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Enzymatic hydrolysis

Fátima Arrutia, Rebeca Fernández, Carlos Menéndez, Ulises A González, Francisco A Riera
In order to exploit industrial discards, protein enzymatic hydrolysis is a currently popular methodology for obtaining bioactive peptides. However, once released, most promising peptides have to be selected from the mixture. In this work, the suitability of pepsin (EC to hydrolyse serum albumin in order to obtain bioactive peptides was assessed. Then, a suitable process to obtain best separation of bioactive peptides was evaluated, using polyethersulfone membranes at different pH values. Serum albumin was easily hydrolysed by pepsin, reaching a DH value of the 65...
December 8, 2017: Scientific Reports
Jitrayut Jitonnom, James R Ketudat-Cairns, Supa Hannongbua
Fructosyltransferases (FTs) act on sucrose by cleaving the β-(2→1) linkage, releasing glucose, and then transferring the fructosyl group to an acceptor molecule. These enzymes are capable of producing prebiotic fructooligosaccharides (FOSs) that are of industrial interest. While several FOS-synthesizing enzymes FTs have been investigated, their catalytic mechanism is not yet fully understood, especially the molecular details of how FOS are enzymatically synthesized from sucrose. Here, we present a comparative quantum mechanics/molecular mechanics (QM/MM) study on the hydrolysis and transfructosylation reactions catalyzed by A...
November 24, 2017: Journal of Molecular Graphics & Modelling
Alice B Nongonierma, Candice Lamoureux, Richard J FitzGerald
Tropical banded crickets (Gryllodes sigillatus) were studied for their ability to yield hydrolysates with dipeptidyl peptidase IV (DPP-IV) inhibitory properties. A cricket protein isolate (CPI) was prepared following extraction of the water soluble proteins from G. sigillatus powder (CP). The extraction yield and purity were 20.90 ± 0.35% and 57.0 ± 2.23%, respectively. Endogenous proteinase activities were detected in the CP, which were linked to the significant protein breakdown seen in this sample. Fifteen CPI hydrolysates (H1-H15) were generated with Protamex™ using a design of experiments (DOE) approach combining three parameters, temperature (40, 50 and 60 °C), enzyme to substrate ratio (E : S, 0...
December 8, 2017: Food & Function
Yining Zeng, Michael E Himmel, Shi-You Ding
Understanding plant cell wall cross-linking chemistry and polymeric architecture is key to the efficient utilization of biomass in all prospects from rational genetic modification to downstream chemical and biological conversion to produce fuels and value chemicals. In fact, the bulk properties of cell wall recalcitrance are collectively determined by its chemical features over a wide range of length scales from tissue, cellular to polymeric architectures. Microscopic visualization of cell walls from the nanometer to the micrometer scale offers an in situ approach to study their chemical functionality considering its spatial and chemical complexity, particularly the capabilities of characterizing biomass non-destructively and in real-time during conversion processes...
2017: Biotechnology for Biofuels
In Yung Sunwoo, Trung Hau Nguyen, Pailin Sukwong, Gwi-Teak Jeong, Sung-Koo Kim
The waste seaweed from Gwangalli beach, Busan, Korea was utilized as biomass for ethanol production. Sagassum fulvellum (brown seaweed, Mojaban in Korean name) comprised 72% of the biomass. The optimal hyper thermal acid hydrolysis conditions were obtained as 8% slurry contents, 138 mM sulfuric acid and 160 °C of treatment temperature for 10 min with a low content of inhibitory compounds. To obtain more monosaccharide, enzymatic saccharification was carried out with Viscozyme L for 48 h. After pretreatment, 34 g/L of monosaccharides were obtained...
December 8, 2017: Journal of Microbiology and Biotechnology
Kristin T Ravndal, Roald Kommedal
A large fraction of the organic substrate in municipal wastewater is particulate. Prior to uptake, particles have to be degraded through potentially a range of intermediates. However, research on intermediate dynamics during particle hydrolysis is limited. In this paper, batch experiments on flocculated and dispersed biomass microcosms using starch as particulate substrate are reported. Overall hydrolysis rate was not significantly different between the two systems. Particle colonization, and increased particle porosity in combination with particle breakup, led to increased substrate availability over time...
December 2017: Water Science and Technology: a Journal of the International Association on Water Pollution Research
Diego Currò
A healthy gut microbiota is necessary for the normal operation of several body functions, including gastrointestinal sensitivity and motility, lipid and glucid metabolism, immune surveillance, and host behavior. In addition, intestinal bacteria contribute to determining the pharmacological properties of several drugs by producing different drug metabolizing enzymes. Areas covered: Four enzymatic processes are discussed: prodrug activation; drug inactivation; drug deconjugation; and hydrolysis of natural glycosides with further metabolism of released aglycones...
