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Glycoside hydrolase

André Busch, Grit Kunert, Natalie Wielsch, Yannick Pauchet
Cellulose is a major component of the primary and secondary cell walls in plants. Cellulose is considered to be the most abundant biopolymer on Earth and represents a large potential source of metabolic energy. Yet, cellulose degradation is rare and mostly restricted to cellulolytic microorganisms. Recently, various metazoans, including leaf beetles, have been found to encode their own cellulases, giving them the ability to degrade cellulose independently of cellulolytic symbionts. Here, we analyzed the cellulosic capacity of the leaf beetle Gastrophysa viridula, which typically feeds on Rumex plants...
May 17, 2018: Insect Molecular Biology
Joana G Desiderato, Danillo O Alvarenga, Milena T L Constancio, Lucia M C Alves, Alessandro M Varani
Cellulose and its associated polymers are structural components of the plant cell wall, constituting one of the major sources of carbon and energy in nature. The carbon cycle is dependent on cellulose- and lignin-decomposing microbial communities and their enzymatic systems acting as consortia. These microbial consortia are under constant exploration for their potential biotechnological use. Herein, we describe the characterization of the genome of Dyella jiangningensis FCAV SCS01, recovered from the metagenome of a lignocellulose-degrading microbial consortium, which was isolated from a sugarcane crop soil under mechanical harvesting and covered by decomposing straw...
May 14, 2018: Genetics and Molecular Biology
Benwei Zhu, Fang Ni, Yun Sun, Xianyu Zhu, Heng Yin, Zhong Yao, Yuguang Du
Carrageenan, a kind of linear sulfated polysaccharides consisting of D-galactose with alternating α-1,3 and β-1,4 linkages, has been widely applied in the food and cosmetic industries as thickening and gelling agents due to excellent properties, such as gel-forming ability and chemical stability. It can be degraded by carrageenases to produce a series of even-numbered carrageenan oligosaccharides, which exhibit various fascinating functions, such as anti-inflammation, anti-coagulation, anti-tumor, and anti-thrombosis effects...
May 15, 2018: Critical Reviews in Biotechnology
David R Rose, Marcia M Chaudet, Kyra Jones
OBJECTIVES: Maltase-glucoamylase and sucrase-isomaltase are enzymes in the brush-border membrane of the small intestinal lumen responsible for the breakdown of postamylase starch polysaccharides to release monomeric glucose. As such, they are critical players in healthy nutrition and their malfunction can lead to severe disorders. METHODS: This review covers investigations of the structures and functions of these enzymes. RESULTS: Each consists of 2 enzyme domains of the glycoside hydrolase family GH31 classification, yet with somewhat differing enzymatic properties...
June 2018: Journal of Pediatric Gastroenterology and Nutrition
Beatriz Trastoy, Erik Klontz, Jared Orwenyo, Alberto Marina, Lai-Xi Wang, Eric J Sundberg, Marcelo E Guerin
Endoglycosidase S (EndoS) is a bacterial endo-β-N-acetylglucosaminidase that specifically catalyzes the hydrolysis of the β-1,4 linkage between the first two N-acetylglucosamine residues of the biantennary complex-type N-linked glycans of IgG Fc regions. It is used for the chemoenzymatic synthesis of homogeneously glycosylated antibodies with improved therapeutic properties, but the molecular basis for its substrate specificity is unknown. Here, we report the crystal structure of the full-length EndoS in complex with its oligosaccharide G2 product...
May 14, 2018: Nature Communications
Ewelina Cholewińska, Katarzyna Ognik, Bartosz Fotschki, Zenon Zduńczyk, Jerzy Juśkiewicz
The aim of the study was to investigate the effect of two forms (CuCO3 (CuS); and Cu nanoparticles (CuNP)) and dosages (standard 6.5 mg/kg (H), half of the standard (L)) of additional dietary Cu administered to growing rats on gastrointestinal and hepatic function and morphology. Copper in the form of CuNP vs CuS caused lower Cu faecal/urinal excretion and increased Cu accumulation in the brain tissue. Hepatic high-grade hydropic degeneration and necrotic lesions were observed only in the CuNP-H animals. In the lower gut, the dietary application of CuNP stifled bacterial enzymatic activity of caecal gut microbiota and resulted in lower SCFA production...
2018: PloS One
Jie Zhou, Zhoukun Li, Han Zhang, Jiale Wu, Xianfeng Ye, Weiliang Dong, Min Jiang, Yan Huang, Zhongli Cui
Here, a novel amylolytic enzyme gene designated coMA was cloned from Corallococcus sp. EGB. The deduced amino acid sequence contained a predicted lipoprotein signal peptide (residues 1-18) and conserved glycoside hydrolase family 13 (GH13) module. The amino acid sequence of CoMA exhibits low sequence identity (10-19%) with cyclodextrin-hydrolyzing enzymes (GH13_20) and is assigned to GH13_36. The most outstanding feature of CoMA is its ability to catalyze the conversion of maltooligosaccharides (≥ G3) and soluble starch to maltose as the sole hydrolysate...
May 11, 2018: Applied and Environmental Microbiology
Varsha Bohra, Nishant A Dafale, Hemant J Purohit
Genome sequence of Paenibacillus polymyxa ND25 isolated from cow rumen is reported for being a potential candidate in hydrolysis of lignocellulosic plant biomass. Draft genome sequence generated 5.73 Mb data containing 4922 putative protein coding genes, of which 140 are annotated for glycoside hydrolases. P. polymyxa ND25 strain comprises diverse lignocellulolytic components, especially 12 cellulase along with 23 hemicellulases and 11 esterases, signifying its potential for lignocellulose hydrolysis. Subsequent enzyme assay exhibited the potential of strain to produce 0...
May 2018: 3 Biotech
Liangkun Long, