanol. The enhanced sugar production increased the ABE yield from 93.8 to 135.0 g/kg pretreated straw. The combined application of xylanase and surfactants has a large potential to improve sugar production from barley straw pretreated with a mild acid and that the hydrolysate showed good fermentability in ABE production. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enzymatically structured emulsions in simulated gastrointestinal environment: impact on interfacial proteolysis and diffusion in intestinal mucus.

Science.gov (United States)

Macierzanka, Adam; Böttger, Franziska; Rigby, Neil M; Lille, Martina; Poutanen, Kaisa; Mills, E N Clare; Mackie, Alan R

2012-12-18

Fundamental knowledge of physicochemical interactions in the gastrointestinal environment is required in order to support rational designing of protein-stabilized colloidal food and pharmaceutical delivery systems with controlled behavior. In this paper, we report on the colloidal behavior of emulsions stabilized with the milk protein sodium caseinate (Na-Cas), and exposed to conditions simulating the human upper gastrointestinal tract. In particular, we looked at how the kinetics of proteolysis was affected by adsorption to an oil-water interface in emulsion and whether the proteolysis and the emulsion stability could be manipulated by enzymatic structuring of the interface. After cross-linking with the enzyme transglutaminase, the protein was digested with use of an in vitro model of gastro-duodenal proteolysis in the presence or absence of physiologically relevant surfactants (phosphatidylcholine, PC; bile salts, BS). Significant differences were found between the rates of digestion of Na-Cas cross-linked in emulsion (adsorbed protein) and in solution. In emulsion, the digestion of a population of polypeptides of M(r) ca. 50-100 kDa was significantly retarded through the gastric digestion. The persistent interfacial polypeptides maintained the original emulsion droplet size and prevented the system from phase separating. Rapid pepsinolysis of adsorbed, non-cross-linked Na-Cas and its displacement by PC led to emulsion destabilization. These results suggest that structuring of emulsions by enzymatic cross-linking of the interfacial protein may affect the phase behavior of emulsion in the stomach and the gastric digestion rate in vivo. Measurements of ζ-potential revealed that BS displaced the remaining protein from the oil droplets during the simulated duodenal phase of digestion. Diffusion of the postdigestion emulsion droplets through ex vivo porcine intestinal mucus was only significant in the presence of BS due to the high negative charge these

Change in digestibility of gamma-irradiated starch by low temperature cooking

International Nuclear Information System (INIS)

Kume, T.; Ishigaki, I.; Rahman, S.

1988-01-01

Combination effect of irradiation and low temperature cooking on starch digestibility has been investigated as a basic research for application of radiosterilization on starch fermentation. The digestion of corn starch by glucoamylase after cooking at low temperature was enhanced by γ-irradiation and the required cooking temperature was decreased from 75-80 0 C to 65 0 C by 25 kGy. Gelatinization of starches except tapioca starch was enhanced by irradiation and it corresponds to the digestibility. The digestibility of potato starch which has a high viscosity was especially enhanced at low temperature cooking because the viscosity was markedly decreased by irradiation. These results show that the irradiation of starches is useful not only for the sterilization of fermentation broth but also for the enhancement of digestion. (orig.) [de

Change in digestibility of gamma-irradiated starch by low temperature cooking

Energy Technology Data Exchange (ETDEWEB)

Kume, T.; Ishigaki, I.; Rahman, S.

1988-04-01

Combination effect of irradiation and low temperature cooking on starch digestibility has been investigated as a basic research for application of radiosterilization on starch fermentation. The digestion of corn starch by glucoamylase after cooking at low temperature was enhanced by ..gamma..-irradiation and the required cooking temperature was decreased from 75-80/sup 0/C to 65/sup 0/C by 25 kGy. Gelatinization of starches except tapioca starch was enhanced by irradiation and it corresponds to the digestibility. The digestibility of potato starch which has a high viscosity was especially enhanced at low temperature cooking because the viscosity was markedly decreased by irradiation. These results show that the irradiation of starches is useful not only for the sterilization of fermentation broth but also for the enhancement of digestion.

Scale-up of ethanol production from winter barley by the EDGE (enhanced dry grind enzymatic) process in fermentors up to 300 liters

Science.gov (United States)

A fermentation process, which was designated the EDGE (enhanced dry grind enzymatic) process, has recently been developed for barley ethanol production. In the EDGE process, in addition to the enzymes normally required for starch hydrolysis, commercial Beta-glucanases were used to hydrolyze (1,3)(1,...

Effect of Hot-Pressing Temperature on the Subsequent Enzymatic Saccharification and Fermentation Performance of SPORL Pretreated Forest Biomass

Science.gov (United States)

Jingzhi Zhang; Andrea Laguna; Craig Clemons; Michael P. Wolcott; Rolland Gleisner; J.Y. Zhu; Xu Zhang

2015-01-01

Methods to increase the energy density ofbiofuel feedstock for shipment are important towards improving supply chain efficiency in upstream processes. Towards this end, densified pretreated lignocellulosic biomass was produced using hot-pressing. The effects offiber hornification induced by hot-pressing on enzymatic digestibilities of lodgepolepine and poplar NE222...

Integrated microreactor for enzymatic reaction automation: An easy step toward the quality control of monoclonal antibodies.

Science.gov (United States)

Ladner, Yoann; Mas, Silvia; Coussot, Gaelle; Bartley, Killian; Montels, Jérôme; Morel, Jacques; Perrin, Catherine

2017-12-15

The main purpose of the present work is to provide a fully integrated miniaturized electrophoretic methodology in order to facilitate the quality control of monoclonal antibodies (mAbs). This methodology called D-PES, which stands for Diffusion-mediated Proteolysis combined with an Electrophoretic Separation, permits to perform subsequently mAb tryptic digestion and electrophoresis separation of proteolysis products in an automated manner. Tryptic digestion conditions were optimized regarding the influence of enzyme concentration and incubation time in order to achieve similar enzymatic digestion efficiency to that obtained with the classical methodology (off-line). Then, the optimization of electrophoretic separation conditions concerning the nature of background electrolyte (BGE), ionic strength and pH was realized. Successful and repeatable electrophoretic profiles of three mAbs digests (Trastuzumab, Infliximab and Tocilizumab), comparable to the off-line digestion profiles, were obtained demonstrating the feasibility and robustness of the proposed methodology. In summary, the use of the proposed and optimized in-line approach opens a new, fast and easy way for the quality control of mAbs. Copyright © 2017 Elsevier B.V. All rights reserved.

A 3-D Model of a Perennial Ryegrass Primary Cell Wall and Its Enzymatic Degradation

Directory of Open Access Journals (Sweden)

Indrakumar Vetharaniam

2014-05-01

Full Text Available We have developed a novel 3-D, agent-based model of cell-wall digestion to improve our understanding of ruminal cell-wall digestion. It offers a capability to study cell walls and their enzymatic modification, by providing a representation of cellulose microfibrils and non-cellulosic polysaccharides and by simulating their spatial and catalytic interactions with enzymes. One can vary cell-wall composition and the types and numbers of enzyme molecules, allowing the model to be applied to a range of systems where cell walls are degraded and to the modification of cell walls by endogenous enzymes. As a proof of principle, we have modelled the wall of a mesophyll cell from the leaf of perennial ryegrass and then simulated its enzymatic degradation. This is a primary, non-lignified cell wall and the model includes cellulose, hemicelluloses (glucuronoarabinoxylans, 1,3;1,4-β-glucans, and xyloglucans and pectin. These polymers are represented at the level of constituent monosaccharides, and assembled to form a 3-D, meso-scale representation of the molecular structure of the cell wall. The composition of the cell wall can be parameterised to represent different walls in different cell types and taxa. The model can contain arbitrary combinations of different enzymes. It simulates their random diffusion through the polymer networks taking collisions into account, allowing steric hindrance from cell-wall polymers to be modelled. Steric considerations are included when target bonds are encountered, and breakdown products resulting from enzymatic activity are predicted.

Períodos de digestão enzimática para o resgate de folículos pré-antrais em ovários de fetos bovinos Periods of enzimatic digestion for rescueing preantral follicles from fetal bovine ovaries

Directory of Open Access Journals (Sweden)

Silvia Ferreira Carambula

1998-12-01

Full Text Available O presente experimento foi delineado com o objetivo de determinar a influência de diferentes períodos de digestão enzimática nos ovários de fetos bovinos, sobre o número total de folículos pré-antrais resgatados e o número de folículos resgatados em cada estágio de desenvolvimento. A presença da membrana basal, envolvendo o complexo formado pelas células da granulosa e o oócito, foi observada através de um estudo histológico, o que permitiu avaliar a integridade morfológica dos folículos pré-antrais isolados. Para isso, ovários de fetos bovinos, entre 150 e 270 dias de gestação, foram obtidos em frigorífico. No laboratório, os ovários foram seccionados em vários fragmentos com uma tesoura cirúrgica e dissociados com pipetas de Pasteur, com diâmetros aproximados de 1000 e 500mim. Após este processo, os fragmentos foram submetidos à digestão enzimática com colagenase em uma concentração de 200UI/ml de TCM199 modificado, por períodos de 20, 30 ou 40 minutos. Os resultados obtidos com esta técnica permitiram determinar que o número de folículos pré-antrais isolados, assim como, os estágios de desen- volvimento em que esses folículos encontravam-se no momento do isolamento, são similares nos três períodos de incubação enzimática. A estrutura morfológica desses folículos, formada pelo oócito, células da granulosa e membrana basal, manteve-se intacta após o isolamento nos três períodos de digestão enzimática.The objective of this experiment was to determine the efficiency and the effect of different enzymatic digestion periods on the number and developmental stages of preantral follicles isolated from bovine fetal ovaries. The presence of basal membrane, surrounding granulosa cells-oocyte complex, was observed, after digestion, through histologycal study. For this, ovaries from bovine fetues, were collected in slaughterhouses, between 150 and 270 days of pregnancy. In the laboratory, the ovaries

Enzymatic Modification of Corn Starch Influences Human Fecal Fermentation Profiles.

Science.gov (United States)

Dura, Angela; Rose, Devin J; Rosell, Cristina M

2017-06-14

Enzymatically modified starches have been widely used in food applications to develop new products, but information regarding digestion and fecal fermentation of these products is sparse. The objective of this study was to determine the fermentation properties of corn starch modified with α-amylase, amyloglucosidase, or cyclodextrin glycosyltransferase and the possible role of hydrolysis products. Samples differed in their digestibility and availability to be fermented by the microbiota, resulting in differences in microbial metabolites produced during in vitro fermentation. The presence or absence of hydrolysis products and gelatinization affected starch composition and subsequent metabolite production by the microbiota. Amyloglucosidase-treated starch led to the greatest production of short- and branched-chain fatty acid production by the microbiota. Results from this study could be taken into consideration to confirm the possible nutritional claims and potential health benefits of these starches as raw ingredients for food development.

Enhancement of anaerobic digestion of Nile perch fish processing ...

African Journals Online (AJOL)

To overcome these limitations, the effects of co-digestion, physical and biological pretreatments on extent of methane yield were investigated. At a loading ratio of 1:1 (inoculum to substrate) with raw FPW, a methane yield of 0.56 m3/kgVS was obtained. Co-digestion of the residue with 10% gVS of brewery wastewater ...

The enhancement of enzymatic hydrolysis of lignocellulosic substrates by the addition of accessory enzymes such as xylanase: is it an additive or synergistic effect?

Directory of Open Access Journals (Sweden)

Saddler Jack N

2011-10-01

Full Text Available Abstract Background We and other workers have shown that accessory enzymes, such as β-glucosidase, xylanase, and cellulase cofactors, such as GH61, can considerably enhance the hydrolysis effectiveness of cellulase cocktails when added to pretreated lignocellulosic substrates. It is generally acknowledged that, among the several factors that hamper our current ability to attain efficient lignocellulosic biomass conversion yields at low enzyme loadings, a major problem lies in our incomplete understanding of the cooperative action of the different enzymes acting on pretreated lignocellulosic substrates. Results The reported work assessed the interaction between cellulase and xylanase enzymes and their potential to improve the hydrolysis efficiency of various pretreated lignocellulosic substrates when added at low protein loadings. When xylanases were added to the minimum amount of cellulase enzymes required to achieve 70% cellulose hydrolysis of steam pretreated corn stover (SPCS, or used to partially replace the equivalent cellulase dose, both approaches resulted in enhanced enzymatic hydrolysis. However, the xylanase supplementation approach increased the total protein loading required to achieve significant improvements in hydrolysis (an additive effect, whereas the partial replacement of cellulases with xylanase resulted in similar improvements in hydrolysis without increasing enzyme loading (a synergistic effect. The enhancement resulting from xylanase-aided synergism was higher when enzymes were added simultaneously at the beginning of hydrolysis. This co-hydrolysis of the xylan also influenced the gross fiber characteristics (for example, fiber swelling resulting in increased accessibility of the cellulose to the cellulase enzymes. These apparent increases in accessibility enhanced the steam pretreated corn stover digestibility, resulting in three times faster cellulose and xylan hydrolysis, a seven-fold decrease in cellulase loading and

Anaerobic digestion of organic by-products from meat-processing industry. The effect of pre-treatments and co-digestion

Energy Technology Data Exchange (ETDEWEB)

Luste, S.

2011-07-01

Anaerobic digestion is a multi-beneficial biological treatment during which micro-organisms degrade organic material producing biogas (i.e. methane) and stabilised end-product (i.e. digestate). Methane is a versatile renewable energy source and digestate can be used as an organic fertiliser and/or soil improver. Because of the increasing consumption and tightening environment and health legislation, production of organic wastes suitable for anaerobic digestion increases. Animal by-products (ABP) from the meat-processing industry are often rendered (contaminated material), used as feedstock (in fur breeding), or composted. However, ABPs studied could not be utilised in fodder or in animal food production and have currently been rendered or directed to composting, despite being mostly considered unsuitable for composting. Many ABPs are energy-rich, wet and pasty materials and suitable for the anaerobic digestion process. Moreover, suitable pre-treatment to hydrolyse solid materials and/or co-digestion of two or several materials may improve the anaerobic digestion with ultimate goal to increase the methane production, stabilisation and reusability of digestate. The case chosen for more detailed research was that of a middle-sized Finnish meat-processing industry. The aim of the thesis was to evaluate the feasibility of different ABPs presently available for treatment as raw material for anaerobic digestion. Another objective was to enhance the anaerobic digestion process via specific pre-treatments and co-digestion cases with the ultimate aim to increase the methane production and the quality of the digestate. The general goal was to observe the overall process from the perspective of real-circumstances in Finland to rise to needs in practice and to produce exploitable information for adopting sustainable development locally and case-specifically into practice via versatile anaerobic digestion technology. The ABPs studied were highly bio-degradable and especially

Performance of thermophilic anaerobic digesters using inoculum mixes with enhanced methanogenic diversity

KAUST Repository

Ghanimeh, Sophia

2017-05-30

BACKGROUND Reportedly, various mixes of seeds were quasi-randomly selected to startup anaerobic digesters. In contrast, this study examines the impact of inoculating thermophilic anaerobic digesters with a designed mix of non-acclimated seeds based on their methanogen composition, using Quantitative Polymerase Chain Reaction (QPCR) of 16S rRNA gene, to achieve high abundance and diversity of methanogens. RESULTS Based on QPCR results, two seed mixes were selected to inoculate two anaerobic digesters: digester (A) was inoculated with a control seed consisting of digestate, manure, and activated sludge; and digester (B) was inoculated with a further methanogen-enriched seed consisting of the control seed with added compost and leachate. Both seed combinations yielded a balanced microflora that is able to achieve a successful startup. However, upon reaching steady state, digester B exhibited lower propionate levels, resulting in lower VFA concentration and increased buffering capacity, indicating greater stability. Acetotrophs and hydrogenotrophs were dominated by Methanosarcinaceae and Methanobacteriales, respectively, in both digesters, exhibiting an average ratio of 66-to-34% in A and 76-to-24% in B during steady state. CONCLUSION The inoculation strategy in digester B resulted in improved stability, lower propionate concentration and 10% higher relative abundance of acetotrophs.

Ex vivo evaluation of biodegradable poly(ε-caprolactone) films in digestive fluids

Energy Technology Data Exchange (ETDEWEB)

Chang, Hao-Ming [Division of General Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei 100, Taiwan, ROC (China); Prasannan, Adhimoorthy [Graduate Institute of Applied Science and Technology, National Taiwan University of Science and Technology, Taipei 106, Taiwan, ROC (China); Tsai, Hsieh-Chih, E-mail: h.c.tsai@mail.ntust.edu.tw [Graduate Institute of Applied Science and Technology, National Taiwan University of Science and Technology, Taipei 106, Taiwan, ROC (China); Jhu, Jheng-Jun [Graduate Institute of Applied Science and Technology, National Taiwan University of Science and Technology, Taipei 106, Taiwan, ROC (China)

2014-09-15

Graphical abstract: The biodegradable poly(ε-caprolactone) (PCL) films in several digestive glands using films formed from a solution of PCL in dichloromethane and ether. The PCL films degraded very differently in different digestive glands while they continued to undergo hydrolysis. - Highlights: • Hydrolytic and protein enzymatic degradation of PCL films were studied in terms of weight loss, and the degraded films were characterized using SEM, TGA, FTIR, and DSC. • Degradation of the PCL films in saliva and gastric juice was followed by superficial erosion, while the degradation of the PCL films immersed in bile and PBS was followed by bulk erosion. • For all degraded PCL films, degradation began in the amorphous regions of the films and the fraction of crystalline PCL increased slightly. - Abstract: Successful application of biodegradable implants in the gastrointestinal tract requires intensive research to understand their degradation behavior. We studied biodegradable poly(ε-caprolactone) (PCL) films in several digestive glands using films formed from a solution of PCL in dichloromethane and ether. Examination of the film morphology showed that solvent evaporation resulted in the formation of a smooth and flat surface. In addition, our results revealed that the PCL films degraded very differently in different digestive glands while they continued to undergo hydrolysis. There was also significant weight loss in the PCL films in saliva and gastric juice, which indicates that the degradation was sensitive to protein enzymes. Deconstruction of the PCL film was first discovered when it was incubated with saliva for 6 weeks. These results indicate that the degradation by saliva was mainly responsible for the enzymatic erosion of the film, which was confirmed by scanning electron microscopy.

Comparison of sodium carbonate pretreatment for enzymatic hydrolysis of wheat straw stem and leaf to produce fermentable sugars.

Science.gov (United States)

Jin, Yongcan; Huang, Ting; Geng, Wenhui; Yang, Linfeng

2013-06-01

The specific characteristics of biomass structure and chemical composition of straw stem and leaf may result in different behavior of pretreatment and enzymatic hydrolysis. In this work, sodium carbonate (SC) was employed as a pretreatment to improve the enzymatic digestibility of wheat straw. The chemical composition and enzymatic hydrolysis of wheat straw stem and leaf (sheath included) were investigated comparatively. Most of the polysaccharides are kept in the solid fractions after SC pretreatment, while the stem has better delignification selectivity than leaf at high temperature. The enzymatic hydrolysis efficiency of wheat straw leaf is significantly higher than that of stem. The maximum total sugar yield from SC pretreated leaf was about 16% higher than stem. The results show that sodium carbonate is of great potential to be used as a pretreatment for the production of bioethanol from straw handling waste in a straw pulp mill with a low feedstock cost. Copyright © 2013 Elsevier Ltd. All rights reserved.

Psychrophilic anaerobic digestion of guinea pig manure in low-cost tubular digesters at high altitude.

Science.gov (United States)

Garfí, Marianna; Ferrer-Martí, Laia; Villegas, Vidal; Ferrer, Ivet

2011-05-01

Guinea pig is one of the most common livestock in rural communities of the Andes. The aim of this research was to study the anaerobic digestion of guinea pig manure in low-cost unheated tubular digesters at high altitude. To this end, the performance of two pilot digesters was monitored during 7 months; and two greenhouse designs were compared. In the dome roof digester the temperature and biogas production were significantly higher than in the shed roof digester. However, the biogas production rate was low (0.04 m(biogas)(3)m(digester)(-3) d(-1)), which is attributed to the low organic loading rate (0.6 kg(VS)m(digester)(-3)d(-1)) and temperature (23°C) of the system, among other factors. In a preliminary fertilization study, the potato yield per hectare was increased by 100% using the effluent as biofertilizer. Improving manure management techniques, increasing the organic loading rate and co digesting other substrates may be considered to enhance the process. Copyright © 2011 Elsevier Ltd. All rights reserved.

Enhancement of Palm Oil Extraction Using Cell Wall Degrading Enzyme Formulation

International Nuclear Information System (INIS)

Silvamany, H.; Jamaliah Md Jahim

2015-01-01

In this recent work, application of aqueous enzymatic process to enhance recovery of palm oil was studied. Experiments were carried out to investigate the structural carbohydrate composition of oil palm mesocarp (Elaeis guineensis) and to analyze the effect of different combination of enzymes on the palm oil recovery and degree of digestibility and the respective correlation. The optimum combination of enzymes comprising of Cellic CTec2 (X 1 ), Cellic HTec2 (X 2 ) and Pectinex Ultra SP-L (X 3 ) for Aqueous Enzymatic Oil Extraction Process (AEOEP), were determined using Simplex Lattice mixture design under fixed parameters. Maximum oil recovery of 88 % was achieved with ratio of enzymes at 0.46: 0.34: 0.2 (X 1 :X 2 :X 3 ), at enzyme loading of 30 mg protein/ 10 g substrate, substrate loading of 50 % w/v, pH 4.8, and 2 hours of incubation at 50 degree Celsius. The conversion of reducing sugar at corresponding condition was measured to evaluate the effectiveness of enzymes in degrading fruit cell wall releasing trapped oil. Moreover, transmission electron microscopy (TEM) was utilized to indicate the increase in cell wall disintegration leading to higher release of oil with enzymatic treatment. (author)

Radiation pretreatments of cellulose materials for the enhancement of enzymatic hydrolysis

International Nuclear Information System (INIS)

Ait Si Mamar, S.; Hadjadj, A.

1990-01-01

The conversion of wheat straw agricultural cellulosic wastes to reducing sugars and glucose has been studied by pretreatments by acid hydrolysis and gamma radiolysis over the dose 0-2 MGy. The pretreatment of cellulosic wastes by gamma radiolysis in the presence of sulfuric acid solution shows that the reducing sugars yield increases with the irradiation dose. The effect of radiation degradation on cellulosic wastes between 0.1 MGy and 2 MGy shows the glucose and reducing sugars yields after enzymatic hydrolysis by cellulase vary with the dose. In the relatively low dose range, up to about 0.5 MGy, the reducing sugars yields vary slightly. For an acid hydrolysis followed by radiation at dose range below 0.5 MGy the reducing sugars yields are practically insensitive to radiation. On the other hand, the pretreatment by radiation in higher dose range from 0.5 to 2 MGy followed by enzymatic hydrolysis is effective for the conversion of cellulosic wastes into glucose. The radiation induced degradation of cellulose into glucose depends on the type of acid hydrolysis and on the enzymatic hydrolysis time by cellulase. Pre-irradiation in air is more effective than in acid solution. (author)

Radiation pretreatments of cellulose materials for the enhancement of enzymatic hydrolysis

Science.gov (United States)

Mamar, S. Ait Si; Hadjadj, A.

The conversion of wheat straw agricultural cellulosic wastes to reduning sugars and glucose has been studied by pretreatments by acid hydrolysis and gamma radiolysis over the dose 0-2 MGy. The pretreatment of cellulosic wastes by gamma radiolysis in the presence of sulfuric acid solution shows that the reducing sugars yield increases with the irradiation dose. The effect of radiation degradation on cellulosic wastes between 0.1 MGy and 2 MGy shows the glucose and reducing sugars yields after enzymatic hydrolysis by cellulase vary with the dose. In the relatively low dose range, up to about 0.5 MGy, the reducing sugars yields vary slightly. For an acid hydrolysis followed by radiation at dose range below 0.5 MGy the reducing sugars yields are practically insensitive to radiation. On the other hand, the pretreatment by radiation in higher dose range from 0.5 to 2 MGy followed by enzymatic hydrolysis is effective for the conversion of cellulosic wastes into glucose. The radiation induced degradation of cellulose into glucose depends on the type of acid hydrolysis and on the enzymatic hydrolysis time by cellulase. Pre-irradiation in air is more effective than in acid solution.

Saliva and gastrointestinal functions of mastication, taste and textural perception, swallowing and initial digestion

DEFF Research Database (Denmark)

Pedersen, Anne Marie Lynge; Sørensen, Christiane E; Proctor, Gordon

2018-01-01

pattern. This review paper provides insight into the mechanisms by which saliva acts in relation to taste, mastication, bolus formation, enzymatic digestion, and swallowing. Also the protective functions of saliva including maintenance of dental and mucosal integrity will be discussed as they indirectly......Saliva exerts multiple functions in relation to the initial digestive processes taking place in the upper parts of the gastrointestinal tract. Ingestion of food and beverages, in turn, is a strong stimulus for secretion of saliva with a differential composition depending on the neuronal stimulation...... influence the digestive process. The final part of this paper focuses on the implications of xerostomia and salivary gland dysfunction on gastrointestinal functions. This article is protected by copyright. All rights reserved....

Effects of alkaline or liquid-ammonia treatment on crystalline cellulose: changes in crystalline structure and effects on enzymatic digestibility

Directory of Open Access Journals (Sweden)

Himmel Michael E

2011-10-01

Full Text Available Abstract Background In converting biomass to bioethanol, pretreatment is a key step intended to render cellulose more amenable and accessible to cellulase enzymes and thus increase glucose yields. In this study, four cellulose samples with different degrees of polymerization and crystallinity indexes were subjected to aqueous sodium hydroxide and anhydrous liquid ammonia treatments. The effects of the treatments on cellulose crystalline structure were studied, in addition to the effects on the digestibility of the celluloses by a cellulase complex. Results From X-ray diffractograms and nuclear magnetic resonance spectra, it was revealed that treatment with liquid ammonia produced the cellulose IIII allomorph; however, crystallinity depended on treatment conditions. Treatment at a low temperature (25°C resulted in a less crystalline product, whereas treatment at elevated temperatures (130°C or 140°C gave a more crystalline product. Treatment of cellulose I with aqueous sodium hydroxide (16.5 percent by weight resulted in formation of cellulose II, but also produced a much less crystalline cellulose. The relative digestibilities of the different cellulose allomorphs were tested by exposing the treated and untreated cellulose samples to a commercial enzyme mixture (Genencor-Danisco; GC 220. The digestibility results showed that the starting cellulose I samples were the least digestible (except for corn stover cellulose, which had a high amorphous content. Treatment with sodium hydroxide produced the most digestible cellulose, followed by treatment with liquid ammonia at a low temperature. Factor analysis indicated that initial rates of digestion (up to 24 hours were most strongly correlated with amorphous content. Correlation of allomorph type with digestibility was weak, but was strongest with cellulose conversion at later times. The cellulose IIII samples produced at higher temperatures had comparable crystallinities to the initial cellulose I

Efficient Anaerobic Digestion of Microalgae Biomass: Proteins as a Key Macromolecule.

Science.gov (United States)

Magdalena, Jose Antonio; Ballesteros, Mercedes; González-Fernandez, Cristina

2018-05-06

Biogas generation is the least complex technology to transform microalgae biomass into bioenergy. Since hydrolysis has been pointed out as the rate limiting stage of anaerobic digestion, the main challenge for an efficient biogas production is the optimization of cell wall disruption/hydrolysis. Among all tested pretreatments, enzymatic treatments were demonstrated not only very effective in disruption/hydrolysis but they also revealed the impact of microalgae macromolecular composition in the anaerobic process. Although carbohydrates have been traditionally recognized as the polymers responsible for the low microalgae digestibility, protease addition resulted in the highest organic matter solubilization and the highest methane production. However, protein solubilization during the pretreatment can result in anaerobic digestion inhibition due to the release of large amounts of ammonium nitrogen. The possible solutions to overcome these negative effects include the reduction of protein biomass levels by culturing the microalgae in low nitrogen media and the use of ammonia tolerant anaerobic inocula. Overall, this review is intended to evidence the relevance of microalgae proteins in different stages of anaerobic digestion, namely hydrolysis and methanogenesis.

Characterisation of selenium compounds in rye seedling biomass using {sup 75}Se-labelling/SDS-PAGE separation/{gamma}-scintillation counting, and HPLC-ICP-MS analysis of a range of enzymatic digests

Energy Technology Data Exchange (ETDEWEB)

Bryszewska, Malgorzata A. [Technical University of Lodz (Poland). Institute of General Food Chemistry; Ambroziak, Wojciech [Technical University of Lodz (Poland). Institute of Fermentation Technology and Microbiology; Rudzinski, Juliusz [Technical University of Lodz (Poland). Institute of Applied Radiation Chemistry; Lewis, D. John [Central Science Laboratory, York (United Kingdom)

2005-07-01

In the present study, selenium-enriched plant biomass was investigated to evaluate the ability of rye seedlings to take up, and assimilate, inorganic selenium. Two different analytical approaches were used. Electrophoretic separation (SDS-PAGE) of proteins extracted from {sup 75}Se-labelled biomass was used to investigate the biotransformation of selenite into organic forms of the element. Ion-pair chromatography coupled with ICP-MS detection was chosen for the analysis of selenium species, enzymatically extracted from the plant biomass. The results of three enzymatic hydrolysis procedures and three sequential enzymatic extractions procedures are compared. The most effective single extraction was proteolysis (using protease type XIV), giving an overall extraction efficiency of 48%. However, for combinations of enzymes, the most effective was cellulase (Trichoderma viride) followed by sequential extraction of the solid pellet using protease type XIV, giving an extraction efficiency of 70%. The complementary data from the electrophoretic fractionation of proteins, and the HPLC separation of Se-species in the proteolytic digests, reveal the existence of large number of selenium-containing compounds in the rye seedling plant biomass. The results showed the complete biotransformation of inorganic selenium into organic forms during germination of the rye seedlings. HPLC-ICP-MS analysis of extracts from the plant biomass did not show the presence of selenate or selenite. At the time of this study, the lack of suitable organic-MS facilities meant that it was not possible to characterise them fully. However, the data does show that a combination of different enzymes, rather than just the commonly-used protease, should be considered when developing an extraction strategy for selenium in different food types to those already reported in the literature. (orig.)

Chemically modified, immobilized trypsin reactor with improved digestion efficiency

NARCIS (Netherlands)

Freije, J.R.; Mulder, P.P.; Werkman, W.; Rieux, L.; Niederlander, H.A G; Verpoorte, Sabeth; Bischoff, Rainer

2005-01-01

Tryptic digestion followed by identification using mass spectrometry is an important step in many proteomic studies. Here, we describe the preparation of immobilized, acetylated trypsin for enhanced digestion efficacy in integrated protein analysis platforms. Complete digestion of cytochrome c was

Hydrothermal treatment and enzymatic hydrolysis of Tamarix ramosissima: evaluation of the process as a conversion method in a biorefinery concept.

Science.gov (United States)

Xiao, Ling-Ping; Shi, Zheng-Jun; Xu, Feng; Sun, Run-Cang

2013-05-01

The present work investigated the effects of hydrothermal treatment (HTT) of Tamarix ramosissima by determination of sugar and inhibitor formation in the liquid fraction, and chemical and morphological changes of the pretreated solid material coupled with an evaluation of enzymatic hydrolysis. HTT was carried out in a batch reactor system at a maximal temperature (TMAX 180-240 °C) and evaluated for severities logRo ranging from 2.40 to 4.17. The liquid fractions were analyzed by HPLC, GPC, and GC-MS. The morphology and composition of the solid residues were characterized using an array of techniques, such as SEM, XRD, BET surface area, and CP/MAS (13)C NMR. Using a variety of tools, we have developed a better understanding of how HTT process affects biomass structure and cellulose properties that impact on its digestibility. These results provided new insights into the factors limiting enzymatic digestibility and mechanism of biomass deconstruction during hydrothermal process. Crown Copyright © 2012. Published by Elsevier Ltd. All rights reserved.

Effect of enzymes on anaerobic digestion of primary sludge and septic tank performance.

Science.gov (United States)

Diak, James; Örmeci, Banu; Kennedy, Kevin J

2012-11-01

Enzyme additives are believed to improve septic tank performance by increasing the hydrolysis and digestion rates and maintaining a healthy microbial population. Previous studies reported mixed results on the effectiveness of enzymes on mesophilic and thermophilic digestion, and it is not clear whether enzymes would be effective under septic tank conditions where there is no heating or mixing, quantities of enzymes added are small, and they can be washed out quickly. In this study, batch reactors and continuous-flow reactors designed and operated as septic tanks were used to evaluate whether enzymatic treatment would increase the hydrolysis and digestion rates in primary sludge. Total solids, volatile solids, total suspended solids, total and soluble chemical oxygen demand, concentrations of protein, carbohydrate, ammonia and volatile acids in sludge and effluent samples were measured to determine the differences in digestion rates in the presence and absence of enzymes. Overall, no significant improvement was observed in enzyme-treated reactors compared with the control reactors.

Synergistic effect of co-digestion to enhance anaerobic degradation of catering waste and orange peel for biogas production.

Science.gov (United States)

Anjum, Muzammil; Khalid, Azeem; Qadeer, Samia; Miandad, Rashid

2017-09-01

Catering waste and orange peel were co-digested using an anaerobic digestion process. Orange peel is difficult to degrade anaerobically due to the presence of antimicrobial agents such as limonene. The present study aimed to examine the feasibility of anaerobic co-digestion of catering waste with orange peel to provide the optimum nutrient balance with reduced inhibitory effects of orange peel. Batch experiments were conducted using catering waste as a potential substrate mixed in varying ratios (20-50%) with orange peel. Similar ratios were followed using green vegetable waste as co-substrate. The results showed that the highest organic matter degradation (49%) was achieved with co-digestion of catering waste and orange peel at a 50% mixing ratio (CF4). Similarly, the soluble chemical oxygen demand (sCOD) was increased by 51% and reached its maximum value (9040 mg l -1 ) due to conversion of organic matter from insoluble to soluble form. Biogas production was increased by 1.5 times in CF4 where accumulative biogas was 89.61 m 3 t -1 substrate compared with 57.35 m 3 t -1 substrate in the control after 80 days. The main reason behind the improved biogas production and degradation is the dilution of inhibitory factors (limonene), with subsequent provision of balanced nutrients in the co-digestion system. The tCOD of the final digestate was decreased by 79.9% in CF4, which was quite high as compared with 68.3% for the control. Overall, this study revealed that orange peel waste is a highly feasible co-substrate for anaerobic digestion with catering waste for enhanced biogas production.

Anaerobic Co-digestion for Enhanced Renewable Energy and Green House Gas Emission Reduction

Energy Technology Data Exchange (ETDEWEB)

Navaratnam, Navaneethan; Zitomer, Daniel

2012-05-01

The need to develop renewable energy is important for replacing fossil fuel, which is limited in quantity and also tends to increase in price over time. The addition of high strength organic wastes in municipal anaerobic digesters is growing and tends to increase renewable energy production. In addition, conversion of wastes to energy significantly reduces uncontrolled greenhouse gas emissions. Co-digestion of municipal sludge with any combination of wastes can result in synergistic, antagonistic or neutral outcomes. The objectives of this study were to identify potential co-digestates, determine synergistic, antagonistic and neutral effects, determine economic benefits, quantify performance of bench scale co-digesters, identify influence of co-digestion on microbial communities and implement appropriate co-digestion, if warranted, after full-scale testing. A market study was used to identify promising co-digestates. Most promising wastes were determined by biochemical methane potential (BMP) and other testing followed by a simple economic analysis. Performance was investigated using bench-scale digesters receiving synthetic primary sludge with and without co-digestates. Denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction (qPCR) analyses were performed on the gene encoding the α subunit of methyl coenzyme M reductase (mcrA) to compare methanogen communities among the digesters. One significant band contributing to the greatest difference in banding patterns was excised, cloned, amplified and sequenced. Full- scale co-digestion was conducted using the most promising co-digestate at South Shore Wastewater Reclamation Facility (Oak Creek, WI). Over 80 wastes were identified from 54 facilities within 160 km of an existing municipal digester. A simple economic comparison identified the greatest benefits for seven co-digestates. Methane production rates of two co- digester systems increased by 105% and 66% in comparison to a control

Comparative efficacy of up to 50% partial fish meal replacement with fermented soybean meal or enzymatically prepared soybean meal on growth performance, nutrient digestibility and fecal microflora in weaned pigs.

Science.gov (United States)

Jeong, Jin Suk; Kim, In Ho

2015-06-01

This study was conducted to determine the comparative efficacy of partial fish meal (FM) replacement (up to 50%) with fermented soybean meal (FSBM; SoELAB, PepSoyGen and Soytide) or enzymatically prepared SBM (HP 300) on growth performance, nutrient digestibility and fecal microflora in weaned pigs. A total of 100 weaned pigs (body weight 6.59 ± 0.29 kg) were used in experimental feeding trials, lasting for up to 6 weeks, and were randomly allotted to five groups with four block replicates of five pigs per pen serving as one block. Dietary treatments were as follows: (i) 100% FM, (ii) 50% FM + 50% SoELAB-54, (iii) 50% FM + 50% PepSoyGen, (iv) 50% FM + 50% Soytide and (v) 50% FM + 50% HP 300. Concerning growth performance, none of the treated SBM preparations demonstrated any significantly different effect compared with FM treatment. With respect to nutrient digestibility, SoELAB and HP 300 treatments demonstrated no significant difference compared with FM treatment. Lastly, none of the SBM preparations demonstrated any significant differences in animal fecal score and all of the differentially treated SBM increased fecal Lactobacillus counts, while maintaining similar Escherichia coli counts compared with FM treatment. © 2014 Japanese Society of Animal Science.

Enzymatic and Structural Characterization of the Major Endopeptidase in the Venus Flytrap Digestion Fluid*

Science.gov (United States)

Risør, Michael W.; Thomsen, Line R.; Sanggaard, Kristian W.; Nielsen, Tania A.; Thøgersen, Ida B.; Lukassen, Marie V.; Rossen, Litten; Garcia-Ferrer, Irene; Guevara, Tibisay; Scavenius, Carsten; Meinjohanns, Ernst; Gomis-Rüth, F. Xavier; Enghild, Jan J.

2016-01-01

Carnivorous plants primarily use aspartic proteases during digestion of captured prey. In contrast, the major endopeptidases in the digestive fluid of the Venus flytrap (Dionaea muscipula) are cysteine proteases (dionain-1 to -4). Here, we present the crystal structure of mature dionain-1 in covalent complex with inhibitor E-64 at 1.5 Å resolution. The enzyme exhibits an overall protein fold reminiscent of other plant cysteine proteases. The inactive glycosylated pro-form undergoes autoprocessing and self-activation, optimally at the physiologically relevant pH value of 3.6, at which the protective effect of the pro-domain is lost. The mature enzyme was able to efficiently degrade a Drosophila fly protein extract at pH 4 showing high activity against the abundant Lys- and Arg-rich protein, myosin. The substrate specificity of dionain-1 was largely similar to that of papain with a preference for hydrophobic and aliphatic residues in subsite S2 and for positively charged residues in S1. A tentative structure of the pro-domain was obtained by homology modeling and suggested that a pro-peptide Lys residue intrudes into the S2 pocket, which is more spacious than in papain. This study provides the first analysis of a cysteine protease from the digestive fluid of a carnivorous plant and confirms the close relationship between carnivorous action and plant defense mechanisms. PMID:26627834

The enzymatic activity of 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (ATIC) is enhanced by NPM-ALK

DEFF Research Database (Denmark)

Boccalatte, Francesco E; Voena, Claudia; Riganti, Chiara

2009-01-01

. A well-defined set of ALK-associated tyrosine phospho-peptides, including metabolic enzymes, kinases, ribosomal and cytoskeletal proteins was identified. Validation studies confirmed that VASP and ATIC associated with NPM-ALK and their phosphorylation required ALK activity. ATIC phosphorylation was also...... documented in cell lines and primary tumors carrying ALK proteins and other tyrosine kinases, including TPR-Met and wild type c-Met. Functional analyses revealed that ALK-mediated ATIC phosphorylation enhanced its enzymatic activity, dampering the methotrexate-mediated transformylase activity inhibition...

Genetic loci simultaneously controlling lignin monomers and biomass digestibility of rice straw.

Science.gov (United States)

Hu, Zhen; Zhang, Guifen; Muhammad, Ali; Samad, Rana Abdul; Wang, Youmei; Walton, Jonathan D; He, Yuqing; Peng, Liangcai; Wang, Lingqiang

2018-02-26

Lignin content and composition are crucial factors affecting biomass digestibility. Exploring the genetic loci simultaneously affecting lignin-relevant traits and biomass digestibility is a precondition for lignin genetic manipulation towards energy crop breeding. In this study, a high-throughput platform was employed to assay the lignin content, lignin composition and biomass enzymatic digestibility of a rice recombinant inbred line population. Correlation analysis indicated that the absolute content of lignin monomers rather than lignin content had negative effects on biomass saccharification, whereas the relative content of p-hydroxyphenyl unit and the molar ratio of p-hydroxyphenyl unit to guaiacyl unit exhibited positive roles. Eight QTL clusters were identified and four of them affecting both lignin composition and biomass digestibility. The additive effects of clustered QTL revealed consistent relationships between lignin-relevant traits and biomass digestibility. Pyramiding rice lines containing the above four positive alleles for increasing biomass digestibility were selected and showed comparable lignin content, decreased syringyl or guaiacyl unit and increased molar percentage of p-hydroxyphenyl unit, the molar ratio of p-hydroxyphenyl unit to guaiacyl unit and sugar releases. More importantly, the lodging resistance and eating/cooking quality of pyramiding lines were not sacrificed, indicating the QTL information could be applied to select desirable energy rice lines.

Effects of Chain Length and Degree of Unsaturation of Fatty Acids on Structure and in Vitro Digestibility of Starch-Protein-Fatty Acid Complexes.

Science.gov (United States)

Zheng, Mengge; Chao, Chen; Yu, Jinglin; Copeland, Les; Wang, Shuo; Wang, Shujun

2018-02-28

The effects of chain length and degree of unsaturation of fatty acids (FAs) on structure and in vitro digestibility of starch-protein-FA complexes were investigated in model systems. Studies with the rapid visco analyzer (RVA) showed that the formation of ternary complex resulted in higher viscosities than those of binary complex during the cooling and holding stages. The results of differential scanning calorimetry (DSC), Raman, and X-ray diffraction (XRD) showed that the structural differences for ternary complexes were much less than those for binary complexes. Starch-protein-FA complexes presented lower in vitro enzymatic digestibility compared with starch-FAs complexes. We conclude that shorter chain and lower unsaturation FAs favor the formation of ternary complexes but decrease the thermal stability of these complexes. FAs had a smaller effect on the ordered structures of ternary complexes than on those of binary complexes and little effect on enzymatic digestibility of both binary and ternary complexes.

Enzymatic production of pectic oligosaccharides from onion skins.

Science.gov (United States)

Babbar, Neha; Baldassarre, Stefania; Maesen, Miranda; Prandi, Barbara; Dejonghe, Winnie; Sforza, Stefano; Elst, Kathy

2016-08-01

Onion skins are evaluated as a new raw material for the enzymatic production of pectic oligosaccharides (POS) with a targeted degree of polymerization (DP). The process is based on a two-stage process consisting of a chelator-based crude pectin extraction followed by a controlled enzymatic hydrolysis. Treatment of the extracted crude onion skin's pectin with various enzymes (EPG-M2, Viscozyme and Pectinase) shows that EPG-M2 is the most appropriate enzyme for tailored POS production. The experiments reveal that the highest amount of DP2 and DP3 is obtained at a time scale of 75-90min with an EPG-M2 concentration of 26IU/mL. At these conditions the production amounts 2.5-3.0% (w/w) d.m for DP2 and 5.5-5.6% (w/w) d.m for DP3 respectively. In contrast, maximum DP4 production of 5.2-5.5% (w/w) d.m. is obtained with 5.2IU/mL at a time scale of 15-30min. Detailed LC-MS analysis reveals the presence of more methylated oligomers compared to acetylated forms in the digests. Copyright © 2016 Elsevier Ltd. All rights reserved.

Starch digestibility and predicted glycemic index of fried sweet potato cultivars

Directory of Open Access Journals (Sweden)

Amaka Odenigbo

2012-07-01

Full Text Available Background: Sweet potato (Ipomoea batatas L. is a very rich source of starch. There is increased interest in starch digestibility and the prevention and management of metabolic diseases.Objective: The aim of this study was to evaluate the levels of starch fractions and predicted glycemic index of different cultivars of sweet potato. Material and Method: French fries produced from five cultivars of sweet potato (‘Ginseng Red’, ‘Beauregard’, ‘White Travis’, ‘Georgia Jet clone #2010’ and ‘Georgia Jet’ were used. The level of total starch (TS, resistant starch (RS, digestible starch (DS, and starch digestion index starch digestion index in the samples were evaluated. In vitro starch hydrolysis at 30, 90, and 120 min were determined enzymatically for calculation of rapidly digestible starch (RDS, predicted glycemic index (pGI and slowly digestible starch (SDS respectively. Results: The RS content in all samples had an inversely significant correlation with pGI (-0.52; P<0.05 while RDS had positive and significant influence on both pGI (r=0.55; P<0.05 and SDI (r= 0.94; P<0.01. ‘White Travis’ and ‘Ginseng Red’ had higher levels of beneficial starch fractions (RS and SDS with low pGI and starch digestion Index (SDI, despite their higher TS content. Generally, all the cultivars had products with low to moderate GI values. Conclusion: The glycemic index of these food products highlights the health promoting characteristics of sweet potato cultivars.

Kinetic study of batch and fed-batch enzymatic saccharification of pretreated substrate and subsequent fermentation to ethanol

Directory of Open Access Journals (Sweden)

Gupta Rishi

2012-03-01

Full Text Available Abstract Background Enzymatic hydrolysis, the rate limiting step in the process development for biofuel, is always hampered by its low sugar concentration. High solid enzymatic saccharification could solve this problem but has several other drawbacks such as low rate of reaction. In the present study we have attempted to enhance the concentration of sugars in enzymatic hydrolysate of delignified Prosopis juliflora, using a fed-batch enzymatic hydrolysis approach. Results The enzymatic hydrolysis was carried out at elevated solid loading up to 20% (w/v and a comparison kinetics of batch and fed-batch enzymatic hydrolysis was carried out using kinetic regimes. Under batch mode, the actual sugar concentration values at 20% initial substrate consistency were found deviated from the predicted values and the maximum sugar concentration obtained was 80.78 g/L. Fed-batch strategy was implemented to enhance the final sugar concentration to 127 g/L. The batch and fed-batch enzymatic hydrolysates were fermented with Saccharomyces cerevisiae and ethanol production of 34.78 g/L and 52.83 g/L, respectively, were achieved. Furthermore, model simulations showed that higher insoluble solids in the feed resulted in both smaller reactor volume and shorter residence time. Conclusion Fed-batch enzymatic hydrolysis is an efficient procedure for enhancing the sugar concentration in the hydrolysate. Restricting the process to suitable kinetic regimes could result in higher conversion rates.

Kinetic study of batch and fed-batch enzymatic saccharification of pretreated substrate and subsequent fermentation to ethanol

Science.gov (United States)

2012-01-01

Background Enzymatic hydrolysis, the rate limiting step in the process development for biofuel, is always hampered by its low sugar concentration. High solid enzymatic saccharification could solve this problem but has several other drawbacks such as low rate of reaction. In the present study we have attempted to enhance the concentration of sugars in enzymatic hydrolysate of delignified Prosopis juliflora, using a fed-batch enzymatic hydrolysis approach. Results The enzymatic hydrolysis was carried out at elevated solid loading up to 20% (w/v) and a comparison kinetics of batch and fed-batch enzymatic hydrolysis was carried out using kinetic regimes. Under batch mode, the actual sugar concentration values at 20% initial substrate consistency were found deviated from the predicted values and the maximum sugar concentration obtained was 80.78 g/L. Fed-batch strategy was implemented to enhance the final sugar concentration to 127 g/L. The batch and fed-batch enzymatic hydrolysates were fermented with Saccharomyces cerevisiae and ethanol production of 34.78 g/L and 52.83 g/L, respectively, were achieved. Furthermore, model simulations showed that higher insoluble solids in the feed resulted in both smaller reactor volume and shorter residence time. Conclusion Fed-batch enzymatic hydrolysis is an efficient procedure for enhancing the sugar concentration in the hydrolysate. Restricting the process to suitable kinetic regimes could result in higher conversion rates. PMID:22433563

Understanding the effects of lignosulfonate on enzymatic saccharification of pure cellulose

Science.gov (United States)

Hongming Lou; Haifeng Zhou; Xiuli Li; Mengxia Wang; J.Y. Zhu; Xueqing Qiu

2014-01-01

The effects of lignosulfonate (LS) on enzymatic saccharification of pure cellulose were studied. Four fractions of LS with different molecular weight (MW) prepared by ultrafiltration of a commercial LS were applied at different loadings to enzymatic hydrolysis of Whatman paper under different pH. Using LS fractions with low MW and high degree of sulfonation can enhance...

The challenges of anaerobic digestion and the role of biochar in optimizing anaerobic digestion.

Science.gov (United States)

Fagbohungbe, Michael O; Herbert, Ben M J; Hurst, Lois; Ibeto, Cynthia N; Li, Hong; Usmani, Shams Q; Semple, Kirk T

2017-03-01

Biochar, like most other adsorbents, is a carbonaceous material, which is formed from the combustion of plant materials, in low-zero oxygen conditions and results in a material, which has the capacity to sorb chemicals onto its surfaces. Currently, research is being carried out to investigate the relevance of biochar in improving the soil ecosystem, digestate quality and most recently the anaerobic digestion process. Anaerobic digestion (AD) of organic substrates provides both a sustainable source of energy and a digestate with the potential to enhance plant growth and soil health. In order to ensure that these benefits are realised, the anaerobic digestion system must be optimized for process stability and high nutrient retention capacity in the digestate produced. Substrate-induced inhibition is a major issue, which can disrupt the stable functioning of the AD system reducing microbial breakdown of the organic waste and formation of methane, which in turn reduces energy output. Likewise, the spreading of digestate on land can often result in nutrient loss, surface runoff and leaching. This review will examine substrate inhibition and their impact on anaerobic digestion, nutrient leaching and their environmental implications, the properties and functionality of biochar material in counteracting these challenges. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effects of surface proteins and lipids on molecular structure, thermal properties, and enzymatic hydrolysis of rice starch

Directory of Open Access Journals (Sweden)

Pan HU

Full Text Available Abstract Rice starches with different amylose contents were treated with sodium dodecyl sulfate (SDS to deplete surface proteins and lipids, and the changes in molecular structure, thermal properties, and enzymatic hydrolysis were evaluated. SDS treatment did not significantly change the molecular weight distribution, crystalline structure, short-range ordered degree, and gelatinization properties of starch, but significantly altered the pasting properties and increased the swelling power of starch. The removal of surface proteins and lipids increased the enzymatic hydrolysis and in vitro digestion of starch. The influences of removing surface proteins and lipids from starch on swelling power, pasting properties, and enzymatic hydrolysis were different among the various starches because of the differences in molecular structures of different starch styles. The aforementioned results indicated that removing the surface proteins and lipids from starch did not change the molecular structure but had significant effects on some functional properties.

Trypsin immobilization in ordered porous polymer membranes for effective protein digestion

International Nuclear Information System (INIS)

Qiao, Juan; Kim, Jin Yong; Wang, Yuan Yuan; Qi, Li; Wang, Fu Yi; Moon, Myeong Hee

2016-01-01

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.

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.

Iron uptake by Caco-2 cells following in vitro digestion: effects of heat treatments of pork meat and pH of the digests.

Science.gov (United States)

Sørensen, Anne D; Bukhave, Klaus

2010-10-01

The present in vitro studies report on iron uptake by Caco-2 cells from pepsin and pepsin+pancreatin-digested pork meat proteins at pH values between 4.6 and 7 mimicking conditions in the duodenum and the proximal jejunum, respectively. Heat treatment of the pork meat resulted in increased iron uptake from pepsin-digested samples to Caco-2 cells at pH 4.6. The major enhancing effects on iron uptake by Caco-2 cells were observed after pepsin digestion in the pH range 4.6-6.0, whereas the pepsin+pancreatin-digested samples resulted in negligible iron uptake in Caco-2 cells at pH 7. Thus, the results emphasize the importance of separating pepsin-digested and pepsin+pancreatin-digested proteins during in vitro studies on iron availability. Furthermore, the present results showed the pH dependency of iron uptake anticipated. The enhancing effect of ascorbic acid was verified by increased iron uptake from pepsin-digested pork meat samples at pH 4.6, while no effect of ascorbic acid was observed at pH 7 in pepsin+pancreatin-digested samples. Copyright © 2010 Elsevier GmbH. All rights reserved.

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.

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

Science.gov (United States)

Pečová, M.; Šebela, M.; Marková, Z.; Poláková, K.; Čuda, J.; Šafářová, K.; Zbořil, R.

2013-03-01

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

Comparative effects of undigested and anaerobically digested ...

African Journals Online (AJOL)

EJIRO

factors investigated. Generally, results in respect of crops treated with digested manure, were quite ... conclusion, digested poultry manure enhanced the growth characteristics of the treated plants for the ... values of animal waste production range from 144 million ..... Ph.D thesis, Department of Crop, Soil and Pest Manage.,.

Mussel-inspired co-deposition to enhance bisphenol A removal in a bifacial enzymatic membrane reactor

DEFF Research Database (Denmark)

Cao, Xiaotong; Luo, Jianquan; Woodley, John M.

2018-01-01

were used as the matrix to further exploit the potential of the biocatalytic membranes. such prepared biocatalytic membranes were enzymatically active on both sides, making it possible to construct a bifacial enzymatic membrane reactor (EMR) for highly efficient micro-pollutants removal (taking...

Enhancement of enzymatic hydrolysis of wheat straw by gamma irradiation–alkaline pretreatment

International Nuclear Information System (INIS)

Yin, Yanan; Wang, Jianlong

2016-01-01

Pretreatment of wheat straw with gamma irradiation and NaOH was performed to enhance the enzymatic hydrolysis of wheat straw for production of reducing sugar. The results showed that the irradiation of wheat straw at 50 kGy decreased the yield of reducing sugar, however, the reducing sugar yield increased with increasing dose from 50 kGy to 400 kGy. The irradiation of wheat straw at 100 kGy can significantly decrease NaOH consumption and treatment time. The reducing sugar yield could reach 72.67% after irradiation at 100 kGy and 2% NaOH treatment for 1 h. The combined pretreatment of wheat straw by gamma radiation and NaOH immersion can increase the solubilization of hemicellulose and lignin as well as the accessible surface area for enzyme molecules. - Highlights: • Pretreatment of wheat straw by gamma radiation and NaOH was investigated. • Irradiation pretreatment can significantly decrease NaOH consumption. • Reducing sugar yield reached 72.67% at 100 kGy and 2% NaOH treatment for 1 h.

Digestive enzyme ratios are good indicators of hatchling yolk reserve and digestive gland maturation in early life stages of cuttlefish Sepia officinalis L.: application of these new tools in ecology and aquaculture.

Science.gov (United States)

Safi, Georges; Martinez, A S; Le Pabic, C; Le Bihan, E; Robin, J P; Koueta, N

2018-01-01

In Sepia officinalis (Linnaeus, 1758), the digestive gland matures during the first month post-hatching, while a shift from intracellular acid to extracellular alkaline digestion occurs. The purpose of this study was to investigate the possibility of using enzymatic ratios for the description of digestive system maturation in early life stages of S. officinalis. Second, it is intended to apply these new tools as eco-physiological indicators for understanding the impact of cuttlefish eggs' life history from different spawning sites of the English Channel on digestive performance of juveniles. An experimental rearing was performed over 35 days after hatching (DAH) on juveniles from wild collected eggs in 2010 and 2011. Four digestive enzyme activities and their ratios [i.e., trypsin, cathepsin, acid (ACP), and alkaline (ALP) phosphatase, ALP/ACP, and trypsin/cathepsin] were studied along with histological features (e.g., internal yolk surface and digestive gland development). The two enzyme ratios were good indicators of digestive system maturation allowing the study of the digestive gland's development. They were highly correlated to juveniles' weight increase and histological features of the gland in early DAH. These ratios described more accurately the shift occurring between the intracellular acid and the extracellular alkaline modes of digestion in S. officinalis and were more specific than separated enzyme activities. Their application as eco-physiological tools revealed that enzyme ratios reflected yolk content and digestive gland development in new hatching juveniles. Finally, ALP/ACP ratio was shown to be a powerful tool to describe growth performance of S. officinalis which is useful for aquaculture optimization.

Electron beam combined with hydrothermal treatment for enhancing the enzymatic convertibility of sugarcane bagasse

International Nuclear Information System (INIS)

Duarte, C.L.; Ribeiro, M.A.; Oikawa, H.; Mori, M.N.; Napolitano, C.M.; Galvão, C.A.

2012-01-01

The use of microbial cellulolytic enzymes is the most efficient process to liberate glucose from cellulose in biomass without the formation of fermentation inhibitors. A combination of pretreatment technologies is an alternative way to increase the access of enzymes to cellulose, and consequently, the conversion yield. In this way, the present study reports on the enzymatic hydrolysis of SCB submitted to three kinds of pretreatment: electron beam processing (EBP), and EBP followed by hydrothermal (TH) and diluted acid (AH) treatment. SCB samples were irradiated using a radiation dynamics electron beam accelerator, and then submitted to thermal and acid (0.1% sulfuric acid) hydrolysis for 40 and 60 min at 180 °C. These samples were submitted to enzymatic hydrolysis (EH) using commercial preparations, including Celluclast 1.5 L and beta-glycosidase. The addition of diluted acid improved TH treatment allowing for a shorter application time. EBP with 50 kGy increased the enzymatic hydrolysis yield of cellulose by 20% after TH and 30% after AH. - Highlights: ► We study the enzymatic hydrolysis of cellulose and hemicellulose in sugarcane bagasse. ► We study the combination of three pretreatments: electron beam processing, EBP followed by hydrothermal and by diluted acid treatment. ► The electron beam processing increased the enzymatic hydrolysis from 8% to 15% with 20 kGy. ► The enzymes used were commercial preparations, as Celluclast 1.5 L and β-glycosidase. ► The EBP with 50 kGy increased on 20% the yield of EH of cellulose after TH and 30% after AH.

Hemicelluloses negatively affect lignocellulose crystallinity for high biomass digestibility under NaOH and H2SO4 pretreatments in Miscanthus

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Xu Ning

2012-08-01

Full Text Available Abstract Background Lignocellulose is the most abundant biomass on earth. However, biomass recalcitrance has become a major factor affecting biofuel production. Although cellulose crystallinity significantly influences biomass saccharification, little is known about the impact of three major wall polymers on cellulose crystallization. In this study, we selected six typical pairs of Miscanthus samples that presented different cell wall compositions, and then compared their cellulose crystallinity and biomass digestibility after various chemical pretreatments. Results A Miscanthus sample with a high hemicelluloses level was determined to have a relatively low cellulose crystallinity index (CrI and enhanced biomass digestibility at similar rates after pretreatments of NaOH and H2SO4 with three concentrations. By contrast, a Miscanthus sample with a high cellulose or lignin level showed increased CrI and low biomass saccharification, particularly after H2SO4 pretreatment. Correlation analysis revealed that the cellulose CrI negatively affected biomass digestion. Increased hemicelluloses level by 25% or decreased cellulose and lignin contents by 31% and 37% were also found to result in increased hexose yields by 1.3-times to 2.2-times released from enzymatic hydrolysis after NaOH or H2SO4 pretreatments. The findings indicated that hemicelluloses were the dominant and positive factor, whereas cellulose and lignin had synergistic and negative effects on biomass digestibility. Conclusions Using six pairs of Miscanthus samples with different cell wall compositions, hemicelluloses were revealed to be the dominant factor that positively determined biomass digestibility after pretreatments with NaOH or H2SO4 by negatively affecting cellulose crystallinity. The results suggested potential approaches to the genetic modifications of bioenergy crops.

Biomonitoring of carcinogenic substances: enzymatic digestion of globin for detecting alkylated amino acids

Science.gov (United States)

Bader, Michael; Rauscher, Dankwart; Geibel, Kurt; Angerer, Juergen

1993-03-01

We report the application of proteases for the total hydrolysis of globin with subsequent determination of amino acids. Optimization of the proteolysis was made with respect to enzyme concentration, time of incubation and type of protease. Ethylene oxide modified globin was used to compare the results of the analysis of the N-terminal amino acid valine after enzymatic cleavage to those obtained from the widely used modified Edman procedure. It is shown that the cleavage is of good reproducibility and yields more alkylated amino acid than the Edman procedure.

Low-heat, mild alkaline pretreatment of switchgrass for anaerobic digestion.

Science.gov (United States)

Jin, Guang; Bierma, Tom; Walker, Paul M

2014-01-01

This study examines the effectiveness of alkaline pretreatment under mild heat conditions (100°C or 212°F) on the anaerobic co-digestion of switchgrass. The effects of alkaline concentration, types of alkaline, heating time and rinsing were evaluated. In addition to batch studies, continuous-feed studies were performed in triplicate to identify potential digester operational problems caused by switchgrass co-digestion while accounting for uncertainty due to digester variability. Few studies have examined anaerobic digestion of switchgrass or the effects of mild heating to enhance alkaline pretreatment prior to biomass digestion. Results indicate that pretreatment can significantly enhance digestion of coarse-ground (≤ 0.78 cm particle size) switchgrass. Energy conversion efficiency as high as 63% was observed, and was comparable or superior to fine-grinding as a pretreatment method. The optimal NaOH concentration was found to be 5.5% (wt/wt alkaline/biomass) with a 91.7% moisture level. No evidence of operational problems such as solids build-up, poor mixing, or floating materials were observed. These results suggest the use of waste heat from a generator could reduce the concentration of alkaline required to adequately pretreat lignocellulosic feedstock prior to anaerobic digestion.

Extended light exposure increases stem digestibility and biomass production of switchgrass

Science.gov (United States)

Zhao, Chunqiao; Hou, Xincun; Zhu, Yi; Yue, Yuesen; Wu, Juying

2017-01-01

Switchgrass is a photoperiod-sensitive energy grass suitable for growing in the marginal lands of China. We explored the effects of extended photoperiods of low-irradiance light (7 μmol·m-2·s-1, no effective photosynthesis) on the growth, the biomass dry weight, the biomass allocation, and, especially, the stem digestibility and cell wall characteristics of switchgrass. Two extended photoperiods (i.e., 18 and 24 h) were applied over Alamo. Extended light exposure (18 and 24 h) resulted in delayed heading and higher dry weights of vegetative organs (by 32.87 and 35.94%, respectively) at the expense of reducing the amount of sexual organs (by 40.05 and 50.87%, respectively). Compared to the control group (i.e., natural photoperiod), the yield of hexoses (% dry matter) in the stems after a direct enzymatic hydrolysis (DEH) treatment significantly increased (by 44.02 and 46.10%) for those groups irradiated during 18 and 24 h, respectively. Moreover, the yield of hexoses obtained via enzymatic hydrolysis increased after both basic (1% NaOH) and acid (1% H2SO4) pretreatments for the groups irradiated during 18 and 24 h. Additionally, low-irradiance light extension (LILE) significantly increased the content of non-structural carbohydrates (NSCs) while notably reducing the lignin content and the syringyl to guaiacyl (S/G) ratio. These structural changes were in part responsible for the observed improved stem digestibility. Remarkably, LILE significantly decreased the cellulose crystallinity index (CrI) of switchgrass by significantly increasing both the arabinose substitution degree in xylan and the content of ammonium oxalate-extractable uronic acids, both favoring cellulose digestibility. Despite this LILE technology is not applied to the cultivation of switchgrass on a large scale yet, we believe that the present work is important in that it reveals important relationships between extended day length irradiations and biomass production and quality. Additionally, this

Enzymatic complex in diets for gold fish fingerlings (Carassius auratusComplexo enzimático na dieta de alevinos de kinguio (Carassius auratus

Directory of Open Access Journals (Sweden)

Arcângelo Augusto Signor

2013-06-01

Full Text Available Aiming to evaluate the enzymatic complex supplementation in diets for goldfish fingerlings (Carassius auratus, 240 fish weighing initially 1,36 ± 0,02g, randomly distributed in 20 tanks with 150L, in four treatments and five replications, with twelve fish in each experimental unit were used. The fish were fed at 8:00 and 11:00 a.m. and 2:00 and 5:00 p.m. with diets containing different inclusion levels (0; 0,033; 0,066 e 0,099% of enzymatic complex (amilase, protease, celulase, lipase, ?-glucanase and phytase, and formulated with 32,36% of digestible protein and 3.023kcal of digestible energy kg-1. There were no differences observed (P>0,05 in the mean final weight, weight gain, total length, standard length, survival and carcass composition. However, the fish apparent feed conversion was impaired by the supplementation of enzymatic complex with 0,099% in diet. The use of enzymatic complex does not provides benefits in the productive performance for goldfish fingerlings. Objetivando avaliar a suplementação de complexo enzimático em dietas de alevinos de kinguios (Carassius auratus, foram utilizados 240 peixes com peso inicial de 1,36 ± 0,02g, distribuídos aleatoriamente, em 20 tanques de 150L, em quatro tratamentos e cinco repetições, com 12 peixes em cada unidade experimental. Os peixes foram arraçoados as 8h, 11h, 14h e 17h, com dietas contendo diferentes níveis de inclusão (0; 0,033; 0,066 e 0,099% de complexo enzimático (amilase, protease, celulase, lipase, b-glucanase e fitase, e formuladas para conter 32,36% de proteína digestível e 3.023kcal de energia digestível kg-1. Não foram observadas diferenças no peso final, ganho de peso, comprimento total, comprimento padrão, sobrevivência e composição de carcaça. No entanto, a conversão alimentar aparente dos peixes alimentados com rações contendo 0,099% do complexo enzimático foi inferior em relação aos alimentados com as demais dietas. Conclui-se que a utiliza

Biological strategies for enhanced hydrolysis of lignocellulosic biomass during anaerobic digestion: Current status and future perspectives.

Science.gov (United States)

Shrestha, Shilva; Fonoll, Xavier; Khanal, Samir Kumar; Raskin, Lutgarde

2017-12-01

Lignocellulosic biomass is the most abundant renewable bioresource on earth. In lignocellulosic biomass, the cellulose and hemicellulose are bound with lignin and other molecules to form a complex structure not easily accessible to microbial degradation. Anaerobic digestion (AD) of lignocellulosic biomass with a focus on improving hydrolysis, the rate limiting step in AD of lignocellulosic feedstocks, has received considerable attention. This review highlights challenges with AD of lignocellulosic biomass, factors contributing to its recalcitrance, and natural microbial ecosystems, such as the gastrointestinal tracts of herbivorous animals, capable of performing hydrolysis efficiently. Biological strategies that have been evaluated to enhance hydrolysis of lignocellulosic biomass include biological pretreatment, co-digestion, and inoculum selection. Strategies to further improve these approaches along with future research directions are outlined with a focus on linking studies of microbial communities involved in hydrolysis of lignocellulosics to process engineering. Copyright © 2017 Elsevier Ltd. All rights reserved.

Enzymatic hydrolsis of pretreated rice straw

Energy Technology Data Exchange (ETDEWEB)

Vlasenko, E.Y.; Shoemaker, S.P. [California Inst. of Food and Agricultural Research, Davis, CA (United States); Ding, H. [California Univ., Davis (Canada). Dept. of Food Science and Technology; Labavitch, J.M. [California Univ., Davis, CA (United States). Dept. of Pomology

1997-02-01

California rice straw is being evaluated as a feedstock for production of power and fuel. This paper examines the initial steps in the process: pretreatment of rice straw and enzymatic hydrolysis of the polysaccharides in the pretreated material to soluble sugars. Rice straw was subjected to three distinct pretreatment procedures: acid-catalyzed steam explosion (Swan Biomass Company), acid hydrolysis (U.S. DOE National Renewable Energy Laboratory), and ammonia fiber explosion or AFEX (Texas A and M University). Standard conditions for each pretreatment were used, but none was optimized for rice straw specifically. Six commercial cellulases, products of Genencor International (USA), Novo (Denmark), Iogen (Canada) and Fermtech (Russia) were used for hydrolysis. The Swan- and the acid-pretreatments effectively removed hemicellulose from rice straw, providing high yields of fermentable sugars. The AFEX-pretreatment was distinctly different from other pretreatments in that it did not significantly solubilize hemicellulose. All three pretreatment procedures substantially increased enzymatic digestibility of rice straw. Three commercial Trichoderma-reesei-derived enzyme preparations: Cellulase 100L (Iogen), Spezyme CP (Genencor), and Al (Fermtech), were more active on pretreated rice straw compared than others tested. Conditions for hydrolysis of rice straw using Cellulase 100L were evaluated. The supplementation of this enzyme preparation with cellobiase (Novozyme 188) significantly improved the parameters of hydrolysis for the Swan- and the acid-pretreated materials, but did not affect the hydrolysis of the AFEX-pretreated rice straw. (Author)

Tunable Enzymatic Activity and Enhanced Stability of Cellulase Immobilized in Biohybrid Nanogels.

Science.gov (United States)

Peng, Huan; Rübsam, Kristin; Jakob, Felix; Schwaneberg, Ulrich; Pich, Andrij

2016-11-14

This paper reports a facile approach for encapsulation of enzymes in nanogels. Our approach is based on the use of reactive copolymers able to get conjugated with enzyme and build 3D colloidal networks or biohybrid nanogels. In a systematic study, we address the following question: how the chemical structure of nanogel network influences the biocatalytic activity of entrapped enzyme? The developed method allows precise control of the enzyme activity and improvement of enzyme resistance against harsh store conditions, chaotropic agents, and organic solvents. The nanogels were constructed via direct chemical cross-linking of water-soluble reactive copolymers poly(N-vinylpyrrolidone-co-N-methacryloxysuccinimide) with proteins such as enhanced green fluorescent protein (EGFP) and cellulase in water-in-oil emulsion. The water-soluble reactive copolymers with controlled amount of reactive succinimide groups and narrow dispersity were synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization. Poly(ethylene glycol) bis(3-aminopropyl) and branched polyethylenimine were utilized as model cross-linkers to optimize synthesis of nanogels with different architectures in the preliminary experiments. Biofluorescent nanogels with different loading amount of EGFP and varying cross-linking densities were obtained. We demonstrate that the biocatalytic activity of cellulase-conjugated nanogels (CNG) can be elegantly tuned by control of their cross-linking degrees. Circular dichroism (CD) spectra demonstrated that the secondary structures of the immobilized cellulase were changed in the aspect of α-helix contents. The secondary structures of cellulase in highly cross-linked nanogels were strongly altered compared with loosely cross-linked nanogels. The fluorescence resonance energy transfer (FRET) based study further revealed that nanogels with lower cross-linking degree enable higher substrate transport rate, providing easier access to the active site of

Evaluation of wet oxidation pretreatment for enzymatic hydrolysis of softwood

DEFF Research Database (Denmark)

Palonen, H.; Thomsen, A.B.; Tenkanen, M.

2004-01-01

The wet oxidation pretreatment (water, oxygen, elevated temperature, and pressure) of softwood (Picea abies) was investigated for enhancing enzymatic hydrolysis. The pretreatment was preliminarily optimized. Six different combinations of reaction time, temperature, and pH were applied......, and the compositions of solid and liquid fractions were analyzed. The solid fraction after wet oxidation contained 58-64% cellulose, 2-16% hemicellulose, and 24-30% lignin. The pretreatment series gave information about the roles of lignin and hemicellulose in the enzymatic hydrolysis. The temperature...

Digestion of thyroglobulin with purified thyroid lysosomes: preferential release of iodoamino acids

International Nuclear Information System (INIS)

Tokuyama, T.; Yoshinari, M.; Rawitch, A.B.; Taurog, A.

1987-01-01

[ 131 I]Thyroglobulin [( 131 I]Tg), prepared by either enzymatic iodination of human goiter Tg in vitro or isolation from the thyroids of rats previously injected with 131 I, was digested with a solubilized enzyme mixture prepared from purified hog thyroid lysosomes. The digestion was performed at 37 C for 24 h under nitrogen at pH 5.0 in the presence of 4 mM dithiothreitol. Under these conditions the release of free [ 131 I] iodoamino acids (MIT, DIT, T4, and T3) was quantitatively very similar to that observed with a standard pronase digestion procedure. To determine whether other amino acids in Tg were released as quantitatively as the iodoamino acids, free amino acids in the lysosomal digest were measured, and total free amino acid release was compared with a similar analysis performed after digestion of [ 131 I]Tg with 6 N HCl. Total amino acid release was much less complete than iodoamino acid release, indicating preferential release of iodoamino acids from Tg by lysosomal digestion. Analysis of the lysosomal digest by HPLC on a size exclusion column indicated that Tg was degraded to peptides with a mol wt less than 4000. Assuming that the in vitro lysosomal digestion system represents a valid model for the physiological proteolytic system that degrades Tg, the results of the present study suggest that a substantial portion of the Tg in the thyroid is not degraded to free amino acids and that peptide fragments of Tg are normally present in the thyroid. In such a case, the fate and possible physiological activity of these fragments require further elucidation

Pretreatments to enhance the digestibility of lignocellulosic biomass

NARCIS (Netherlands)

Hendriks, A.T.W.M.; Zeeman, G.

2009-01-01

Lignocellulosic biomass represents a rather unused source for biogas and ethanol production. Many factors, like lignin content, crystallinity of cellulose, and particle size, limit the digestibility of the hemicellulose and cellulose present in the lignocellulosic biomass. Pretreatments have as a

Biogas production enhancement by soya sludge amendment in cattle dung digesters

Energy Technology Data Exchange (ETDEWEB)

Satyanarayan, Shanta; Ramakant; Shivayogi [WWT Division, NEERI, Nagpur 400 020 (India)

2010-09-15

Biogas energy production from cattle dung is an economically feasible and eco-friendly in nature. But dependence only on cattle dung is a limiting factor. Rich nitrogen containing substrate addition to extra carbohydrate digester like cattle dung could improve the biogas production. Detailed performance of the digesters at different ratios of cattle dung and soya sludge has been discussed in this paper considering the cold countries climate. Soya sludge substrate not only has high nitrogen content of 4.0-4.8% but it also has high percentage of volatile solids content in the range of 97.8-98.8%. Soya sludge addition also improved the manurial value of the digested slurry and also improved the dewater-ability of the sludge. Results indicated an increment of 27.0% gas production at 25.0% amendment of soya sludge in non-homogenized cattle dung (NCD) digester. The amount of gas production increased to 46.4% in case of homogenized cattle dung (HCD) with respect to NCD feed at the same amendment. (author)

Performance of thermophilic anaerobic digesters using inoculum mixes with enhanced methanogenic diversity

KAUST Repository

Ghanimeh, Sophia; El-Fadel, Mutasem; Saikaly, Pascal

2017-01-01

Reportedly, various mixes of seeds were quasi-randomly selected to startup anaerobic digesters. In contrast, this study examines the impact of inoculating thermophilic anaerobic digesters with a designed mix of non-acclimated seeds based

ENZYMATIC HYDROLYSIS OF SWITCHGRASS AND COASTAL BERMUDA GRASS PRETREATED USING DIFFERENT CHEMICAL METHODS

Directory of Open Access Journals (Sweden)

Jiele Xu

2011-06-01

Full Text Available To investigate the effects of biomass feedstock and pretreatment method on the enzyme requirement during hydrolysis, swichgrass and coastal Bermuda grass pretreated using H2SO4, NaOH, and Ca(OH2 at the optimal conditions were subjected to enzymatic hydrolysis using two enzyme combinations: NS 50013 + NS 50010 and Cellic CTec + Cellic HTec. The enzyme loadings were optimized, and correlations between feedstock property, pretreatment strategy, and enzyme usage were evaluated. The results show that pretreatment methods resulting in greater lignin contents in the pretreated biomass were generally associated with higher enzyme requirements. More sugars could be recovered from alkaline-pretreated biomass during enzymatic hydrolysis due to the better carbohydrate preservation achieved at mild pretreatment temperatures. The cellulase enzyme, Cellic CTec, was more efficient in catalyzing the hydrolysis of coastal Bermuda grass, a feedstock more digestible than the pretreated swichgrass, following pretreatment with NaOH or Ca(OH2.

Effect of alkali lignins with different molecular weights from alkali pretreated rice straw hydrolyzate on enzymatic hydrolysis.

Science.gov (United States)

Li, Yun; Qi, Benkun; Luo, Jianquan; Wan, Yinhua

2016-01-01

This study investigated the effect of alkali lignins with different molecular weights on enzymatic hydrolysis of lignocellulose. Different alkali lignins fractions, which were obtained from cascade ultrafiltration, were added into the dilute acid pretreated (DAP) and alkali pretreated (AP) rice straws respectively during enzymatic hydrolysis. The results showed that the addition of alkali lignins enhanced the hydrolysis and the enhancement for hydrolysis increased with increasing molecular weights of alkali lignins, with maximum enhancement being 28.69% for DAP and 20.05% for AP, respectively. The enhancement was partly attributed to the improved cellulase activity, and filter paper activity increased by 18.03% when adding lignin with highest molecular weight. It was found that the enhancement of enzymatic hydrolysis was correlated with the adsorption affinity of cellulase on alkali lignins, and the difference in surface charge and hydrophobicity of alkali lignins were responsible for the difference in affinity between cellulase and lignins. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enhancement of biogas production in anaerobic co-digestion by ultrasonic pretreatment

International Nuclear Information System (INIS)

Zou, Shuzhen; Wang, Xiaojiao; Chen, Yuanlin; Wan, Haiwen; Feng, Yongzhong

2016-01-01

Highlights: • Ultrasonic pretreatment changed physical structure of samples. • Ultrasonic pretreatment improved biogas production via changing environment before and during anaerobic digestion process. • The main factors affecting biogas production differ in different pretreated samples. - Abstract: This paper optimized the anaerobic digestion (AD) pretreatment process and identified the influence of pretreatment on the co-digestion of maize straw (MS) and dairy manure (DM). In the study, ultrasonic was used to pretreat MS and DM prior to digestion, with power intensities of 0, 189.39, 284.09, and 378.79 kJ at 0, 20, 30, and 40 min, respectively. Changes in the surface structures of MS and DM were observed by scanning electron microscopy (SEM), and factor analysis was used to analyze the main factors affecting biogas production in the AD process. The result showed that the structure of DM was distributed and that the structure of MS became more roughness following the ultrasonic pretreatment (UP). The highest total biogas production of co-digestion (240.32 mL/g VS_f_e_d) was obtained when MS was pretreated for 30 min without DM pretreatment (MS_3_0DM_0). This was significantly higher than that of the untreated sample (CK) (141.65 mL/g VS_f_e_d). The cellulose activity (CA), reducing sugar (RS) content, volatile fatty acid (VFA) content and pH in the digester feed, and their maximum and minimum values in the AD process was affected by UP. Factor 1 of MS_3_0DM_0 was determined by RS content, pH and VFA content that they had the most influence on biogas production on days 6, 18, 24 and 30. Factor 2 of it was determined by CA, and it had most influence on days 0, 12, 36 and 42 in the AD process, The result of the factor analysis indicated that the main factors affecting biogas production were affected by UP and they differ according to the different digestion stages. This research concluded that UP improved total biogas production via changing the initial

Quality assessment of digested sludges produced by advanced stabilization processes.

Science.gov (United States)

Braguglia, C M; Coors, A; Gallipoli, A; Gianico, A; Guillon, E; Kunkel, U; Mascolo, G; Richter, E; Ternes, T A; Tomei, M C; Mininni, G

2015-05-01

The European Union (EU) Project Routes aimed to discover new routes in sludge stabilization treatments leading to high-quality digested sludge, suitable for land application. In order to investigate the impact of different enhanced sludge stabilization processes such as (a) thermophilic digestion integrated with thermal hydrolysis pretreatment (TT), (b) sonication before mesophilic/thermophilic digestion (UMT), and (c) sequential anaerobic/aerobic digestion (AA) on digested sludge quality, a broad class of conventional and emerging organic micropollutants as well as ecotoxicity was analyzed, extending the assessment beyond the parameters typically considered (i.e., stability index and heavy metals). The stability index was improved by adding aerobic posttreatment or by operating dual-stage process but not by pretreatment integration. Filterability was worsened by thermophilic digestion, either alone (TT) or coupled with mesophilic digestion (UMT). The concentrations of heavy metals, present in ranking order Zn > Cu > Pb > Cr ~ Ni > Cd > Hg, were always below the current legal requirements for use on land and were not removed during the processes. Removals of conventional and emerging organic pollutants were greatly enhanced by performing double-stage digestion (UMT and AA treatment) compared to a single-stage process as TT; the same trend was found as regards toxicity reduction. Overall, all the digested sludges exhibited toxicity to the soil bacterium Arthrobacter globiformis at concentrations about factor 100 higher than the usual application rate of sludge to soil in Europe. For earthworms, a safety margin of factor 30 was generally achieved for all the digested samples.

Pretreatments and enzymatic hydrolysis of sugarcane bagasse aiming at the enhancement of the yield of glucose and xylose

Directory of Open Access Journals (Sweden)

A. de A. Guilherme

Full Text Available ABSTRACT This work studied the enzymatic hydrolysis of sugarcane bagasse aiming at the production of glucose and xylose. The bagasse was subjected to two different pretreatments: combined acid and alkalinepretreatment and hydrogen peroxidepretreatment. The enzymatic hydrolysis was optimized and a kinetic study was carried out in a stirred tank reactor (STR in batch mode. Optimal conditions were obtained by subjecting the bagasse to the hydrogen peroxide pretreatment followed by enzymatic hydrolysis. The addition of xylanases to the enzymatic mixture improved the production of fermentable sugars by 48%.

An enzymatic deglycosylation scheme enabling identification of core fucosylated N-glycans and O-glycosylation site mapping of human plasma proteins

DEFF Research Database (Denmark)

Hägglund, Per; Matthiesen, R.; Elortza, F.

2007-01-01

and N-acetyl-β-glucosaminidase) are also included. The two strategies were here applied to identify 103 N-glycosylation sites in the Cohn IV fraction of human plasma. In addition, Endo D/H digestion uniquely enabled identification of 23 fucosylated N-glycosylation sites. Several O-glycosylated peptides......, thereby reducing the complexity and facilitating glycosylation site determinations. Here, we have used two different enzymatic deglycosylation strategies for N-glycosylation site analysis. (1) Removal of entire N-glycan chains by peptide- N-glycosidase (PNGase) digestion, with concomitant deamidation...... of the released asparagine residue. The reaction is carried out in H218O to facilitate identification of the formerly glycosylated peptide by incorporatation of 18O into the formed aspartic acid residue. (2) Digestion with two endo-β- N-acetylglucosaminidases (Endo D and Endo H) that cleave the glycosidic bond...

Comparison of HPLC-RI, LC/MS-MS and enzymatic assays for the analysis of residual lactose in lactose-free milk.

Science.gov (United States)

Trani, A; Gambacorta, G; Loizzo, P; Cassone, A; Fasciano, C; Zambrini, A V; Faccia, M

2017-10-15

Lactose intolerance is the decreased ability to digest lactose, and the population involved is rapidly increasing all over the world. Different procedures have been reported in the literature to quantify lactose in dairy products, but the official method of analysis is based on enzymatic assay. In this paper, the effectiveness of two enzymatic kits in detecting residual lactose in lactose-free milk was investigated, and a comparison with two alternative chromatographic methods was done. The investigation used several samples of UHT milk containing different levels of lactose, and the results highlighted the inadequacy of the enzymatic assays and of the HPLC-RI method to analyse lactose-free milk. An LC-MS/MS method using the formate adduct was developed, and it allowed quantitation of lactose and lactulose in all samples at a high level of precision and repeatability. Copyright © 2017 Elsevier Ltd. All rights reserved.

Enhanced biogas recovery by applying post-digestion in large-scale centralized biogas plants

DEFF Research Database (Denmark)

Angelidaki, Irini; Hejnfelt, Anette; Ellegaard, L.

2006-01-01

industry to generate biogas, which is used for electricity and thermal energy. A total of 20 such plants are currently active in Denmark, most of which were included in the investigation. From the plants, samples were obtained from various steps of the process. Samples were analysed and the residual biogas......The main objective of this study was to investigate the degradation efficiency of centralized biogas plants and provide guidance for the design of more efficient digester and post-digestion systems. These centralized biogas plants in Denmark digest manure together with organic waste from the food...... potential determined by batch post-digestion at various temperature levels. Results were correlated with plant characteristics and production statistics in order to judge the efficiency of various digestion concepts. A simplified model based on a two-step biogas production process was developed...

Urease immobilized polymer hydrogel: Long-term stability and enhancement of enzymatic activity.

Science.gov (United States)

Kutcherlapati, S N Raju; Yeole, Niranjan; Jana, Tushar

2016-02-01

A method has been developed in which an enzyme namely urease was immobilized inside hydrogel matrix to study the stability and enzymatic activity in room temperature (∼27-30°C). This urease coupled hydrogel (UCG) was obtained by amine-acid coupling reaction and this procedure is such that it ensured the wider opening of mobile flap of enzyme active site. A systematic comparison of urea-urease assay and the detailed kinetic data clearly revealed that the urease shows activity for more than a month when stored at ∼27-30°C in case of UCG whereas it becomes inactive in case of free urease (enzyme in buffer solution). The aqueous microenvironment inside the hydrogel, unusual morphological features and thermal behaviour were believed to be the reasons for unexpected behaviour. UCG displayed enzyme activity at basic pH and up to 60°C. UCG showed significant enhancement in activity against thermal degradation compared to free urease. In summary, this method is a suitable process to stabilize the biomacromolecules in standard room temperature for many practical uses. Copyright © 2015 Elsevier Inc. All rights reserved.

Production of fermentable sugars by combined chemo-enzymatic hydrolysis of cellulosic material for bioethanol production

Directory of Open Access Journals (Sweden)

M. Idrees

2014-06-01

Full Text Available To change the recalcitrant nature of the lignocellulosic material for maximum hydrolysis yield, a comprehensive study was done by using sulphuric acid as an exclusive catalyst for the pretreatment process. The enzymatic digestibility of the biomass [Water Hyacinth: Eichhornia crassipes] after pretreatment was determined by measuring the hydrolysis yield of the pretreated material obtained from twenty four different pretreatment conditions. These included different concentrations of sulphuric acid (0.0, 1.0, 2.0 and 3.0%, at two different temperatures (108 and 121 ºC for different residence times (1.0, 2.0 and 3.0h.The highest reducing sugar yield (36.65 g/L from enzymatic hydrolysis was obtained when plant material was pretreated at 121 ºC for 1.0 h residence time using 3.0% (v/v sulphuric acid and at 1:10 (w/v solid to liquid ratio. The total reducing sugars obtained from the two-stage process (pretreatment + enzymatic hydrolysis was 69.6g/L. The resulting sugars were fermented into ethanol by using Saccharomyces cerevisiae. The ethanol yield from the enzymatic hydrolyzate was 95.2% of the theoretical yield (0.51g/g glucose, as determined by GS-MS, and nearly 100% since no reducing sugars were detected in the fermenting media by TLC and DNS analysis.

Fat content affects bioaccessibility and efficiency of enzymatic hydrolysis of lutein esters added to milk and yogurt

OpenAIRE

Xavier, Ana Augusta Odorissi; Mercadante, Adriana Zerlotti; Garrido-Fernández, Juan; Pérez-Gálvez, Antonio

2014-01-01

© 2014 Elsevier Ltd. Addition of lutein to dairy products is an alternative that widens the range of foods which could be lutein sources. However, bioaccessibility is an essential aspect to be considered during the development of products with added bioactive substances. We evaluated the in vitro bioaccessibility of lutein esters added to milk and yogurt with different fat contents, and determined the efficiency of enzymatic hydrolysis of the esters during digestion. Bioaccessibility of lutei...

Homology of yeast photoreactivating gene fragment with human genomic digests

International Nuclear Information System (INIS)

Meechan, P.J.; Milam, K.M.; Cleaver, J.E.

1984-01-01

Enzymatic photoreactivation of UV-induced DNA lesions has been demonstrated for a variety of prokaryotic and eukaryotic organisms. Its presence in placental mammals, however, has not been clearly established. The authors attempted to resolve this question by assaying for the presence (or absence) of sequences in human DNA complimentary to a fragment of the photoreactivating gene from S. cerevisiae that has recently been cloned. In another study, DNA from human, chick E. coli and yeast cells was digested with either HindIII of BglII, electrophoresed on a 0.5% agarose gel, transferred (Southern blot) to a nylon membrane and probed for homology against a Sau3A restriction fragment from S. cerevisiae that compliments phr/sup -/ cells. Hybridization to human DNA digests was observed only under relatively non-stringent conditions indicating the gene is not conserved in placental mammals. These results are correlated with current literature data concerning photoreactivating enzymes

Impact of α-amylase combined with hydrochloric acid hydrolysis on structure and digestion of waxy rice starch.

Science.gov (United States)

Li, Hongyan; Zhu, Yanqiao; Jiao, Aiquan; Zhao, Jianwei; Chen, Xiaoming; Wei, Benxi; Hu, Xiuting; Wu, Chunsen; Jin, Zhengyu; Tian, Yaoqi

2013-04-01

The structure and in vitro digestibility of native waxy rice starch by the combined hydrolysis of α-amylase and hydrochloric acid were investigated in this study. The combined hydrolysis technique generated higher hydrolysis rate and extent than the enzymatic hydrolysis. The granular appearance and chromatograph profile demonstrated that α-amylase and hydrochloric acid exhibited different patterns of hydrolysis. The rise in the ratio of absorbance 1047/1022cm(-1), the melting temperature range (Tc-To), and the melting enthalpy (ΔH) were observed during the combined hydrolysis. These results suggest that α-amylase simultaneously cleaves the amorphous and crystalline regions, whereas the amorphous regions of starch granules are preferentially hydrolyzed during the acid hydrolysis. Furthermore, the combined hydrolysis increased rapidly digestible starch (RDS) while decreased slowly digestible starch (SDS) and resistant starch (RS), indicating that the hydrolysis mode affected the digestion property of native waxy rice starch. Copyright © 2013 Elsevier B.V. All rights reserved.

Simultaneous addition of zero-valent iron and activated carbon on enhanced mesophilic anaerobic digestion of waste-activated sludge.

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Wang, Tongyu; Qin, Yujie; Cao, Yan; Han, Bin; Ren, Junyi

2017-10-01

The performance of biogas generation and sludge degradation was studied under different zero-valent iron/activated carbon (ZVI/AC) ratios in detail in mesophilic anaerobic digestion of sludge. A good enhancement of methane production was obtained at the 10:1 ZVI/AC ratio, and the cumulative methane production was 132.1 mL/g VS, 37.6% higher than the blank. The methane content at the 10:1 ZVI/AC ratio reached 68.8%, which was higher than the blank (55.2%) and the sludge-added AC alone (59.6%). For sludge degradation, the removal efficiencies of total chemical oxygen demand (TCOD), proteins, and polysaccharides were all the highest at the 10:1 ZVI/AC ratio. The concentration of available phosphorus (AP) decreased after anaerobic digestion process. On the other hand, the concentrations of available nitrogen (AN) and available potassium (AK) increased after the anaerobic digestion process and showed a gradually decreasing trend with increasing ZVI/AC ratio. The concentrations of AN and AK were 2303.1-4200.3 and 274.7-388.3 mg/kg, showing a potential for land utilization.

Enhancing enzymatic hydrolysis of coconut husk through Pseudomonas aeruginosa AP 029/GLVIIA rhamnolipid preparation.

Science.gov (United States)

de Araújo, Cynthia Kérzia Costa; de Oliveira Campos, Alan; de Araújo Padilha, Carlos Eduardo; de Sousa Júnior, Francisco Canindé; do Nascimento, Ruthinéia Jéssica Alves; de Macedo, Gorete Ribeiro; Dos Santos, Everaldo Silvino

2017-08-01

This work investigated the influence of chemical (Triton X-100) and biological surfactant preparation (rhamnolipids) in coconut husk hydrolysis that was subjected to pretreatment with acid-alkali or alkaline hydrogen peroxide. The natural and pretreated biomass was characterized using the National Renewable Energy Laboratory protocol analysis as well as X-ray diffraction and scanning electron microscopy. The results demonstrated that in terms of the total reducing sugars, there was no significant difference between the hydrolysis using Triton X-100 and rhamnolipids, regardless of the pretreatment. A cellulosic conversion value as high as 33.0% was obtained in experiments with rhamnolipids. The coconut husk was observed to be a potential biomass that could produce second generation ethanol, and the rhamnolipid preparation can be used to support for the enzymatic hydrolysis, enhancing the advantage of cellulose conversion into glucose over chemical surfactants because it is an environmentally friendly approach. Copyright © 2017 Elsevier Ltd. All rights reserved.

Slowly digestible properties of lotus seed starch-glycerine monostearin complexes formed by high pressure homogenization.

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Chen, Bingyan; Jia, Xiangze; Miao, Song; Zeng, Shaoxiao; Guo, Zebin; Zhang, Yi; Zheng, Baodong

2018-06-30

Starch-lipid complexes were prepared using lotus seed starch (LS) and glycerin monostearate (GMS) via a high-pressure homogenization process, and the effect of high pressure homogenization (HPH) on the slow digestion properties of LS-GMS was investigated. The digestion profiles showed HPH treatment reduced the digestive rate of LS-GMS, and the extent of this change was dependent on homogenized pressure. Scanning electron microscopy displayed HPH treatment change the morphology of LS-GMS, with high pressure producing more compact block-shape structure to resist enzyme digestion. The results of Gel-permeation chromatography and Small-angle X-ray scattering revealed high homogenization pressure impacted molecular weight distribution and semi-crystalline region of complexes, resulting in the formation of new semi-crystalline with repeat unit distance of 16-18 nm and molecular weight distribution of 2.50-2.80 × 10 5  Da, which displayed strong enzymatic resistance. Differential scanning calorimeter results revealed new semi-crystalline lamellar may originate from type-II complexes that exhibited a high transition temperature. Copyright © 2018 Elsevier Ltd. All rights reserved.

Preparation and Enzymatic Degradation of Porous Crosslinked Polylactides of Biomass Origin

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Yuya Kido

2014-06-01

Full Text Available To understand the enzymatic degradation behavior of crosslinked polylactide (PLA, the preparation and enzymatic degradation of both thermoplastic (linear and crosslinked PLAs that have pore structures with different dimensions were carried out. The porous structures of the linear PLA samples were of micro and nanoporous nature, and prepared by batch foaming with supercritical CO2 and compared with the porous structures of crosslinked PLA (Lait-X created by the salt leaching method. The surface and cross-sectional morphologies of the porous structures were investigated by using scanning electron microscopy. The morphological analysis of porous Lait-X showed a rapid loss of physical features within 120 h of exposure to proteinase-K enzymatic degradation at 37 °C. Due to the higher affinity for water, enhanced enzymatic activity as compared to the linear PLA porous structures in the micro and nanoporous range was observed.

Rethinking the starch digestion hypothesis for AMY1 copy number variation in humans.

Science.gov (United States)

Fernández, Catalina I; Wiley, Andrea S

2017-08-01

Alpha-amylase exists across taxonomic kingdoms with a deep evolutionary history of gene duplications that resulted in several α-amylase paralogs. Copy number variation (CNV) in the salivary α-amylase gene (AMY1) exists in many taxa, but among primates, humans appear to have higher average AMY1 copies than nonhuman primates. Additionally, AMY1 CNV in humans has been associated with starch content of diets, and one known function of α-amylase is its involvement in starch digestion. Thus high AMY1 CNV is considered to result from selection favoring more efficient starch digestion in the Homo lineage. Here, we present several lines of evidence that challenge the hypothesis that increased AMY1 CNV is an adaptation to starch consumption. We observe that α- amylase plays a very limited role in starch digestion, with additional steps required for starch digestion and glucose metabolism. Specifically, we note that α-amylase hydrolysis only produces a minute amount of free glucose with further enzymatic digestion and glucose absorption being rate-limiting steps for glucose availability. Indeed α-amylase is nonessential for starch digestion since sucrase-isomaltase and maltase-glucoamylase can hydrolyze whole starch granules while releasing glucose. While higher AMY1 CN and CNV among human populations may result from natural selection, existing evidence does not support starch digestion as the major selective force. We report that in humans α-amylase is expressed in several other tissues where it may have potential roles of evolutionary significance. © 2017 Wiley Periodicals, Inc.

The in vitro digestibility of beef varies with its inherent ultimate pH.

Science.gov (United States)

Farouk, Mustafa M; Wu, Guojie; Frost, Deborah A; Clerens, Stefan; Knowles, Scott O

2014-11-01

Animal carcasses and cuts of meat are usually differentiated and valued according to size and compositional attributes. An underappreciated variable of red meat is its inherent ultimate pH (pHu) value, which affects organoleptic and processing characteristics. This study tests the hypothesis that high pHu aged meat would be more digestible than low pHu unaged (fresh) meat. Longissimus dorsi muscles collected from 59 bull carcasses had pHu values of 5.6-6.9. These were aged for 21 days at -1.5 °C, then raw and cooked (72 °C) samples were enzymatically digested at 37 °C with pepsin (pH 1.9 for 90 min) followed by pancreatin (pH 8.0 for an additional 120 min) to simulate conditions in the stomach and small intestine, respectively. Meat proteins and peptides in the digests were separated by 1D SDS PAGE. Regardless of pHu, ageing or cooking, most sarcoplasmic and myofibrillar proteins were rapidly digested by pepsin, with concomitant release of products identified by LC-MS/MS as mainly myosin-1, -2 and -7, α-actinin-2 or -3 and tropomyosin beta and alpha chains. These products were resistant to further digestion for the entire 210 min duration of the incubation. In terms of rate and extent of digestibility of these resistant products, high pHu > low pHu (P unaged (P meat samples increased with increasing pHu (P meat was highly digestible but could be further differentiated on the basis of its pHu and the ease of digestibility of proteins. Specific carcasses or cuts could be targeted to consumer groups in order to provide benefits and add value.

Enhanced methane yield by co-digestion of sewage sludge with micro-algae and catering waste leachate.

Science.gov (United States)

2018-04-04

The co-digestion of different wastes is a promising concept to improve methane generation during anaerobic process. However, the anaerobic co-digestion of catering waste leachate with algal biomass and sewage sludge has not been studied to date. This work investigated the methane generation by the anaerobic co-digestion of different mixtures of catering waste leachate, micro-algal biomass, and sewage sludge. Co-digestion of waste mixture containing equal ratios of three substrates had 39.31% higher methane yield than anaerobic digestion of raw sludge. This was possibly due to a proliferation of methanogens during the co-digestion period induced by multi-phase digestion of different wastes with different degrees of digestibility. Therefore, co-digestion of catering waste leachate, micro-algal biomass, and sewage sludge appears to be an efficient technology for energy conversion from waste resources. The scientific application of this co-digestion technology with these three substrates may play a role in solving important environmental issues of waste management.

Enzymological Characterization of Atm, the First Laccase from Agrobacterium sp. S5-1, with the Ability to Enhance In Vitro digestibility of Maize Straw.

Directory of Open Access Journals (Sweden)

Wei Si

Full Text Available Laccase is an enzyme that catalyzes oxidation of phenolic compounds, diamines and aromatic amines. In this study, a novel laccase-like gene (atm in a ligninolyitic isolate Agrobacterium sp. S5-1 from soil humus was identified and heterologously expressed in Escherichia coli. Atm exhibited its maximal activity at pH 4.5 and at 50°C. This enzyme was tolerant to high temperature, a broad range of pH, heavy metal ions (Co3+, Mn2+, Cu2+ and Ni2+, 20 mM and all tested organic solvents. Furthermore, Atm significantly (p<0.05 increased dry matter digestibility of maize straw from 23.44% to 27.96% and from 29.53% to 37.10% after 8 or 24 h of digestion and improved acid detergent fiber digestibility from 5.81% to 10.33% and from 12.80% to 19.07% after 8 or 24 h of digestion, respectively. The combination of Atm and fibrolytic enzymes significantly (p<0.05 enhanced neutral detergent fiber digestibility from 19.02% to 24.55% after 24 h of digestion respectively. Results showed treatment with Atm effectively improved in vitro digestibility of maize straw, thus suggesting that Atm has an application potential for bioconversion of lignin rich agricultural byproducts into animal feed and cellulosic ethanol.

Vitellin- and hemoglobin-digesting enzymes in Rhipicephalus (Boophilus) microplus larvae and females.

Science.gov (United States)

Estrela, Andréia Bergamo; Seixas, Adriana; Teixeira, Vivian de Oliveira Nunes; Pinto, Antônio Frederico Michel; Termignoni, Carlos

2010-12-01

The aim of the present study was to address the involvement of Rhipicephalus microplus larval cysteine endopeptidase (RmLCE) in protein digestion in R. microplus larvae and adult females. In this work, an improved purification protocol for native RmLCE was developed. Partial amino acid sequence of the purified enzyme indicates that it is the same enzyme as Boophilus microplus cathepsin-L1 (BmCL1). When vitellin (Vt) degradation by egg and larval enzymes was analyzed, stage-specific differences for RmLCE activity in comparison to vitellin-degrading cysteine endopeptidase (VTDCE) were observed. RmLCE is also able to degrade host hemoglobin (Hb). In agreement, an acidic cysteine endopeptidase activity was detected in larval gut. It was shown that cysteine and aspartic endopeptidases are involved in Vt and Hb digestion in R. microplus larvae and females. Interestingly, we observed that the aspartic endopeptidase Boophilus yolk cathepsin (BYC) is associated with a cysteine endopeptidase activity, in larvae. Synergic hemoglobin digestion by BYC and RmLCE was observed and indicates the presence of an Hb-degrading enzymatic cascade involving these enzymes. Our results suggest that RmLCE/BmCL1 has a continued role in vitellin and hemoglobin digestion during tick development. Copyright © 2010 Elsevier Inc. All rights reserved.

Photoelectrochemical enzymatic biosensors.

Science.gov (United States)

Zhao, Wei-Wei; Xu, Jing-Juan; Chen, Hong-Yuan

2017-06-15

Enzymatic biosensors have been valuable bioanalytical devices for analysis of diverse targets in disease diagnosis, biological and biomedical research, etc. Photoelectrochemical (PEC) bioanalysis is a recently emerged method that promptly becoming a subject of new research interests due to its attractive potential for future bioanalysis with high sensitivity and specificity. PEC enzymatic biosensors integrate the inherent sensitivities of PEC bioanalysis and the selectivity of enzymes and thus share their both advantages. Currently, PEC enzymatic biosensors have become a hot topic of significant research and the recent impetus has grown rapidly as demonstrated by increased research papers. Given the pace of advances in this area, this review will make a thorough discussion and survey on the fundamentals, sensing strategies, applications and the state of the art in PEC enzymatic biosensors, followed by future prospects based on our own opinions. We hope this work could provide an accessible introduction to PEC enzymatic biosensors for any scientist. Copyright © 2016 Elsevier B.V. All rights reserved.

Comparing two enhancing methods for improving kitchen waste anaerobic digestion: bentonite addition and autoclaved de-oiling pretreatment

DEFF Research Database (Denmark)

Zhang, Duojiao; Duan, Na; Tian, Hailin

2018-01-01

The effects of different enhancement methods, including adding bentonite (1.25%, w/w, wet substrate) and autoclaved de-oiling pretreatment (121 °C, 30 minutes), on the anaerobic digestion of kitchen waste (KW) were comparably studied. Mesophilic continuous stirred tank reactors were used under...... different organic loading rates (OLRs) of 1.11 to 1.84 gVS (volatile solid)L−1d−1 and two different hydraulic retention times (HRTs) (20 d and 25 d). In this study, two enhancement methods and extending HRT could prevent volatile fatty acids (VFA) accumulation and obtain a high methane production at low OLR...... design and process evaluation of a CSTR biogas plant treating with KW based on the laboratory experiment was stated....

Harvesting microalgae using activated sludge can decrease polymer dosing and enhance methane production via co-digestion in a bacterial-microalgal process

DEFF Research Database (Denmark)

Wágner, Dorottya Sarolta; Radovici, Maria; Smets, Barth F.

2016-01-01

, there is the potential to produce energy by co-digesting the two types of biomass. We present an innovative approach to recover microalgal biomass via a two-step flocculation using bacterial biomass after the destabilisation of microalgae with conventional cationic polymer. A short solids retention time (SRT) enhanced...

NUTRIENT DIGESTIBILITY AND PERFORMANCE IN BROILER CONSUMING DIETS FORMULATED WITH DIFFERENT FIBER LEVELS AND SUPLEMENTED WITH ENZYMATIC COMPLEX DIGESTIBILIDADE DOS NUTRIENTES E DESEMPENHO DE FRANGOS DE CORTE CONSUMINDO DIETAS FORMULADAS COM DIFERENTES NÍVEIS DE FIBRA E SUPLEMENTADAS COM ENZIMAS EXÓGENAS

Directory of Open Access Journals (Sweden)

Bárbara Karolina Ratier da Silva

2008-12-01

Full Text Available The effects of fiber level and the enzymatic supplementation in diets for broilers were evaluated in two trials. The experimental treatments were diets with high and low fiber level and with enzymatic supplementation (α-galactosidase, cellulase, amylase e protease. In the first trial (14 to 19 days of birds age, 190 broilers were randomly distributed in a 2x2 factorial arrangement (fiber x enzymes, placed in metabolic cages between (9-10 birds/cage, 5 cages per treatment. It was evaluate the metabolizability (MET of the dry matter (DM, the crude protein (CP and the energy (E, the total apparent digestibility coefficient (TDC for fat (F and acid detergent fiber (ADF, the apparent metabolisable energy (AME and the performance. Four replicates of each treatment were maintained to use in the second trial. In the 21st day age, after adaptation to the intake of the diets with indicator, the broilers were slaughtered and the ileal content was collected to evaluation of the ileal apparent digestibility coefficient (IDC for DM, CP, E, F and the weight of the caeca. The high fiber level resulted in lower MET of DM, E, CP, and lower TDC of F and ADF (P<0.05, as well as lower IDC of DM and E (P<0.01. Enzymatic supplementation reduced the fiber effect on MET of DM, E, CP and on the TDC of ADF (P<0.05. Higher empty weight of the caeca of the birds on the high fiber diet was observed (P<0.01, as well as lower weight gain (P<0.03 and worst feed conversion ratio (P<0.01. The performance of broilers was more related with ileal apparent digestibility then with the total apparent digestibility of the nutrients and the energy.

KEY WORDS: Caeca, ileal digestibility, metabolizability, performance, total digestibility. Avaliaram-se os efeitos do nível de fibra e da suplementação enzimática em dietas de frangos de corte em dois experimentos. Os tratamentos experimentais foram as dietas formuladas com alta ou baixa fibra e suplementação enzimática (�

The complexities of hydrolytic enzymes from the termite digestive system.

Science.gov (United States)

Saadeddin, Anas

2014-06-01

The main challenge in second generation bioethanol production is the efficient breakdown of cellulose to sugar monomers (hydrolysis). Due to the recalcitrant character of cellulose, feedstock pretreatment and adapted hydrolysis steps are needed to obtain fermentable sugar monomers. The conventional industrial production process of second-generation bioethanol from biomass comprises several steps: thermochemical pretreatment, enzymatic hydrolysis and sugar fermentation. This process is undergoing continuous optimization in order to increase the bioethanol yield and reduce the economic cost. Therefore, the discovery of new enzymes with high lignocellulytic activity or new strategies is extremely important. In nature, wood-feeding termites have developed a sophisticated and efficient cellulose degrading system in terms of the rate and extent of cellulose hydrolysis and exploitation. This system, which represents a model for digestive symbiosis has attracted the attention of biofuel researchers. This review describes the termite digestive system, gut symbionts, termite enzyme resources, in vitro studies of isolated enzymes and lignin degradation in termites.

Epigallocatechin gallate incorporation into lignin enhances the alkaline delignification and enzymatic saccharification of cell walls

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Elumalai Sasikumar

2012-08-01

Full Text Available Abstract Background Lignin is an integral component of the plant cell wall matrix but impedes the conversion of biomass into biofuels. The plasticity of lignin biosynthesis should permit the inclusion of new compatible phenolic monomers such as flavonoids into cell wall lignins that are consequently less recalcitrant to biomass processing. In the present study, epigallocatechin gallate (EGCG was evaluated as a potential lignin bioengineering target for rendering biomass more amenable to processing for biofuel production. Results In vitro peroxidase-catalyzed polymerization experiments revealed that both gallate and pyrogallyl (B-ring moieties in EGCG underwent radical cross-coupling with monolignols mainly by β–O–4-type cross-coupling, producing benzodioxane units following rearomatization reactions. Biomimetic lignification of maize cell walls with a 3:1 molar ratio of monolignols and EGCG permitted extensive alkaline delignification of cell walls (72 to 92% that far exceeded that for lignified controls (44 to 62%. Alkali-insoluble residues from EGCG-lignified walls yielded up to 34% more glucose and total sugars following enzymatic saccharification than lignified controls. Conclusions It was found that EGCG readily copolymerized with monolignols to become integrally cross-coupled into cell wall lignins, where it greatly enhanced alkaline delignification and subsequent enzymatic saccharification. Improved delignification may be attributed to internal trapping of quinone-methide intermediates to prevent benzyl ether cross-linking of lignin to structural polysaccharides during lignification, and to the cleavage of ester intra-unit linkages within EGCG during pretreatment. Overall, our results suggest that apoplastic deposition of EGCG for incorporation into lignin would be a promising plant genetic engineering target for improving the delignification and saccharification of biomass crops.

Effect of hydrothermal pre-treatment (HTP) on poultry slaughterhouse waste (PSW) sludge for the enhancement of the solubilization, physical properties, and biogas production through anaerobic digestion.

Science.gov (United States)

Park, Seyong; Yoon, Young-Man; Han, Seong Kuk; Kim, Daegi; Kim, Ho

2017-06-01

This study is an assessment of the hydrothermal pre-treatment (HTP) of poultry slaughterhouse waste (PSW) sludge for the enhancement of the solubilization, physical properties, and biogas production through anaerobic digestion. This assessment was carried out to ascertain the optimal HTP temperature. The solubilization and physical properties efficacy was investigated by capillary suction time (CST), time to filter (TTF), and particle size. In addition, the anaerobic digestion was investigated through biochemical methane potential (BMP) tests and subsequent statistical analysis using the modified Gompertz model. HTP was found to have improved the solubilization of the PSW sludge with increasing HTP temperature. In addition, the results of the CST, TTF, and particle size decreased with increasing HTP temperature. These results of the assessment that was conducted in this study confirm that the HTP process indeed modifies the physical properties of PSWs to enhance the solubilization of organic solids. Nevertheless, the results of the BMP tests and the modified Gompertz model analysis show that the optimal HTP temperature of PSWs for anaerobic digestion is 190°C. These findings show that to achieve high conversion efficiency, an accurately designed pre-treatment step must be included in the overall anaerobic digestion process for wastewater treatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

Fat digestion and absorption in spice-pretreated rats.

Science.gov (United States)

Prakash, Usha N S; Srinivasan, Krishnapura

2012-02-01

A few common spices are known to stimulate secretion of bile with higher amount of bile acids which play a major role in digestion and absorption of dietary lipids. It would be appropriate to verify if these spices enable efficient digestion and absorption during high-fat intake. In this context, dietary ginger (0.05%), piperine (0.02%), capsaicin (0.015%), and curcumin (0.5%) were examined for their influence on bile secretion, digestive enzymes of pancreas and absorption of dietary fat in high-fat (30%) fed Wistar rats for 8 weeks. These spices enhanced the activity of pancreatic lipase, amylase, trypsin and chymotrypsin by 22-57%, 32-51%, 63-81% and 12-38%, respectively. Dietary intake of spices along with high-fat enhanced fat absorption. These dietary spices increased bile secretion with higher bile acid content. Stimulation of lipid mobilisation from adipose tissue was suggested by the decrease in perirenal adipose tissue weight by dietary capsaicin and piperine. This was also accompanied by prevention of the accumulation of triglyceride in liver and serum in high-fat fed rats. Activities of key lipogenic enzymes in liver were reduced which was accompanied by an increased activity of hormone-sensitive lipase. Thus, dietary ginger and other spice compounds enhance fat digestion and absorption in high-fat fed situation through enhanced secretion of bile salts and a stimulation of the activity pancreatic lipase. At the same time, the energy expenditure is facilitated by these spices to prevent the accumulation of absorbed fat. Copyright © 2011 Society of Chemical Industry.

High-solids anaerobic co-digestion of sewage sludge and food waste in comparison with mono digestions: stability and performance.

Science.gov (United States)

Dai, Xiaohu; Duan, Nina; Dong, Bin; Dai, Lingling

2013-02-01

System stability and performance of high-solids anaerobic co-digestion of dewatered sludge (DS) and food waste (FW) in comparison with mono digestions were investigated. System stability was improved in co-digestion systems with co-substrate acting as a diluting agent to toxic chemicals like ammonia or Na(+). For high-solids digestion of DS, the addition of FW not only improved system stability but also greatly enhanced volumetric biogas production. For high-solids digestion of FW, the addition of DS could reduce Na(+) concentration and help maintain satisfactory stability during the conversion of FW into biogas. System performances of co-digestion systems were mainly determined by the mixing ratios of DS and FW. Biogas production and volatile solids (VSs) reduction in digestion of the co-mixture of DS and FW increased linearly with higher ratios of FW. A kinetic model, which aimed to forecast the performance of co-digestion and to assist reactor design, was developed from long-term semi-continuous experiments. Maximum VS reduction for DS and FW was estimated to be 44.3% and 90.3%, respectively, and first order constant k was found to be 0.17d(-1) and 0.50 d(-1), respectively. Experimental data of co-digestion were in good conformity to the predictions of the model. Copyright © 2012 Elsevier Ltd. All rights reserved.

Enhanced biogas production from anaerobic co-digestion of pig slurry and horse manure with mechanical pre-treatment.

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Lopes, Madalena; Baptista, Patrícia; Duarte, Elizabeth; Moreira, António L N

2018-01-02

Enhanced biogas production from anaerobic co-digestion of pig slurry and horse manure with mechanical pre-treatment. In this study, co-digestion of horse manure and pig slurry was investigated in a continuously stirred tank reactor, with a mechanical pre-treatment. Experiments were conducted at 37°C, with hydraulic retention times of 23 days and increasing shares of horse manure, corresponding to different horse manure to pig slurry ratios (HM:PS) equal to 0:100, 10:90, 13:87 and 20:80, in terms of percentage of inlet volatile solids (%VS inlet). The results show that the best synergetic effect between the microbial consortia of pig slurry and the high Carbon to Nitrogen ratio (C/N) of horse manure is obtained for the mixture of 20:80%VS inlet, yielding the highest specific methane production (SMP = 142.6 L kg TCOD -1 ) and the highest soluble chemical oxygen demand (SCOD) reduction (68.5%), due to the high volatile dissolved solids content and soluble chemical oxygen demand to total chemical oxygen demand ratio (SCOD/TCOD). Thus, co-digestion of horse manure and pig slurry is shown to be a promising approach for biogas production and as a waste treatment solution. Furthermore, the analysis provides a methodology for the pre-treatment of these substrates and to investigate into the best combination for improved biogas production.

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

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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.

Evaluation of anaerobic digestion processes for short sludge-age waste activated sludge combined with anammox treatment of digestate liquor.

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Ge, Huoqing; Batstone, Damien; Keller, Jurg

2016-01-01

The need to reduce energy input and enhance energy recovery from wastewater is driving renewed interest in high-rate activated sludge treatment (i.e. short hydraulic and solids retention times (HRT and SRT, respectively)). This process generates short SRT activated sludge stream, which should be highly degradable. However, the evaluation of anaerobic digestion of short SRT sludge has been limited. This paper assesses anaerobic digestion of short SRT sludge digestion derived from meat processing wastewater under thermophilic and mesophilic conditions. The thermophilic digestion system (55°C) achieved 60 and 68% volatile solids destruction at 8 day and 10 day HRT, respectively, compared with 50% in the mesophilic digestion system (35°C, 10 day HRT). The digestion effluents from the thermophilic (8-10 day HRT) and mesophilic systems were stable, as assessed by residual methane potentials. The ammonia rich sludge dewatering liquor was effectively treated by a batch anammox process, which exhibited comparable nitrogen removal rate as the tests using a control synthetic ammonia solution, indicating that the dewatering liquor did not have inhibiting/toxic effects on the anammox activity.

Enhanced primary treatment of concentrated black water and kitchen residues within DESAR concept using two types of anaerobic digesters.

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Kujawa-Roeleveld, K; Elmitwalli, T; Zeeman, G

2006-01-01

Anaerobic digestion of concentrated domestic wastewater streams--black or brown water, and solid fraction of kitchen waste is considered as a core technology in a source separation based sanitation concept (DESAR--decentralised sanitation and reuse). A simple anaerobic digester can be implemented for an enhanced primary treatment or, in some situations, as a main treatment. Two reactor configurations were extensively studied; accumulation system (AC) and UASB septic tank at 15, 20 and 25 degrees C. Due to long retention times in an AC reactor, far stabilisation of treated medium can be accomplished with methanisation up to 60%. The AC systems are the most suitable to apply when the volume of waste to be treated is minimal and when a direct reuse of a treated medium in agriculture is possible. Digested effluent contains both liquid and solids. In a UASB septic tank, efficient separation of solids and liquid is accomplished. The total COD removal was above 80% at 25 degrees C. The effluent contains COD and nutrients, mainly in a soluble form. The frequency of excess sludge removal is low and sludge is well stabilised due to a long accumulation time.

Ultrasonic sludge disintegration for enhanced methane production in anaerobic digestion: effects of sludge hydrolysis efficiency and hydraulic retention time.

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Kim, Dong-Jin; Lee, Jonghak

2012-01-01

Hydrolysis of waste activated sludge (WAS) has been regarded as the rate limiting step of anaerobic sludge digestion. Therefore, in this study, the effect of ultrasound and hydraulic residence time during sludge hydrolysis was investigated with the goal of enhancing methane production from anaerobic digestion (AD). WAS was ultrasonically disintegrated for hydrolysis, and it was semi-continuously fed to an anaerobic digesters at various hydraulic retention times (HRTs). The results of these experiments showed that the solids and chemical oxygen demand (COD) removal efficiencies when using ultrasonically disintegrated sludge were higher during AD than the control sludge. The longer the HRT, the higher the removal efficiencies of solids and COD, while methane production increased with lower HRT. Sludge with 30% hydrolysis produced 7 × more methane production than the control sludge. The highest methane yields were 0.350 m(3)/kg volatile solids (VS)(add) and 0.301 m(3)/kg COD(con) for 16 and 30% hydrolyzed sludge, respectively. In addition, we found that excess ultrasound irradiation may inhibit AD since the 50% hydrolyzed sludge produced lower methane yields than 16 and 30% hydrolyzed sludge.

Biocolloids with ordered urease multilayer shells as enzymatic reactors.

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Lvov, Y; Caruso, F

2001-09-01

The preparation of biocolloids with organized enzyme-containing multilayer shells for exploitation as colloidal enzymatic nanoreactors is described. Urease multilayers were assembled onto submicrometer-sized polystyrene spheres by the sequential adsorption of urease and polyelectrolyte, in a predetermined order, utilizing electrostatic interactions for layer growth. The catalytic activity of the biocolloids increased proportionally with the number of urease layers deposited on the particles, demonstrating that biocolloid particles with tailored enzymatic activities can be produced. It was further found that precoating the latex spheres with nanoparticles (40-nm silica or 12-nm magnetite) enhanced both the stability (with respect to adsorption) and enzymatic activity of the urease multilayers. The presence of the magnetite nanoparticle coating also provided a magnetic function that allowed the biocolloids to be easily and rapidly separated with a permanent magnet. The fabrication of such colloids opens new avenues for the application of bioparticles and represents a promising route for the creation of complex catalytic particles.

The effects of xylitol and sorbitol on lysozyme- and peroxidase-related enzymatic and candidacidal activities.

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Kim, Bum-Soo; Chang, Ji-Youn; Kim, Yoon-Young; Kho, Hong-Seop

2015-07-01

To investigate whether xylitol and sorbitol affect enzymatic and candidacidal activities of lysozyme, the peroxidase system, and the glucose oxidase-mediated peroxidase system. Xylitol and sorbitol were added to hen egg-white lysozyme, bovine lactoperoxidase, glucose oxidase-mediated peroxidase, and whole saliva in solution and on hydroxyapatite surfaces. The enzymatic activities of lysozyme, peroxidase, and glucose oxidase-mediated peroxidase were determined by the turbidimetric method, the NbsSCN assay, and production of oxidized o-dianisidine, respectively. Candidacidal activities were determined by comparing colony forming units using Candida albicans ATCC strains 10231, 11006, and 18804. While xylitol and sorbitol did not affect the enzymatic activity of hen egg-white lysozyme both in solution and on hydroxyapatite surfaces, they did inhibit the enzymatic activity of salivary lysozyme significantly in solution, but not on the surfaces. Xylitol and sorbitol enhanced the enzymatic activities of both bovine lactoperoxidase and salivary peroxidase significantly in a dose-dependent manner in solution, but not on the surfaces. Sorbitol, but not xylitol, inhibited the enzymatic activity of glucose oxidase-mediated peroxidase significantly. Both xylitol and sorbitol did not affect candidacidal activities of hen egg-white lysozyme, the bovine lactoperoxidase system, or the glucose oxidase-mediated bovine lactoperoxidase system. Xylitol and sorbitol inhibited salivary lysozyme activity, but enhanced both bovine lactoperoxidase and salivary peroxidase activities significantly in solution. Xylitol and sorbitol did not augment lysozyme- and peroxidase-related candidacidal activities. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enzymatic Hydrolysis Does Not Reduce the Biological Reactivity of Soybean Proteins for All Allergic Subjects.

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Panda, Rakhi; Tetteh, Afua O; Pramod, Siddanakoppalu N; Goodman, Richard E

2015-11-04

Many soybean protein products are processed by enzymatic hydrolysis to attain desirable functional food properties or in some cases to reduce allergenicity. However, few studies have investigated the effects of enzymatic hydrolysis on the allergenicity of soybean products. In this study the allergenicity of soybean protein isolates (SPI) hydrolyzed by Alcalase, trypsin, chymotrypsin, bromelain, or papain was evaluated by IgE immunoblots using eight soybean-allergic patient sera. The biological relevance of IgE binding was evaluated by a functional assay using a humanized rat basophilic leukemia (hRBL) cell line and serum from one subject. Results indicated that hydrolysis of SPI by the enzymes did not reduce the allergenicity, and hydrolysis by chymotrypsin or bromelain has the potential to increase the allergenicity of SPI. Two-dimensional (2D) immunoblot and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of the chymotrypsin-hydrolyzed samples indicated fragments of β-conglycinin protein are responsible for the apparent higher allergenic potential of digested SPI.

Enzymatically-Mediated Co-Production of Cellulose Nanocrystals and Fermentable Sugars

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Dawit Beyene

2017-10-01

Full Text Available Cellulose nanocrystals (CNCs can be extracted from cellulosic materials through the degradation of non-crystalline cellulose domains in the feedstock via acid hydrolysis. However, the sugars released from the hydrolysis process cannot be easily recovered from the acid waste stream. In this study, cellulases were used to preferentially degrade non-crystalline domains with the objectives of recovering sugars and generating a feedstock with concentrated CNC precursors for a more efficient acid hydrolysis process. Filter paper and wood pulp substrates were enzyme-treated for 2–10 h to recover 20–40 wt % glucose. Substantial xylose yield (6–12 wt % was generated from wood pulp. CNC yields from acid hydrolysis of cellulases-treated filter paper, and wood pulp improved by 8–18% and 58–86%, respectively, when compared with the original substrate. It was thought that CNC precursors accumulated in the cellulases-treated feedstock due to enzymatic digestion of the more accessible non-crystalline celluloses. Therefore, acid hydrolysis from enzyme-treated feedstock will require proportionally less water and reagents resulting in increased efficiency and productivity in downstream processes. This study demonstrates that an enzymatically-mediated process allows recovery of fermentable sugars and improves acid hydrolysis efficiency for CNC production.

Application of multi-enzymatic hydrolysis for improving the efficiency of the biogas production in solid waste fermentation process in Ostróda WWTP

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Lipiński, Kamil; Umiejewska, Katarzyna

2017-11-01

Biomass fermentation is one of the important sources of renewable energy in EU. Application of multi-enzymatic hydrolysis process enables a significant increase in efficiency of biogas production. The main goal of the paper is to present the results of the pilot scale research performed in WWTP in óstroda. The fixed combination of three enzymes was continiously introduced: amylase, lipase and protease. Research aimed at verifying the impact of enzyme dose on sludge digestion process and on the amount of biogas produced. Statistical analysis of the research results allows to determine the influence of dosing the enzymes in mesophilic digestion on the biogas production.

Protocols for the analytical characterization of therapeutic monoclonal antibodies. II - Enzymatic and chemical sample preparation.

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Bobaly, Balazs; D'Atri, Valentina; Goyon, Alexandre; Colas, Olivier; Beck, Alain; Fekete, Szabolcs; Guillarme, Davy

2017-08-15

The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. These approaches make the sample more suitable for chromatographic and mass spectrometric analysis. Structural elucidation and quality control (QC) analysis of biopharmaceutics are usually performed at intact, subunit and peptide levels. In this paper, general sample preparation approaches used to attain peptide, subunit and glycan level analysis are overviewed. Protocols are described to perform tryptic proteolysis, IdeS and papain digestion, reduction as well as deglycosylation by PNGase F and EndoS2 enzymes. Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). The described protocols may help analysts to develop sample preparation methods in the field of therapeutic protein analysis. Copyright © 2017 Elsevier B.V. All rights reserved.

Monitoring the progress of non-enzymatic glycation in vitro

International Nuclear Information System (INIS)

Shaw, S.M.; Crabbe, M.J.

1994-01-01

The progress of in vitro non-enzymatic glycation of bovine serum albumin was followed by using 14 C-glucose and a nitroblue tetrazolium assay, absorption and fluorescence spectroscopy, SDS gel electrophoresis and protease digestion. The number of adducts detectable using both 14 C-tracers and a fructosamine assay remained low at physiological glucose concentrations, fewer than five adducts being detectable. When glucose concentrations > 1.0 M were used the number of adducts was found to greatly exceed the number of lysyl residues available in BSA, indicative of cross-linking between Maillard products. Incubation of BSA with glucose concentrations of up to 160 mM for one month produced no observable increase in molecular weight by SDS gel electrophoresis, showing that at physiological glucose concentrations, increases in molecular weight were minimal for short incubation periods. Increases in absorption were proportial to both the glucose concentration and the incubation time. Several absorption peaks, at 370, 488 and 554 nm, were consistent in appearance throughout the course of each incubation. Fluorescence spectroscopy of the modified proteins showed a disappearance of the fluorescence associated with peptide bonds and aromatic residues and the appearance of a broad peak at longer wavelengths due to the wide range of absorptive/fluorescent wavelengths of the developing Maillard products. Protease digestion gave similar patterns with non-glycated and glycated protein, suggesting that glycation did not block digestion sites, and that partial digestion did not cause significant further exposure of susceptible sites. Our results show that while glycation ultimately results in protein conformational changes and the formation of large molecular weight species, these occur at a relatively late stage in the maturation of protein Maillard products, after ≥ nine months of incubation with glucose concentration of ≥ 20 mM. Monitoring of AGE maturation in vitro is better

Elucidating the role of ferrous ion cocatalyst in enhancing dilute acid pretreatment of lignocellulosic biomass

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Wei Hui

2011-11-01

Full Text Available Abstract Background Recently developed iron cocatalyst enhancement of dilute acid pretreatment of biomass is a promising approach for enhancing sugar release from recalcitrant lignocellulosic biomass. However, very little is known about the underlying mechanisms of this enhancement. In the current study, our aim was to identify several essential factors that contribute to ferrous ion-enhanced efficiency during dilute acid pretreatment of biomass and to initiate the investigation of the mechanisms that result in this enhancement. Results During dilute acid and ferrous ion cocatalyst pretreatments, we observed concomitant increases in solubilized sugars in the hydrolysate and reducing sugars in the (insoluble biomass residues. We also observed enhancements in sugar release during subsequent enzymatic saccharification of iron cocatalyst-pretreated biomass. Fourier transform Raman spectroscopy showed that major peaks representing the C-O-C and C-H bonds in cellulose are significantly attenuated by iron cocatalyst pretreatment. Imaging using Prussian blue staining indicated that Fe2+ ions associate with both cellulose/xylan and lignin in untreated as well as dilute acid/Fe2+ ion-pretreated corn stover samples. Analyses by scanning electron microscopy and transmission electron microscopy revealed structural details of biomass after dilute acid/Fe2+ ion pretreatment, in which delamination and fibrillation of the cell wall were observed. Conclusions By using this multimodal approach, we have revealed that (1 acid-ferrous ion-assisted pretreatment increases solubilization and enzymatic digestion of both cellulose and xylan to monomers and (2 this pretreatment likely targets multiple chemistries in plant cell wall polymer networks, including those represented by the C-O-C and C-H bonds in cellulose.

Enhancing the enzymatic hydrolysis of corn stover by an integrated wet-milling and alkali pretreatment.

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He, Xun; Miao, Yelian; Jiang, Xuejian; Xu, Zidong; Ouyang, Pingkai

2010-04-01

An integrated wet-milling and alkali pretreatment was applied to corn stover prior to enzymatic hydrolysis. The effects of NaOH concentration in the pretreatment on crystalline structure, chemical composition, and reducing-sugar yield of corn stover were investigated, and the mechanism of increasing reducing-sugar yield by the pretreatment was discussed. The experimental results showed that the crystalline structure of corn stover was disrupted, and lignin was removed, while cellulose and hemicellulose were retained in corn stover by the pretreatment with 1% NaOH in 1 h. The reducing-sugar yield from the pretreated corn stovers increased from 20.2% to 46.7% when the NaOH concentration increased from 0% to 1%. The 1% NaOH pretreated corn stover had a holocellulose conversion of 55.1%. The increase in reducing-sugar yield was related to the crystalline structure disruption and delignification of corn stover. It was clarified that the pretreatment significantly enhanced the conversion of cellulose and hemicellulose in the corn stover to sugars.

Effect of the molecular structure of lignin-based polyoxyethylene ether on enzymatic hydrolysis efficiency and kinetics of lignocelluloses.

Science.gov (United States)

Lin, Xuliang; Qiu, Xueqing; Zhu, Duming; Li, Zihao; Zhan, Ningxin; Zheng, Jieyi; Lou, Hongming; Zhou, Mingsong; Yang, Dongjie

2015-10-01

Effect of the molecular structure of lignin-based polyoxyethylene ether (EHL-PEG) on enzymatic hydrolysis of Avicel and corn stover was investigated. With the increase of PEG contents and molecular weight of EHL-PEG, glucose yield of corn stover increased. EHL-PEG enhanced enzymatic hydrolysis of corn stover significantly at buffer pH 4.8-5.5. Glucose yield of corn stover at 20% solid content increased from 32.8% to 63.8% by adding EHL-PEG, while that with PEG4600 was 54.2%. Effect of EHL-PEG on enzymatic hydrolysis kinetics of cellulose film was studied by quartz crystal microbalance with dissipation monitoring (QCM-D) and atomic force microscopy (AFM). An enhancing mechanism of EHL-PEG on enzymatic hydrolysis kinetics of cellulose was proposed. Cellulase aggregates dispersed by EHL-PEG excavated extensive cavities into the surface of cellulose film, making the film become more loose and exposed. After the maximum enzymatic hydrolysis rate, the film was mainly peeled off layer by layer until equilibrium. Copyright © 2015 Elsevier Ltd. All rights reserved.

A systematic quantification of carbonic anhydrase transcripts in the mouse digestive system

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Parkkila Seppo

2007-03-01

Full Text Available Abstract Background Carbonic anhydrases (CAs are physiologically important enzymes which participate in many gastrointestinal processes such as acid and bicarbonate secretion and metabolic pathways including gluconeogenesis and ureagenesis. The genomic data suggests that there are thirteen enzymatically active members of the mammalian CA isozyme family. In the present study, we systematically examined the mRNA expression levels of all known CA isozymes by quantitative real-time PCR in eight tissues of the digestive system of male and female mice. Results The CAs expressed in all tissues were Car5b, Car7, and Car15, among which Car5b showed moderate and Car7 and Car15 extremely low expression levels. Car3, Car12, Car13, and Car14 were detected in seven out of eight tissues and Car2 and Car4 were expressed in six tissues. Importantly, Car1, Car3, and Car13 showed very high expression levels in certain tissues as compared to the other CAs, suggesting that these low activity isozymes may also participate in physiological processes other than CA catalysis and high expression levels are required to fulfil their functions in the body. Conclusion A comprehensive mRNA expression profile of the 13 enzymatically active CAs in the murine gastrointestinal tract was produced in the present study. It contributes to a deeper understanding of the distribution of CA isozymes and their potential roles in the mouse digestive system.

Enrichment of pasta with faba bean does not impact glycemic or insulin response but can enhance satiety feeling and digestive comfort when dried at very high temperature.

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Greffeuille, Valérie; Marsset-Baglieri, Agnès; Molinari, Nicolas; Cassan, Denis; Sutra, Thibault; Avignon, Antoine; Micard, Valérie

2015-09-01

Enrichment of durum wheat pasta with legume flour enhances their protein and essential amino acid content, especially lysine content. However, despite its nutritional potential, the addition of a legume alters the rheological properties of pasta. High temperature drying of pasta reduces this negative effect by strengthening its protein network. The aim of our study was to determine if these changes in the pasta structure alter its in vitro carbohydrate digestibility, in vivo glycemic, insulin and satiety responses. We also investigated if high temperature drying of pasta can reduce the well-known digestive discomfort associated with the consumption of legume grains. Fifteen healthy volunteers consumed three test meals: durum wheat pasta dried at a low temperature (control), and pasta enriched with 35% faba bean dried at a low and at a very high temperature. When enriched with 35% legume flour, pasta maintained its nutritionally valuable low glycemic and insulin index, despite its weaker protein network. Drying 35% faba bean pasta at a high temperature strengthened its protein network, and decreased its in vitro carbohydrate digestion with no further decrease in its in vivo glycemic or insulin index. Drying pasta at a very high temperature reduced digestive discomfort and enhanced self-reported satiety, and was not associated with a modification of energy intake in the following meal.

Enhanced dewaterability of sludge during anaerobic digestion with thermal hydrolysis pretreatment: New insights through structure evolution.

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Zhang, Jingsi; Li, Ning; Dai, Xiaohu; Tao, Wenquan; Jenkinson, Ian R; Li, Zhuo

2017-12-19

Comprehensive insights into the sludge digestate dewaterability were gained through porous network structure of sludge. We measured the evolution of digestate dewaterability, represented by the solid content of centrifugally dewatered cake, in high-solids sequencing batch digesters with and without thermal hydrolysis pretreatment (THP). The results show that the dewaterability of the sludge after digestion was improved by 3.5% (±0.5%) for unpretreated sludge and 5.1% (±0.4%) for thermally hydrolyzed sludge. Compared to the unpretreated sludge digestate, thermal hydrolysis pretreatment eventually resulted in an improvement of dewaterability by 4.6% (±0.5%). Smaller particle size and larger surface area of sludge were induced by thermal hydrolysis and anaerobic digestion treatments. The structure strength and compactness of sludge, represented by elastic modulus and fractal dimension respectively, decreased with increase of digestion time. The porous network structure was broken up by thermal hydrolysis pretreatment and was further weakened during anaerobic digestion, which correspondingly improved the dewaterability of digestates. The logarithm of elastic modulus increased linearly with fractal dimension regardless of the pretreatment. Both fractal dimension and elastic modulus showed linear relationship with dewaterability. The rheological characterization combined with the analysis of fractal dimension of sewage sludge porous network structure was found applicable in quantitative evaluation of sludge dewaterability, which depended positively on both thermal hydrolysis and anaerobic digestion. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of iron salt type and dosing mode on Fenton-based pretreatment of rice straw for enzymatic hydrolysis.

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Gan, Yu-Yan; Zhou, Si-Li; Dai, Xiao; Wu, Han; Xiong, Zi-Yao; Qin, Yuan-Hang; Ma, Jiayu; Yang, Li; Wu, Zai-Kun; Wang, Tie-Lin; Wang, Wei-Guo; Wang, Cun-Wen

2018-06-15

Fenton-based processes with four different iron salts in two different dosing modes were used to pretreat rice straw (RS) samples to increase their enzymatic digestibility. The composition analysis shows that the RS sample pretreated by the dosing mode of iron salt adding into H 2 O 2 has a much lower hemicellulose content than that pretreated by the dosing mode of H 2 O 2 adding into iron salt, and the RS sample pretreated by the chloride salt-based Fenton process has a much lower lignin content and a slightly lower hemicellulose content than that pretreated by the sulphate salt-based Fenton process. The higher concentration of reducing sugar observed on the RS sample with lower lignin and hemicellulose contents justifies that the Fenton-based process could enhance the enzymic hydrolysis of RS by removing hemicellulose and lignin and increasing its accessibility to cellulase. FeCl 3 ·6H 2 O adding into H 2 O 2 is the most efficient Fenton-based process for RS pretreatment. Copyright © 2018 Elsevier Ltd. All rights reserved.

Energetic and biochemical valorization of cork boiling wastewater by anaerobic digestion.

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Marques, Isabel Paula; Gil, Luís; La Cara, Francesco

2014-01-01

In addition to energy benefits, anaerobic digestion offers other interesting advantages. The cork industry is of great environmental, economic and social significance in the western Mediterranean region, with Portugal being the world-leading producer and exporter. Cork boiling wastewater (CBW) is a toxic and recalcitrant organic effluent produced by this sector, which constitutes a serious environmental hazard. However, there is no documented research on anaerobic treatment/valorization performed with this effluent. The work presented here was developed with the aim to use the anaerobic digestion process to convert the CBW polluting organic load into an energy carrier gas and valuable molecules for industry. No lag phases were observed and a methane yield of 0.126 to 0.142 m(3) kg(-1) chemical oxygen demand (COD)added was registered in the mesophilic consortium experiments carried out in batch flasks at 37 ± 1°C. Anaerobic digestion can be advantageously connected to ultrafiltration or electrochemical processes, due to the following: 1) reduction of ellagic acid content and consequent decrease of CBW viscosity; and 2) increase in conductivity after the anaerobic process, avoiding the electrolyte application of the electrochemical process. The improvement of several CBW biochemical features shows that anaerobic digestion may provide additionally useful molecules. The rise in concentration of some of these compounds, belonging to the benzoic acid family (gallic, protocatechuic, vanillic and syringic acids), is responsible for the increase of antiradical activity of the phenolic fraction. Additionally, some enzymatic activity was also observed and while the laccase activity increased in the digested effluent by anaerobiosis, xylanase was formed in the process. The multidisciplinary approach adopted allowed the valorization of CBW in terms of energy and valuable biomolecules. By exploiting the anaerobic digestion process potential, a novel methodology to toxic

Engineering Digestion: Multiscale Processes of Food Digestion.

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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. © 2016 Institute of Food Technologists®

Real-time monitoring of peptic and tryptic digestions of bovine {beta}-casein using quartz crystal microbalance

Energy Technology Data Exchange (ETDEWEB)

Huenerbein, Andreas [Institute of Pharmaceutics and Biopharmaceutics, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-Str. 4, 06120 Halle (Saale) (Germany)]. E-mail: andreas.huenerbein@pharmazie.uni-halle.de; Schmelzer, Christian E.H. [Institute of Pharmaceutics and Biopharmaceutics, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-Str. 4, 06120 Halle (Saale) (Germany); Neubert, Reinhard H.H. [Institute of Pharmaceutics and Biopharmaceutics, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-Str. 4, 06120 Halle (Saale) (Germany)

2007-02-12

In this study peptic and tryptic digestions of bovine {beta}-casein were investigated using quartz crystal microbalance (QCM). {beta}-Casein, which was used as a model protein, was immobilized on the surface of the QCM sensor where its degradation caused shifts in the resonant frequency. Atomic force microscopy was applied for the characterization of the protein layer. Different pH-values for peptic or tryptic digestions were chosen to visualize their effect on enzyme activity. Lower frequency shifts were observed at pH-values deviating from those at the maximum enzyme activity. In the case of the peptic digestion the frequency shift at pH 4 was more than 10 times smaller than those at pH 2. The frequency shifts for tryptic digestions at pH 5.4 and pH 6.4 were about two thirds compared to that obtained for the digestion at pH 7.4. The identification of peptides using MALDI-ToF mass spectrometry was used for verification of the proteolyses of the immobilized protein. Furthermore, it was shown that the QCM technique allows close observation of the effect of different pH-values on the immobilized casein layer. All in all, QCM facilitates the monitoring of the progress of enzymatic reactions in real-time.

Real-time monitoring of peptic and tryptic digestions of bovine β-casein using quartz crystal microbalance

International Nuclear Information System (INIS)

Huenerbein, Andreas; Schmelzer, Christian E.H.; Neubert, Reinhard H.H.

2007-01-01

In this study peptic and tryptic digestions of bovine β-casein were investigated using quartz crystal microbalance (QCM). β-Casein, which was used as a model protein, was immobilized on the surface of the QCM sensor where its degradation caused shifts in the resonant frequency. Atomic force microscopy was applied for the characterization of the protein layer. Different pH-values for peptic or tryptic digestions were chosen to visualize their effect on enzyme activity. Lower frequency shifts were observed at pH-values deviating from those at the maximum enzyme activity. In the case of the peptic digestion the frequency shift at pH 4 was more than 10 times smaller than those at pH 2. The frequency shifts for tryptic digestions at pH 5.4 and pH 6.4 were about two thirds compared to that obtained for the digestion at pH 7.4. The identification of peptides using MALDI-ToF mass spectrometry was used for verification of the proteolyses of the immobilized protein. Furthermore, it was shown that the QCM technique allows close observation of the effect of different pH-values on the immobilized casein layer. All in all, QCM facilitates the monitoring of the progress of enzymatic reactions in real-time

Fed batch enzymatic saccharification of food waste improves the sugar concentration in the hydrolysates and eventually the ethanol fermentation by Saccharomyces cerevisiae H058

Directory of Open Access Journals (Sweden)

Shoubao Yan

2012-04-01

Full Text Available The enzymatic hydrolysis of food waste by commercially available enzymes and the subsequent ethanol fermentation of the hydrolysates by Saccharomyces cerecisiae H058 were studied in this work. The optimum batch enzymatic conditions were found to be saccharification pH of 4.5, temperature of 55!, glucoamylase concentration of 120 u/g, α-amylase concentration of 10 u/g, solid-liquid ratio of 1: 0.75 (w/w. Fed batch hydrolysis process was started with a solid-liquid ratio of 1: 1 (w/w, with solid food waste added at time lapse of 2 h to get a final solid-liquid ratio of 1: 0.5 (w/w. After 4 h of reaction, the reducing sugar concentration reached 194.43 g/L with a enzymatic digestibility of 93.12%. Further fermentation of the batch and fed batch enzymatic hydrolysates, which contained reducing sugar concentration of 131.41 and 194.43 g/L respectively, was performed using Saccharomyces cerevisiae H058, 62.93 and 90.72 g/L ethanol was obtained within 48 h.

Combination of Superheated Steam with Laccase Pretreatment Together with Size Reduction to Enhance Enzymatic Hydrolysis of Oil Palm Biomass

Directory of Open Access Journals (Sweden)

Nur Fatin Athirah Ahmad Rizal

2018-04-01

Full Text Available The combination of superheated steam (SHS with ligninolytic enzyme laccase pretreatment together with size reduction was conducted in order to enhance the enzymatic hydrolysis of oil palm biomass into glucose. The oil palm empty fruit bunch (OPEFB and oil palm mesocarp fiber (OPMF were pretreated with SHS and ground using a hammer mill to sizes of 2, 1, 0.5 and 0.25 mm before pretreatment using laccase to remove lignin. This study showed that reduction of size from raw to 0.25 mm plays important role in lignin degradation by laccase that removed 38.7% and 39.6% of the lignin from OPEFB and OPMF, respectively. The subsequent saccharification process of these pretreated OPEFB and OPMF generates glucose yields of 71.5% and 63.0%, which represent a 4.6 and 4.8-fold increase, respectively, as compared to untreated samples. This study showed that the combination of SHS with laccase pretreatment together with size reduction could enhance the glucose yield.

Digestive enzymes of the Californian two-spot octopus, Octopus bimaculoides (Pickford and McConnaughey, 1949).

Science.gov (United States)

Ibarra-García, Laura Elizabeth; Tovar-Ramírez, Dariel; Rosas, Carlos; Campa-Córdova, Ángel Isidro; Mazón-Suástegui, José Manuel

2018-01-01

Octopus bimaculoides is an important commercially fished species in the California Peninsula with aquaculture potential; however, to date limited information is available regarding its digestive physiology. The objective of this study was focused on biochemically characterizing the main enzymes involved in the digestion of O. bimaculoides. Optimum pH, temperature and thermostability were determined for amylases, lipases, trypsin and chymotrypsin; optimum pH and protease inhibitor effect were assessed for acidic and alkaline proteases, and the effect of divalent ions on trypsin and chymotrypsin activity was evaluated in enzymatic extracts from the digestive (DG) and salivary glands (SG) and crop gastric juices (GJ). High amylase activity was detected in GD and GJ whereas this activity is very low in other cephalopods. Salivary glands had the greatest activity in most of the enzyme groups, showing the importance of this organ in digestion. Optimum pH was different depending on the organ and enzyme analyzed. The optimum pH in DG was 3 showing the predominance of acidic proteases in the digestion process. All enzymes were resistant and stable at high temperatures in contrast with other marine species. Trypsin and chymotrypsin activity were highly incremented with the presence of Mg 2+ , Co 2+ , Cu 2+ and Zn 2+ in some tissues. The inhibitor assay showed the importance of serine proteases, metalloproteases and aspartic proteases in the digestive process of this species. This study is the first in assessing the main digestive enzymes of O. bimaculoides and in remarking the importance of other digestive enzyme groups besides proteases in octopuses. Copyright © 2017 Elsevier Inc. All rights reserved.

Improvement in Saccharification Yield of Mixed Rumen Enzymes by Identification of Recalcitrant Cell Wall Constituents Using Enzyme Fingerprinting.

Science.gov (United States)

Badhan, Ajay; Wang, Yu-Xi; Gruninger, Robert; Patton, Donald; Powlowski, Justin; Tsang, Adrian; McAllister, Tim A

2015-01-01

Identification of recalcitrant factors that limit digestion of forages and the development of enzymatic approaches that improve hydrolysis could play a key role in improving the efficiency of meat and milk production in ruminants. Enzyme fingerprinting of barley silage fed to heifers and total tract indigestible fibre residue (TIFR) collected from feces was used to identify cell wall components resistant to total tract digestion. Enzyme fingerprinting results identified acetyl xylan esterases as key to the enhanced ruminal digestion. FTIR analysis also suggested cross-link cell wall polymers as principal components of indigested fiber residues in feces. Based on structural information from enzymatic fingerprinting and FTIR, enzyme pretreatment to enhance glucose yield from barley straw and alfalfa hay upon exposure to mixed rumen-enzymes was developed. Prehydrolysis effects of recombinant fungal fibrolytic hydrolases were analyzed using microassay in combination with statistical experimental design. Recombinant hemicellulases and auxiliary enzymes initiated degradation of plant structural polysaccharides upon application and improved the in vitro saccharification of alfalfa and barley straw by mixed rumen enzymes. The validation results showed that microassay in combination with statistical experimental design can be successfully used to predict effective enzyme pretreatments that can enhance plant cell wall digestion by mixed rumen enzymes.

Enhanced anaerobic treatment of CSTR-digested effluent from chicken manure: The effect of ammonia inhibition.

Science.gov (United States)

Liu, Zhan-Guang; Zhou, Xue-Fei; Zhang, Ya-Lei; Zhu, Hong-Guang

2012-01-01

The effect of ammonia inhibition was evaluated during the enhanced anaerobic treatment of digested effluent from a 700m(3) chicken-manure continuous stirred tank reactor (CSTR). A 12.3L internal circulation (IC) reactor inoculated with an anaerobic granular sludge and operated at 35±1°C was employed for the investigation. With a corresponding organic loading rate of 1.5-3.5kg-COD/m(3)d over a hydraulic retention time of 1.5d, a maximum volumetric biogas production rate of 1.2m(3)/m(3)d and TCOD (total COD) removal efficiency ranging from 70% to 80% was achieved. However, the continual increase in the influent TAN content led to ammonia inhibition in the methanogenesis system. The SCOD/TAN (soluble COD/total ammonia nitrogen) ratio was presented to be the key controlling factor for the anaerobic treatment of semi-digested chicken manure, and further validation through shock loading and ammonia inhibition experiments was conducted. The threshold value of the SCOD/TAN ratio was determined to be 2.4 (corresponding to a TAN of 1250mg/L) at an influent pH of 8.5-9. Copyright © 2011 Elsevier Ltd. All rights reserved.

Effects of dietary yeastβ-glucan on nutrient digestibility and serum proifles in pre-ruminant Holstein calves

Institute of Scientific and Technical Information of China (English)

MA Tao; TU Yan; ZHANG Nai-feng; GUO Jiang-peng; DENG Kai-dong; ZHOU Yi; YUN Qiang; DIAO Qi-yu

2015-01-01

This study aimed to investigate the effects of dietary supplementation of yeastβ-glucan on the nutrient digestibility and serum proifles in pre-ruminant Holstein calves. Forty-two neonatal Holstein calves ((39.6±4.2) kg) were randomly al otted to six groups, and each was offered one of the fol owing diets:a basal diet (control) or the basal diet supplemented with 25, 50, 75, 100 or 200 mg of yeastβ-glucan kg–1 feed (dry matter basis). The basal diet consisted of a milk replacer and a starter feed. The trial lasted for 56 d. Two digestibility trials were conducted from d 14 to 20 and from d 42 to 48. Blood samples were col ected on d 0, 14, 28 and 42 for serum proifle analyses. On d 56, three calves from each group were slaughtered, and intestinal samples were col ected to assess the vil ous height, crypt depth and mucosal thickness. Although feed intake was not affected by dietary treatment (P>0.05), the average daily gain (ADG) and gain-to-feed ratios were higher (P0.05). Compared with the control group, supplementation of yeastβ-glucan decreased (P0.05). The supplementation of yeastβ-glucan stimu-lated the enzymatic activity of alkaline phosphatase (ALP) (P<0.05) compared with the control group. The lysozyme (LYZ) concentration increased quadratical y (P<0.05) with increasing yeastβ-glucan levels. The results suggested that dietary supplementation of yeastβ-glucan at 75 mg kg–1 feed improved nutrient digestibility, enhanced immunity by increasing the immunoglobulin concentration and stimulating ALP, and exerted no adverse effects on metabolism in pre-ruminant calves.

Comparison of the suitability of alkaline or enzymatic sample pre-treatment for characterization of silver nanoparticles in human tissue by single particle ICP-MS

DEFF Research Database (Denmark)

Vidmar, Janja; Buerki-Thurnherr, Tina; Löschner, Katrin

2018-01-01

and their size are required for studying NP accumulation in placental tissue. In the present study, we applied and compared two sample preparation techniques, alkaline and enzymatic treatment, followed by single particle ICP-MS (spICP-MS) analysis, for characterizing AgNPs spiked to human placental tissue. Both...... sample preparation approaches are currently used for AgNPs in biological tissues but have not been directly compared yet. We showed that the method using enzymatic tissue treatment followed by spICP-MS is efficient for determination of mass and number concentration and size distribution of AgNPs in human...... placental tissues. Properties of the AgNPs were preserved during enzymatic digestion and comparable with the primary particles. The matrix effect on the determination of Ag sensitivity and transport efficiency in spICP-MS analysis was systematically evaluated as well. The method was applied to human...

Rapid tryptic mapping using enzymatically active mass spectrometer probe tips

Energy Technology Data Exchange (ETDEWEB)

Dogruel, D.; Williams, P.; Nelson, R.W. [Arizona State Univ., Tempe, AZ (United States)

1995-12-01

A method has been developed for rapid, sensitive, and accurate tryptic mapping of polypeptides using matrix-assisted laser desorption/ionization time-of-flight mass analysis. The technique utilizes mass spectrometer probe tips which have been activated through the covalent immobilization of trypsin. The enzymatically active probe tips were used for the tryptic mapping of chicken egg lysozyme and the results compared with those obtained using either free trypsin or agarose-immobilized trypsin. A significant increase in the overall sensitivity of the process was observed using the active probe tips, as well as the production of more characteristic proteolytic fragments and the elimination of background signals due to the autolysis of the trypsin. Further, probe tip digestions were found to be rapid and convenient. 19 refs., 6 figs., 2 tabs.

Aerobic treatment of swine manure to enhance anaerobic digestion and microalgal cultivation.

Science.gov (United States)

Bekoe, Dominic; Wang, Lijun; Zhang, Bo; Scott Todd, Matthew; Shahbazi, Abolghasem

2018-02-01

Aerobic treatment of swine manure was coupled with anaerobic digestion and microalgal cultivation. A 14-day aerobic treatment reduced the total solid content of swine manure by >15%. Ammonia and carbon dioxide were stripped by the air supplied, and this off-gas was further used to aerate the culture of Chlorella vulgaris. The microalgal growth rates in Bristol medium and the wastewater with the off-gas increased from 0.08 to 0.22 g/L/d and from 0.15 to 0.24 g/L/d, respectively. Meanwhile, the aerobically treated swine manure showed a higher methane yield during anaerobic digestion. The experimental results were used to establish a demonstration unit consisting of a 100 L composter, a 200 L anaerobic digester, a 60 L tubular photobioreactor, and a 300 L micro-open raceway pond.

Thermal pre-treatment of primary and secondary sludge at 70 °C prior to anaerobic digestion

DEFF Research Database (Denmark)

Skiadas, Ioannis; Gavala, Hariklia N.; Lu, J.

2005-01-01

In general, mesophilic anaerobic digestion of sewage sludge is more widely used compared tothermophilic digestion, mainly because of the lower energy requirements and higher stability of the process. However, the thermophilic anaerobic digestion process is usually characterised by accelerated...... studyinvestigates the effect of the pre-treatment at 70 °C on thermophilic (55 °C) anaerobic digestion of primaryand secondary sludge in continuously operated digesters. Thermal pre-treatment of primary and secondarysludge at 70 °C enhanced the removal of organic matter and the methane production during...... the subsequentanaerobic digestion step at 55 °C. It also greatly contributed to the destruction of pathogens present inprimary sludge. Finally it results in enhanced microbial activities of the subsequent anaerobic stepsuggesting that the same efficiencies in organic matter removal and methane recovery could be obtained...

Prospects for methane production and nutrient recycling from lipid extracted residues and whole Nannochloropsis salina using anaerobic digestion

International Nuclear Information System (INIS)

Bohutskyi, Pavlo; Chow, Steven; Ketter, Ben; Betenbaugh, Michael J.; Bouwer, Edward J.

2015-01-01

Highlights: • Semi-continuous AD of untreated and enzymatically pretreated lipid extracted algae. • Coupled biodiesel and methane process yields 40% more energy than biodiesel alone. • Thermal pretreatment (150–170 °C) of whole algae was more effective than enzymatic. • Addition of 5% of AD effluent was optimal to support high growth of Nannochloropsis. • AD effluent can partly replace chemical fertilizer for algal growth. - Abstract: Sustainable mass production of algal biofuels requires a reduction in nutrient demand and efficient conversion into fuels of all biomass including lipid-extracted algal residues (LEA). This study evaluated methane production, nutrient recovery and recycling from untreated and enzymatically pretreated Nannochloropsis LEA using semi-continuous anaerobic digestion (AD). Additionally, this process was compared to methane generation from whole Nannochloropsis alga (WA) and thermally pretreated WA. The methane production from untreated LEA and WA reached up to 0.22 L and 0.24 L per gram of biomass volatile solids (VS), respectively, corresponding to only 36–38% of the theoretical potential. Additionally, observed VS reduction was only 40–50% confirming biomass recalcitrance to biodegradation. While enzymatic treatment hydrolyzed up to 65% of the LEA polysaccharides, the methane production increased by only 15%. Alternatively, WA thermal pretreatment at 150–170 °C enhanced methane production up to 40%. Overall, an integrated process of lipid conversion into biodiesel coupled with LEA conversion into methane generates nearly 40% more energy compared to methane production from WA, and about 100% more energy than from biodiesel alone. Additionally, the AD effluent contained up to 60–70% of the LEA phosphorus content, 30–50% of the nitrogen, sulfur, calcium and boron, 20% of the iron and cobalt, and 10% of manganese, zinc and copper, which can partially replace chemical fertilizers during algal cultivation. Consequently

Processing biogas planet digestates into value-added products -BIOVIRTA

Energy Technology Data Exchange (ETDEWEB)

Paavola, T.; Rintala, J. (MTT Agrifood Research Finland, Jokioinen (Finland)), Email: teija.paavola@mtt.fi; Sahltroem, L.; Maunuksela, L.; Torniainen, M. (Finnish Food Safety Authority, EVIRA, Helsinki (Finland)), Email: leena.sahlstrom@evira.fi; Kaparaju, P.; Rintala, J. (Univ. of Jyvaeskylae (Finland)), Email: jukka.rintala@jyu.fi; Vikman, M.; Kapanen, A. (VTT Technical Research Centre of Finland, Espoo (Finland)), Email: minna.vikman@vtt.fi

2010-10-15

The objective of BIOVIRTA project is to develop technologies and practices with which digestates, originating from anaerobic digestion of different organic wastes and by-products, can be refined to value-added and safe products for various end-uses. It is expected that the operational preconditions for biogas plants will be significantly enhanced when the end-products are proven safe and applicable. Selection of the raw materials for anaerobic codigestion is the main operational strategy that could influence the nutrient content in the digestate. This has been clearly established in the laboratory and full-scale studies with various digestates originating from different raw materials, e.g. rendering and slaughterhouse byproducts. The nutrient content in the digestate also affects the opportunities to produce refined digestate products. In this project, the possibilities for several processing technologies, e.g. mechanical separation and stripping, have been intensively evaluated for the production of different digestate products. Their mass balances have also been estimated. The feasibility for the use of the digestate products has been assessed based on their chemical and hygienic quality and for various end-uses, including as organic fertiliser and/or soil improver in crop production. The preliminary results of these field-experiments showed that the yield of barley fertilised with digestate products was comparable to inorganic fertilisers. (orig.)

Relationship between Molecular Structure Characteristics of Feed Proteins and Protein Digestibility and Solubility

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Mingmei Bai

2016-08-01

Full Text Available 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 (p<0.001 and solubility (p = 0.002. These results demonstrate that the molecular structure characteristics of feed proteins 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.

Enhancement of certain agro-cellulosic wastes through radiation processing for livestock feed

International Nuclear Information System (INIS)

Farag, D.M.H.; Zakaria, S.

1999-01-01

A study was undertaken with air-dried peanut hulls, sugarcane bagasse, corn stalks, concobs, banana leaves, wheat straw and broad bean straw to determine the effect of radiation treatment up to 3 MGY on their chemical composition and the effect of enzymatic hydrolysis on treated wastes. The effect of gamma irradiation on crude protein, fat and ash of these wastes was negligible. While radiation treatment reduced the content of crude fiber in all these wastes with corresponding increases in dry matter digestibility in vitro, as a function of radiation dose. When peanut hulls, sugarcane bagasse and wheat straw were exposed to gamma rays at 0, 1, 2 and 3 MGy a linear reduction in fiber components (neutral detergent fiber, acid detergent fiber, and lignin), increased in glucose yield enzymatic hydrolysis and dry matter digestibility were observed for peanut hulls, sugarcane bagasse and wheat straw. The findings confirm that irradiation of fibrous vegetable materials could convert them into a valuable feed supplement

Barley grain constituents, starch composition, and structure affect starch in vitro enzymatic hydrolysis.

Science.gov (United States)

Asare, Eric K; Jaiswal, Sarita; Maley, Jason; Båga, Monica; Sammynaiken, Ramaswami; Rossnagel, Brian G; Chibbar, Ravindra N

2011-05-11

The relationship between starch physical properties and enzymatic hydrolysis was determined using ten different hulless barley genotypes with variable carbohydrate composition. The ten barley genotypes included one normal starch (CDC McGwire), three increased amylose starches (SH99250, SH99073, and SB94893), and six waxy starches (CDC Alamo, CDC Fibar, CDC Candle, Waxy Betzes, CDC Rattan, and SB94912). Total starch concentration positively influenced thousand grain weight (TGW) (r(2) = 0.70, p starch concentration (r(2) = -0.80, p hydrolysis of pure starch (r(2) = -0.67, p starch concentration (r(2) = 0.46, p starch (RS) in meal and pure starch samples. The rate of starch hydrolysis was high in pure starch samples as compared to meal samples. Enzymatic hydrolysis rate both in meal and pure starch samples followed the order waxy > normal > increased amylose. Rapidly digestible starch (RDS) increased with a decrease in amylose concentration. Atomic force microscopy (AFM) analysis revealed a higher polydispersity index of amylose in CDC McGwire and increased amylose genotypes which could contribute to their reduced enzymatic hydrolysis, compared to waxy starch genotypes. Increased β-glucan and dietary fiber concentration also reduced the enzymatic hydrolysis of meal samples. An average linkage cluster analysis dendrogram revealed that variation in amylose concentration significantly (p starch concentration in meal and pure starch samples. RS is also associated with B-type granules (5-15 μm) and the amylopectin F-III (19-36 DP) fraction. In conclusion, the results suggest that barley genotype SH99250 with less decrease in grain weight in comparison to that of other increased amylose genotypes (SH99073 and SH94893) could be a promising genotype to develop cultivars with increased amylose grain starch without compromising grain weight and yield.

ETHANOL ORGANOSOLV PRETREATMENT OF BAMBOO FOR EFFICIENT ENZYMATIC SACCHARIFICATION

Directory of Open Access Journals (Sweden)

Zhiqiang Li,

2012-06-01

Full Text Available Bamboo is a potential lignocellulosic biomass for the production of bioethanol because of its high cellulose and hemicelluloses content. In this research, ethanol organosolv pretreatment with dilute sulfuric acid as the catalyst was studied in order to enhance enzymatic saccharification of moso bamboo. The addition of 2% (w/w bamboo dilute sulfuric acid in 75% ethanol had a particularly strong effect on fractionation of bamboo. It yielded a solids fraction containing 83.4% cellulose in the treated substrate. The cellulose conversion to glucose yield reached 77.1 to 83.4% after enzymatic hydrolysis of the solids fraction for 48 h at an enzyme loading of 15 FPU cellulase/g cellulose and 30 IU β-glucosidase/g cellulose. The enzymatic hydrolysis rate was significantly accelerated as the ethanol organosolv pretreatment time increased, reaching the highest enzymatic glucose yield of 83.4% after 48 h at 50 °C. The concentrations of fermentation inhibitors such as HMF (5-hydroxy-2-methyl furfural and furfural were 0.96 g/L and 4.38 g/L in the spent liquor after the ethanol organosolv pretreatment, which were slightly lower than the concentrations quantified during H2SO4-water treatment. Spent liquor was diluted with water, and more than 87.2% of lignin in raw bamboo was recovered as ethanol organosolv lignin through the filtration process.

Anaerobic co-digestion of municipal organic wastes and pre-treatment to enhance biogas production from waste.

Science.gov (United States)

Li, Chenxi; Champagne, Pascale; Anderson, Bruce C

2014-01-01

Co-digestion and pre-treatment have been recognized as effective, low-cost and commercially viable approaches to reduce anaerobic digestion process limitations and improve biogas yields. In our previous batch-scale study, fat, oil, and grease (FOG) was investigated as a suitable potential co-substrate, and thermo-chemical pre-treatment (TCPT) at pH = 10 and 55 °C improved CH4 production from FOG co-digestions. In this project, co-digestions with FOG were studied in bench-scale two-stage thermophilic semi-continuous flow co-digesters with suitable TCPT (pH = 10, 55 °C). Overall, a 25.14 ± 2.14 L/d (70.2 ± 1.4% CH4) biogas production was obtained, which was higher than in the two-stage system without pre-treatment. The results could provide valuable fundamental information to support full-scale investigations of anaerobic co-digestion of municipal organic wastes.

Boosting methane generation by co-digestion of sludge with fruit and vegetable waste: Internal environment of digester and methanogenic pathway.

Science.gov (United States)

Di Maria, Francesco; Barratta, Martino

2015-09-01

The effects of anaerobic co-digestion of waste-mixed sludge with fruit and vegetable waste (FVW) on the methane generation of a mesophilic digester was investigated. Organic loading rates (OLR) were 1.46kgVS/m(3)day, 2.1kgVS/m(3)day and 2.8kgVS/m(3)day. Increase in the OLR due to FVW co-digestion caused modification of the internal environment of the digester, mainly in terms of N-NH4 (mg/L). Corresponding microbial populations were investigated by metagenomic high-throughput sequencing. Maximum specific bio-methane generation of 435 NLCH4 per kgVS feed was achieved for an OLR of 2.1kgVS/m(3)day, which corresponded to a biomethane generation per kgVS removed of about 1700 NLCH4. In these conditions the methanogenic pathway was dominated by aceticlastic Methanosaeta and hydrogenotrophic/aceticlastic Methanoscarcinae. Ammonia concentration in the digester resulted a key parameter for enhancing syntrophic acetate oxidation, enabling a balanced aceticlastic and hydrogenotrophic/aceticlastic methanogenic pathway. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effects of processing method and solute interactions on pepsin digestibility of cooked proso millet flour.

Science.gov (United States)

Gulati, Paridhi; Sabillón, Luis; Rose, Devin J

2018-07-01

Previous studies have reported a substantial decline in in vitro digestibility of proso millet protein upon cooking. In this study, several processing techniques and cooking solutions were tested with the objective of preventing the loss in pepsin digestibility. Proso millet flour was subjected to the following processing techniques: high pressure processing (200 and 600 MPa for 5 and 20 min); germination (96 h); fermentation (48 h); roasting (dry heating); autoclaving (121 °C, 3 h), and treatment with transglutaminase (160 mg/g protein, 37 °C, 2 h). To study the interaction of millet proteins with solutes, millet flour was heated with sucrose (3-7 M); NaCl (2-6 M); and CaCl 2 (0.5-3 M). All processing treatments failed to prevent the loss in pepsin digestibility except germination and treatment with transglutaminase, which resulted in 23 and 39% increases in digestibility upon cooking, respectively, when compared with unprocessed cooked flours. Heating in concentrated solutions of sucrose and NaCl were effective in preventing the loss in pepsin digestibility, an effect that was attributed to a reduction in water activity (a w ). CaCl 2 was also successful in preventing the loss in digestibility but its action was similar to chaotrops like urea. Thus, a combination of enzymatic modification and cooking of millet flour with either naturally low a w substances or edible sources of chaotropic ions may be useful in processing of proso millet for development of novel foods without loss in digestibility. However, more research is required to determine optimum processing conditions. Copyright © 2018 Elsevier Ltd. All rights reserved.

Liquid nitrogen pretreatment of eucalyptus sawdust and rice hull for enhanced enzymatic saccharification.

Science.gov (United States)

Castoldi, Rafael; Correa, Vanesa G; de Morais, Gutierrez Rodrigues; de Souza, Cristina G M; Bracht, Adelar; Peralta, Rosely A; Peralta-Muniz Moreira, Regina F; Peralta, Rosane M

2017-01-01

In this work, liquid nitrogen was used for the first time in the pretreatment of plant biomasses for purposes of enzymatic saccharification. After treatment (cryocrushing), the initial rates of the enzymatic hydrolysis of eucalyptus sawdust and rice hull were increased more than ten-fold. Cryocrushing did not modify significantly the contents of cellulose, hemicellulose and lignin in both eucalyptus sawdust and rice hulls. However, substantial disorganization of the lignocellulosic materials in consequence of the pretreatment could be observed by electron microscopy. Cryocrushing was highly efficient in improving the saccharification of the holocellulose component of the plant biomasses (from 4.3% to 54.1% for eucalyptus sawdust and from 3.9% to 40.6% for rice hull). It is important to emphasize that it consists in a simple operation with low requirements of water and chemicals, no corrosion, no release of products such as soluble phenolics, furfural and hydroxymethylfurfural and no waste generation. Copyright © 2016 Elsevier Ltd. All rights reserved.

Feeding indices and enzymatic activities of carob moth Ectomyelois ceratoniae (Zeller (Lepidoptera: pyrallidae on two commercial pistachio cultivars and an artificial diet

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Naeimeh Teimouri

2015-01-01

Full Text Available Feeding indices and enzymatic activities of Ectomyelois ceratoniae (Zeller were studied in a growth chamber under controlled conditions (29 ± 2 °C, relative humidity of 70 ± 5% and a photoperiod of 16:8 (L:D hours on two commercial Pistachio cultivars (Akbari and Kalequchi and an artificial diet. Feeding indices of E. ceratoniae larvae differed significantly on three hosts (P < 0.05. The relative consumption rate was calculated to be 5.36 ± 0.009, 11.10 ± 1.49 and 10.631 ± 0.599 (mg/mg/day on artificial diet, Akbari and Kalequchi cultivars, respectively. Carob moth larvae reared on Akbari cultivar showed the highest efficiency of conversion of digested food (ECD (5.64 ± 0.43. The highest amount of efficiency of conversion of ingested food (ECI was obtained on artificial diet but approximate digestibility (AD was the lowest on this diet. The highest enzymatic activities of alpha-amylase, general proteases and lipase were observed in the midgut of larvae reared on artificial diet. Total protein and lipid value were highest in larvae that were reared on artificial diet.

Hydrodynamic cavitation as a strategy to enhance the efficiency of lignocellulosic biomass pretreatment

DEFF Research Database (Denmark)

Terán Hilares, Ruly; Ramos, Lucas; da Silva, Silvio Silvério

2018-01-01

to accelerate certain chemical reactions. The application of cavitation energy to enhance the efficiency of lignocellulosic biomass pretreatment is an interesting strategy proposed for integration in biorefineries for the production of bio-based products. Moreover, the use of an HC-assisted process...... was demonstrated as an attractive alternative when compared to other conventional pretreatment technologies. This is not only due to high pretreatment efficiency resulting in high enzymatic digestibility of carbohydrate fraction, but also, by its high energy efficiency, simple configuration, and construction...... of systems, besides the possibility of using on the large scale. This paper gives an overview regarding HC technology and its potential for application on the pretreatment of lignocellulosic biomass. The parameters affecting this process and the perspectives for future developments in this area are also...

Methane emissions from digestate at an agricultural biogas plant.

Science.gov (United States)

Baldé, Hambaliou; VanderZaag, Andrew C; Burtt, Stephen D; Wagner-Riddle, Claudia; Crolla, Anna; Desjardins, Raymond L; MacDonald, Douglas J

2016-09-01

Methane (CH4) emissions were measured over two years at an earthen storage containing digestate from a mesophilic biodigester in Ontario, Canada. The digester processed dairy manure and co-substrates from the food industry, and destroyed 62% of the influent volatile solids (VS). Annual average emissions were 19gCH4m(-3)d(-1) and 0.27gCH4kg(-1)VSd(-1). About 76% of annual emissions occurred from June to October. Annual cumulative emissions from digestate corresponded to 12% of the CH4 produced within the digester. A key contributor to CH4 emissions was the sludge layer in storage, which contained as much VS as the annual discharge from the digester. These findings suggest that digestate management provides an opportunity to further enhance the benefits of biogas (i.e. reducing CH4 emissions compared to undigested liquid manure, and producing renewable energy). Potential best practices for future study include complete storage emptying, solid-liquid separation, and storage covering. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

Digestive Diseases

Science.gov (United States)

... cells and provide energy. This process is called digestion. Your digestive system is a series of hollow organs joined ... are also involved. They produce juices to help digestion. There are many types of digestive disorders. The ...

Enhancing recovery of ammonia from swine manure anaerobic digester effluent using gas-permeable membrane technology.

Science.gov (United States)

Dube, P J; Vanotti, M B; Szogi, A A; García-González, M C

2016-03-01

Gas-permeable membrane technology is useful to recover ammonia from manure. In this study, the technology was enhanced using aeration instead of alkali chemicals to increase pH and the ammonium (NH4(+)) recovery rate. Digested effluents from covered anaerobic swine lagoons containing 1465-2097 mg NH4(+)-N L(-1) were treated using submerged membranes (0.13 cm(2) cm(-3)), low-rate aeration (120 mL air L-manure(-1) min(-1)) and nitrification inhibitor (22 mg L(-1)) to prevent nitrification. The experiment included a control without aeration. The pH of the manure with aeration rose from 8.6 to 9.2 while the manure without aeration decreased from 8.6 to 8.1. With aeration, 97-99% of the NH4(+) was removed in about 5 days of operation with 96-98% recovery efficiency. In contrast, without aeration it took 25 days to treat the NH4(+). Therefore, the recovery of NH4(+) was five times faster with the low-rate aeration treatment. This enhancement could reduce costs by 70%. Published by Elsevier Ltd.

The effect of harvest time, dry matter content and mechanical pretreatments on anaerobic digestion and enzymatic hydrolysis of miscanthus

DEFF Research Database (Denmark)

Nielsen, Susanne Frydendal; Hjorth, Maibritt; Baby, Sanmohan

2016-01-01

-milling, extrusion or grinding and accumulated methane production and enzymatically-accessible sugars were measured. Accumulated methane production was studied using sigmoid curves that allowed comparison among the treatments of the rate of the methane production and ultimate methane yield. The green biomass gave...

Anaerobic digestion and co-digestion of slaughterhouse waste (SHW): influence of heat and pressure pre-treatment in biogas yield.

Science.gov (United States)

Cuetos, M J; Gómez, X; Otero, M; Morán, A

2010-10-01

Mesophilic anaerobic digestion (34+/-1 degrees C) of pre-treated (for 20 min at 133 degrees C, >3 bar) slaughterhouse waste and its co-digestion with the organic fraction of municipal solid waste (OFMSW) have been assessed. Semi-continuously-fed digesters worked with a hydraulic retention time (HRT) of 36 d and organic loading rates (OLR) of 1.2 and 2.6 kg VS(feed)/m(3)d for digestion and co-digestion, respectively, with a previous acclimatization period in all cases. It was not possible to carry out an efficient treatment of hygienized waste, even less so when OFMSW was added as co-substrate. These digesters presented volatile fatty acids (VFA), long chain fatty acids (LCFA) and fats accumulation, leading to instability and inhibition of the degradation process. The aim of applying a heat and pressure pre-treatment to promote splitting of complex lipids and nitrogen-rich waste into simpler and more biodegradable constituents and to enhance biogas production was not successful. These results indicate that the temperature and the high pressure of the pre-treatment applied favoured the formation of compounds that are refractory to anaerobic digestion. The pre-treated slaughterhouse wastes and the final products of these systems were analyzed by FTIR and TGA. These tools verified the existence of complex nitrogen-containing polymers in the final effluents, confirming the formation of refractory compounds during pre-treatment. (c) 2010 Elsevier Ltd. All rights reserved.

Chewing bread: impact on alpha-amylase secretion and oral digestion.