December 6, 2017: Expert Review of Clinical Pharmacology
Xinghao Yang, Xiao Xiao, Dan Liu, Ribang Wu, Cuiling Wu, Jiang Zhang, Jiafeng Huang, Binqiang Liao, Hailun He
Collagenases are the most important group of commercially-produced enzymes. However, even though biological resources are abundant in the sea, very few of these commercially popular enzymes are from marine sources, especially from marine bacteria. We optimized the production of marine collagenases by Pseudoalteromonas sp. SJN2 and investigated the antioxidant activities of the hydrolysates. Media components and culture conditions associated with marine collagenase production by Pseudoalteromonas sp. SJN2 were optimized by statistical methods, namely Plackett-Burman design and response surface methodology (RSM)...
December 4, 2017: Marine Drugs
Feng Wang, Qing He, Kaixuan Su, Tiandi Wei, Sujuan Xu, Lichuan Gu
The Asp-His-His and Asp-His-His-associated (DHH/DHHA1) domain-containing phosphodiesterases (PDEs) that catalyze degradation of cyclic di-adenosine monophosphate (c-di-AMP) could be subdivided into two subfamilies based on the final product is 5'-phosphadenylyl-adenosine (5'-pApA) or AMP. In previous study we revealed that Rv2837c, a stand-alone DHH/DHHA1 PDE, employs a 5'-pApA internal flipping mechanism to produce AMPs. However, why the membrane-bound DHH/DHHA1 PDE can only degrade c-di-AMP to 5'-pApA remains obscure...
December 4, 2017: Biochemical Journal
Charlotte Jendresen, Michael R Daws, Lars N G Nilsson
INTRODUCTION: Reporter cells expressing a chimeric receptor that activates a reporter can be used for screening ligand-mediated signal transduction. In this study, we used reporter cells harboring an NFAT/lacZ construct that express β-galactosidase when the chimeric receptor is stimulated. A colorimetric β-galactosidase substrate, chlorophenol-red β-d-galactopyranoside (CPRG), was used to detect enzymatic activity. Sub-optimal conditions have unfortunately extensively been reported with such reporter-based β-galactosidase assays...
December 1, 2017: Journal of Pharmacological and Toxicological Methods
Ga-Hui Oh, Joo-Eun Kim, Young-Joon Park
The purpose of this study was to develop a hydrolysis-resistant optimized oral formulation of tenofovir disoproxil (TD) using a stabilizer. To develop a stabilized TD tablet bioequivalent to the commercial TD fumarate (TDF, Viread®) tablet, TD free base was prepared and its degradation profile and stability were investigated. The TD tablet showed antiviral activity, but its absorption was limited in the intestinal tract because of premature degradation. The drug subjected to severe conditions for the stress test was catalyzed under neutral, basic, oxidative, and thermolytic conditions, whereas it was comparatively stable under acidic, photolytic, and humid states...
December 4, 2017: Drug Development and Industrial Pharmacy
Xiangmin Miao, Zhiyuan Cheng, Haiyan Ma, Zongbing Li, Ning Xue, Po Wang
A novel strategy was developed for microRNA-155 (miRNA-155) detection based on the fluorescence quenching of positively charged gold nanoparticles ((+)AuNPs) to Ag nanoclusters (AgNCs). In the designed system, DNA-stabilized Ag nanoclusters (DNA/AgNCs) were introduced as fluorescent probes, and DNA-RNA heteroduplexes were formed upon the addition of target miRNA-155. Meanwhile, the (+)AuNPs could be electrostatically adsorbed on the negatively charged single-stranded DNA (ssDNA) or DNA-RNA heteroduplexes to quench the fluorescence signal...
December 4, 2017: Analytical Chemistry
Marta Bevilacqua, Giulia Praticò, Johanne Plesner, Maja Molloy, Thomas Skov, Flemming H Larsen
Protein hydrolysates are of great interest in the food industry due to their nutritional and functional properties, but their use often implies solubilization in water and therefore hamper the use of plant proteins with inherent low water solubility. Protein solubility in water can be modified by enzymatic hydrolysis, but during this process several collateral properties of the protein hydrolysates changes. It is therefore important to determine the end-point of the process and to monitor its development. In this feasibility study, we demonstrated the potential of different spectroscopic techniques (1H NMR and IR) coupled with chemometrics analysis in monitoring the hydrolysis of five different industrial grade plant proteins by the enzyme Alcalase...
December 2017: Food Research International
Anna S Dotsenko, Gleb S Dotsenko, Olga V Senko, Nikolay A Stepanov, Ilya V Lyagin, Elena N Efremenko, Alexander V Gusakov, Ivan N Zorov, Ekaterina A Rubtsova
The pretreatment of softwood and hardwood samples (spruce and hornbeam wood) with 1-butyl-3-methylimidazolium chloride ([Bmim]Cl) was undertaken for further simultaneous enzymatic saccharification of renewable non-food lignocellulosic biomass and microbial fermentation of obtained sugars to ethanol and fumaric acid. A multienzyme cocktail based on cellulases and yeast or fungus cells producing ethanol and fumaric acid were the main objects of [Bmim]Cl influence studies. A complex effect of lignocellulosic biomass pretreatment with [Bmim]Cl on various aspects of the process (both action of cellulases and microbial conversion of hydrolysates to target products) was revealed...