Science.gov (United States)

Joubert, Marianne; Septier, Chantal; Brignot, Hélène; Salles, Christian; Panouillé, Maud; Feron, Gilles; Tournier, Carole

2017-02-22

During chewing, saliva helps in preparing the food bolus by agglomerating the formed particles, and it initiates enzymatic food breakdown. However, limited information is actually available on the adaptation of saliva composition during the oral processing of complex foods, especially for foods that are sensitive to salivary enzymes. We addressed this question in the context of starch-based products and salivary alpha-amylase. The objectives were two-fold: (1) to determine if salivary alpha-amylase secretion can be modulated by the bread type and (2) to evaluate the contribution of the oral phase in bread enzymatic breakdown. Mouthfuls of three different wheat breads (industrial, artisan and whole-meal breads) were chewed by twelve subjects. Saliva samples were collected at rest and at different times corresponding to 33, 66 and 100% of the individual's chewing sequence. Alpha-amylase activity and total protein content were determined for all saliva samples that were collected. Additionally, the salivary maltose concentration was measured as a marker of bread enzymatic digestion. Boluses were collected at the swallowing time to evaluate the saliva uptake. Chewing industrial bread induced higher saliva uptake than the other breads despite a similar chewing duration. The evolution of salivary amylase activity tended to depend on the type of bread and was highly influenced by a large degree of inter- and intra-subject variability. The protein and maltose concentration steadily increased during chewing as a result of bread breakdown. The salivary protein concentration was mainly affected by the release of the water-soluble proteins of the bread. The salivary maltose concentration was found to be significantly lower for the whole-meal bread. When considering the weight of the mouthful, enzymatic breakdown was found to be most efficient for the breads ranking from industrial > artisan > whole-meal.

Co-digestion of municipal sludge and external organic wastes for enhanced biogas production under realistic plant constraints.

Science.gov (United States)

Tandukar, Madan; Pavlostathis, Spyros G

2015-12-15

A bench-scale investigation was conducted to select external organic wastes and mixing ratios for co-digestion with municipal sludge at the F. Wayne Hill Water Resources Center (FWHWRC), Gwinnett County, GA, USA to support a combined heat and power (CHP) project. External wastes were chosen and used subject to two constraints: a) digester retention time no lower than 15 d; and b) total biogas (methane) production not to exceed a specific target level based on air permit constraints on CO2 emissions. Primary sludge (PS), thickened waste activated sludge (TWAS) and digested sludge collected at the FWHWRC, industrial liquid waste obtained from a chewing gum manufacturing plant (GW) and dewatered fat-oil-grease (FOG) were used. All sludge and waste samples were characterized and their ultimate digestibility was assessed at 35 °C. The ultimate COD to methane conversion of PS, TWAS, municipal sludge (PS + TWAS; 40:60 w/w TS basis), GW and FOG was 49.2, 35.2, 40.3, 72.7, and 81.1%, respectively. Co-digestion of municipal sludge with GW, FOG or both, was evaluated using four bench-scale, mesophilic (35 °C) digesters. Biogas production increased significantly and additional degradation of the municipal sludge between 1.1 and 30.7% was observed. Biogas and methane production was very close to the target levels necessary to close the energy deficit at the FWHWRC. Co-digestion resulted in an effluent quality similar to that of the control digester fed only with the municipal sludge, indicating that co-digestion had no adverse effects. Study results prove that high methane production is achievable with the addition of concentrated external organic wastes to municipal digesters, at acceptable higher digester organic loadings and lower retention times, allowing the effective implementation of CHP programs at municipal wastewater treatment plants, with significant cost savings. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effect of in Vitro Gastrointestinal Digestion on Encapsulated and Nonencapsulated Phenolic Compounds of Carob (Ceratonia siliqua L.) Pulp Extracts and Their Antioxidant Capacity.

Science.gov (United States)

Ydjedd, Siham; Bouriche, Sihem; López-Nicolás, Rubén; Sánchez-Moya, Teresa; Frontela-Saseta, Carmen; Ros-Berruezo, Gaspar; Rezgui, Farouk; Louaileche, Hayette; Kati, Djamel-Edine

2017-02-01

To determine the effect of in vitro gastrointestinal digestion on the release and antioxidant capacity of encapsulated and nonencapsulated phenolics carob pulp extracts, unripe and ripe carob pulp extracts were microencapsulated with polycaprolactone via double emulsion/solvent evaporation technique. Microcapsules' characterization was performed using scanning electron microscopy and Fourier transform infrared spectrometry analysis. Total phenolics and flavonoids content and antioxidant activities (ORAC, DPPH, and FRAP) were evaluated after each digestion step. The release of phenolic acids and flavonoids was measured along the digestion process by HPLC-MS/MS analysis. The most important phenolics and flavonoids content as well as antioxidant activities were observed after gastric and intestinal phases for nonencapsulated and encapsulated extracts, respectively. The microencapsulation of carob polyphenols showed a protective effect against pH changes and enzymatic activities along digestion, thereby promoting a controlled release and targeted delivery of the encapsulated compound, which contributed to an increase in its bioaccessibility in the gut.

Effective of Microwave-KOH Pretreatment on Enzymatic Hydrolysis of Bamboo

Science.gov (United States)

Zhiqiang Li; Zehui Jiang; Yan Yu; Zhiyong Cai

2012-01-01

Bamboo, with its advantages of fast growth, short renovation, easy propagation and rich in cellulose and hemicellulose, is a potential feedstock for bioethanol or other biofuels production. The objective of this study was to examine the fea- sibility of microwave assistant KOH pretreatments to enhance enzymatic hydrolysis of bamboo. Pretreatment was car- ried out by...

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 (pdigestibility at 28 h and solubility. Furthermore, the α-helix-to-β-sheet ratio can be used to predict the nutritional value of feed proteins.

Ammonia Fiber Expansion Pretreatment and Enzymatic Hydrolysis on Two Different Growth Stages of Reed Canarygrass

Science.gov (United States)

Bradshaw, Tamika C.; Alizadeh, Hasan; Teymouri, Farzaneh; Balan, Venkatesh; Dale, Bruce E.

Plant materials from the vegetative growth stage of reed canarygrass and the seed stage of reed canarygrass are pretreated by ammonia fiber expansion (AFEX) and enzymatically hydrolyzed using 15 filter paper units (FPU) cellulase/g glucan to evaluate glucose and xylose yields. Percent conversions of glucose and xylose, effects of temperature and ammonia loading, and hydrolysis profiles are analyzed to determine the most effective AFEX treatment condition for each of the selected materials. The controls used in this study were untreated samples of each biomass material. All pretreatment conditions tested enhanced enzyme digestibility and improved sugar conversions for reed canarygrass compared with their untreated counterparts. Based on 168 h hydrolysis results using 15 FPU Spezyme CP cellulase/g glucan the most effective AFEX treatment conditions were determined as: vegetative growth stage of reed canarygrass—100°C, 60% moisture content, 1.2∶1 kg ammonia/kg of dry matter (86% glucose and 78% xylose) and seed stage of reed canarygrass—100°C, 60% moisture content, 0.8∶1 kg ammonia/kg of dry matter (89% glucose and 81% xylose). Supplementation by commercial Multifect 720 xylanase along with cellulase further increased both glucose and xylose yields by 10-12% at the most effective AFEX conditions.

Respiration and enzymatic activities as indicators of stabilization of sewage sludge composting.

Science.gov (United States)

Nikaeen, Mahnaz; Nafez, Amir Hossein; Bina, Bijan; Nabavi, BiBi Fatemeh; Hassanzadeh, Akbar

2015-05-01

The objective of this work was to study the evolution of physico-chemical and microbial parameters in the composting process of sewage sludge (SS) with pruning wastes (PW) in order to compare these parameters with respect to their applicability in the evaluation of organic matter (OM) stabilization. To evaluate the composting process and organic matter stability, different microbial activities were compared during composting of anaerobically digested SS with two volumetric ratios, 1:1 and 3:1 of PW:SS and two aeration techniques including aerated static piles (ASP) and turned windrows (TW). Dehydrogenase activity, fluorescein diacetate hydrolysis, and specific oxygen uptake rate (SOUR) were used as microbial activity indices. These indices were compared with traditional parameters, including temperature, pH, moisture content, organic matter, and C/N ratio. The results showed that the TW method and 3:1 (PW:SS) proportion was superior to the ASP method and 1:1 proportion, since the former accelerate the composting process by catalyzing the OM stabilization. Enzymatic activities and SOUR, which reflect microbial activity, correlated well with temperature fluctuations. Based on these results it appears that SOUR and the enzymatic activities are useful parameters to monitor the stabilization of SS compost. Copyright © 2015 Elsevier Ltd. All rights reserved.

ZVI Addition in Continuous Anaerobic Digestion Systems Dramatically Decreases P Recovery Potential: Dynamic Modelling

DEFF Research Database (Denmark)

Puyol, D.; Flores Alsina, Xavier; Segura, Y.

2017-01-01

The objective of this study is to show the preliminary results of a (dynamic) mathematical model describing the effects of zero valent iron (ZVI) addition during the anaerobic digestion of waste activated sludge from wastewater treatment systems. A modified version of the Anaerobic Digestion Model...... No. 1 (ADM1) upgraded with an improved physico-chemical description, ZVI corrosion, propionate uptake enhancement and multiple mineral precipitation is used as a modelling platform. The proposed approach is tested against two case studies which correspond to two lab scale anaerobic digesters (AD2, AD....... Simulations demonstrate that the model is capable to satisfactorily reproduce the dynamics of hydrolysis, acetogenesis, acidogenesis, nutrient release, pH and methanogenesis in the control anaerobic digester (AD1). This study also evidences the enhancement of methane production by the influence of ZVI...

Modeling of anaerobic digestion of complex substrates

International Nuclear Information System (INIS)

Keshtkar, A. R.; Abolhamd, G.; Meyssami, B.; Ghaforian, H.

2003-01-01

A structured mathematical model of anaerobic conversion of complex organic materials in non-ideally cyclic-batch reactors for biogas production has been developed. The model is based on multiple-reaction stoichiometry (enzymatic hydrolysis, acidogenesis, aceto genesis and methano genesis), microbial growth kinetics, conventional material balances in the liquid and gas phases for a cyclic-batch reactor, liquid-gas interactions, liquid-phase equilibrium reactions and a simple mixing model which considers the reactor volume in two separate sections: the flow-through and the retention regions. The dynamic model describes the effects of reactant's distribution resulting from the mixing conditions, time interval of feeding, hydraulic retention time and mixing parameters on the process performance. The model is applied in the simulation of anaerobic digestion of cattle manure under different operating conditions. The model is compared with experimental data and good correlations are obtained

Biogas from mesophilic anaerobic digestion of cow dung using gelatin as additive

Science.gov (United States)

Salam, Bodius; Rahman, Md Mizanur; Sikder, Md Asif R.; Islam, Majedul

2017-06-01

A research work was conducted to investigate the enhanced production ability of biogas from mesophilic anaerobic digestions of cow dung (CD) using gelatin as additive. Five laboratory scale digesters were constructed to digest cow dung, where one set up was used for digestion of cow dung without additive and the other set up were used for digestion with additive. Gelatin additive was added in the slurry of amount 0.29, 0,57, 0.85 and 1.14% (wt.). The digesters were made of glass conical flask of 1-liter capacity each. Cow dung was used 335 gm and water was used 365 gm in each experiment. In the slurry, total solid content was maintained 8% (wt.) for all the observations. The digesters were fed on batch basis. The digesters were operated at ambient temperatures of 26 - 35°C. The total gas yield was obtained about 14.4 L/kg CD for digestion without additive and about 65% more biogas for digestion with 0.29% gelatin additive. The retention time for digestion without additive was 38 days and with additive retention time varied between 24 and 52 days.

Roles of magnetite and granular activated carbon in improvement of anaerobic sludge digestion.

Science.gov (United States)

Peng, Hong; Zhang, Yaobin; Tan, Dongmei; Zhao, Zhiqiang; Zhao, Huimin; Quan, Xie

2018-02-01

Granular activated carbon (GAC) or magnetite could promote methane production from organic wastes, but their roles in enhancing anaerobic sludge digestion have not been clarified. GAC, magnetite and their combination were complemented into sludge digesters, respectively. Experimental results showed that average methane production increased by 7.3% for magnetite, 13.1% for GAC, and 20% for the combination of magnetite and GAC, and the effluent TCOD of the control, magnetite, GAC and magnetite-GAC digesters on day 56 were 53.2, 49.6, 48.0 and 46.6 g/L, respectively. Scanning electron microscope (SEM), nitrogen adsorption, Fourier transform infrared spectroscopy (FTIR) and microbial analysis indicated that magnetite enriched iron-reducing bacteria responsible for sludge hydrolysis while GAC enhanced syntrophic metabolism between iron-reducing bacteria and methanogens due to its high electrical conductivity and large surface area. Supplementing magnetite and GAC together into an anaerobic digester simultaneously accelerated sludge hydrolysis and methane production, resulting in better sludge digestion performance. Copyright © 2017 Elsevier Ltd. All rights reserved.

A novel fluorescent biosensor for Adenosine Triphosphate detection based on the polydopamine nanospheres integrating with enzymatic recycling amplification.

Science.gov (United States)

Ji, Xiaoting; Yi, Bingqing; Xu, Yujuan; Zhao, Yanan; Zhong, Hua; Ding, Caifeng

2017-07-01

Based on the protective performance of polydopamine nanospheres (PDANSs) for DNA against nuclease digestion and the specific recognition characteristic of aptamer, we have developed an enzymatic recycling signal amplification method for highly sensitive and selective detection of adenosine triphosphate (ATP). Fluorescence measurements were carried out to verify the DNA polymerase and exonuclease III (Exo III) assisted target recycling process and fluorescence signal amplification. In the absence of the ATP, initially, the signal DNA-PDANSs complex was in the "off" state due to the efficient fluorescence quenching of 6-carboxyfluorescein (FAM) adjacent to the surface of PDANSs. Due to the binding of the aptamer by ATP, it trigger DNA polymerase and Exo III assisted target recycling process by the product of release, the complex would change into the "on" state as a result of the dissociation of the FAM from the surface of PDANSs, thus providing greatly enhanced fluorescence emission intensity. The method allows quantitative detection of ATP in the range of 20-600nM with a detection limit of 8.32nM. This biosensor requires no complex operations, and is a new high efficiency method for ATP detection. Copyright © 2017 Elsevier B.V. All rights reserved.

Capillary gel electrophoresis-coupled aptamer enzymatic cleavage protection strategy for the simultaneous detection of multiple small analytes.

Science.gov (United States)

Perrier, Sandrine; Zhu, Zhenyu; Fiore, Emmanuelle; Ravelet, Corinne; Guieu, Valérie; Peyrin, Eric

2014-05-06

This novel, multi small-analyte sensing strategy is the result of combining the target-induced aptamer enzymatic protection approach with the CGE-LIF (capillary gel electrophoresis with laser-induced fluorescence) technique. The implemented assay principle is based on an analysis of the phosphodiesterase I (PDE I)-mediated size variation of a fluorescein-labeled aptamer (FApt), the enzyme catalyzing the removal of nucleotides from DNA in the 3' to 5' direction. In the absence of the target, the unfolded aptamer was enzymatically cleaved into short DNA fragments. Upon target binding, the DNA substrate was partially protected against enzymatic hydrolysis. The amount of bound aptamer remaining after the exonuclease reaction was proportional to the concentration of the target. The CGE technique, which was used to determine the separation of FApt species from DNA digested products, permitted the quantification of adenosine (A), ochratoxin A (O), and tyrosinamide (T) under the same optimized enzymatic conditions. This assay strategy was subsequently applied to the simultaneous detection of A, O, and T in a single capillary under buffered conditions using corresponding FApt probes of different lengths (23, 36, and 49 nucleotides, respectively). Additionally, the detection of these three small molecules was successfully achieved in a complex medium (diluted, heat-treated human serum) showing a good recovery. It is worth noting that the multiplexed analysis was accomplished for targets with different charge states by using aptamers possessing various structural features. This sensing platform constitutes a rationalized and reliable approach with an expanded potential for a high-throughput determination of small analytes in a single capillary.

Effect of depth beating on the fiber properties and enzymatic saccharification efficiency of softwood kraft pulp.

Science.gov (United States)

Gao, Wenhua; Xiang, Zhouyang; Chen, Kefu; Yang, Rendang; Yang, Fei

2015-01-01

Commercial bleached softwood kraft pulp was mechanically fibrillated by a PFI-mill with beating revolution from 5000 to 30,000 r. The extent of fibrillating on the pulp was evaluated by beating degree, fiber morphological properties (fiber length, width, coarseness and curls index), water retention value (WRV) and physical properties of paper made from the pulp. Depth beating process significantly affected the pulp fibrillations as showed by the decreased fiber length and width as well as the SEM analysis, but the effects were limited after beating revolution of 15,000. Depth beating process also improved the total internal pore and inter-fibril surface areas as shown by the increased WRV values. Substrate enzymatic digestibility (SED) of beaten pulp at 5000 revolutions could reach 95% at cellulase loading of 15 FPU/g of glucan. After the enzymatic hydrolysis, the size of the pulp residues was reduced to micro-scale, and a relative uniform size distribution of the residues appeared at 10,000 r beating revolution. Copyright © 2015 Elsevier Ltd. All rights reserved.

Thermophilic anaerobic co-digestion of poultry litter and thin stillage.

Science.gov (United States)

Sharma, Deepak; Espinosa-Solares, Teodoro; Huber, David H

2013-05-01

The purpose of this study was to test whether the performance of a thermophilic CSTR digester that has been stabilized on poultry litter will be enhanced or diminished by the addition of thin stillage as co-substrate. Replicate laboratory digesters, derived from a stable pilot-scale digester, were operated with increasing ratios (w/w) of thin stillage/poultry litter feedstock. After a period of adaptation to 20% and 40% thin stillage, digester performance showed increases in biogas, percent methane and COD removal, as well as a decrease in volatile acids. Peak performance occurred with 60% thin stillage. However, 80% thin stillage caused significant reduction of performance, including declines of methanogenic activity and COD removal. In conclusion, supplementing the thermophilic digestion of poultry litter with thin stillage improved the bioenergy (methane) output, but thin stillage became inhibitory at high concentrations. Copyright © 2013 Elsevier Ltd. All rights reserved.

Enzymatic Production of Monoclonal Stoichiometric Single-Stranded DNA Oligonucleotides

Science.gov (United States)

Ducani, Cosimo; Kaul, Corinna; Moche, Martin; Shih, William M.; Högberg, Björn

2013-01-01

Single-stranded oligonucleotides are important as research tools as probes for diagnostics and gene therapy. Today, production of oligonucleotides is done via solid-phase synthesis. However, the capabilities of current polymer chemistry are limited in comparison to what can be produced in biological systems. The errors in synthetic DNA increases with oligonucleotide length, and sequence diversity can often be a problem. Here, we present the Monoclonal Stoichiometric (MOSIC) method for enzymatic DNA oligonucleotide production. Using this method, we amplify oligonucleotides from clonal templates followed by digestion of a cutter-hairpin, resulting in pools of monoclonal oligonucleotides with precisely controlled relative stoichiometric ratios. We present data where MOSIC oligonucleotides, 14–378 nt long, were prepared either by in vitro rolling-circle amplification, or by amplification in Escherichia coli in the form of phagemid DNA. The formation of a DNA crystal and folding of DNA nanostructures confirmed the scalability, purity and stoichiometry of the produced oligonucleotides. PMID:23727986

Biogas Improvement by Adding Australian Zeolite During the Anaerobic Digestion of C:N Ratio Adjusted Swine Manure

DEFF Research Database (Denmark)

Wijesinghe, D. Thushari N.; Dassanayake, Kithsiri B.; Sommer, Sven G.

2018-01-01

Abstract: Maintenance of the ideal carbon: nitrogen (C:N) ratio with a minimum level of TAN is a key challenge for achieving maximum potential CH4 production through the anaerobic digestion process of agricultural waste such as swine manure. Biogas production can be enhanced by adding zeolite...... into the anaerobic digestion medium. However, the effects of zeolite addition to C:N ratio adjusted feedstock, on the digester performance is unknown. The objectives of this study were to investigate the effect of Australian zeolite on anaerobic digestion of swine manure with a C:N ratio adjusted to 30...... and to determine the optimal zeolite application rate to achieve the best performance. The Australian zeolite significantly enhanced CH4 production and reduced the lag phase of anaerobic digestion in batch production. The optimal addition rate of zeolite was appeared to be around 40 g/L. The better digester...

Kinetic modelling of enzymatic starch hydrolysis

NARCIS (Netherlands)

Bednarska, K.A.

2015-01-01

Kinetic modelling of enzymatic starch hydrolysis – a summary

K.A. Bednarska

The dissertation entitled ‘Kinetic modelling of enzymatic starch hydrolysis’ describes the enzymatic hydrolysis and kinetic modelling of liquefaction and saccharification of wheat starch.

Continuous Quadrupole Magnetic Separation of Islets during Digestion Improves Purified Porcine Islet Viability.

Science.gov (United States)

Weegman, Bradley P; Kumar Sajja, Venkata Sunil; Suszynski, Thomas M; Rizzari, Michael D; Scott Iii, William E; Kitzmann, Jennifer P; Mueller, Kate R; Hanley, Thomas R; Kennedy, David J; Todd, Paul W; Balamurugan, Appakalai N; Hering, Bernhard J; Papas, Klearchos K

2016-01-01

Islet transplantation (ITx) is an emerging and promising therapy for patients with uncontrolled type 1 diabetes. The islet isolation and purification processes require exposure to extended cold ischemia, warm-enzymatic digestion, mechanical agitation, and use of damaging chemicals for density gradient separation (DG), all of which reduce viable islet yield. In this paper, we describe initial proof-of-concept studies exploring quadrupole magnetic separation (QMS) of islets as an alternative to DG to reduce exposure to these harsh conditions. Three porcine pancreata were split into two parts, the splenic lobe (SPL) and the combined connecting/duodenal lobes (CDL), for paired digestions and purifications. Islets in the SPL were preferentially labeled using magnetic microparticles (MMPs) that lodge within the islet microvasculature when infused into the pancreas and were continuously separated from the exocrine tissue by QMS during the collection phase of the digestion process. Unlabeled islets from the CDL were purified by conventional DG. Islets purified by QMS exhibited significantly improved viability (measured by oxygen consumption rate per DNA, p < 0.03) and better morphology relative to control islets. Islet purification by QMS can reduce the detrimental effects of prolonged exposure to toxic enzymes and density gradient solutions and substantially improve islet viability after isolation.

Continuous Quadrupole Magnetic Separation of Islets during Digestion Improves Purified Porcine Islet Viability

Directory of Open Access Journals (Sweden)

Bradley P. Weegman

2016-01-01

Full Text Available Islet transplantation (ITx is an emerging and promising therapy for patients with uncontrolled type 1 diabetes. The islet isolation and purification processes require exposure to extended cold ischemia, warm-enzymatic digestion, mechanical agitation, and use of damaging chemicals for density gradient separation (DG, all of which reduce viable islet yield. In this paper, we describe initial proof-of-concept studies exploring quadrupole magnetic separation (QMS of islets as an alternative to DG to reduce exposure to these harsh conditions. Three porcine pancreata were split into two parts, the splenic lobe (SPL and the combined connecting/duodenal lobes (CDL, for paired digestions and purifications. Islets in the SPL were preferentially labeled using magnetic microparticles (MMPs that lodge within the islet microvasculature when infused into the pancreas and were continuously separated from the exocrine tissue by QMS during the collection phase of the digestion process. Unlabeled islets from the CDL were purified by conventional DG. Islets purified by QMS exhibited significantly improved viability (measured by oxygen consumption rate per DNA, p<0.03 and better morphology relative to control islets. Islet purification by QMS can reduce the detrimental effects of prolonged exposure to toxic enzymes and density gradient solutions and substantially improve islet viability after isolation.

A comprehensive review on food waste anaerobic digestion: Research updates and tendencies.

Science.gov (United States)

Ren, Yuanyuan; Yu, Miao; Wu, Chuanfu; Wang, Qunhui; Gao, Ming; Huang, Qiqi; Liu, Yu

2018-01-01

Anaerobic digestion has been practically applied in agricultural and industrial waste treatment and recognized as an economical-effective way for food waste disposal. This paper presented an overview on the researches about anaerobic digestion of food waste. Technologies (e.g., pretreatment, co-digestion, inhibition and mitigation, anaerobic digestion systems, etc.) were introduced and evaluated on the basis of bibliometric analysis. Results indicated that ethanol and aerobic prefermentation were novel approaches to enhance substrates hydrolysis and methane yield. With the promotion of resource recovery, more attention should be paid to biorefinery technologies which can produce more useful products toward zero emissions. Furthermore, a technological route for food waste conversion based on anaerobic digestion was proposed. Copyright © 2017 Elsevier Ltd. All rights reserved.

Determination of total dietary fiber (CODEX definition) by enzymatic-gravimetric method and liquid chromatography: collaborative study.

Science.gov (United States)

McCleary, Barry V; DeVries, Jonathan W; Rader, Jeanne I; Cohen, Gerald; Prosky, Leon; Mugford, David C; Champ, Martine; Okuma, Kazuhiro

2010-01-01

A method for the determination of total dietary fiber (TDF), as defined by the CODEX Alimentarius, was validated in foods. Based upon the principles of AOAC Official Methods 985.29, 991.43, 2001.03, and 2002.02, the method quantitates high- and low-molecular-weight dietary fiber (HMWDF and LMWDF, respectively). In 2007, McCleary described a method of extended enzymatic digestion at 37 degrees C to simulate human intestinal digestion followed by gravimetric isolation and quantitation of HMWDF and the use of LC to quantitate low-molecular-weight soluble dietary fiber (LMWSDF). The method thus quantitates the complete range of dietary fiber components from resistant starch (by utilizing the digestion conditions of AOAC Method 2002.02) to digestion resistant oligosaccharides (by incorporating the deionization and LC procedures of AOAC Method 2001.03). The method was evaluated through an AOAC collaborative study. Eighteen laboratories participated with 16 laboratories returning valid assay data for 16 test portions (eight blind duplicates) consisting of samples with a range of traditional dietary fiber, resistant starch, and nondigestible oligosaccharides. The dietary fiber content of the eight test pairs ranged from 11.57 to 47.83%. Digestion of samples under the conditions of AOAC Method 2002.02 followed by the isolation and gravimetric procedures of AOAC Methods 985.29 and 991.43 results in quantitation of HMWDF. The filtrate from the quantitation of HMWDF is concentrated, deionized, concentrated again, and analyzed by LC to determine the LMWSDF, i.e., all nondigestible oligosaccharides of degree of polymerization > or =3. TDF is calculated as the sum of HMWDF and LMWSDF. Repeatability standard deviations (Sr) ranged from 0.41 to 1.43, and reproducibility standard deviations (S(R)) ranged from 1.18 to 5.44. These results are comparable to other official dietary fiber methods, and the method is recommended for adoption as Official First Action.

Impact of mechanical, chemical and enzymatic pre-treatments on the methane yield from the anaerobic digestion of switchgrass

International Nuclear Information System (INIS)

Frigon, Jean-Claude; Mehta, Punita; Guiot, Serge R.

2012-01-01

The conversion of cellulosic crops into biofuels, including methane, is receiving a lot of attention lately. Panicum vergatum, or switchgrass, is a warm season perennial grass well adapted to grow in North America. Different pre-treatments were tested in 0.5 l batch reactors, at 35 °C, in order to enhance the methane production from switchgrass, including temperature, sonication, alkalinization and autoclaving. The methane production on the basis of volatile solids (VS) added to the fermentation were 112.4 ± 8.4, 132.5 ± 9.7 and 139.8 ml g −1 after 38 days of incubation for winter harvested switchgrass (WHS) after grinding, grinding with alkalinization, and grinding with alkalinization and autoclaving, respectively. The methane production was higher for fresh summer harvested switchgrass (SHS), with a production of 256.6 ± 8.2 ml g −1 VS after mulching, alkalinization and autoclaving. The methane production from SHS was improved by 29 and 42% when applying lignin (LiP) or manganese peroxidase (MnP), at 202.1 ± 9.8 and 222.9 ± 22.5 ml g −1 VS, respectively. The combination of an alkali pre-treatment with the MnP increased the methane production furthermore at 297.7 ml g −1 VS. The use of pectinases without chemical pre-treatment showed promising yields at 287.4 and 239.5 ml g −1 VS for pectate-lyase and poly-galacturonase, respectively. An estimation of the methane yield per hectare of crop harvested resulted in net energy production of 29.8, 49.7 and 78.1 GJ for winter harvested switchgrass, mulched and pretreated summer harvested switchgrass, respectively. Switchgrass represents an interesting candidate as a lignocellulosic crop for methane production. -- Highlights: ► Switchgrass is a model energy crops for biofuels production. ► This study evaluated different pre-treatments to enhance methane production. ► Pre-treatments increase significantly the methane produced from switchgrass. ► Enzymatic pre-treatments were superior to physical and

Effects of coffee processing residues on anaerobic microorganisms and corresponding digestion performance.

Science.gov (United States)

Rojas-Sossa, Juan Pablo; Murillo-Roos, Mariana; Uribe, Lidieth; Uribe-Lorio, Lorena; Marsh, Terence; Larsen, Niels; Chen, Rui; Miranda, Alberto; Solís, Kattia; Rodriguez, Werner; Kirk, Dana; Liao, Wei

2017-12-01

The objective of this study was to delineate the effects of different coffee processing residues on the anaerobic microbes and corresponding digestion performance. The results elucidated that mucilage-rich feed enhanced the accumulation of methanogens, which consequently led to better digestion performance of biogas production. Fifty percent more methane and up to 3 times more net energy (heat and electricity) output were achieved by the digestion of the mucilage-rich feed (M3). The microbial community and statistical analyses further elucidated that different residues in the feed had significant impact on microbial distribution and correspondingly influenced the digestion performance. Copyright © 2017 Elsevier Ltd. All rights reserved.

Digestion of isolated legume cells in a stomach-duodenum model: three mechanisms limit starch and protein hydrolysis.

Science.gov (United States)

Bhattarai, Rewati R; Dhital, Sushil; Wu, Peng; Chen, Xiao Dong; Gidley, Michael J

2017-07-19

Retention of intact plant cells to the end of the small intestine leads to transport of entrapped macronutrients such as starch and protein for colonic microbial fermentation, and is a promising mechanism to increase the content of resistant starch in diets. However, the effect of gastro-intestinal bio-mechanical processing on the intactness of plant cells and the subsequent resistance to enzymatic digestion of intracellular starch and protein are not well understood. In this study, intact cells isolated from legume cotyledons are digested in a laboratory model which mimics the mechanical and biochemical conditions of the rat stomach and duodenum. The resulting digesta are characterised in terms of cell (wall) integrity as well as intracellular starch and protein hydrolysis. The cells remained essentially intact in the model with negligible (ca. 2-3%) starch or protein digestion; however when the cells were mechanically broken and digested in the model, the hydrolysis was increased to 45-50% suggesting that intact cellular structures could survive the mixing regimes in the model stomach and duodenum sufficiently to prevent digestive enzyme access. Apart from intact cell walls providing effective barrier properties, they also limit digestibility by restricting starch gelatinisation during cooking, and significant non-specific binding of α-amylase is observed to both intact and broken cell wall components, providing a third mechanism hindering starch hydrolysis. The study suggests that the preservation of intactness of plant cells, such as from legumes, could be a viable approach to achieve the targeted delivery of resistant starch to the colon.

Steam explosion pretreatment for enhancing biogas production of late harvested hay.

Science.gov (United States)

Bauer, Alexander; Lizasoain, Javier; Theuretzbacher, Franz; Agger, Jane W; Rincón, María; Menardo, Simona; Saylor, Molly K; Enguídanos, Ramón; Nielsen, Paal J; Potthast, Antje; Zweckmair, Thomas; Gronauer, Andreas; Horn, Svein J

2014-08-01

Grasslands are often abandoned due to lack of profitability. Extensively cultivating grassland for utilization in a biogas-based biorefinery concept could mend this problem. Efficient bioconversion of this lignocellulosic biomass requires a pretreatment step. In this study the effect of different steam explosion conditions on hay digestibility have been investigated. Increasing severity in the pretreatment induced degradation of the hemicellulose, which at the same time led to the production of inhibitors and formation of pseudo-lignin. Enzymatic hydrolysis showed that the maximum glucose yields were obtained under pretreatment at 220 °C for 15 min, while higher xylose yields were obtained at 175 °C for 10 min. Pretreatment of hay by steam explosion enhanced 15.9% the methane yield in comparison to the untreated hay. Results indicate that hay can be effectively converted to methane after steam explosion pretreatment. Copyright © 2014 Elsevier Ltd. All rights reserved.

Improving the enzymatic hydrolysis of thermo-mechanical fiber from Eucalyptus urophylla by a combination of hydrothermal pretreatment and alkali fractionation.

Science.gov (United States)

Sun, Shaoni; Cao, Xuefei; Sun, Shaolong; Xu, Feng; Song, Xianliang; Sun, Run-Cang; Jones, Gwynn Lloyd

2014-01-01

The recalcitrance of lignocellulosic biomass is a major limitation for its conversion into biofuels by enzymatic hydrolysis. The use of a pretreatment technology is an essential step to diminish biomass recalcitrance for bioethanol production. In this study, a two-step pretreatment using hydrothermal pretreatment at various temperatures and alkali fractionation was performed on eucalyptus fiber. The detailed chemical composition, physicochemical characteristics, and morphology of the pretreated fibers in each of the fractions were evaluated to advance the performance of eucalyptus fiber in enzymatic digestibility. The hydrothermal pretreatment (100 to 220°C) significantly degraded hemicelluloses, resulting in an increased crystallinity of the pretreated fibers. However, as the pretreatment temperature reached 240°C, partial cellulose was degraded, resulting in a reduced crystallinity of cellulose. As compared to the hydrothermal pretreatment alone, a combination of hydrothermal and alkali treatments significantly removed hemicelluloses and lignin, resulting in an improved enzymatic hydrolysis of the cellulose-rich fractions. As compared with the raw fiber, the enzymatic hydrolysis rate increased 1.1 to 8.5 times as the hydrothermal pretreatment temperature increased from 100 to 240°C. Interestingly, after a combination of hydrothermal pretreatment and alkali fractionation, the enzymatic hydrolysis rate increased 3.7 to 9.2 times. Taking into consideration the consumption of energy and the production of xylo-oligosaccharides and lignin, an optimum pretreatment condition was found to be hydrothermal pretreatment at 180°C for 30 min and alkali fractionation with 2% NaOH at 90°C for 2.5 h, in which 66.3% cellulose was converted into glucose by enzymatic hydrolysis. The combination of hydrothermal pretreatment and alkali fractionation was a promising method to remove hemicelluloses and lignin as well as overcome the biomass recalcitrance for enzymatic hydrolysis

Comparison of different pretreatment strategies for enzymatic hydrolysis of wheat and barley straw

DEFF Research Database (Denmark)

Rosgaard, Lisa; Pedersen, Sven; Meyer, Anne Boye Strunge

2007-01-01

In biomass-to-ethanol processes a physico-chemical pretreatment of the lignocellulosic biomass is a critical requirement for enhancing the accessibility of the cellulose substrate to enzymatic attack. This report evaluates the efficacy on barley and wheat straw of three different pretreatment pro...

Lignosulfonate To Enhance Enzymatic Saccharification of Lignocelluloses: Role of Molecular Weight and Substrate Lignin

Science.gov (United States)

Haifeng Zhou; Hongming Lou; Dongjie Yang; J.Y. Zhu; Xueqing Qiu

2013-01-01

This study conducted an investigation of the effect of lignosulfonate (LS) on enzymatic saccharification of lignocelluloses. Two commercial LSs and one laboratory sulfonated kraft lignin were applied to Whatman paper, dilute acid and SPORL (sulfite pretreatment to overcome recalcitrance of lignocelluloses) pretreated aspen, and kraft alkaline and SPORL pretreated...

Lab-scale demonstration of recuperative thickening technology for enhanced biogas production and dewaterability in anaerobic digestion processes.

Science.gov (United States)

Cobbledick, Jeffrey; Aubry, Nicholas; Zhang, Victor; Rollings-Scattergood, Sasha; Latulippe, David R

2016-05-15

There is growing interest in the use of high performance anaerobic digestion (AD) processes for the production of biogas at wastewater treatment facilities to offset the energy demands associated with wastewater treatment. Recuperative thickening (RT) is a promising technique which involves recycling a portion of the digested solids back to the incoming feed. In general there exists a significant number of knowledge gaps in the field of RT because the studies that have been conducted to date have almost exclusively occurred in pilot plant or full scale trials; this approach greatly limits the amount of process optimization that can be done in a given trial. In this work, a detailed and comprehensive study of RT was conducted at the lab scale; two custom designed digesters (capacity = 1.5 L) were operated in parallel with one acting as a 'control' digester and the other operating under a semi-batch RT mode. There was no significant change in biogas methane composition for the two digesters, however the RT digester had an average biogas productivity over two times higher than the control one. It was found that the recycling of the polymer flocculant back into the RT digester resulted in a significant improvement in dewatering performance. At the highest polymer concentration tested, the capillary suction time (CST) values for flocculated samples for the RT digester were over 6 times lower than the corresponding values for the control digester. Thus, there exists an opportunity to decrease the overall consumption of polymer flocculants through judicious selection of the dose of polymer flocculant that is used both for the thickening and end-stage dewatering steps in RT processes. Copyright © 2016 Elsevier Ltd. All rights reserved.

Co-digestion of pig slaughterhouse waste with sewage sludge.

Science.gov (United States)

Borowski, Sebastian; Kubacki, Przemysław

2015-06-01

Slaughterhouse wastes (SHW) are potentially very attractive substrates for biogas production. However, mono-digestion of these wastes creates great technological problems associated with the inhibitory effects of ammonia and fatty acids on methanogens as well as with the foaming in the digesters. In the following study, the co-digestion of slaughterhouse wastes with sewage sludge (SS) was undertaken. Batch and semi-continuous experiments were performed at 35°C with municipal sewage sludge and pig SHW composed of meat tissue, intestines, bristles and post-flotation sludge. In batch assays, meat tissue and intestinal wastes gave the highest methane productions of 976 and 826 dm(3)/kg VS, respectively, whereas the methane yield from the sludge was only 370 dm(3)/kg VS. The co-digestion of sewage sludge with 50% SHW (weight basis) provided the methane yield exceeding 600 dm(3)/kg VS, which was more than twice as high as the methane production from sewage sludge alone. However, when the loading rate exceeded 4 kg VS/m(3) d, a slight inhibition of methanogenesis was observed, without affecting the digester stability. The experiments showed that the co-digestion of sewage sludge with large amount of slaughterhouse wastes is feasible, and the enhanced methane production does not affect the digester stability. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enhancing anaerobic digestion of waste activated sludge by pretreatment: effect of volatile to total solids.

Science.gov (United States)

Wang, Xiao; Duan, Xu; Chen, Jianguang; Fang, Kuo; Feng, Leiyu; Yan, Yuanyuan; Zhou, Qi

2016-01-01

In this study the effect of volatile to total solids (VS/TS) on anaerobic digestion of waste activated sludge (WAS) pretreated by alkaline, thermal and thermal-alkaline strategies was studied. Experimental results showed that the production of methane from sludge was increased with VS/TS. When anaerobic digesters were fed with sludge pretreated by the thermal-alkaline method, the average methane yield was improved from 2.8 L/d at VS/TS 0.35 to 4.7 L/d at VS/TS 0.56. Also, the efficiency of VS reduction during sludge anaerobic digestion varied between 18.9% and 45.6%, and increased gradually with VS/TS. Mechanism investigation of VS/TS on WAS anaerobic digestion suggested that the general activities of anaerobic microorganisms, activities of key enzymes related to sludge hydrolysis, acidification and methanogenesis, and the ratio of Archaea to Bacteria were all increased with VS/TS, showing good agreement with methane production.

Anaerobic co-digestion of the organic fraction of municipal solid waste with FOG waste from a sewage treatment plant: Recovering a wasted methane potential and enhancing the biogas yield

International Nuclear Information System (INIS)

Martin-Gonzalez, L.; Colturato, L.F.; Font, X.; Vicent, T.

2010-01-01

Anaerobic digestion is applied widely to treat the source collected organic fraction of municipal solid wastes (SC-OFMSW). Lipid-rich wastes are a valuable substrate for anaerobic digestion due to their high theoretical methane potential. Nevertheless, although fat, oil and grease waste from sewage treatment plants (STP-FOGW) are commonly disposed of in landfill, European legislation is aimed at encouraging more effective forms of treatment. Co-digestion of the above wastes may enhance valorisation of STP-FOGW and lead to a higher biogas yield throughout the anaerobic digestion process. In the present study, STP-FOGW was evaluated as a co-substrate in wet anaerobic digestion of SC-OFMSW under mesophilic conditions (37 o C). Batch experiments carried out at different co-digestion ratios showed an improvement in methane production related to STP-FOGW addition. A 1:7 (VS/VS) STP-FOGW:SC-OFMSW feed ratio was selected for use in performing further lab-scale studies in a 5 L continuous reactor. Biogas yield increased from 0.38 ± 0.02 L g VS feed -1 to 0.55 ± 0.05 L g VS feed -1 as a result of adding STP-FOGW to reactor feed. Both VS reduction values and biogas methane content were maintained and inhibition produced by long chain fatty acid (LCFA) accumulation was not observed. Recovery of a currently wasted methane potential from STP-FOGW was achieved in a co-digestion process with SC-OFMSW.

Combined subcritical water and enzymatic hydrolysis for reducing sugar production from coconut husk

Science.gov (United States)

Muharja, Maktum; Junianti, Fitri; Nurtono, Tantular; Widjaja, Arief

2017-05-01

Coconut husk wastes are abundantly available in Indonesia. It has a potential to be used into alternative renewable energy sources such as hydrogen using enzymatic hydrolysis followed by a fermentation process. Unfortunately, enzymatic hydrolysis is hampered by the complex structure of lignocellulose, so the cellulose component is hard to degrade. In this study, Combined Subcritical Water (SCW) and enzymatic hydrolysis are applied to enhance fermentable, thereby reducing production of sugar from coconut husk. There were two steps in this study, the first step was coconut husk pretreated by SCW in batch reactor at 80 bar and 150-200°C for 60 minutes reaction time. Secondly, solid fraction from the results of SCW was hydrolyzed using the mixture of pure cellulose and xylanase enzymes. Analysis was conducted on untreated and SCW-treated by gravimetric assay, liquid fraction after SCW and solid fraction after enzymatic hydrolysis using DNS assay. The maximum yield of reducing sugar (including xylose, arabinose glucose, galactose, mannose) was 1.254 gr per 6 gr raw material, representing 53.95% of total sugar in coconut husk biomass which was obtained at 150°C 80 bar for 60 minutes reaction time of SCW-treated and 6 hour of enzymatic hydrolysis using mixture of pure cellulose and xylanase enzymes (18.6 U /gram of coconut husk).

Enzymatic activity of the cellulolytic complex produced by trichoderma reesei. Enzymatic hydrolysis of cellulose

International Nuclear Information System (INIS)

Alfonsel Jaen, M.; Negro, M.J.; Saez, R.; Martin Moreno, C.

1986-01-01

The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reese QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass from Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars productions, have been selected. Previous studies on enzymatic hydrolysis of O. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (author). 10 figs.; 10 refs

Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose

International Nuclear Information System (INIS)

Alfonsel J, M.; Negro A, M. J.; Saez A, R.; Martin M, C.

1986-01-01

The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs

Enhancement options for the utilisation of nitrogen rich animal by-products in anaerobic digestion.

Science.gov (United States)

Resch, Christoph; Wörl, Alexander; Waltenberger, Reinhold; Braun, Rudolf; Kirchmayr, Roland

2011-02-01

This study focuses on the enhancement of an Austrian anaerobic digestion plant at a slaughterhouse site which exclusively uses animal by-products as substrate. High ammonia concentrations from protein degradation cause severe inhibitions of anaerobic microorganisms. For improving the current situation the COD:TKN ratio is widened by (a) ammonia stripping directly out of the process and (b) addition of a C source to the substrate. Different OLR and HRT were tested in continuous experiments to simulate new operating conditions. The results show that the addition of carbon cannot improve fermentation capacity. The reduction of ammonia boosts the degradation: After reduction of TKN from 7.5 to 4.0 g kg(-1) the initially high VFA concentration decreased and the COD degradation was improved by 55.5%. Hence, the implementation of the new N reduction process facilitates either the increase of the OLR by 61% or the reduction of the HRT by 25%. Copyright © 2010 Elsevier Ltd. All rights reserved.