November 23, 2017: Bioresource Technology
Weiguang Li, Peng Ji, Qinzheng Zhou, Chengyao Hua, Chao Han
Enzymatic hydrolysis is considered an efficient and environmental strategy for the degradation of organic waste materials. Compared to mesophilic cellulases, thermostable cellulases with considerable activity are more advantageous in waste paper hydrolysis, particularly in terms of their participation in synergistic action. In this study, the synergistic effect of two different types of thermostable Chaetomium thermophilum cellulases, the endoglucanase CTendo45 and the cellobiohydrolase CtCel6, on five common kinds of waste papers was investigated...
November 30, 2017: Molecular Biotechnology
P R Heinen, A Bauermeister, L F Ribeiro, J M Messias, P Z Almeida, L A B Moraes, C G Vargas-Rechia, A H C de Oliveira, R J Ward, E X F Filho, M K Kadowak, J A Jorge, M L T M Polizeli
The present study describes the one-step purification and biochemical characterization of an endo-1,4-β-xylanase from Aspergillus tamarii Kita. Extracellular xylanase was purified to homogeneity 7.43-fold through CM-cellulose. Enzyme molecular weight and pI were estimated to be 19.5kDa and 8.5, respectively. The highest activity of the xylanase was obtained at 60°C and it was active over a broad pH range (4.0-9.0), with maximal activity at pH 5.5. The enzyme was thermostable at 50°C, retaining more than 70% of its initial activity for 480min...
November 27, 2017: International Journal of Biological Macromolecules
Christin Cürten, Nico Anders, Niels Juchem, Nina Ihling, Kristina Volkenborn, Andreas Knapp, Karl-Erich Jaeger, Jochen Büchs, Antje C Spiess
In contrast to biochemical reactions, which are often carried out under automatic control and maintained overnight, the automation of chemical analysis is usually neglected. Samples are either analyzed in a rudimentary fashion using in situ techniques, or aliquots are withdrawn and stored to facilitate more precise offline measurements, which can result in sampling and storage errors. Therefore, in this study, we implemented automated reaction control, sampling, and analysis. As an example, the activities of xylanases on xylotetraose and soluble xylan were examined using high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD)...
November 28, 2017: Analytical and Bioanalytical Chemistry
Nancy Gisela González Garza, Janice Azucena Chuc Koyoc, Jorge Ariel Torres Castillo, Eduardo Alejandro García Zambrano, David Betancur Ancona, Luis Chel Guerrero, Sugey Ramona Sinagawa García
Moringa oleifera (Moringaceae) is a specie of significant importance because of its multiple nutraceutical properties, that has led to increase in its consumption. The seeds contain a high percentage of protein (37.48%). However, little is known about the bioactive properties of these proteins and peptides, especially those generated by enzymatic hydrolysis. The objective of this study was to evaluate the biofunctional properties of total hydrolysates (TH) and peptide fractions from protein isolates of moringa seeds...
December 2017: Journal of Food Science and Technology
Priyanka Dash, Goutam Ghosh
In this study, the effect of enzymatic hydrolysis of globulin fraction of C. moschata (CMH), C. lanatus (CLH) and L. siceraria (LSH) on antioxidant capacity, functional properties, structural and micro-structural properties, as well as amino acid compositions were evaluated. All the hydrolysates exhibited significant antioxidant properties. The essential amino acids content in LSH (92.7 mg/g) was higher than CMH (79.9 mg/g) and CLH (70.5 mg/g). Water absorption capacity (5 g/g), heat stability (89%), emulsifying activity index (98...
December 2017: Journal of Food Science and Technology
Guilherme C Q Pereira, Danilo S Braz, Marcelo Hamaguchi, Thaddeus C Ezeji, Rubens Maciel Filho, Adriano P Mariano
This work proposes a strategy, from a process design standpoint, for pulp companies to enter the Brazilian ethanol market. The flexible plant converts eucalyptus-derived glucose to either ethanol or butanol (according to market conditions) and xylose only to butanol production. Depending on the biomass pretreatment technology, Monte Carlo simulations showed that the Net Present Value (NPV) of the flexible plant increases by 20-28% in relation to an ethanol-dedicated plant. Whereas the lower costs of the steam explosion technology turns the investment more attractive (NPV = 184 MMUSD; IRR = 29%), the organosolv technology provides better flexibility to the plant...
November 10, 2017: Bioresource Technology
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