Structural changes in lignin during organosolv pretreatment of Liriodendron tulipifera and the effect on enzymatic hydrolysis

International Nuclear Information System (INIS)

Koo, Bon-Wook; Min, Byeong-Cheol; Gwak, Ki-Seob; Lee, Soo-Min; Choi, Joon-Weon; Yeo, Hwanmyeong; Choi, In-Gyu

2012-01-01

Although organosolv pretreatment removed substantial amounts of lignin and xylan, the yield of glucan which is a major sugar source for fermentation to ethanol is more than 90% in most conditions of the organosolv pretreatment. Relative lignin contents of all pretreated biomass were more than 200 g kg −1 , however enzymatic conversions were increased dramatically comparing to untreated biomass. Therefore the correlation between lignin and enzymatic hydrolysis could not be explained just by lignin content, and other changes resulting from lignin removal affected enzymatic hydrolysis. Results on enzymatic conversion and sugar recovery suggested that the critical temperature improving enzymatic hydrolysis significantly was between 120 °C and 130 °C. Microscopic analysis using Field emission scanning electron microscopy (FE-SEM) showed that structural lignin changes happened through organosolv pretreatment. Lignins were isolated from lignin carbohydrate complex (LCC) at the initial stage and then migrated to the surface of biomass. The isolated and migrated lignins were finally redistributed onto surface. These structural changes formed droplets on surface and increased pore volume in pretreated biomass. The increase in pore volume also increased available surface area and enzyme adsorption at initial stage, and thus enzymatic conversion increased significantly through organosolv pretreatment. It was verified that the droplets were mainly composed of lignin and the lignin droplets inhibited enzymatic hydrolysis through adsorption with cellulase. -- Highlights: ► Just lignin contents cannot explain a correlation with enzymatic hydrolysis. ► Several changes resulted from lignin removal must affect enzymatic hydrolysis. ► Droplets are formed by structural changes in lignin during organosolv pretreatment. ► Formation of the lignin droplet increases the pore volume in biomass. ► The increase in pore volume enhances the enzymatic hydrolysis.

Enzymatic Refining and Cellulose Nanofiber Addition in Papermaking Processes from Recycled and Deinked Slurries

Directory of Open Access Journals (Sweden)

Marc Delgado-Aguilar

2015-07-01

Full Text Available Recycling and deinking processes cause fiber damage because of hornification phenomena and increased external fibrillation. Mechanical refining has been used for many years to enhance the mechanical properties of paper. Biorefining of pulp using enzymes is receiving increasing interest for energy reduction at the refining step of the paper-making process. Moreover, enzymes have also been used for the enhancement of mechanical properties without affecting the drainage rate. As an alternative to mechanical refining treatment, a combination of an enzymatic treatment and cellulose nanofibril (CNF addition was explored to enhance the mechanical properties of paper. The tests were carried out on a deinked pulp (DIP suspension made of 50% old newspapers (ONP and 50% old magazines (OMG. Various enzyme charges and CNF amounts were added to the mixture of ONP and OMG. All pulps (treated and untreated were characterized from a morphological point of view, and the paper sheets made thereof were mechanically characterized. The combination of the enzymatic treatment with the addition of 3% CNF provided sufficient tensile strength for the paper to be used in high-performance applications.

Enzymatically crosslinked silk-hyaluronic acid hydrogels.

Science.gov (United States)

Raia, Nicole R; Partlow, Benjamin P; McGill, Meghan; Kimmerling, Erica Palma; Ghezzi, Chiara E; Kaplan, David L

2017-07-01

In this study, silk fibroin and hyaluronic acid (HA) were enzymatically crosslinked to form biocompatible composite hydrogels with tunable mechanical properties similar to that of native tissues. The formation of di-tyrosine crosslinks between silk fibroin proteins via horseradish peroxidase has resulted in a highly elastic hydrogel but exhibits time-dependent stiffening related to silk self-assembly and crystallization. Utilizing the same method of crosslinking, tyramine-substituted HA forms hydrophilic and bioactive hydrogels that tend to have limited mechanics and degrade rapidly. To address the limitations of these singular component scaffolds, HA was covalently crosslinked with silk, forming a composite hydrogel that exhibited both mechanical integrity and hydrophilicity. The composite hydrogels were assessed using unconfined compression and infrared spectroscopy to reveal of the physical properties over time in relation to polymer concentration. In addition, the hydrogels were characterized by enzymatic degradation and for cytotoxicity. Results showed that increasing HA concentration, decreased gelation time, increased degradation rate, and reduced changes that were observed over time in mechanics, water retention, and crystallization. These hydrogel composites provide a biologically relevant system with controllable temporal stiffening and elasticity, thus offering enhanced tunable scaffolds for short or long term applications in tissue engineering. Copyright © 2017 Elsevier Ltd. All rights reserved.

Performance of dry anaerobic technology in the co-digestion of rural organic solid wastes in China

International Nuclear Information System (INIS)

Yang, Tianxue; Li, Yingjun; Gao, Jixi; Huang, Caihong; Chen, Bin; Zhang, Lieyu; Wang, Xiaowei; Zhao, Ying; Xi, Beidou; Li, Xiang

2015-01-01

The dry anaerobic co-digestion of LW (livestock waste), OFHW (organic fraction of household waste), and AR (agricultural residue) was evaluated in terms of pH stability, organic removal rate, and methane yield. The total quantity of the solids involved in the digestion was adjusted to 25%. All the reactors were inoculated by 20% (in dry weight) of the municipal sludge. The dynamic changes in the pH values of the LW-AR-OFHW mixture co-digestions underwent four stages and differed from those of wet anaerobic digestion. The decrease in VS (volatile solids), volume, and weight of the LW-AR-OFHW mixtures was higher than those in AR and OFHW. The VS, volume, and weight reductions in LW-AR-OFHW co-digestion were 54.7%, 82.2%, and 72.7%, respectively. However, the VS, volume, and weight reductions in AR were only 11.1%, 20.5%, and 19.8%, respectively, and those in OFHW were only 27.4%, 45.0%, and 40.9%, respectively. The LW-AR-OFHW mixture co-digestions enhanced the methane production of the co-digester (256 m 3 /ton VS), whereas AR and OFHW produced only 12 and 93 m 3 methane/ton VS, respectively. - Highlights: • The pH values dynamic of LW-AR-OFHW differed from wet anaerobic digestion. • The degradation effect of LW-AR-OFHW was better than those of AR and OFHW. • The LW-AR-OFHW mixture co-digestions enhanced the methane production.

Enhanced biohydrogen production from oat straw co-digested with cow dung / sewage sludge by combined aerobic digestion and anaerobic fermentation

Directory of Open Access Journals (Sweden)

Loretta Li

2016-03-01

Full Text Available Hydrogen was produced from oat straw by combined aerobic and anaerobic fermentation with fungi and cow dung. With aerobic pre-digestion, the maximum hydrogen production rate reached 133 ml/g volatile suspended solids per hour. The maximum hydrogen yield was 71.5 ml/g straw in 6 days by biological process. The lignocellulosic conversion of oak-straw waste was 39%, with the complex component converting 68% of the hemi-cellulose and 61% of the cellulose, but only 34% of lignin conversion. Aerobic pre-digestion by Trichoderma viride and Saccharomyces cerevisiae was significantly effective for lignin degradation.  Combining aerobic and anaerobic fermentation is a promising low-cost efficient and environmentally friendly method, compared with hydrogen fermentation, not only for hydrogen production, but also for converting straw biomass.

Thermal pre-treatment of primary and secondary sludge at 70ºC prior to anaerobic digestion

DEFF Research Database (Denmark)

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

2005-01-01

. The present study investigates the effect of the pre-treatment at 70 degrees C on thermophilic (55 degrees C) anaerobic digestion of primary and secondary sludge in continuously operated digesters. Thermal pre-treatment of primary and secondary sludge at 70 degrees C enhanced the removal of organic matter...... and the methane production during the subsequent anaerobic digestion step at 55 degrees C. It also greatly contributed to the destruction of pathogens present in primary sludge. Finally it results in enhanced microbial activities of the subsequent anaerobic step suggesting that the same efficiencies in organic...... matter removal and methane recovery could be obtained at lower HRTs....

Carbon capture and biogas enhancement by carbon dioxide enrichment of anaerobic digesters treating sewage sludge or food waste.

Science.gov (United States)

Bajón Fernández, Y; Soares, A; Villa, R; Vale, P; Cartmell, E

2014-05-01

The increasing concentration of carbon dioxide (CO2) in the atmosphere and the stringent greenhouse gases (GHG) reduction targets, require the development of CO2 sequestration technologies applicable for the waste and wastewater sector. This study addressed the reduction of CO2 emissions and enhancement of biogas production associated with CO2 enrichment of anaerobic digesters (ADs). The benefits of CO2 enrichment were examined by injecting CO2 at 0, 0.3, 0.6 and 0.9 M fractions into batch ADs treating food waste or sewage sludge. Daily specific methane (CH4) production increased 11-16% for food waste and 96-138% for sewage sludge over the first 24h. Potential CO2 reductions of 8-34% for sewage sludge and 3-11% for food waste were estimated. The capacity of ADs to utilise additional CO2 was demonstrated, which could provide a potential solution for onsite sequestration of CO2 streams while enhancing renewable energy production. Copyright © 2014 Elsevier Ltd. All rights reserved.

[Non-enzymatic glycosylation of dietary protein in vitro].

Science.gov (United States)

Bednykh, B S; Evdokimov, I A; Sokolov, A I

2015-01-01

Non-enzymatic glycosylation of proteins, based on discovered by Mayarn reaction of carbohydrate aldehyde group with a free amino group of a protein molecule, is well known to experts in biochemistry of food industry. Generated brown solid in some cases give the product marketable qualities--crackling bread--in others conversely, worsen the product. The biological effects of far-advanced products of non-enzymatic protein glycosylation reaction have not been studied enough, although it was reported previously that they are not split by digestive enzymes and couldn't be absorbed by animals. The objective of this work was to compare the depth of glycosylation of different food proteins of animal and vegetable origin. The objects of the study were proteins of animal (casein, lactoglobulin, albumin) and vegetable (soy isolate, proteins of rice flour, buckwheat, oatmeal) origin, glucose and fructose were selected as glycosylation agents, exposure 15 days at 37 degrees C. Lactoglobulin was glycosylated to a lesser extent among the proteins of animal origin while protein of oatmeal was glycosylated in the least degree among vegetable proteins. Conversely, such proteins as casein and soya isolate protein bound rather large amounts of carbohydrates. Fructose binding with protein was generally higher than the binding of glucose. The only exception was a protein of oatmeal. When of glucose and fructose simultaneously presented in the incubation medium, glucose binding usually increased while binding of fructose, in contrast, reduced. According to the total amount of carbohydrate (mcg), which is able to attach a protein (mg) the studied food proteins located in the following order: albumin (38) > soy protein isolate (23) > casein (15,) > whey protein rice flour protein (6) > protein from buckwheat flour (3) > globulin (2) > protein of oatmeal (0.3). The results obtained are to be used to select the optimal combination of proteins and carbohydrates, in which the glycosylation

Contrast agent enhanced pQCT of articular cartilage

Energy Technology Data Exchange (ETDEWEB)

Kallioniemi, A S [Department of Physics, University of Kuopio, POB 1627, 70211 Kuopio (Finland); Jurvelin, J S [Department of Physics, University of Kuopio, POB 1627, 70211 Kuopio (Finland); Nieminen, M T [Department of Diagnostic Radiology, POB 50, 90029 OYS, Oulu University Hospital, Oulu (Finland); Lammi, M J [Department of Anatomy, Institute of Biomedicine, University of Kuopio, POB 1627, 70211 Kuopio (Finland); Toeyraes, J [Department of Physics, University of Kuopio, POB 1627, 70211 Kuopio (Finland)

2007-02-21

The delayed gadolinium enhanced MRI of cartilage (dGEMRIC) technique is the only non-invasive means to estimate proteoglycan (PG) content in articular cartilage. In dGEMRIC, the anionic paramagnetic contrast agent gadopentetate distributes in inverse relation to negatively charged PGs, leading to a linear relation between T{sub 1,Gd} and spatial PG content in tissue. In the present study, for the first time, contrast agent enhanced peripheral quantitative computed tomography (pQCT) was applied, analogously to dGEMRIC, for the quantitative detection of spatial PG content in cartilage. The suitability of two anionic radiographic contrast agents, gadopentetate and ioxaglate, to detect enzymatically induced PG depletion in articular cartilage was investigated. First, the interrelationships of x-ray absorption, as measured with pQCT, and the contrast agent solution concentration were investigated. Optimal contrast agent concentrations for the following experiments were selected. Second, diffusion rates for both contrast agents were investigated in intact (n = 3) and trypsin-degraded (n 3) bovine patellar cartilage. The contrast agent concentration of the cartilaginous layer was measured prior to and 2-27 h after immersion. Optimal immersion time for the further experiments was selected. Third, the suitability of gadopentetate and ioxaglate enhanced pQCT to detect the enzymatically induced specific PG depletion was investigated by determining the contrast agent concentrations and uronic acid and water contents in digested and intact osteochondral samples (n = 16). After trypsin-induced PG loss (-70%, p < 0.05) the penetration of gadopentetate and ioxaglate increased (p < 0.05) by 34% and 48%, respectively. Gadopentetate and ioxaglate concentrations both showed strong correlation (r = -0.95, r -0.94, p < 0.01, respectively) with the uronic acid content. To conclude, contrast agent enhanced pQCT provides a technique to quantify PG content in normal and experimentally

Contrast agent enhanced pQCT of articular cartilage

Science.gov (United States)

Kallioniemi, A. S.; Jurvelin, J. S.; Nieminen, M. T.; Lammi, M. J.; Töyräs, J.

2007-02-01

The delayed gadolinium enhanced MRI of cartilage (dGEMRIC) technique is the only non-invasive means to estimate proteoglycan (PG) content in articular cartilage. In dGEMRIC, the anionic paramagnetic contrast agent gadopentetate distributes in inverse relation to negatively charged PGs, leading to a linear relation between T1,Gd and spatial PG content in tissue. In the present study, for the first time, contrast agent enhanced peripheral quantitative computed tomography (pQCT) was applied, analogously to dGEMRIC, for the quantitative detection of spatial PG content in cartilage. The suitability of two anionic radiographic contrast agents, gadopentetate and ioxaglate, to detect enzymatically induced PG depletion in articular cartilage was investigated. First, the interrelationships of x-ray absorption, as measured with pQCT, and the contrast agent solution concentration were investigated. Optimal contrast agent concentrations for the following experiments were selected. Second, diffusion rates for both contrast agents were investigated in intact (n = 3) and trypsin-degraded (n = 3) bovine patellar cartilage. The contrast agent concentration of the cartilaginous layer was measured prior to and 2-27 h after immersion. Optimal immersion time for the further experiments was selected. Third, the suitability of gadopentetate and ioxaglate enhanced pQCT to detect the enzymatically induced specific PG depletion was investigated by determining the contrast agent concentrations and uronic acid and water contents in digested and intact osteochondral samples (n = 16). After trypsin-induced PG loss (-70%, p < 0.05) the penetration of gadopentetate and ioxaglate increased (p < 0.05) by 34% and 48%, respectively. Gadopentetate and ioxaglate concentrations both showed strong correlation (r = -0.95, r = -0.94, p < 0.01, respectively) with the uronic acid content. To conclude, contrast agent enhanced pQCT provides a technique to quantify PG content in normal and experimentally

Contrast agent enhanced pQCT of articular cartilage

International Nuclear Information System (INIS)

Kallioniemi, A S; Jurvelin, J S; Nieminen, M T; Lammi, M J; Toeyraes, J

2007-01-01

The delayed gadolinium enhanced MRI of cartilage (dGEMRIC) technique is the only non-invasive means to estimate proteoglycan (PG) content in articular cartilage. In dGEMRIC, the anionic paramagnetic contrast agent gadopentetate distributes in inverse relation to negatively charged PGs, leading to a linear relation between T 1,Gd and spatial PG content in tissue. In the present study, for the first time, contrast agent enhanced peripheral quantitative computed tomography (pQCT) was applied, analogously to dGEMRIC, for the quantitative detection of spatial PG content in cartilage. The suitability of two anionic radiographic contrast agents, gadopentetate and ioxaglate, to detect enzymatically induced PG depletion in articular cartilage was investigated. First, the interrelationships of x-ray absorption, as measured with pQCT, and the contrast agent solution concentration were investigated. Optimal contrast agent concentrations for the following experiments were selected. Second, diffusion rates for both contrast agents were investigated in intact (n = 3) and trypsin-degraded (n 3) bovine patellar cartilage. The contrast agent concentration of the cartilaginous layer was measured prior to and 2-27 h after immersion. Optimal immersion time for the further experiments was selected. Third, the suitability of gadopentetate and ioxaglate enhanced pQCT to detect the enzymatically induced specific PG depletion was investigated by determining the contrast agent concentrations and uronic acid and water contents in digested and intact osteochondral samples (n = 16). After trypsin-induced PG loss (-70%, p < 0.05) the penetration of gadopentetate and ioxaglate increased (p < 0.05) by 34% and 48%, respectively. Gadopentetate and ioxaglate concentrations both showed strong correlation (r = -0.95, r -0.94, p < 0.01, respectively) with the uronic acid content. To conclude, contrast agent enhanced pQCT provides a technique to quantify PG content in normal and experimentally degraded

Cooking temperature is a key determinant of in vitro meat protein digestion rate: investigation of underlying mechanisms.

Science.gov (United States)

Bax, Marie-Laure; Aubry, Laurent; Ferreira, Claude; Daudin, Jean-Dominique; Gatellier, Philippe; Rémond, Didier; Santé-Lhoutellier, Véronique

2012-03-14

The present study aimed to evaluate the digestion rate and nutritional quality of pig muscle proteins in relation to different meat processes (aging, mincing, and cooking). Under our experimental conditions, aging and mincing had little impact on protein digestion. Heat treatments had different temperature-dependent effects on the meat protein digestion rate and degradation potential. At 70 °C, the proteins underwent denaturation that enhanced the speed of pepsin digestion by increasing enzyme accessibility to protein cleavage sites. Above 100 °C, oxidation-related protein aggregation slowed pepsin digestion but improved meat protein overall digestibility. The digestion parameters defined here open new insights on the dynamics governing the in vitro digestion of meat protein. However, the effect of cooking temperature on protein digestion observed in vitro needs to be confirmed in vivo.

Lignosulfonate-mediated cellulase adsorption: enhanced enzymatic saccharification of lignocellulose through weakening nonproductive binding to lignin

Science.gov (United States)

Zhaojiang Wang; JY Zhu; Yingjuan Fu; Menghua Qin; Zhiyong Shao; Jungang Jiang; Fang Yang

2013-01-01

Thermochemical pretreatment of lignocellulose is crucial to bioconversion in the fields of biorefinery and biofuels. However, the enzyme inhibitors in pretreatment hydrolysate make solid substrate washing and hydrolysate detoxification indispensable prior to enzymatic hydrolysis. Sulfite pretreatment to overcome recalcitrance of lignocelluloses (SPORL) is a relatively...

Pretreatment of Reed by Wet Oxidation and Subsequent Utilization of the Pretreated Fibers for Ethanol Production

DEFF Research Database (Denmark)

Szijarto, Nora; Kádár, Zsófia; Varga, Eniko

2009-01-01

lignocelluloses usually do. In the present study, wet oxidation was investigated as the pretreatment method to enhance the enzymatic digestibility of reed cellulose to soluble sugars and thus improve the convertibility of reed to ethanol. The most effective treatment increased the digestibility of reed cellulose...... of cellulose to glucose was 82.4%. Simultaneous saccharification and fermentation of pretreated solids resulted in a final ethanol concentration as high as 8.7 g/L, yielding 73% of the theoretical....

Biomass Enzymatic Saccharification Is Determined by the Non-KOH-Extractable Wall Polymer Features That Predominately Affect Cellulose Crystallinity in Corn

Science.gov (United States)

Wu, Leiming; Wang, Hongwu; Wu, Zhiliang; Li, Ming; Huang, Pengyan; Feng, Shengqiu; Chen, Peng; Zheng, Yonglian; Peng, Liangcai

2014-01-01

Corn is a major food crop with enormous biomass residues for biofuel production. Due to cell wall recalcitrance, it becomes essential to identify the key factors of lignocellulose on biomass saccharification. In this study, we examined total 40 corn accessions that displayed a diverse cell wall composition. Correlation analysis showed that cellulose and lignin levels negatively affected biomass digestibility after NaOH pretreatments at pbiomass saccharification after pretreatments with NaOH and H2SO4 at three concentrations. Notably, despite that the non-KOH-extractable residues covered 12%–23% hemicelluloses and lignin of total biomass, their wall polymer features exhibited the predominant effects on biomass enzymatic hydrolysis including Ara substitution degree of xylan (reverse Xyl/Ara) and S/G ratio of lignin. Furthermore, the non-KOH-extractable polymer features could significantly affect lignocellulose crystallinity at pbiomass digestibility. Hence, this study could suggest an optimal approach for genetic modification of plant cell walls in bioenergy corn. PMID:25251456

Enzymatic modification of egg lecithin to improve properties.

Science.gov (United States)

Asomaning, Justice; Curtis, Jonathan M

2017-04-01

This research studied the enzymatic modification of egg yolk phospholipids and its effect on physicochemical properties. Egg yolk lipids were extracted with food grade ethanol and egg phospholipids (ePL) produced by deoiling with acetone. Vegetable oils were used to interesterify ePL utilizing Lipozyme®: sn-1,3 specific lipase. The enzymatic interesterification resulted in a single phase liquid product, whereas simple blending of the ePL and vegetable oil resulted in a product with two phases. In addition solid fat content decreased by 50% at -10°C and 94% at 35°C when compared with egg yolk lipids extract. A decrease in melting temperature resulted from the interesterification process. Interesterification improved emulsion stability index when used as an emulsifier in oil-in-water emulsion and compared to the native and soy lecithin. Enzyme reusability test showed retention of 63% activity after 10 cycles. Overall, the properties of native egg phospholipids were significantly enhanced in a potentially useful manner through interesterification. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effect of Acylglycerol Composition and Fatty Acyl Chain Length on Lipid Digestion in pH-Stat Digestion Model and Simulated In Vitro Digestion Model.

Science.gov (United States)

Qi, Jin F; Jia, Cai H; Shin, Jung A; Woo, Jeong M; Wang, Xiang Y; Park, Jong T; Hong, Soon T; Lee, K-T

2016-02-01

In this study, a pH-stat digestion model and a simulated in vitro digestion model were employed to evaluate the digestion degree of lipids depending on different acylglycerols and acyl chain length (that is, diacylglycerol [DAG] compared with soybean oil representing long-chain triacylglycerol compared with medium-chain triacylglycerol [MCT]). In the pH-stat digestion model, differences were observed among the digestion degrees of 3 oils using digestion rate (k), digestion half-time (t1/2 ), and digestion extent (Φmax). The results showed the digestion rate order was MCT > soybean oil > DAG. Accordingly, the order of digestion half-times was MCT digestion model, digestion rates (k') and digestion half-times (t'1/2 ) were also obtained and the results showed a digestion rate order of MCT (k' = 0.068 min(-1) ) > soybean oil (k' = 0.037 min(-1) ) > DAG (k' = 0.024 min(-1) ). Consequently, the order of digestion half-times was MCT (t'1/2 = 10.20 min) digested faster than soybean oil, and that soybean oil was digested faster than DAG. © 2015 Institute of Food Technologists®

Comparison of the role that entropy has played in processes of non-enzymatic and enzymatic catalysis

International Nuclear Information System (INIS)

Dixon Pineda, Manuel Tomas

2012-01-01

The function that entropy has played is compared in processes of non-enzymatic and enzymatic catalysis. The processes followed are showed: the kinetics of the acid hydrolysis of 3-pentyl acetate and cyclopentyl acetate catalyzed by hydrochloric acid and enzymatic hydrolysis of ethyl acetate and γ-butyrolactone catalyzed by pig liver esterase. The activation parameters of Eyring were determined for each process and interpreted the contribution of the entropy of activation for catalysis in this type of model reactions. (author) [es

Oil-in-water emulsions stabilised by cellulose ethers: stability, structure and in vitro digestion.

Science.gov (United States)

Borreani, Jennifer; Espert, María; Salvador, Ana; Sanz, Teresa; Quiles, Amparo; Hernando, Isabel

2017-04-19

The effect of cellulose ethers in oil-in-water emulsions on stability during storage and on texture, microstructure and lipid digestibility during in vitro gastrointestinal digestion was investigated. All the cellulose ether emulsions showed good physical and oxidative stability during storage. In particular, the methylcellulose with high methoxyl substituents (HMC) made it possible to obtain emulsions with high consistency which remained almost unchanged during gastric digestion, and thus could enhance fullness and satiety perceptions at gastric level. Moreover, the HMC emulsion slowed down lipid digestion to a greater extent than a conventional protein emulsion or the emulsions stabilised by the other cellulose ethers. Therefore, HMC emulsions could be used in weight management to increase satiation capacity and decrease lipid digestion.

Enzymatic Hydrolysis of Pretreated Fibre Pressed Oil Palm Frond by using Sacchariseb C6

Science.gov (United States)

Hashim, F. S.; Yussof, H. W.; Zahari, M. A. K. M.; Rahman, R. A.; Illias, R. M.

2017-06-01

Enzymatic hydrolysis becomes a prominent technology for conversion of cellulosic biomass to its glucose monomers that requires an action of cellulolytic enzymes in a sequential and synergistic manner. In this study, the effect of agitation speed, glucan loading, enzyme loading, temperature and reaction time on the production of glucose from fibre pressed oil palm frond (FPOPF) during enzymatic hydrolysis was screened by a half factorial design 25-1 using Response Surface Methodology (RSM). The FPOPF sample was first delignified by alkaline pretreatment at 4.42 (w/v) sodium hydroxide for an hour prior to enzymatic hydrolysis using commercial cellulase enzyme, Sacchariseb C6. The effect of enzymatic hydrolysis on the structural of FPOPF has been evaluated by Scanning Electron Microscopy (SEM) analysis. Characterization of raw FPOPF comprised of 4.5 extractives, 40.7 glucan, 26.1 xylan, 26.2 lignin and 1.8 ash, whereas for pretreated FPOPF gave 0.3 extractives, 61.4 glucan, 20.4 xylan, 13.3 lignin and 1.3 ash. From this study, it was found that the best enzymatic hydrolysis condition yielded 33.01 ± 0.73 g/L of glucose when performed at 200 rpm of agitation speed, 60 FPU/mL of enzyme loading, 4 (w/w) of glucan loading, temperature at 55 □ and 72 hours of reaction time. The model obtained was significant with p-value enzymatic hydrolysis from pretreated FPOPF produce high amount of glucose that enhances it potential for industrial application. This glucose can be further used to produce high-value products.

Comparison of Enzymatic and Ultrasonic Extraction of Albumin from Defatted Pumpkin (Cucurbita pepo Seed Powder

Directory of Open Access Journals (Sweden)

Gia Loi Tu

2015-01-01

Full Text Available In this study, ultrasound- and enzyme-assisted extractions of albumin (water-soluble protein group from defatted pumpkin (Cucurbita pepo seed powder were compared. Both advanced extraction techniques strongly increased the albumin yield in comparison with conventional extraction. The extraction rate was two times faster in the ultrasonic extraction than in the enzymatic extraction. However, the maximum albumin yield was 16 % higher when using enzymatic extraction. Functional properties of the pumpkin seed albumin concentrates obtained using the enzymatic, ultrasonic and conventional methods were then evaluated. Use of hydrolase for degradation of cell wall of the plant material did not change the functional properties of the albumin concentrate in comparison with the conventional extraction. The ultrasonic extraction enhanced water-holding, oil-holding and emulsifying capacities of the pumpkin seed albumin concentrate, but slightly reduced the foaming capacity, and emulsion and foam stability.

Pancreatic islet isolation by mechanical-enzymatic separation, stationary collagenase digestion and dextran discontinuous density gradient purification: experimental study in dogs Isolamento das ilhotas pancreáticas pela separação mecânica-enzimática digestão estacionária com colagenase e purificação com gradiente de densidade descontínua de dextran: estudo experimental em cães

Directory of Open Access Journals (Sweden)

Jaques Waisberg

2002-04-01

Full Text Available The prospects for allotransplantation of pancreatic islets in man depend on the development of methods that provide sufficient quantities of pancreatic islets from a single donor, which are capable, when transplanted, of achieve the normalization of carbohydrate metabolism. Objective: Evaluate the efficacy of the isolation of Langerhans islets from dogs, by means of mechanical-enzymatic separation technique with stationary digestion using collagenase, and purification with a discontinuous dextran density gradient. Methods: The counting of islet numbers and evaluation of their sizes was accomplished by staining with diphenylthiocarbazone and using stereoscopic microscopes equipped with eyepiece reticule for the measurement of average diameters of stained islets. Results: The results disclosed that the average number of islets isolated was 81032.20 ± 24736.79 and the average number of islets isolated per kg of body weight was 6938.70 ± 1392.43. The average number of islets isolated per kg of body weight showed significant correlation with body weight and weight of the pancreas resected. Conclusion: The number of islets isolated, of a single donor, by mechanical-enzymatic separation, stationary collagenase digestion and discontinuous dextran density gradient purification can be sufficient to success of pancreatic islets transplant in dogs.A perspectiva do alotransplante de ilhotas pancreáticas no homem está na dependência do desenvolvimento de métodos que propiciem quantidades suficientes de ilhotas pancreáticas, originadas de doador único, capazes de, quando transplantadas, levarem à normalização do metabolismo dos hidratos de carbono. Objetivo: Avaliar, em cães, a eficácia do isolamento das ilhotas de Langerhans por meio da técnica de separação mecânica-enzimática, digestão estacionária com colagenase e purificação pelo gradiente de densidade descontínua de dextran. Métodos: A contagem do número e avaliação do tamanho

Selective Osmotic Shock (SOS)-Based Islet Isolation for Microencapsulation.

Science.gov (United States)

Enck, Kevin; McQuilling, John Patrick; Orlando, Giuseppe; Tamburrini, Riccardo; Sivanandane, Sittadjody; Opara, Emmanuel C

2017-01-01

Islet transplantation (IT) has recently been shown to be a promising alternative to pancreas transplantation for reversing diabetes. IT requires the isolation of the islets from the pancreas, and these islets can be used to fabricate a bio-artificial pancreas. Enzymatic digestion is the current gold standard procedure for islet isolation but has lingering concerns. One such concern is that it has been shown to damage the islets due to nonselective tissue digestion. This chapter provides a detailed description of a nonenzymatic method that we are exploring in our lab as an alternative to current enzymatic digestion procedures for islet isolation from human and nonhuman pancreatic tissues. This method is based on selective destruction and protection of specific cell types and has been shown to leave the extracellular matrix (ECM) of islets intact, which may thus enhance islet viability and functionality. We also show that these SOS-isolated islets can be microencapsulated for transplantation.

Effects of disintegration on anaerobic degradation of sewage excess sludge in downflow stationary fixed film digesters.

Science.gov (United States)

Engelhart, M; Krüger, M; Kopp, J; Dichtl, N

2000-01-01

The effects of mechanical disintegration on anaerobic digestibility of sewage excess sludge in downflow stationary fixed film (DSFF) digesters were investigated on laboratory scale. Mechanical pretreatment using a high pressure homogenizer led to significantly enhanced concentrations of soluble proteins and carbohydrates in the feed sludge. Using DSFF digesters with two different tubular plastic media as support material it was shown that a stable digestion process could be achieved at hydraulic retention times (HRT) down to 5 days. Compared to conventional digesters at 10 d and 15 d HRT respectively, the degradation of volatile solids was enhanced up to 25%, also resulting in a higher specific biogas production. Further investigations on degradation of soluble proteins and carbohydrates showed that a slowly degradable fraction of carbohydrates was released via disintegration. Using the distribution of chain length and the concentrations of volatile fatty acids as process parameters, the dependability on the HRT and the degree of disintegration (the release of soluble COD) predominated the effects of specific surface area of the support media.

Transforming anaerobic digestion with the Model T of digesters

Energy Technology Data Exchange (ETDEWEB)

Martin, J.F.; Ciotola, R.; Castano, J.; Eger, C.; Schlea, D. [Ohio State Univ., Columbus, OH (United States). Ecological Engineering Program

2010-07-01

Most livestock farmers in the United States do not take advantage of anaerobic digester technology because of the high cost and large scale. These limitations therefore reduce the production of renewable energy from farmlands. In order to expand anaerobic digestion methods and improve environmental quality, affordable and smaller-scale digesters should be developed to enable most livestock farmers to convert manure to methane. Doing so would improve their economic efficiency and environmental sustainability. This paper provided an analogy to the development of the Model T to better explain the need and potential for this technology. A modified fixed-dome digester was installed on the Ohio State University dairy in Columbus, Ohio. The digester was unheated, buried, had a volume of 1 m{sup 3} and received diluted dairy manure as feedstock. Biogas was produced at digester temperatures as low 10 degrees C during colder ambient temperatures. Water quality also improved. Results from the first year of operation will be analyzed to improve performance and enable future development of this technology.

Headway on co-digestion of wastes

International Nuclear Information System (INIS)

Gokhale, Yogeshwar

2013-01-01

Full text: Many wastewater treatment plants (WWTPs) in Australia produce biogas either directly from the wastewater or from anaerobic digestion of the primary and/or secondary sludge, which in turn is used to create energy. Some WWTPs produce electricity from the biogas to either support the treatment plant or to export the electricity to the grid or both. High-strength organic wastes such as fats, oil and grease, food waste, commercial (restaurant) waste and brewery waste are attractive biogas sources that can be realised through co-digestion with the sludge from wastewater treatment. Co-digestion of high-strength waste can be a tricky business due to the varying nature of the waste, special handling requirements, and potential digester process issues like foaming. However, experiences over the past decade have helped identify mitigation measures and advanced designs to reduce these risks. Several WWTPs in the US accept fats, oil and grease (FOG) as a feedstock for co-digestion, and CH2M Hill has been involved in various capacities on some of those projects. The Douglas L. Smith Middle Basin Wastewater Treatment Plant (WWTP) in Johnson County, Kansas, includes an environmentally friendly approach for the treatment of FOG wastes from local restaurants and industrial sources. FOG waste receipts are handled using a separate onsite liquid receiving facility, and the FOG tanks and pipes are heated to minimise clogging. Co-digestion of FOG enhanced the gas production to fuel a 2.1 megawatt biogas co-generation system. Other CH2M Hill FOG co-digestion projects are FOG addition to the incinerator at the Hampton Roads Sanitation District in Virginia; FOG co-digestion at the Pinellas County in Florida; and FOG co-digestion and co-generation at the Essex Junction in Vermont. This experience was recently expanded to the co-digestion of other high-strength organic waste for Yarra Valley Water (YVW) and City West Water (CWW) in Victoria. The potential high-strength waste

Anaerobic Digestion: Process

DEFF Research Database (Denmark)

Angelidaki, Irini; Batstone, Damien J.

2011-01-01

Organic waste may degrade anaerobically in nature as well as in engineered systems. The latter is called anaerobic digestion or biogasification. Anaerobic digestion produces two main outputs: An energy-rich gas called biogas and an effluent. The effluent, which may be a solid as well as liquid...... 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...

Cooking Chicken Breast Reduces Dialyzable Iron Resulting from Digestion of Muscle Proteins

Directory of Open Access Journals (Sweden)

Aditya S. Gokhale

2014-01-01

Full Text Available The purpose of this research was to study the effect of cooking chicken breast on the production of dialyzable iron (an in vitro indicator of bioavailable iron from added ferric iron. Chicken breast muscle was cooked by boiling, baking, sautéing, or deep-frying. Cooked samples were mixed with ferric iron and either extracted with acid or digested with pepsin and pancreatin. Total and ferrous dialyzable iron was measured after extraction or digestion and compared to raw chicken samples. For uncooked samples, dialyzable iron was significantly enhanced after both extraction and digestion. All cooking methods led to markedly reduced levels of dialyzable iron both by extraction and digestion. In most cooked, digested samples dialyzable iron was no greater than the iron-only (no sample control. Cooked samples showed lower levels of histidine and sulfhydryls but protein digestibility was not reduced, except for the sautéed sample. The results showed that, after cooking, little if any dialyzable iron results from digestion of muscle proteins. Our research indicates that, in cooked chicken, residual acid-extractable components are the most important source of dialyzable iron.

Cooking Chicken Breast Reduces Dialyzable Iron Resulting from Digestion of Muscle Proteins.

Science.gov (United States)

Gokhale, Aditya S; Mahoney, Raymond R

2014-01-01

The purpose of this research was to study the effect of cooking chicken breast on the production of dialyzable iron (an in vitro indicator of bioavailable iron) from added ferric iron. Chicken breast muscle was cooked by boiling, baking, sautéing, or deep-frying. Cooked samples were mixed with ferric iron and either extracted with acid or digested with pepsin and pancreatin. Total and ferrous dialyzable iron was measured after extraction or digestion and compared to raw chicken samples. For uncooked samples, dialyzable iron was significantly enhanced after both extraction and digestion. All cooking methods led to markedly reduced levels of dialyzable iron both by extraction and digestion. In most cooked, digested samples dialyzable iron was no greater than the iron-only (no sample) control. Cooked samples showed lower levels of histidine and sulfhydryls but protein digestibility was not reduced, except for the sautéed sample. The results showed that, after cooking, little if any dialyzable iron results from digestion of muscle proteins. Our research indicates that, in cooked chicken, residual acid-extractable components are the most important source of dialyzable iron.

Critical evaluation of pressurized microwave-assisted digestion efficiency using nitric acid oxidizing systems (M7)

International Nuclear Information System (INIS)

Matusiewicz, H.

2002-01-01

Full text: The possibilities of enhancement of a medium-pressure microwave-assisted digestion system for sample preparation in trace element analysis of biological material was investigated. Based on optimal digestion conditions for oxidizing systems with nitric acid, different digestion procedures were examined to minimize residual carbon. The substitution of nitric acid and the addition of hydrogen peroxide and ozone to nitric acid was evaluated. The residual carbon content of the digestate was determined coulometrically. Addition of hydrogen peroxide during organic oxidation reactions does not lower the resolved carbon in the solution. Ozone was tested as an additional, potentially non-contaminating, digestion/oxidation system to the nitric acid used in the sample preparation method. (author)

Radiation pre-treatment of cellulose materials for the enhancement of enzymatic hydrolysis

International Nuclear Information System (INIS)

Ardica, S.; Calderaro, E.; Cappadona, C.

1984-01-01

The effect of γ-ray pre-irradiation of cotton cellulose on glucose production on enzymatic hydrolysis by cellulase has been investigated. Pure cotton has been irradiated in air, in water and in acetate buffer solution over the dose range 10 3 to 10 6 Gy. Pre-irradiation in water or in acetate buffer solution is more effective than in air. The results show that the glucose yield is not always an increasing function of dose, and that for some dose levels, the glucose yields from the irradiated samples are lower than those from non-irradiated samples. (author)

Anaerobic digestion apparatus and process. Procede et installation de digestion anaerobie

Energy Technology Data Exchange (ETDEWEB)

De Baere, L.

1989-05-09

This invention concerns a process for the anaerobic digestion of apparently solid organic matter. The matter is mixed and kneaded with an inoculant to form an apparently solid mass having a water content which varies from around 55 wt % to around 75 wt %. This mass is then introduced into a digestor, where it is digested for a period of around less than 50 days. The biogas produced during the anaerobic digestion stage is recovered, said biogas being a byproduct of the digestion process. The digested mass is extracted, and at least a third, by weight, of that mass is recycled to act as the inoculant. The non-recycled digested mass is removed.

High-throughput microplate technique for enzymatic hydrolysis of lignocellulosic biomass.

Science.gov (United States)

Chundawat, Shishir P S; Balan, Venkatesh; Dale, Bruce E

2008-04-15

Several factors will influence the viability of a biochemical platform for manufacturing lignocellulosic based fuels and chemicals, for example, genetically engineering energy crops, reducing pre-treatment severity, and minimizing enzyme loading. Past research on biomass conversion has focused largely on acid based pre-treatment technologies that fractionate lignin and hemicellulose from cellulose. However, for alkaline based (e.g., AFEX) and other lower severity pre-treatments it becomes critical to co-hydrolyze cellulose and hemicellulose using an optimized enzyme cocktail. Lignocellulosics are appropriate substrates to assess hydrolytic activity of enzyme mixtures compared to conventional unrealistic substrates (e.g., filter paper, chromogenic, and fluorigenic compounds) for studying synergistic hydrolysis. However, there are few, if any, high-throughput lignocellulosic digestibility analytical platforms for optimizing biomass conversion. The 96-well Biomass Conversion Research Lab (BCRL) microplate method is a high-throughput assay to study digestibility of lignocellulosic biomass as a function of biomass composition, pre-treatment severity, and enzyme composition. The most suitable method for delivering milled biomass to the microplate was through multi-pipetting slurry suspensions. A rapid bio-enzymatic, spectrophotometric assay was used to determine fermentable sugars. The entire procedure was automated using a robotic pipetting workstation. Several parameters that affect hydrolysis in the microplate were studied and optimized (i.e., particle size reduction, slurry solids concentration, glucan loading, mass transfer issues, and time period for hydrolysis). The microplate method was optimized for crystalline cellulose (Avicel) and ammonia fiber expansion (AFEX) pre-treated corn stover. Copyright 2008 Wiley Periodicals, Inc.

Combined electrical-alkali pretreatment to increase the anaerobic hydrolysis rate of waste activated sludge during anaerobic digestion

International Nuclear Information System (INIS)

Zhen, Guangyin; Lu, Xueqin; Li, Yu-You; Zhao, Youcai

2014-01-01

Highlights: • Combined electrical-alkali pretreatment for improving sludge anaerobic digestion was proposed. • Combined process enhanced the cell lysis, biopolymers releases, and thus sludge disintegration. • Increased solubilization of sludge increased the anaerobic hydrolysis rate. • Increased solubilization does not always induce an improved anaerobic digestion efficiency. - Abstract: Pretreatment can be used prior to anaerobic digestion to improve the efficiency of waste activated sludge (WAS) digestion. In this study, electrolysis and a commonly used pretreatment method of alkaline (NaOH) solubilization were integrated as a pretreatment method for promoting WAS anaerobic digestion. Pretreatment effectiveness of combined process were investigated in terms of disintegration degree (DD SCOD ), suspended solids (TSS and VSS) removals, the releases of protein (PN) and polysaccharide (PS), and subsequent anaerobic digestion as well as dewaterability after digestion. Electrolysis was able to crack the microbial cells trapped in sludge gels and release the biopolymers (PN and PS) due to the cooperation of alkaline solubilization, enhancing the sludge floc disintegration/solubilization, which was confirmed by scanning electron microscopy (SEM) analysis. Biochemical methane potential (BMP) assays showed the highest methane yield was achieved with 5 V plus pH 9.2 pretreatment with up to 20.3% improvement over the non-pretreated sludge after 42 days of mesophilic operation. In contrast, no discernible improvements on anaerobic degradability were observed for the rest of pretreated sludges, probably due to the overmuch leakage of refractory soluble organics, partial chemical mineralization of solubilized compounds and sodium inhibition. The statistical analysis further indicated that increased solubilization induced by electrical-alkali pretreatment increased the first-order anaerobic hydrolysis rate (k hyd ), but had no, or very slight enhancement on WAS ultimate

Enhanced Enzymatic Hydrolysis and Structural Features of Corn Stover by NaOH and Ozone Combined Pretreatment

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Wenhui Wang

2018-05-01

Full Text Available A two-step pretreatment using NaOH and ozone was performed to improve the enzymatic hydrolysis, compositions and structural characteristics of corn stover. Comparison between the unpretreated and pretreated corn stover was also made to illustrate the mechanism of the combined pretreatment. A pretreatment with 2% (w/w NaOH at 80 °C for 2 h followed by ozone treatment for 25 min with an initial pH 9 was found to be the optimal procedure and the maximum efficiency (91.73% of cellulose enzymatic hydrolysis was achieved. Furthermore, microscopic observation of changes in the surface structure of the samples showed that holes were formed and lignin and hemicellulose were partially dissolved and removed. X-ray Diffraction (XRD, Fourier Transform Infrared Spectroscopy (FTIR and Cross-Polarization Magic Angle Spinning Carbon-13 Nuclear Magnetic Resonance (CP/MAS 13C-NMR were also used to characterize the chemical structural changes after the combined pretreatment. The results were as follows: part of the cellulose I structure was destroyed and then reformed into cellulose III, the cellulose crystal indices were also changed; a wider space between the crystal layer was observed; disruption of hydrogen bonds in cellulose and disruption of ester bonds in hemicellulose; cleavage of bonds linkage in lignin-carbohydrate complexes; removal of methoxy in lignin and hemicellulose. As a result, all these changes effectively reduced recalcitrance of corn stover and promoted subsequent enzymatic hydrolysis of cellulose.

Enhanced methane production of Chlorella vulgaris and Chlamydomonas reinhardtii by hydrolytic enzymes addition

International Nuclear Information System (INIS)

Mahdy, Ahmed; Mendez, Lara; Ballesteros, Mercedes; González-Fernández, Cristina

2014-01-01

Highlights: • Methane production of microalgae biomass is hampered by their cell wall. • Pretreatment should be designed in accordance to the microalgae specie. • Fresh Chlamydomonas reinhardtii exhibited high anaerobic biodegradability. • Chlorella vulgaris anaerobic biodegradability was enhanced by 50% using protease pretreatment. - Abstract: The effect of enzymatic hydrolysis on microalgae organic matter solubilisation and methane production was investigated in this study. Even though both biomasses, Chlamydomonas reinhardtii and Chlorella vulgaris, exhibited similar macromolecular distribution, their cell wall composition provided different behaviors. The addition of carbohydrolase (Viscozyme) and protease (Alcalase) resulted in high carbohydrates and protein solubilisation on both biomasses (86–96%). Despite the high carbohydrate solubilisation with the carbohydrolase, methane production was enhanced by 14% for C. vulgaris, while hydrolyzed C. reinhardtii did not show any improvement. The addition of protease to C. reinhardtii increased methane production by 1.17-fold. The low enhancement achieved together with the inherent high biodegradability of this biomass would not justify the cost associated to the enzyme addition. On the other hand, C. vulgaris hydrolyzed with the protease resulted in 86% anaerobic biodegradability compared to 54% of the raw biomass. Therefore, the application of protease prior anaerobic digestion of C. vulgaris could be a promising approach to decrease the energetic input required for cell wall disruption

Starch Origin and Thermal Processing Affect Starch Digestion in a Minipig Model of Pancreatic Exocrine Insufficiency.

Science.gov (United States)

Mößeler, Anne; Vagt, Sandra; Beyerbach, Martin; Kamphues, Josef

2015-01-01

Although steatorrhea is the most obvious symptom of pancreatic exocrine insufficiency (PEI), enzymatic digestion of protein and starch is also impaired. Low praecaecal digestibility of starch causes a forced microbial fermentation accounting for energy losses and meteorism. To optimise dietetic measures, knowledge of praecaecal digestibility of starch is needed but such information from PEI patients is rare. Minipigs fitted with an ileocaecal fistula with (n = 3) or without (n = 3) pancreatic duct ligation (PL) were used to estimate the rate of praecaecal disappearance (pcD) of starch. Different botanical sources of starch (rice, amaranth, potato, and pea) were fed either raw or cooked. In the controls (C), there was an almost complete pcD (>92%) except for potato starch (61.5%) which was significantly lower. In PL pcD of raw starch was significantly lower for all sources of starch except for amaranth (87.9%). Thermal processing increased pcD in PL, reaching values of C for starch from rice, potato, and pea. This study clearly underlines the need for precise specification of starch used for patients with specific dietetic needs like PEI. Data should be generated in suitable animal models or patients as tests in healthy individuals would not have given similar conclusions.

Starch Origin and Thermal Processing Affect Starch Digestion in a Minipig Model of Pancreatic Exocrine Insufficiency

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Anne Mößeler

2015-01-01

Full Text Available Although steatorrhea is the most obvious symptom of pancreatic exocrine insufficiency (PEI, enzymatic digestion of protein and starch is also impaired. Low praecaecal digestibility of starch causes a forced microbial fermentation accounting for energy losses and meteorism. To optimise dietetic measures, knowledge of praecaecal digestibility of starch is needed but such information from PEI patients is rare. Minipigs fitted with an ileocaecal fistula with (n=3 or without (n=3 pancreatic duct ligation (PL were used to estimate the rate of praecaecal disappearance (pcD of starch. Different botanical sources of starch (rice, amaranth, potato, and pea were fed either raw or cooked. In the controls (C, there was an almost complete pcD (>92% except for potato starch (61.5% which was significantly lower. In PL pcD of raw starch was significantly lower for all sources of starch except for amaranth (87.9%. Thermal processing increased pcD in PL, reaching values of C for starch from rice, potato, and pea. This study clearly underlines the need for precise specification of starch used for patients with specific dietetic needs like PEI. Data should be generated in suitable animal models or patients as tests in healthy individuals would not have given similar conclusions.

Environmental impacts of anaerobic digestion and the use of anaerobic residues as soil amendment

Energy Technology Data Exchange (ETDEWEB)

Mosey, F.E. [VFA Services Ltd., Herts (United Kingdom)

1996-01-01

This paper defines the environmental role of anaerobic digestion within the overall objective of recovering energy from renewable biomass resources. Examples and opportunities for incorporating anaerobic digestion into biomass-to-energy schemes are discussed, together with environmental aspects of anaerobic digestion plants. These include visual, public amenity, pathogens and public health, odor control, and gaseous emissions. Digestate disposal and the benefits of restrictions on recycling organic wastes and biomass residues back to the land are discussed, particularly as they relate to American and European codes of practice and environmental legislation. The paper concludes that anaerobic digestion, if performed in purpose-designed reactors that efficiently recover and use biogas, is an environmentally benign process that can enhance energy recovery and aid the beneficial land use of plant residues in many biomass-to-energy schemes.

Effect of mixing digested slurry on the rate of biogas production from dairy manure in batch fermenter

Energy Technology Data Exchange (ETDEWEB)

Kalia, A.K.; Singh, S.P.

2001-09-01

Forty kilograms of pure cattle dung and cattle dung mixed with 10% digested slurry obtained from a field biogas plant was batch fermented in horizontal biogas digesters for 15 weeks under field conditions with mean ambient temperature 20-23{sup o}C. Compared to 821 l of biogas from digester I, containing cattle dung alone, 1457 l of biogas was obtained from digester II, containing cattle dung mixed with 10% digested slurry. Mixing of slurry not only speeded up the gas production but also enhanced its rate from 108 l/kg dry matter to 158 l/kg dry matter. It also resulted in 36.1% distraction of total volatile solid in digester II, compared to 23.93% observed in digester I. Mixing digested slurry is recommended for raising biogas production from cattle dung in dry fermenters. (author)

Bioethanol production: an integrated process of low substrate loading hydrolysis-high sugars liquid fermentation and solid state fermentation of enzymatic hydrolysis residue.

Science.gov (United States)

Chu, Qiulu; Li, Xin; Ma, Bin; Xu, Yong; Ouyang, Jia; Zhu, Junjun; Yu, Shiyuan; Yong, Qiang

2012-11-01

An integrated process of enzymatic hydrolysis and fermentation was investigated for high ethanol production. The combination of enzymatic hydrolysis at low substrate loading, liquid fermentation of high sugars concentration and solid state fermentation of enzymatic hydrolysis residue was beneficial for conversion of steam explosion pretreated corn stover to ethanol. The results suggested that low substrate loading hydrolysis caused a high enzymatic hydrolysis yield; the liquid fermentation of about 200g/L glucose by Saccharomyces cerevisiae provided a high ethanol concentration which could significantly decrease cost of the subsequent ethanol distillation. A solid state fermentation of enzymatic hydrolysis residue was combined, which was available to enhance ethanol production and cellulose-to-ethanol conversion. The results of solid state fermentation demonstrated that the solid state fermentation process accompanied by simultaneous saccharification and fermentation. Copyright © 2012 Elsevier Ltd. All rights reserved.

Enhanced biogas yield by thermo-alkali solubilization followed by co-digestion of intestine waste from slaughterhouse with food waste.

Science.gov (United States)

Porselvam, S; Soundara Vishal, N; Srinivasan, S V

2017-10-01

Intestine waste generated from slaughterhouse (IWS) is difficult to degrade in anaerobic process due to the presence of high protein and lipid contents. However, anaerobic co-digestion helps to increase the degradation of IWS by the addition of carbon-rich food waste (FW). To increase the biogas yield, thermo-alkali pretreatment may be more viable method for the anaerobic digestion of protein and lipid rich wastes. In the present study, Thermo-alkali pretreatment of intestine waste from slaughterhouse and food waste alone and mixing of IWS and FW with different ratios (1:1-1:3) on VS basis have been studied. To study the effect of Thermo-alkali pretreatment on solubilization of substrate, the substrate was mixed with alkali solutions (NaOH and KOH) at different concentrations of 1, 2, 3, 4 and 5% solutions. The results revealed that the maximum solubilization was observed to be 94.7% and 90.1% at KOH (1:3 and 5%) and NaOH (1:3 and 5%), respectively. Based on the study, enhancement in biogas yield by 16% (IWS), 11.5% (FW), 12.2% (1:1), 18.11% (1:2) and 22.5% (1:3) in KOH pretreated waste when compared with NaOH pretreated waste.

Supplementing with non-glycoside hydrolase proteins enhances enzymatic deconstruction of plant biomass.

Science.gov (United States)

Su, Xiaoyun; Zhang, Jing; Mackie, Roderick I; Cann, Isaac K O

2012-01-01

The glycoside hydrolases (GH) of Caldicellulosiruptor bescii are thermophilic enzymes, and therefore they can hydrolyze plant cell wall polysaccharides at high temperatures. Analyses of two C. bescii glycoside hydrolases, CbCelA-TM1 and CbXyn10A with cellulase and endoxylanase activity, respectively, demonstrated that each enzyme is highly thermostable under static incubation at 70°C. Both enzymes, however, rapidly lost their enzymatic activities when incubated at 70°C with end-over-end shaking. Since crowding conditions, even at low protein concentrations, seem to influence enzymatic properties, three non-glycoside hydrolase proteins were tested for their capacity to stabilize the thermophilic proteins at high temperatures. The three proteins investigated were a small heat shock protein CbHsp18 from C. bescii, a histone MkHistone1 from Methanopyrus kandleri, and bovine RNase A, from a commercial source. Fascinatingly, each of these proteins increased the thermostability of the glycoside hydrolases at 70°C during end-over-end shaking incubation, and this property translated into increases in hydrolysis of several substrates including the bioenergy feedstock Miscanthus. Furthermore, MkHistone1 and RNase A also altered the initial products released from the cello-oligosaccharide cellopentaose during hydrolysis with the cellodextrinase CbCdx1A, which further demonstrated the capacity of the three non-GH proteins to influence hydrolysis of substrates by the thermophilic glycoside hydrolases. The non-GH proteins used in the present report were small proteins derived from each of the three lineages of life, and therefore expand the space from which different polypeptides can be tested for their influence on plant cell wall hydrolysis, a critical step in the emerging biofuel industry.

Supplementing with non-glycoside hydrolase proteins enhances enzymatic deconstruction of plant biomass.

Directory of Open Access Journals (Sweden)

Xiaoyun Su

Full Text Available The glycoside hydrolases (GH of Caldicellulosiruptor bescii are thermophilic enzymes, and therefore they can hydrolyze plant cell wall polysaccharides at high temperatures. Analyses of two C. bescii glycoside hydrolases, CbCelA-TM1 and CbXyn10A with cellulase and endoxylanase activity, respectively, demonstrated that each enzyme is highly thermostable under static incubation at 70°C. Both enzymes, however, rapidly lost their enzymatic activities when incubated at 70°C with end-over-end shaking. Since crowding conditions, even at low protein concentrations, seem to influence enzymatic properties, three non-glycoside hydrolase proteins were tested for their capacity to stabilize the thermophilic proteins at high temperatures. The three proteins investigated were a small heat shock protein CbHsp18 from C. bescii, a histone MkHistone1 from Methanopyrus kandleri, and bovine RNase A, from a commercial source. Fascinatingly, each of these proteins increased the thermostability of the glycoside hydrolases at 70°C during end-over-end shaking incubation, and this property translated into increases in hydrolysis of several substrates including the bioenergy feedstock Miscanthus. Furthermore, MkHistone1 and RNase A also altered the initial products released from the cello-oligosaccharide cellopentaose during hydrolysis with the cellodextrinase CbCdx1A, which further demonstrated the capacity of the three non-GH proteins to influence hydrolysis of substrates by the thermophilic glycoside hydrolases. The non-GH proteins used in the present report were small proteins derived from each of the three lineages of life, and therefore expand the space from which different polypeptides can be tested for their influence on plant cell wall hydrolysis, a critical step in the emerging biofuel industry.

Enzymatic saccharification of high pressure assist-alkali pretreated cotton stalk and structural characterization.

Science.gov (United States)

Du, Shuang-kui; Su, Xia; Yang, Weihua; Wang, Yanqin; Kuang, Meng; Ma, Lei; Fang, Dan; Zhou, Dayun

2016-04-20

Cotton stalk is a potential biomass for bioethanol production, while the conversion of direct saccharification or biotransformation of cotton stalk is extremely low due to the recalcitrant nature of lignocellulose. To enhance the enzymatic conversion of cotton stalks, the enzymatic saccharification parameters of high pressure assist-alkali pretreatment (HPAP) cotton stalk were optimized in the present study. Results indicated that a maximum reducing sugar yield of 54.7g/100g dry biomass cellulose was achieved at a substrate concentration of 2%, 100rpm agitation, 0.6g/g enzyme loading, 40°C hydrolysis temperature, 50h saccharification time, and pH 5.0. Scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy were used to identify structural changes in native, pretreated biomass and hydrolyzed residues. Structural analysis revealed large part of amorphous cellulose and partial crystalline cellulose in the HPAP cotton stalk were hydrolyzed during enzymatic treatment. HPAP cotton stalk can be used as a potential feed stock for bioethanol production. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enzymatic digestibility and ethanol fermentability of AFEX-treated starch-rich lignocellulosics such as corn silage and whole corn plant

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Thelen Kurt D

2010-06-01

Full Text Available Abstract Background Corn grain is an important renewable source for bioethanol production in the USA. Corn ethanol is currently produced by steam liquefaction of starch-rich grains followed by enzymatic saccharification and fermentation. Corn stover (the non-grain parts of the plant is a potential feedstock to produce cellulosic ethanol in second-generation biorefineries. At present, corn grain is harvested by removing the grain from the living plant while leaving the stover behind on the field. Alternatively, whole corn plants can be harvested to cohydrolyze both starch and cellulose after a suitable thermochemical pretreatment to produce fermentable monomeric sugars. In this study, we used physiologically immature corn silage (CS and matured whole corn plants (WCP as feedstocks to produce ethanol using ammonia fiber expansion (AFEX pretreatment followed by enzymatic hydrolysis (at low enzyme loadings and cofermentation (for both glucose and xylose using a cellulase-amylase-based cocktail and a recombinant Saccharomyces cerevisiae 424A (LNH-ST strain, respectively. The effect on hydrolysis yields of AFEX pretreatment conditions and a starch/cellulose-degrading enzyme addition sequence for both substrates was also studied. Results AFEX-pretreated starch-rich substrates (for example, corn grain, soluble starch had a 1.5-3-fold higher enzymatic hydrolysis yield compared with the untreated substrates. Sequential addition of cellulases after hydrolysis of starch within WCP resulted in 15-20% higher hydrolysis yield compared with simultaneous addition of hydrolytic enzymes. AFEX-pretreated CS gave 70% glucan conversion after 72 h of hydrolysis for 6% glucan loading (at 8 mg total enzyme loading per gram glucan. Microbial inoculation of CS before ensilation yielded a 10-15% lower glucose hydrolysis yield for the pretreated substrate, due to loss in starch content. Ethanol fermentation of AFEX-treated (at 6% w/w glucan loading CS hydrolyzate (resulting

Enzymatic digestibility and ethanol fermentability of AFEX-treated starch-rich lignocellulosics such as corn silage and whole corn plant

Science.gov (United States)

2010-01-01

Background Corn grain is an important renewable source for bioethanol production in the USA. Corn ethanol is currently produced by steam liquefaction of starch-rich grains followed by enzymatic saccharification and fermentation. Corn stover (the non-grain parts of the plant) is a potential feedstock to produce cellulosic ethanol in second-generation biorefineries. At present, corn grain is harvested by removing the grain from the living plant while leaving the stover behind on the field. Alternatively, whole corn plants can be harvested to cohydrolyze both starch and cellulose after a suitable thermochemical pretreatment to produce fermentable monomeric sugars. In this study, we used physiologically immature corn silage (CS) and matured whole corn plants (WCP) as feedstocks to produce ethanol using ammonia fiber expansion (AFEX) pretreatment followed by enzymatic hydrolysis (at low enzyme loadings) and cofermentation (for both glucose and xylose) using a cellulase-amylase-based cocktail and a recombinant Saccharomyces cerevisiae 424A (LNH-ST) strain, respectively. The effect on hydrolysis yields of AFEX pretreatment conditions and a starch/cellulose-degrading enzyme addition sequence for both substrates was also studied. Results AFEX-pretreated starch-rich substrates (for example, corn grain, soluble starch) had a 1.5-3-fold higher enzymatic hydrolysis yield compared with the untreated substrates. Sequential addition of cellulases after hydrolysis of starch within WCP resulted in 15-20% higher hydrolysis yield compared with simultaneous addition of hydrolytic enzymes. AFEX-pretreated CS gave 70% glucan conversion after 72 h of hydrolysis for 6% glucan loading (at 8 mg total enzyme loading per gram glucan). Microbial inoculation of CS before ensilation yielded a 10-15% lower glucose hydrolysis yield for the pretreated substrate, due to loss in starch content. Ethanol fermentation of AFEX-treated (at 6% w/w glucan loading) CS hydrolyzate (resulting in 28 g/L ethanol

ESR studies on the influence of physiological dissolution and digestion media on the lipid phase characteristics of SEDDS and SEDDS pellets.

Science.gov (United States)

Abdalla, Ahmed; Mäder, Karsten

2009-02-09

The aim of the current study is the evaluation of a recently optimized SEDDS, composed of Solutol HS15 and medium chain glycerides, and self-emulsifying pellets by means of ESR. Tempol-benzoate (TB)-loaded SEDDS were produced and electron spin resonance (ESR) spectroscopy was used to evaluate the diluted self-emulsifying mixtures. Moreover, ESR in vitro digestion experiments were carried out to have an insight on the characteristics of the different phases formed during the digestion process and to evaluate the distribution and the localization of TB in these phases. In addition, self-emulsifying pellets were produced using nitroxide-loaded SEDDS and the microenvironment within the pellets during release process was monitored in an online process using ESR spectroscopy. After dilution of nitroxide-loaded SEDDS, the percent of TB localized in the lipophilic compartment was decreasing with increasing the surfactant fraction in the mixture. Moreover, it was found that different phases with variable viscosity and polarity were produced as a result of the enzymatic digestion of SEDDS in physiologically relevant media. This change in lipid composition has largely affected the distribution and the localization of the spin probe during the digestion process. A rapid increase in the mobility of the spin probe inside the pellets was noticed after exposure to the release media. Additionally, TB was localized within the self-emulsifying mixture environment for the time of the experiment. ESR is considered a powerful non-invasive tool to assess the microenvironment of the diluted SEDDS and to monitor in vitro digestion process. Digestion induces a change in lipid composition which can affect the solubilization capacity of the administered drug. Therefore, monitoring in vitro digestion process using ESR spectroscopy will help in providing greater understanding of the interaction between the administered drug and the digested lipid vehicles.

Comparison of Enzymatic and Ultrasonic Extraction of Albumin from Defatted Pumpkin (Cucurbita pepo)
Seed Powder.

Science.gov (United States)

Tu, Gia Loi; Bui, Thi Hoang Nga; Tran, Thi Thu Tra; Ton, Nu Minh Nguyet; Man Le, Van Viet

2015-12-01

In this study, ultrasound- and enzyme-assisted extractions of albumin (water-soluble protein group) from defatted pumpkin ( Cucurbita pepo ) seed powder were compared. Both advanced extraction techniques strongly increased the albumin yield in comparison with conventional extraction. The extraction rate was two times faster in the ultrasonic extraction than in the enzymatic extraction. However, the maximum albumin yield was 16% higher when using enzymatic extraction. Functional properties of the pumpkin seed albumin concentrates obtained using the enzymatic, ultrasonic and conventional methods were then evaluated. Use of hydrolase for degradation of cell wall of the plant material did not change the functional properties of the albumin concentrate in comparison with the conventional extraction. The ultrasonic extraction enhanced water-holding, oil-holding and emulsifying capacities of the pumpkin seed albumin concentrate, but slightly reduced the foaming capacity, and emulsion and foam stability.

PHARMACOTHERAPY OF THE DIGESTIVE DISEASES AMONG TEENAGERS. CURRENT STATE OF THE ISSUE

Directory of Open Access Journals (Sweden)

E.A. Vishneva

2009-01-01

Full Text Available Deterioration of the children's health, especially as long as they study in school is one of the key issues today. The article highlights the results of the «Healthy school» program implementation, which was developed by the «Quality of life» charity fund involving Solvay Pharma. Much attention was paid to the pharmacoepidemiological study of the digestive diseases among teenagers — conformance evaluation of the given nosology pharmacotherapy at the outpatient stage to the existing standards of the evidence based medicine. The researcher described in detail nonconformances to the generally accepted recommendations identified during the study — nonprescription and/or unjustified application of the medications. They also provided the structure of the enzymatic drugs application.Key words: pharmacoepidemiology, gastrointestinal disturbances, pharmacotherapy, children.

Cassava Pulp as a Biofuel Feedstock of an Enzymatic Hydrolysis Proces

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Djuma’ali Djuma’ali

2013-03-01

Full Text Available Cassava pulp, a low cost solid byproduct of cassava starch industry, has been proposed as a high potential ethanolic fermentation substrate due to its high residual starch level, low ash content and small particle size of the lignocellulosic fibers. As the economic feasibility depends on complete degradation of the polysaccharides to fermentable glucose, the comparative hydrolytic potential of cassava pulp by six commercial enzymes were studied. Raw cassava pulp (12% w/v, particle size <320 μm hydrolyzed by both commercial pectinolytic (1 and amylolytic (2 enzymes cocktail, yielded 70.06% DE. Hydrothermal treatment of cassava pulp enhanced its susceptibility to enzymatic cleavageas compared to non-hydrothermal treatment raw cassava pulp. Hydrothermal pretreatment has shown that a glucoamylase (3 was the most effective enzyme for hydrolysis process of cassava pulp at temperature 65 °C or 95 °C for 10 min and yielded approximately 86.22% and 90.18% DE, respectively. Enzymatic pretreatment increased cassava pulp vulnerability to cellulase attacks. The optimum conditions for enzymatic pretreatment of 30% (w/v cassava pulp by a potent cellulolytic/ hemicellulolytic enzyme (4 was achieves at 50 °C for 3, meanwhile for liquefaction and saccharification by a thermo-stable α-amylase (5 was achieved at 95 °C for 1 and a glucoamylase (3 at 50 °C for 24 hours, respectively, yielded a reducing sugar level up to 94,1% DE. The high yield of glucose indicates the potential use of enzymatic-hydrothermally treated cassava pulp as a cheap substrate for ethanol production.

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

International Nuclear Information System (INIS)

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

2010-01-01

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 o C and initial total solid loading (TSL) ranged from 1.2% to 6.0%. Batch co-digestion experiments were performed at 35 ± 1 o 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.

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.

Comparison of chemical, electrophoretic and in vitro digestion methods for predicting fish meal nutritive quality

DEFF Research Database (Denmark)

Bassompierre, M.; Larsen, K.L.; Zimmermann, W.

1998-01-01

utilization, NPU, biological value, BV, and true digestibility, TD), capillary electrophoresis (sodium dodecyl sulphate-capillary gel electrophoresis, SDS-CGE)) and an in vitro enzymatic assay (trinitrobenzene sulphonic acid-based closed system with rainbow trout enzyme extract) were used for further...... different quantities of water-soluble protein (wsp). A low-temperature-dried FM was employed as a reference. Acquired chemical data for each of the FMs included amino acid analysis and proximal composition (protein, fat, ash, ammonia, titration, salt, moisture). Biological methods in rat (net protein...... comparisons with FM wsp content. A high correlation (R = 0.97; P titration volume was observed. In contrast to BV and NPU (R = 0.98; P

Enhanced hydrolysis and methane yield by applying microaeration pretreatment to the anaerobic co-digestion of brown water and food waste

International Nuclear Information System (INIS)

Lim, Jun Wei; Wang, Jing-Yuan

2013-01-01

Highlights: ► Microaeration pretreatment was effective for brown water and food waste mixture. ► The added oxygen was consumed fully by facultative microorganisms. ► Enhanced solubilization, acidification and breakdown of SCFAs to acetate. ► Microaeration pretreatment improved methane yield by 10–21%. ► Nature of inoculum influenced the effects of microaeration. - Abstract: Microaeration has been used conventionally for the desulphurization of biogas, and recently it was shown to be an alternative pretreatment to enhance hydrolysis of the anaerobic digestion (AD) process. Previous studies on microaeration pretreatment were limited to the study of substrates with complex organic matter, while little has been reported on its effect on substrates with higher biodegradability such as brown water and food waste. Due to the lack of consistent microaeration intensities, previous studies were not comparable and thus inconclusive in proving the effectiveness of microaeration to the overall AD process. In this study, the role of microaeration pretreatment in the anaerobic co-digestion of brown water and food waste was evaluated in batch-tests. After a 4-day pretreatment with 37.5 mL-O 2 /L R -d added to the liquid phase of the reactor, the methane production of substrates were monitored in anaerobic conditions over the next 40 days. The added oxygen was consumed fully by facultative microorganisms and a reducing environment for organic matter degradation was maintained. Other than higher COD solubilization, microaeration pretreatment led to greater VFA accumulation and the conversion of other short chain fatty acids to acetate. This could be due to enhanced activities of hydrolytic and acidogenic bacteria and the degradation of slowly biodegradable compounds under microaerobic conditions. This study also found that the nature of inoculum influenced the effects of microaeration as a 21% and 10% increase in methane yield was observed when pretreatment was applied

Enhanced hydrolysis and methane yield by applying microaeration pretreatment to the anaerobic co-digestion of brown water and food waste

Energy Technology Data Exchange (ETDEWEB)

Lim, Jun Wei [Residues and Resource Reclamation Centre, Nanyang Environment and Water Research Institute, Nanyang Technological University, 06-08 CleanTech One, 1 Cleantech Loop, Singapore 637141 (Singapore); School of Civil and Environmental Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore 639798 (Singapore); Wang, Jing-Yuan, E-mail: jywang@ntu.edu.sg [Residues and Resource Reclamation Centre, Nanyang Environment and Water Research Institute, Nanyang Technological University, 06-08 CleanTech One, 1 Cleantech Loop, Singapore 637141 (Singapore); School of Civil and Environmental Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore 639798 (Singapore)

2013-04-15

Highlights: ► Microaeration pretreatment was effective for brown water and food waste mixture. ► The added oxygen was consumed fully by facultative microorganisms. ► Enhanced solubilization, acidification and breakdown of SCFAs to acetate. ► Microaeration pretreatment improved methane yield by 10–21%. ► Nature of inoculum influenced the effects of microaeration. - Abstract: Microaeration has been used conventionally for the desulphurization of biogas, and recently it was shown to be an alternative pretreatment to enhance hydrolysis of the anaerobic digestion (AD) process. Previous studies on microaeration pretreatment were limited to the study of substrates with complex organic matter, while little has been reported on its effect on substrates with higher biodegradability such as brown water and food waste. Due to the lack of consistent microaeration intensities, previous studies were not comparable and thus inconclusive in proving the effectiveness of microaeration to the overall AD process. In this study, the role of microaeration pretreatment in the anaerobic co-digestion of brown water and food waste was evaluated in batch-tests. After a 4-day pretreatment with 37.5 mL-O{sub 2}/L{sub R}-d added to the liquid phase of the reactor, the methane production of substrates were monitored in anaerobic conditions over the next 40 days. The added oxygen was consumed fully by facultative microorganisms and a reducing environment for organic matter degradation was maintained. Other than higher COD solubilization, microaeration pretreatment led to greater VFA accumulation and the conversion of other short chain fatty acids to acetate. This could be due to enhanced activities of hydrolytic and acidogenic bacteria and the degradation of slowly biodegradable compounds under microaerobic conditions. This study also found that the nature of inoculum influenced the effects of microaeration as a 21% and 10% increase in methane yield was observed when pretreatment was

Increased saccharification yields from aspen biomass upon treatment with enzymatically generated peracetic acid.

Science.gov (United States)

Duncan, Shona; Jing, Qing; Katona, Adrian; Kazlauskas, Romas J; Schilling, Jonathan; Tschirner, Ulrike; Aldajani, Waleed Wafa

2010-03-01

The recalcitrance of lignocellulosic biomass to enzymatic release of sugars (saccharification) currently limits its use as feedstock for biofuels. Enzymatic hydrolysis of untreated aspen wood releases only 21.8% of the available sugars due primarily to the lignin barrier. Nature uses oxidative enzymes to selectively degrade lignin in lignocellulosic biomass, but thus far, natural enzymes have been too slow for industrial use. In this study, oxidative pretreatment with commercial peracetic acid (470 mM) removed 40% of the lignin (from 19.9 to 12.0 wt.% lignin) from aspen and enhanced the sugar yields in subsequent enzymatic hydrolysis to about 90%. Increasing the amount of lignin removed correlated with increasing yields of sugar release. Unfortunately, peracetic acid is expensive, and concentrated forms can be hazardous. To reduce costs and hazards associated with using commercial peracetic acid, we used a hydrolase to catalyze the perhydrolysis of ethyl acetate generating 60-70 mM peracetic acid in situ as a pretreatment to remove lignin from aspen wood. A single pretreatment was insufficient, but multiple cycles (up to eight) removed up to 61.7% of the lignin enabling release of >90% of the sugars during saccharification. This value corresponds to a predicted 581 g of fermentable sugars from 1 kg of aspen wood. Improvements in the enzyme stability are needed before the enzymatically generated peracetic acid is a commercially viable alternative.

Enzymatic characterization of peptidic materials isolated from aqueous solutions of ammonium cyanide (pH 9) and hydrocyanic acid (pH 6) exposed to ionizing radiation.

Science.gov (United States)

Niketic, V; Draganić, Z; Nesković, S; Draganić, I

1982-01-01

The enzymatic digestion of some radiolytically produced peptidic materials was examined. The substrates were compounds isolated from 0.1 molar solutions of NH4CN (pH 9) and HCN (pH 6), after their exposure to gamma rays from a 60Co source (15-20 Mrad doses). Commercial proteolytic enzymes pronase and aminopeptidase M were used. The examined materials were of composite nature and proteolytic action was systematically observed after their subsequent purification. In some fractions the effect was found to be positive with up to 30% of peptide bonds cleaved with respect to the amino acid content. These findings support our previous conclusions on the free radical induced formation of peptidic backbones without the intervention of amino acids. Some side effects were also noted which might be of interest in observations on enzymatic cleavage of other composite peptidic materials of abiotic origin.

Methane Production and Kinetic Modeling for Co-digestion of Manure with Lignocellulosic Residues

DEFF Research Database (Denmark)

Awais, Muhammad; Alvarado-Morales, Merlin; Tsapekos, Panagiotis

2016-01-01

L of CH4 g-1 of VS day-1, respectively. Moreover, the positive effects of coupled biological reactions in the reaction mixture of co-digestion were explained using the synergistic effect value (η). The η value was calculated using estimated and experimental methane yields. Furthermore, in MG co...... of different physicochemical characteristics with CM, the overall biodegradability compared to single-substrate digestion is improved and the methane yield is enhanced....

Investigations into Improving Dewaterability at a Bio-P/Anaerobic Digestion Plant.

Science.gov (United States)

Alm, Rebecca; Sealock, Adam W; Nollet, Yabing; Sprouse, George

2016-11-01

  Metropolitan Council Environmental Services has observed poorer than expected dewatering performance at its Empire Plant. This plant has both anaerobic digestion and enhanced biological phosphorus removal in its treatment train. A research program using pilot-scale anaerobic digesters investigated potential solutions to the plant's poor dewaterability. The dewaterability goal was to increase the cake solids from 12% total solids (TS) to 16% TS or higher. This research investigated 20 different reactor conditions including chemical, feed sludge, and digested sludge treatments. At the pilot scale, unaerated storage of waste activated sludge prior to thickening and addition of ferric chloride to digestion was found to achieve dewatered cake solids of nearly 17% TS with the added benefit of reducing polymer demand. Issues including the amount of chemical required and the resulting volatile solids destruction influence the viability of the process change, so a full-scale pilot and financial analysis is recommended before making permanent process changes.

PRELIMINARY HIGH PERFORMANCE CAPILLARY ELECTROPHORESIS (HPCE) STUDIES OF ENZYMATIC DEGRADATION OF HYALURONIC ACID BY HYALURONIDASE IN THE PRESENCE OF POLYVALENT METAL IONS.

Science.gov (United States)

Urbaniak, Bartosz; Plewa, Szymon; Kokot, Zenon Jozef

2017-01-01

The aim of this study was, at first, to examine the influence of metal ions on digestion process of hyaluronic acid by hyaluronidase (HAse) using high performance capillary electrophoresis (HPCE) method. The influence of copper(H), zinc(Il), manganese(II) ions on enzymatic degradation of HA by hyaluronidase enzyme (HA-se) were investigated. Secondly, the kinetic parameters, V(max), K(m), k(cat), and k (cat),/K(m) were determined to estimate the impact of these metal ions (Me) on digestion process of hyaluronic acid (HA). The two different HA-Me mole ratios were analyzed. The examined data were always compared to the digestion process of pure HA solution by hyaluronidase, to exhibit the differences in the digestion process of pure hyaluronan as well as the hyaluronan in the presence of metal ions. It was observed that all of the investigated metal ions have influenced the hyaluronic acid degradation process. The most important conclusion was a decrease of the kinetic parameters both K,, and V,. In the result, it can be assumed that in all of the studied samples with metal ions addition, the uncompetitive mechanism of enzyme inhibition occurred. The results of this study may give new insight into foregoing knowledge about hyaluronic acid behavior. Due to the fact that our study was carried out only for three different metal ions in two concentrations, it is necessary to continue further research comprising wider range of metal ions and their concentrations.

Prediction of in vivo neutral detergent fiber digestibility and digestion rate of potentially digestible neutral detergent fiber: comparison of models.

Science.gov (United States)

Huhtanen, P; Seppälä, A; Ahvenjärvi, S; Rinne, M

2008-10-01

Eleven 1-pool, seven 2-pool, and three 3-pool models were compared in fitting gas production data and predicting in vivo NDF digestibility and effective first-order digestion rate of potentially digestible NDF (pdNDF). Isolated NDF from 15 grass silages harvested at different stages of maturity was incubated in triplicate in rumen fluid-buffer solution for 72 h to estimate the digestion kinetics from cumulative gas production profiles. In vivo digestibility was estimated by the total fecal collection method in sheep fed at a maintenance level of feeding. The concentration of pdNDF was estimated by a 12-d in situ incubation. The parameter values from gas production profiles and pdNDF were used in a 2-compartment rumen model to predict pdNDF digestibility using 50 h of rumen residence time distributed in a ratio of 0.4:0.6 between the non-escapable and escapable pools. The effective first-order digestion rate was computed both from observed in vivo and model-predicted pdNDF digestibility assuming the passage kinetic model described above. There were marked differences between the models in fitting the gas production data. The fit improved with increasing number of pools, suggesting that silage pdNDF is not a homogenous substrate. Generally, the models predicted in vivo NDF digestibility and digestion rate accurately. However, a good fit of gas production data was not necessarily translated into improved predictions of the in vivo data. The models overestimating the asymptotic gas volumes tended to underestimate the in vivo digestibility. Investigating the time-related residuals during the later phases of fermentation is important when the data are used to estimate the first-order digestion rate of pdNDF. Relatively simple models such as the France model or even a single exponential model with discrete lag period satisfied the minimum criteria for a good model. Further, the comparison of feedstuffs on the basis of parameter values is more unequivocal than in the case

Process technology for multi-enzymatic reaction systems

DEFF Research Database (Denmark)

Xue, Rui; Woodley, John M.

2012-01-01

In recent years, biocatalysis has started to provide an important green tool in synthetic organic chemistry. Currently, the idea of using multi-enzymatic systems for industrial production of chemical compounds becomes increasingly attractive. Recent examples demonstrate the potential of enzymatic...... synthesis and fermentation as an alternative to chemical-catalysis for the production of pharmaceuticals and fine chemicals. In particular, the use of multiple enzymes is of special interest. However, many challenges remain in the scale-up of a multi-enzymatic system. This review summarizes and discusses...... the technology options and strategies that are available for the development of multi-enzymatic processes. Some engineering tools, including kinetic models and operating windows, for developing and evaluating such processes are also introduced....

Digested disorder

Science.gov (United States)

DeForte, Shelly; Reddy, Krishna D; Uversky, Vladimir N

2013-01-01

The current literature on intrinsically disordered proteins is overwhelming. To keep interested readers up to speed with this literature, we continue a “Digested Disorder” project and represent a series of reader’s digest type articles objectively representing the research papers and reviews on intrinsically disordered proteins. The only 2 criteria for inclusion in this digest are the publication date (a paper should be published within the covered time frame) and topic (a paper should be dedicated to any aspect of protein intrinsic disorder). The current digest issue covers papers published during the period of April, May, and June of 2013. The papers are grouped hierarchically by topics they cover, and for each of the included paper a short description is given on its major findings. PMID:28516028

The adsorption of α-amylase on barley proteins affects the in vitro digestion of starch in barley flour.

Science.gov (United States)

Yu, Wenwen; Zou, Wei; Dhital, Sushil; Wu, Peng; Gidley, Michael J; Fox, Glen P; Gilbert, Robert G

2018-02-15

The conversion of barley starch to sugars is a complex enzymic process. Most previous work concerned the biotechnical aspect of in situ barley enzymes. However, the interactions among the macromolecular substrates and their effects on enzymic catalysis has been little examined. Here, we explore the mechanisms whereby interactions of protein and starch in barley flour affect the kinetics of enzymatic hydrolysis of starch in an in vitro system, using digestion rate data and structural analysis by confocal microscopy. The degradation kinetics of both uncooked barley flour and of purified starches are found to be two-step sequential processes. Barley proteins, especially the water-soluble component, are found to retard the digestion of starch degraded by α-amylase: the enzyme binds with water-insoluble protein and with starch granules, leading to reduced starch hydrolysis. These findings are of potential industrial value in both the brewing and food industries. Copyright © 2017 Elsevier Ltd. All rights reserved.

125I-labeled cortisol radioimmunoassay in which serum binding protein are enzymatically denatured

International Nuclear Information System (INIS)

Hasler, M.J.; Painter, K.; Niswender, G.D.

1976-01-01

We report an iodine-125 radioimmunoassay for cortisol in biological fluids, in which interfering binding proteins are enzymatically denatured. An antiserum to cortisol-3-carboxymethyloxime-bovine serum albumin, extremely low cross-reacting with other corticosteroids, was raised in rabbits. A cortisol-3-carboxymethyloxime tyrosine methyl ester derivative was synthesized and labeled with iodine-125 by standard radioiodination techniques. To eliminate the need for extraction and recovery procedures, we digested interfering binding with a proteolytic enzyme, which then was heat-inactivated before adding the labeled derivative and the premixed, preincubated antiserum complex. There was quantitative analytical recovery of esogenous cortisol added to sera from a normal man, a normal woman, and a pregnant woman. Values for the same samples agreed after extraction and chromatographic purification and agreed well with values obtained by other techniques by independent reference laboratories. The five-step assay can be done in 6 h or less

Anaerobic digestion and co-digestion of slaughterhouse wastes

Directory of Open Access Journals (Sweden)

Sonia Castellucci

2013-09-01

Full Text Available The use of renewable energy is becoming increasingly necessary in order to address the global warming problem and, as a consequence, has become an high priority for many countries. Biomass is a clean and renewable energy source with growing potential to replace conventional fossil fuels. Among biomass, residual and waste ones represent a great resource for energy generation since they permit both to eliminate a possible waste and to produce energy. In the present work, the case of slaughterhouse wastes (SHWs has been investigated. Anaerobic digestion is nowadays considered as one of the most important and sustainable conversion technology exploiting organic matter and biodegradable wastes. Biogas results from this bio-chemical process and mainly consists of methane and carbon dioxide, leading to produce thermal energy and/or electricity. In this paper, the European Regulations on animal by-products (ABPs are described, and some previous study on anaerobic digestion and co-digestion of ABPs - more precisely SHWs - are considered and compared in order to fix a starting point for future tests on their co-digestion in a micro-scale pilot digester. This is to define optimal feed ratio values which ensure an increasing content of methane in the outgoing biogas.

Characterization of the Micromorphology and Topochemistry of Poplar Wood during Mild Ionic Liquid Pretreatment for Improving Enzymatic Saccharification

Directory of Open Access Journals (Sweden)

Sheng Chen

2017-01-01

Full Text Available Ionic liquids (ILs as designer solvents have been applied in biomass pretreatment to increase cellulose accessibility and therefore improve the enzymatic hydrolysis. We investigated the characterization of the micromorphology and the topochemistry of poplar wood during 1-ethyl-3-methylimidazolium acetate pretreatment with mild conditions (90 °C for 20 and 40 min by multiple microscopic techniques (FE-SEM, CLSM, and CRM. Chemical composition analysis, XRD, cellulase adsorption isotherm, and enzymatic hydrolysis were also performed to monitor the variation of substrate properties. Our results indicated that the biomass conversion was greatly enhanced (from 20.57% to 73.64% due to the cell wall deconstruction and lignin dissolution (29.83% lignin was removed after incubation for 40 min, rather than the decrystallization or crystallinity transformation of substrates. The mild ILs pretreatment, with less energy input, can not only enhance enzymatic hydrolysis, but also provide a potential approach as the first step in improving the sequential pretreatment effectiveness in integrated methods. This study provides new insights on understanding the ILs pretreatment with low temperature and short duration, which is critical for developing individual and/or combined pretreatment technologies with reduced energy consumption.

Noncanonical substrate preference of lambda exonuclease for 5'-nonphosphate-ended dsDNA and a mismatch-induced acceleration effect on the enzymatic reaction.

Science.gov (United States)

Wu, Tongbo; Yang, Yufei; Chen, Wei; Wang, Jiayu; Yang, Ziyu; Wang, Shenlin; Xiao, Xianjin; Li, Mengyuan; Zhao, Meiping

2018-04-06

Lambda exonuclease (λ exo) plays an important role in the resection of DNA ends for DNA repair. Currently, it is also a widely used enzymatic tool in genetic engineering, DNA-binding protein mapping, nanopore sequencing and biosensing. Herein, we disclose two noncanonical properties of this enzyme and suggest a previously undescribed hydrophobic interaction model between λ exo and DNA substrates. We demonstrate that the length of the free portion of the substrate strand in the dsDNA plays an essential role in the initiation of digestion reactions by λ exo. A dsDNA with a 5' non-phosphorylated, two-nucleotide-protruding end can be digested by λ exo with very high efficiency. Moreover, we show that when a conjugated structure is covalently attached to an internal base of the dsDNA, the presence of a single mismatched base pair at the 5' side of the modified base may significantly accelerate the process of digestion by λ exo. A detailed comparison study revealed additional π-π stacking interactions between the attached label and the amino acid residues of the enzyme. These new findings not only broaden our knowledge of the enzyme but will also be very useful for research on DNA repair and in vitro processing of nucleic acids.

Increasing the biogas yield of manure by wet explosion of the digested fiber fraction

DEFF Research Database (Denmark)

Biswas, Rajib; Uellendahl, Hinrich; Ahring, Birgitte Kiær

digested manure fibers from the effluent of an anaerobic digester for enhancing biogas production and exploring the untapped biomass potential. The increase in methane yield of the digested manure fibers was investigated by applying the WEx treatment under 5 different process conditions. The pretreatment......Increasing the biodegradability of the lignocellulosic fiber fraction of manure can ensure higher methane productivity in biogas plants, leading to process profitability and thus larger production of renewable energy. A new pretreatment method, wet explosion (WEx), was investigated to treat...... condition of 180 ºC and a retention time of 10 minutes without addition of chemicals was found to be optimal, resulting in 136% increase in methane yield as compared to the untreated digested manure fibers....

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

International Nuclear Information System (INIS)

Bayr, S.; Ojanperä, M.; Kaparaju, P.; Rintala, J.

2014-01-01

Highlights: • Rendering wastes’ mono-digestion and co-digestion with potato pulp were studied. • CSTR process with OLR of 1.5 kg VS/m 3 d, HRT of 50 d was unstable in mono-digestion. • Free NH 3 inhibited mono-digestion of rendering wastes. • CSTR process with OLR of 1.5 kg VS/m 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 4 -N and/or free NH 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 3 d and hydraulic retention time (HRT) of 50 d was unstable and resulted in methane yields of 450 dm 3 /kg VS 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 3 /kg VS 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

Influence of Bifidobacterium bifidum on release of minerals from bread with differing bran content.

Science.gov (United States)

Nalepa, Beata; Siemianowska, Ewa; Skibniewska, Krystyna Anna

2012-01-01

Bread is considered an important source of minerals; however, the presence of fiber and phytic acid reduces bioavailability of minerals from cereal products. It is well established that activity of microorganisms in human gut increases the amount of nutrients released during digestion. The aim of this study was to determine the influence of Bifidobacterium bifidum on release of some minerals from bread using an in vitro process of enzymatic digestion. White bread and with addition of 15, 30, or 45% of bran was baked in a bakery by traditional methods, with addition of yeasts and rye leaven, from flour made of wheat, Tonacja variety. Concentrations of calcium, magnesium, manganese, zinc, copper, and iron were determined by atomic absorption spectrometry. Bread was enzymatically digested in vitro without and with the addition of Bifidobacterium bifidum KD6 (inoculum 10(6) CFU/cm(3)) and percentages of minerals released were determined. The concentration of minerals released during enzymatic digestion varied depending upon the element, quantity of bran, and presence of bacteria. Increase in bran content decreased release of elements. Bifidobacterium bifidum KD6 enhanced amounts of magnesium and zinc released from all types of bread, while manganese and copper rose only from white bread with 15% bran addition. Bacteria decreased amounts of calcium and iron released from bread. Data indicate that diets rich in beneficial bacteria (probiotics) but not balanced with minerals might increase mineral deficiency.

Next-generation non-starch polysaccharide-degrading, multi-carbohydrase complex rich in xylanase and arabinofuranosidase to enhance broiler feed digestibility.

Science.gov (United States)

Cozannet, Pierre; Kidd, Michael T; Montanhini Neto, Roberto; Geraert, Pierre-André

2017-08-01

This study was carried out to evaluate the effect of a multi-carbohydrase complex (MCC) rich in xylanase (Xyl) and arabinofuranosidase (Abf) on overall broiler feed digestibility in broilers. Energy utilization and digestibility of dry matter (DM), organic matter (OM), protein, starch, fat, and insoluble and soluble fibers were measured using the mass-balance method. The experiment was carried out on 120 broilers (3-week-old chickens). Broilers were distributed over 8 treatments to evaluate the effect of the dietary arabinoxylan content and nutrient density with and without MCC (Rovabio® Advance). The graded content of arabinoxylan (AX) was obtained using different raw materials (wheat, rye, barley, and dried distillers' wheat). Diet-energy density was modified with added fat. Measurements indicated that nutrient density and AX content had a significant effect on most digestibility parameters. Apparent metabolizable energy (AME) was significantly increased (265 kcal kg-1) by MCC. The addition of MCC also resulted in significant improvement in the digestibility of all evaluated nutrients, with average improvements of 3.0, 3.3, 3.2, 3.0, 6.2, 2.9, 5.8, and 3.8% units for DM, OM, protein, starch, fat, insoluble and soluble fibers, and energy utilization, respectively. The interaction between MCC and diet composition was significant for the digestibility of OM, fat, protein, and energy. Nutrient digestibility and diet AME were negatively correlated with AX content (P digestible nutrient (i.e., starch, protein, fat, insoluble and soluble fibers) content with and without MCC (R2 = 0.87; RSD = 78 kcal kg-1). This study confirms that the presence of AX in wheat-based diets and wheat-based diets with other cereals and cereal by-products reduces nutrient digestibility in broiler chickens. Furthermore, the dietary addition of MCC, which is rich in Xyn and Abf, reduced deleterious effect of fiber and improved overall nutrient digestibility in broiler diets. © 2017 Poultry

Anaerobic Digestion and its Applications

Science.gov (United States)

Anaerobic digestion is a natural biological process. The initials "AD" may refer to the process of anaerobic digestion, or the built systems of anaerobic digesters. While there are many kinds of digesters, the biology is basically the same for all. Anaerobic digesters are built...

Enhanced sensitivity in non-enzymatic glucose detection by improved growth kinetics of Ni-based nanostructures

Science.gov (United States)

Urso, M.; Pellegrino, G.; Strano, V.; Bruno, E.; Priolo, F.; Mirabella, S.

2018-04-01

Ni-based nanostructures are attractive catalytic materials for many electrochemical applications, among which are non-enzymatic sensing, charge storage, and water splitting. In this work, we clarify the synthesis kinetics of Ni(OH)2/NiOOH nanowalls grown by chemical bath deposition at room temperature and at 50 °C. We applied the results to non-enzymatic glucose sensing, reaching a highest sensitivity of 31 mA cm-2mM-1. Using scanning electron microscopy, x-ray diffraction analysis and Rutherford backscattering spectrometry we found that the growth occurs through two regimes: first, a quick random growth leading to disordered sheets of Ni oxy-hydroxide, followed by a slower growth of well-aligned sheets of Ni hydroxide. A high growth temperature (50 °C), leading mainly to well-aligned sheets, offers superior electrochemical properties in terms of charge storage, charge carrier transport and catalytic action, as confirmed by cyclic voltammetry and electrochemical impedance spectroscopy analyses. The reported results on the optimization and application of low-cost synthesis of these Ni-based nanostructures have a large potential for application in catalysis, (bio)sensing, and supercapacitors areas.

Digested disorder

Science.gov (United States)

Reddy, Krishna D; DeForte, Shelly; Uversky, Vladimir N

2014-01-01

The current literature on intrinsically disordered proteins grows fast. To keep interested readers up to speed with this literature, we continue a “Digested Disorder” project and represent a new issue of reader’s digest of the research papers and reviews on intrinsically disordered proteins. The only 2 criteria for inclusion in this digest are the publication date (a paper should be published within the covered time frame) and topic (a paper should be dedicated to any aspect of protein intrinsic disorder). The current digest issue covers papers published during the third quarter of 2013; i.e., during the period of June, July, and September of 2013. Similar to previous issues, the papers are grouped hierarchically by topics they cover, and for each of the included paper a short description is given on its major findings. PMID:28232877

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...

Increased release of fermentable sugars from elephant grass by enzymatic hydrolysis in the presence of surfactants

International Nuclear Information System (INIS)

Menegol, Daiane; Scholl, Angélica Luisi; Fontana, Roselei Claudete; Dillon, Aldo José Pinheiro; Camassola, Marli

2014-01-01

Highlights: • Milling is an attractive method to enhance the enzymatic hydrolysis of biomass. • Surfactants improve the efficiency of lignocellulose enzymatic hydrolysis. • Pretreatment with NaOH, smaller particle size and Tween 80® were more efficient. - Abstract: In the search for renewable energy sources, elephant grass is an alternative substrate for ethanol production, but this substrate must be hydrolyzed by cellulases and xylanases to liberate fermentable sugars. During enzymatic hydrolysis, cellulase activity is reduced by the irreversible adsorption of cellulase onto cellulose, decreasing the rate of hydrolysis. Adding surfactants during hydrolysis can improve the process. The effects of Tween® and Triton® surfactants on the enzymatic hydrolysis of elephant grass were evaluated in this context. The data indicate that pretreatment with sodium hydroxide, along with a smaller particle size (0.075–0.152 mm) and the use of Tween 80®, increased the efficiency of releasing reducing sugars from pretreated elephant grass biomass. Thus, it is possible to reduce grinding costs in second-generation ethanol production through the use of surfactants, as they allow efficient hydrolysis of larger biomass particles

Peptide profiling and the bioactivity character of yogurt in the simulated gastrointestinal digestion.

Science.gov (United States)

Jin, Yan; Yu, Yang; Qi, Yanxia; Wang, Fangjun; Yan, Jiaze; Zou, Hanfa

2016-06-01

This study investigated the relationship between peptide profiles and the bioactivity character of yogurt in simulated gastrointestinal trials. A total of 250, 434 and 466 peptides were identified by LC-MS/MS analyses of yogurt, gastric digest and pancreatic digest. Forty peptides of yogurt survived in gastrointestinal digestion. κ-CN and β-CN contributed the diversity of peptides during the fermentation process and gastrointestinal digestion, respectively. The favorite of κ-CN by lactic acid bacteria complemented gut digestion by hydrolyzing κ-CN, the low abundance milk proteins. The potential bioactivities were evaluated by in vitro ACE and DPP-IV inhibition assays. The ACE inhibition rate of the pancreatic digests was ~4 - and ~2 - fold greater than that of yogurt and the gastric digests. The ACE inhibitory peptides generated during gastrointestinal digestion improved the ACE inhibitory activity of the gastric and pancreatic digests. The DPP-IV inhibition rate of the pancreatic digest was ~6 - and ~3 - fold greater than that of yogurt and the gastric digest. The numbers of potential DPP-IV inhibitory peptides were positively correlated to the DPP-IV inhibitory activity of the gastric and pancreatic digests. The present study describes the characters and bioactivities of peptides from yogurt in a simulated gastrointestinal digestion. The number of peptides identified from yogurt and gastrointestinal digests by LC-MS/MS increased in the simulated gastrointestinal trials. The in vitro ACE and DPP-IV inhibition bioactivities revealed that the bioactivity of yogurt was enhanced during gastrointestinal digestion. The correlation between peptides and bioactivity in vitro indicated that not only the peptides amount but also the proportion of peptides with high bioactivities contributed to increased bioactivity during gastrointestinal digestion. The study of peptides identified from yogurt and digests revealed that the number of released peptides was not determined

Comparison of artificial digestion and Baermann's methods for detection of Trichinella spiralis pre-encapsulated larvae in muscles with low-level infections.

Science.gov (United States)

Jiang, Peng; Wang, Zhong-Quan; Cui, Jing; Zhang, Xi

2012-01-01

Artificial digestion method is widely used for the detection of Trichinella larvae (mainly the mature larvae, e.g., encapsulated larvae in encapsulated Trichinella) in meat. The previous studies demonstrated that Trichinella spiralis pre-encapsulated larvae (PEL) at 14-18 days postinfection (dpi) had the infectivity to new hosts. However, to our knowledge, there is no report on the detection methods of PEL in meat. The purpose of this study was to compare the efficiency of artificial digestion and Baermann's methods for detection of T. spiralis PEL in meat, and to test the factors affecting the sensitivity of the two methods. Forty-five male Kunming mice were randomly divided into 3 groups (15 mice per group), and each group of mice was orally inoculated with 20, 10, or 5 muscle larvae of T. spiralis, respectively. All infected mice were slaughtered at 18 dpi, and the muscles were minced. The digestion method that was recommended by International Commission on Trichinellosis and Baermann's method were used to detect the PEL in the infected mice. The detection rate of PEL in both mice infected with 20 muscle larvae by digestion and Baermann's method was 100% (15/15); the detection rates of PEL in mice infected with 10 larvae by the two methods just mentioned were 93.33% (14/15) and 100% (15/15), respectively; when the mice infected with 5 larvae were tested, the different detection rate of PEL was achieved by using digestion method (63.33%) and Baermann's method (100%). Additionally, the number of PEL collected from the mice infected with 20, 10, or 5 larvae by Baermann's method was greater than that by digestion methods. The mortality of PEL increased along with the prolongation of digestion duration, because the PEL were not resistant to enzymatic digestion. The results revealed that the Baermann's method is superior to the digestion methods for detection of T. spiralis PEL in muscle samples with low-level infections.

Lessons from Digestive-Tract Symbioses Between Bacteria and Invertebrates.

Science.gov (United States)

Graf, Joerg

2016-09-08

In most animals, digestive tracts harbor the greatest number of bacteria in the animal that contribute to its health: by aiding in the digestion of nutrients, provisioning essential nutrients and protecting against colonization by pathogens. Invertebrates have been used to enhance our understanding of metabolic processes and microbe-host interactions owing to experimental advantages. This review describes how advances in DNA sequencing technologies have dramatically altered how researchers investigate microbe-host interactions, including 16S rRNA gene surveys, metagenome experiments, and metatranscriptome studies. Advantages and challenges of each of these approaches are described herein. Hypotheses generated through omics studies can be directly tested using site-directed mutagenesis, and findings from transposon studies and site-directed experiments are presented. Finally, unique structural aspects of invertebrate digestive tracts that contribute to symbiont specificity are presented. The combination of omics approaches with genetics and microscopy allows researchers to move beyond correlations to identify conserved mechanisms of microbe-host interactions.

Integrated Bioethanol Fermentation/Anaerobic Digestion for Valorization of Sugar Beet Pulp

Directory of Open Access Journals (Sweden)

Joanna Berlowska

2017-08-01

Full Text Available Large amounts of waste biomass are generated in sugar factories from the processing of sugar beets. After diffusion with hot water to draw the sugar from the beet pieces, a wet material remains called pulp. In this study, waste sugar beet pulp biomass was enzymatically depolymerized, and the obtained hydrolyzates were subjected to fermentation processes. Bioethanol, biomethane, and biohydrogen were produced directly from the substrate or in combined mode. Stillage, a distillery by-product, was used as a feedstock for anaerobic digestion. During biosynthesis of ethanol, most of the carbohydrates released from the sugar beet pulp were utilized by a co-culture of Saccharomyces cerevisiae Ethanol Red, and Scheffersomyces stipitis LOCK0047 giving 12.6 g/L of ethanol. Stillage containing unfermented sugars (mainly arabinose, galactose and raffinose was found to be a good substrate for methane production (444 dm3 CH4/kg volatile solids (VS. Better results were achieved with this medium than with enzymatic saccharified biomass. Thermal pre-treatment and adjusting the pH of the inoculum resulted in higher hydrogen production. The largest (p < 0.05 hydrogen yield (252 dm3 H2/kg VS was achieved with sugar beet stillage (SBS. In contrast, without pre-treatment the same medium yielded 35 dm3 H2/kg VS. However, dark fermentation of biohydrogen was more efficient when sugar beet pulp hydrolyzate was used.

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.

D-Amino acid oxidase bio-functionalized platforms: Toward an enhanced enzymatic bio-activity

Science.gov (United States)

Herrera, Elisa; Valdez Taubas, Javier; Giacomelli, Carla E.

2015-11-01

The purpose of this work is to study the adsorption process and surface bio-activity of His-tagged D-amino acid oxidase (DAAO) from Rhodotorula gracilis (His6-RgDAAO) as the first step for the development of an electrochemical bio-functionalized platform. With such a purpose this work comprises: (a) the His6-RgDAAO bio-activity in solution determined by amperometry, (b) the adsorption mechanism of His6-RgDAAO on bare gold and carboxylated modified substrates in the absence (substrate/COO-) and presence of Ni(II) (substrate/COO- + Ni(II)) determined by reflectometry, and (c) the bio-activity of the His6-RgDAAO bio-functionalized platforms determined by amperometry. Comparing the adsorption behavior and bio-activity of His6-RgDAAO on these different solid substrates allows understanding the contribution of the diverse interactions responsible for the platform performance. His6-RgDAAO enzymatic performance in solution is highly improved when compared to the previously used pig kidney (pk) DAAO. His6-RgDAAO exhibits an amperometrically detectable bio-activity at concentrations as low as those expected on a bio-functional platform; hence, it is a viable bio-recognition element of D-amino acids to be coupled to electrochemical platforms. Moreover, His6-RgDAAO bio-functionalized platforms exhibit a higher surface activity than pkDAAO physically adsorbed on gold. The platform built on Ni(II) modified substrates present enhanced bio-activity because the surface complexes histidine-Ni(II) provide with site-oriented, native-like enzymes. The adsorption mechanism responsible of the excellent performance of the bio-functionalized platform takes place in two steps involving electrostatic and bio-affinity interactions whose prevalence depends on the degree of surface coverage.

Digested disorder, Quarterly intrinsic disorder digest (October-November-December, 2013).

Science.gov (United States)

DeForte, Shelly; Reddy, Krishna D; Uversky, Vladimir N

2015-01-01

This is the 4th issue of the Digested Disorder series that represents reader's digest of the scientific literature on intrinsically disordered proteins. The only 2 criteria for inclusion in this digest are the publication date (a paper should be published within the covered time frame) and topic (a paper should be dedicated to any aspect of protein intrinsic disorder). The current digest issue covers papers published during the fourth quarter of 2013; i.e. during the period of October, November, and December of 2013. Similar to previous issues, the papers are grouped hierarchically by topics they cover, and for each of the included paper a short description is given on its major findings.

assesment of mesophilic co assesment of mesophilic co-digestion

African Journals Online (AJOL)

eobe

(fossil) and its attendant pollution menace have provided the avenue to consider alternative ... that anaerobic co-digestion of cow dung with lemon grass produced a high quality biogas. .... for wealth creation as they are harnessed as feed ... enhanced methane yield production [15, 16, 17, 18,. 19]. .... The values given by the.

Cerebral ischemia is exacerbated by extracellular nicotinamide phosphoribosyltransferase via a non-enzymatic mechanism.

Directory of Open Access Journals (Sweden)

Bing Zhao

Full Text Available Intracellular nicotinamide phosphoribosyltransferase (iNAMPT in neuron has been known as a protective factor against cerebral ischemia through its enzymatic activity, but the role of central extracellular NAMPT (eNAMPT is not clear. Here we show that eNAMPT protein level was elevated in the ischemic rat brain after middle-cerebral-artery occlusion (MCAO and reperfusion, which can be traced to at least in part from blood circulation. Administration of recombinant NAMPT protein exacerbated MCAO-induced neuronal injury in rat brain, while exacerbated oxygen-glucose-deprivation (OGD induced neuronal injury only in neuron-glial mixed culture, but not in neuron culture. In the mixed culture, NAMPT protein promoted TNF-α release in a time- and concentration-dependent fashion, while TNF-α neutralizing antibody protected OGD-induced, NAMPT-enhanced neuronal injury. Importantly, H247A mutant of NAMPT with essentially no enzymatic activity exerted similar effects on ischemic neuronal injury and TNF-α release as the wild type protein. Thus, eNAMPT is an injurious and inflammatory factor in cerebral ischemia and aggravates ischemic neuronal injury by triggering TNF-α release from glia cells, via a mechanism not related to NAMPT enzymatic activity.

Noncanonical substrate preference of lambda exonuclease for 5′-nonphosphate-ended dsDNA and a mismatch-induced acceleration effect on the enzymatic reaction

Science.gov (United States)

Yang, Yufei; Chen, Wei; Wang, Jiayu; Yang, Ziyu; Wang, Shenlin; Xiao, Xianjin; Li, Mengyuan

2018-01-01

Abstract Lambda exonuclease (λ exo) plays an important role in the resection of DNA ends for DNA repair. Currently, it is also a widely used enzymatic tool in genetic engineering, DNA-binding protein mapping, nanopore sequencing and biosensing. Herein, we disclose two noncanonical properties of this enzyme and suggest a previously undescribed hydrophobic interaction model between λ exo and DNA substrates. We demonstrate that the length of the free portion of the substrate strand in the dsDNA plays an essential role in the initiation of digestion reactions by λ exo. A dsDNA with a 5′ non-phosphorylated, two-nucleotide-protruding end can be digested by λ exo with very high efficiency. Moreover, we show that when a conjugated structure is covalently attached to an internal base of the dsDNA, the presence of a single mismatched base pair at the 5′ side of the modified base may significantly accelerate the process of digestion by λ exo. A detailed comparison study revealed additional π–π stacking interactions between the attached label and the amino acid residues of the enzyme. These new findings not only broaden our knowledge of the enzyme but will also be very useful for research on DNA repair and in vitro processing of nucleic acids. PMID:29490081

Lactose hydrolysis in an enzymatic membrane reactor

Energy Technology Data Exchange (ETDEWEB)

Mertens, B; Huyghebaert, A

1987-10-01

The enzymatic hydrolysis of lactose in whey permeate with subsequent recuperation of Saccharomyces lactis lactase by means of ultrafiltration was investigated. In whey permeate, S. lactis lactase shows maximal activity at pH 6.5; the optimal temperature was found to be 45/sup 0/C and is limited by strong thermal inactivation beyond this temperature. High activity combined with acceptable thermal inactivation (< 10% after 5 h incubation) was established at 30/sup 0/C. S. lactis lactase also displays considerable activity at low temperature (5/sup 0/C). Enzyme stability is reduced drastically by demineralisation: addition of low concentrations of manganese ions (10/sup -3/ M) considerably enhances stability. Using a DDS Lab-Unit 35 fitted with GR61PP polysulphon membranes (cut-off: 20.000), pilot scale experiments were carried out (pH 6.5; 30/sup 0/C) in which whey permeate was hydrolyzed to a degree of hydrolysis of 82% minimum. Enzyme recuperation amounted to 96.5% per batch, all enzyme activity loss being due to thermal inactivation. Microbiological examination of the enzymatic membrane reactor showed that growth of mcicroorganisms can largely be suppressed by working at lower temperature (5/sup 0/C). Eventually, 50 ppm H/sub 2/O/sub 2/ or sterile filtration will adequately solve microbiological problems without affecting enzyme activity.

Digested disorder: Quarterly intrinsic disorder digest (April-May-June, 2013).

Science.gov (United States)

DeForte, Shelly; Reddy, Krishna D; Uversky, Vladimir N

2013-01-01

The current literature on intrinsically disordered proteins is overwhelming. To keep interested readers up to speed with this literature, we continue a "Digested Disorder" project and represent a series of reader's digest type articles objectively representing the research papers and reviews on intrinsically disordered proteins. The only 2 criteria for inclusion in this digest are the publication date (a paper should be published within the covered time frame) and topic (a paper should be dedicated to any aspect of protein intrinsic disorder). The current digest issue covers papers published during the period of April, May, and June of 2013. The papers are grouped hierarchically by topics they cover, and for each of the included paper a short description is given on its major findings.

Enzymatic hydrolysis of pretreated soybean straw

International Nuclear Information System (INIS)

Xu Zhong; Wang Qunhui; Jiang Zhaohua; Yang Xuexin; Ji Yongzhen

2007-01-01

In order to produce lactic acid, from agricultural residues such as soybean straw, which is a raw material for biodegradable plastic production, it is necessary to decompose the soybean straw into soluble sugars. Enzymatic hydrolysis is one of the methods in common use, while pretreatment is the effective way to increase the hydrolysis rate. The optimal conditions of pretreatment using ammonia and enzymatic hydrolysis of soybean straw were determined. Compared with the untreated straw, cellulose in straw pretreated by ammonia liquor (10%) soaking for 24 h at room temperature increased 70.27%, whereas hemicellulose and lignin in pretreated straw decreased to 41.45% and 30.16%, respectively. The results of infrared spectra (IR), scanning electron microscope (SEM) and X-ray diffraction (XRD) analysis also showed that the structure and the surface of the straw were changed through pretreatment that is in favor of the following enzymatic hydrolysis. maximum enzymatic hydrolysis rate of 51.22% was achieved at a substrate concentration of 5% (w/v) at 50 deg. C and pH 4.8 using cellulase (50 fpu/g of substrate) for 36 h

Implications of the Maillard reaction on bovine alpha-lactalbumin and its proteolysis during in vitro infant digestion.

Science.gov (United States)

Joubran, Yousef; Moscovici, Alice; Portmann, Reto; Lesmes, Uri

2017-06-21

This study investigated the functionality and digestibility of Maillard reaction products (MRPs) of alpha-lactalbumin (α-la), a major whey protein and component of infant formulas. The impact of different carbohydrates (glucose, galactose or galacto-oligosaccharides (GOS)) and heating duration was studied. SDS-PAGE, UV and color measurements monitored reaction extent, which varied between carbohydrates whereby galactose reacted more readily than glucose. Surface hydrophobicity and antioxidant capacity were found to be significantly (p Maillard conjugation, with GOS-based MRPs elevating antioxidant capacity ∼50-fold compared to α-la. In addition, the digestive proteolysis of MRPs was evaluated using an infant in vitro gastro-duodenal model. SDS-PAGE analyses of digesta revealed Maillard conjugation generally increased α-la's susceptibility to proteolysis. Interestingly, GOS-based MRPs presented an optimization challenge, since heating for 12 h delayed proteolysis, while extended heating resulted in the highest susceptibility to proteolysis. Proteomic analyses further demonstrated the differences in enzymatic cleavage patterns and helped identify bioactive peptides rendered bioaccessible during the digestion of α-la or its MRPs. Bioinformatic mining of the proteomic data using PeptideRanker also gave rise to two potentially novel bioactive peptides, FQINNKIW and GINYWLAHKALCS. Finally, antioxidant capacity of luminal contents, measured by DPPH, revealed Maillard conjugation increased the antioxidant capacity of both gastric and duodenal digesta. Overall, this work draws a link between the Maillard reaction, digestive proteolysis and the bioaccessibility of bioactive peptides and antioxidant species in the infant alimentary canal. This could help rationally process infant formulas towards improved nutritional and extra-nutritional benefits.

Role of enzymatic and non enzymatic antioxidant in ameliorating salinity induced damage in nostoc muscorum

International Nuclear Information System (INIS)

Hend, A.; Abeer, A.; Allah, A.

2015-01-01

Presence of high salt concentration in the growth medium adversely affected the plant growth and productivity by altering its metabolic activities. Experiments were conducted on cyanobacteriaum Nostoc muscorum grown in nitrogen free medium supplemented with 250 mM NaCl to evaluate the salt stress induced changes in growth, antioxidants and lipid composition. Salt stress significantly reduced the growth and physio-biochemical attributes. Salt stress increased malonaldehyde content thereby causing alterations in the lipid fraction. Significant reduction in polyunsaturated fatty acids including phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI) and phosphatidylserine (PS) was observed. Where as diacylglycerol, sterol ester and non-esterified fatty acids were increased. Activities of antioxidant enzymes and contents of non-enzymatic antioxidants including glutathione enhanced due to salt stress. An increase in accumulation of proline was also observed. Hence increased activity of antioxidants and altered fatty acid composition was observed in salt stressed Nostoc muscorum. (author)

Enzymatic approaches to rare sugar production.

Science.gov (United States)

Zhang, Wenli; Zhang, Tao; Jiang, Bo; Mu, Wanmeng

Rare sugars have recently attracted much attention because of their potential applications in the food, nutraceutical, and pharmaceutical industries. A systematic strategy for enzymatic production of rare sugars, named Izumoring, was developed >10years ago. The strategy consists of aldose-ketose isomerization, ketose C-3 epimerization, and monosaccharide oxidation-reduction. Recent development of the Izumoring strategy is reviewed herein, especially the genetic approaches to the improvement of rare sugar-producing enzymes and the applications of target-oriented bioconversion. In addition, novel non-Izumoring enzymatic approaches are also summarized, including enzymatic condensation, phosphorylation-dephosphorylation cascade reaction, aldose epimerization, ulosonic acid decarboxylation, and biosynthesis of rare disaccharides. Copyright © 2017 Elsevier Inc. All rights reserved.

Analysis of pyrimidine dimer content of isolated DNA by nuclease digestion

International Nuclear Information System (INIS)

Farland, W.H.; Sutherland, B.M.

1980-01-01

Isolated DNA is highly susceptible to degradation by exogenous nucleases. Complete digestion is possible with a number of well-characterized enzymes from a variety of sources. Treatment of DNA with a battery of enzymes including both phosphodiesterase and phosphatase activities yields a mixture of nucleosides and inorganic phosphate (P/sub i/) as a final product. Unlike native DNA, ultraviolet-irradiated DNA is resistant to complete digestion. Setlow et al. demonstrated that the structural changes in the DNA responsible for the nuclease resistance were the formation of cyclobutyl pyrimidine dimers, the major photoproduct in UV-irradiated DNA. Using venom phosphodiesterase, they demonstrated that UV irradiation of DNA affected both the rate and extent of enzymatic hydrolysis. In addition, it was demonstrated that the major nuclease-resistant product of this hydrolysis was an oligonucleotide containing dimerized pyrimidines. Treatment of the DNA to split the dimers, either photochemically or photoenzymatically, rendered the polymer more susceptible to hydrolysis by the phosphodiesterase. The specificity of photoreactivating enzyme for pyrimidine dimers lends support to the role of these structures in conferring nuclease resistance to UV-irradiated DNA. The nuclease resistance of DNA containing dimers has been the basis of several assays for the measurement of these photoproducts. Sutherland and Chamberlin reported the development of a rapid and sensitive assay for dimers in 32 P-labeled DNA

Repeated inoculation of cattle rumen with bison rumen contents alters the rumen microbiome and improves nitrogen digestibility in cattle

OpenAIRE

Ribeiro, Gabriel O.; Oss, Daniela B.; He, Zhixiong; Gruninger, Robert J.; Elekwachi, Chijioke; Forster, Robert J.; Yang, WenZhu; Beauchemin, Karen A.; McAllister, Tim A.

2017-01-01

Future growth in demand for meat and milk, and the socioeconomic and environmental challenges that farmers face, represent a ?grand challenge for humanity?. Improving the digestibility of crop residues such as straw could enhance the sustainability of ruminant production systems. Here, we investigated if transfer of rumen contents from bison to cattle could alter the rumen microbiome and enhance total tract digestibility of a barley straw-based diet. Beef heifers were adapted to the diet for ...

Bacterial cellulose production from cotton-based waste textiles: enzymatic saccharification enhanced by ionic liquid pretreatment.

Science.gov (United States)

Hong, Feng; Guo, Xiang; Zhang, Shuo; Han, Shi-fen; Yang, Guang; Jönsson, Leif J

2012-01-01

Cotton-based waste textiles were explored as alternative feedstock for production of bacterial cellulose (BC) by Gluconacetobacter xylinus. The cellulosic fabrics were treated with the ionic liquid (IL) 1-allyl-3-methylimidazolium chloride ([AMIM]Cl). [AMIM]Cl caused 25% inactivation of cellulase activity at a concentration as low as of 0.02 g/mL and decreased BC production during fermentation when present in concentrations higher than 0.0005 g/mL. Therefore, removal of residual IL by washing with hot water was highly beneficial to enzymatic saccharification as well as BC production. IL-treated fabrics exhibited a 5-7-fold higher enzymatic hydrolysis rate and gave a seven times larger yield of fermentable sugars than untreated fabrics. BC from cotton cloth hydrolysate was obtained at an yield of 10.8 g/L which was 83% higher than that from the culture grown on glucose-based medium. The BC from G. xylinus grown on IL-treated fabric hydrolysate had a 79% higher tensile strength than BC from glucose-based culture medium which suggests that waste cotton pretreated with [AMIM]Cl has potential to serve as a high-quality carbon source for BC production. Copyright © 2011 Elsevier Ltd. All rights reserved.

Development of optimal enzymatic and microbial conversion systems for biofuel production

Science.gov (United States)

Aramrueang, Natthiporn

materials can range from 470 to 533 L ethanol/metric ton (d.b.) Sugar beet leaves contain nearly equal amounts of cellulose (13%), hemicellulose (16%), and pectin (17%). The potential ethanol yield of sugar beet leaves is 340 L ethanol/metric ton (d.b.). As remaining unused in great quantities during the production of sugar beet as a sugar and energy crop, sugar beet leaves was studied as a potential feedstock for the production of biofuel and valuable products. The enzymatic hydrolysis of sugar beet leaves was optimized for fermentable sugar production. Optimization of enzyme usage was performed to make the biorefinery process more cost- and energy-effective. In this research, response surface methodology was used to study the effects of enzyme loadings during the hydrolysis of sugar beet leaves at 10% total solids content, using a mix of cellulases, hemicellulases, and pectinases. The effects of enzyme loadings were studied with a five-level rotatable central composite design for maximum conversion of sugar beet leaves to fermentable sugars. The last part of this study investigated biogas production through the anaerobic digestion of microalgae as they have received much attention as another potential biofuel feedstock. Anaerobic digestion of Spirulina ( Arthrospira platensis) was conducted in batch reactors for the study of the kinetics and, in continuous stirred tank reactors (CSTR), for the study of the two important operating parameters: hydraulic retention time (HRT) and organic loading rate (OLR). The kinetics study on methane production from batch experiments shows first order kinetics and a reaction rate constant of 0.382 d-1. The maximum biogas and methane yields for Spirulina are 0.514 L/gVS and 0.360 L CH4/gVS, respectively. The methane content of the biogas is 68%. During the continuous anaerobic digestion in CSTR for OLR in the range of 1.0-4.0 gVS/L/d, biogas and methane yields are in the ranges of 0.276-0.502 L/ gVS and 0.163-0.342 L CH4/gVS, respectively

Long-term anaerobic digestion of food waste stabilized by trace elements

International Nuclear Information System (INIS)

Zhang Lei; Jahng, Deokjin

2012-01-01

Highlights: ► Korean food waste was found to contain low level of trace elements. ► Stable anaerobic digestion of food waste was achieved by adding trace elements. ► Iron played an important role in anaerobic digestion of food waste. ► Cobalt addition further enhanced the process performance in the presence of iron. - Abstract: The purpose of this study was to examine if long-term anaerobic digestion of food waste in a semi-continuous single-stage reactor could be stabilized by supplementing trace elements. Contrary to the failure of anaerobic digestion of food waste alone, stable anaerobic digestion of food waste was achieved for 368 days by supplementing trace elements. Under the conditions of OLR (organic loading rates) of 2.19–6.64 g VS (volatile solid)/L day and 20–30 days of HRT (hydraulic retention time), a high methane yield (352–450 mL CH 4 /g VS added ) was obtained, and no significant accumulation of volatile fatty acids was observed. The subsequent investigation on effects of individual trace elements (Co, Fe, Mo and Ni) showed that iron was essential for maintaining stable methane production. These results proved that the food waste used in this study was deficient in trace elements.

Synergistic pretreatment of waste activated sludge using CaO_2 in combination with microwave irradiation to enhance methane production during anaerobic digestion

International Nuclear Information System (INIS)

Wang, Jie; Li, Yongmei

2016-01-01

Highlights: • CaO_2/MW pretreatment synergistically enhanced WAS solubilization and CH_4 production. • MW irradiation facilitated more "·OH generation from CaO_2. • The optimal pretreatment condition for methane production was determined. • The growths of both hydrogenotrophic and acetate-utilizing methanogens were promoted. • The dewaterability of WAS was improved considerably by CaO_2/MW treatment. - Abstract: To investigate the effects of combined calcium peroxide (CaO_2) and microwave pretreatment on anaerobic digestion of waste activated sludge, lab-scale experiments were conducted to measure the solubilization, biodegradation, and dewaterability of the waste activated sludge. Additionally, the synergistic effects between CaO_2 and microwave were studied, and the microbial activity and methanogenic archaea community structure were analyzed. Combined pretreatment considerably facilitated the solubilization and subsequent anaerobic digestion of the waste activated sludge. The optimal pretreatment condition was CaO_2 (0.1 g/gVSS)/microwave (480 W, 2 min) for methane production during the subsequent anaerobic digestion process. Under this condition, 80.2% higher CH_4 accumulation yield was achieved after 16 d of anaerobic digestion when compared with the control. The synergistic effects of CaO_2/microwave pretreatment resulted from the different mechanisms of CaO_2 and microwave treatments. Further, microwave irradiation increased "·OH generation from CaO_2 and significantly alleviated the inhibitory effect of CaO_2 on methanogens. The activities of hydrolytic enzymes and acid-forming enzymes in the waste activated sludge were improved after CaO_2 (0.1 g/gVSS)/microwave (480 W, 2 min) pretreatment. Methanogenesis enzyme activity was also higher after CaO_2 treatment (0.1 g/gVSS)/microwave (480 W, 2 min) following a lag period. Illumina MiSeq sequencing analysis indicated that acetate-utilizing methanogen (Methanosaeta sp.) and H_2/CO_2-utilizing

Digested disorder: Quarterly intrinsic disorder digest (July-August-September, 2013).

Science.gov (United States)

Reddy, Krishna D; DeForte, Shelly; Uversky, Vladimir N

2014-01-01

The current literature on intrinsically disordered proteins grows fast. To keep interested readers up to speed with this literature, we continue a "Digested Disorder" project and represent a new issue of reader's digest of the research papers and reviews on intrinsically disordered proteins. The only 2 criteria for inclusion in this digest are the publication date (a paper should be published within the covered time frame) and topic (a paper should be dedicated to any aspect of protein intrinsic disorder). The current digest issue covers papers published during the third quarter of 2013; i.e., during the period of June, July, and September of 2013. Similar to previous issues, the papers are grouped hierarchically by topics they cover, and for each of the included paper a short description is given on its major findings.

Lytic process studies on anaerobic digestion of organic wastes. Etude des activites lytiques intervenant au cours de la digestion anaerobie des dechets organiques; Rapport final

Energy Technology Data Exchange (ETDEWEB)

Durecu, S.; Thauront, J. (PEC Engineering, 95 - Cergy Pontoise (France). Service de Recherche et Developpement); Festino, C.; Aubart, C.; Reisinger, O. (Nancy-1 Univ., 54 - Vandoeuvre-les-Nancy (France). Lab. d' Ecologie Microbienne)

1990-01-01

To improve anaerobic digestion of pig manure, solubilization of the solid fraction was studied as the rate limiting step in the biomethanation process in an experimental completely mixed digester. The performance of conventional digesters should anaerobic microflora were inefficient in degrading complex biopolymers such as plant fibers. For pectin or cellulose, the use of digestible co-substrates accelerated methanation by increasing the yield of methane and a doubling of the apparent first order solubilization rate constant (Kp = 0.090/d). Lignin should methanation by decreasing methane yield and reducing the rate constant (Kp = 0.035/d). This inhibition was unrelated to volatile fatty acid accumulation. Nine strains of pectinolytic and/or cellulolytic bacteria were isolated. Chitin, a structural constituent of many final species, was effectively solubilized dining anaerobic digestion of pig manure. Seven strains of chitinolytic bacteria were isolated by high chitnese activity. The mycolytic power of fermenting manure processes acting through lytic microflora has been shown to be an effective antagonist of soil borne phytopathogenic fungi, as well as a fertilizer. In greenhouse trails, this compiled fraction demonstrated its ability to control flux unit. Keratin enhanced methane production, and increased H{sub 2}S nearly six-fold. Bacterial strains able to solubilize keratin were also used in autoclawed feather meal to extract the amino acids. (KJD)

Enzymatic hydrolysis of biomimetic bacterial cellulose-hemicellulose composites.

Science.gov (United States)

Penttilä, Paavo A; Imai, Tomoya; Hemming, Jarl; Willför, Stefan; Sugiyama, Junji

2018-06-15

The production of biofuels and other chemicals from lignocellulosic biomass is limited by the inefficiency of enzymatic hydrolysis. Here a biomimetic composite material consisting of bacterial cellulose and wood-based hemicelluloses was used to study the effects of hemicelluloses on the enzymatic hydrolysis with a commercial cellulase mixture. Bacterial cellulose synthesized in the presence of hemicelluloses, especially xylan, was found to be more susceptible to enzymatic hydrolysis than hemicellulose-free bacterial cellulose. The reason for the easier hydrolysis could be related to the nanoscale structure of the substrate, particularly the packing of cellulose microfibrils into ribbons or bundles. In addition, small-angle X-ray scattering was used to show that the average nanoscale morphology of bacterial cellulose remained unchanged during the enzymatic hydrolysis. The reported easier enzymatic hydrolysis of bacterial cellulose produced in the presence of wood-based xylan offers new insights to overcome biomass recalcitrance through genetic engineering. Copyright © 2018 Elsevier Ltd. All rights reserved.

Selective ultrasound-enhanced enzymatic hydrolysis of oleuropein to its aglycon in olive (Olea europaea L.) leaf extracts.

Science.gov (United States)

Delgado-Povedano, María Del Mar; Priego-Capote, Feliciano; Luque de Castro, María Dolores

2017-04-01

Hydrolysis of oleuropein, the main phenol in olive (Olea europaea L.) leaf extracts, to oleuropein aglycon and other subsequent products in the hydrolytic pathway can be catalyzed by different enzymes. Three of the most used hydrolases were assayed to catalyze the process, and β-glucosidase from Aspergillus niger was selected. Acceleration of the enzymatic hydrolysis by ultrasound (US) was studied using a Box-Behnken design (duty cycle, amplitude, cycle time) and an oleuropein standard, and the optimum US conditions for achieving maximum yield of oleuropein aglycon were 0.5s/s duty cycle, 50% amplitude and 45s cycle. The method was applied to obtain oleuropein aglycon from commercial and laboratory extracts from olive leaves, which may have a pharmacological use as deduced by its healthy properties. The kinetics of the US-assisted enzymatic hydrolysis was monitored by analysis of the target compounds using liquid chromatography-tandem mass spectrometry. Copyright © 2016 Elsevier Ltd. All rights reserved.

Evaluation of the pepsin digestibility assay for predicting amino acid digestibility of meat and bone meals.

Science.gov (United States)

Davis, T M; Parsons, C M; Utterback, P L; Kirstein, D

2015-05-01

Sixteen meat and bone meal (MBM) samples were obtained and selected from various company plants to provide a wide range in pepsin nitrogen digestibility values. Pepsin digestibility was determined using either 0.02 or 0.002% pepsin. Amino acid (AA) digestibility of the 16 MBM samples was then determined using a precision-fed cecectomized rooster assay. The 0.02% pepsin digestibility values were numerically higher than the 0.002% pepsin values. The values varied from 77 to 93% for 0.02% pepsin and from 67 to 91% for 0.002% pepsin. The rooster AA digestibility results showed a wide range of values among MBM samples mostly due to the 4 samples having lowest and highest AA digestibility. A precision-fed broiler chick ileal AA digestibility assay confirmed that there were large differences in AA digestibility among the MBM samples having the lowest and highest rooster digestibility values. Correlation analyses between pepsin and AA digestibility values showed that the correlation values (r) were highly significant (P digestibility values were not included in the correlation analyses, the correlation coefficient values (r) were generally very low and not significant (P > 0.05). The results indicated that the pepsin nitrogen digestibility assay is only useful for detecting large differences in AA digestibility among MBM. There also was no advantage for using 0.02 versus 0.002% pepsin. © 2015 Poultry Science Association Inc.

Thermo-chemical pretreatment and enzymatic hydrolysis for enhancing saccharification of catalpa sawdust.

Science.gov (United States)

Jin, Shuguang; Zhang, Guangming; Zhang, Panyue; Li, Fan; Fan, Shiyang; Li, Juan

2016-04-01

To improve the reducing sugar production from catalpa sawdust, thermo-chemical pretreatments were examined and the chemicals used including NaOH, Ca(OH)2, H2SO4, and HCl. The hemicellulose solubilization and cellulose crystallinity index (CrI) were significantly increased after thermo-alkaline pretreatments, and the thermo-Ca(OH)2 pretreatment showed the best improvement for reducing sugar production comparing to other three pretreatments. The conditions of thermo-Ca(OH)2 pretreatment and enzymatic hydrolysis were systematically optimized. Under the optimal conditions, the reducing sugar yield increased by 1185.7% comparing to the control. This study indicates that the thermo-Ca(OH)2 pretreatment is ideal for the saccharification of catalpa sawdust and that catalpa sawdust is a promising raw material for biofuel. Copyright © 2016 Elsevier Ltd. All rights reserved.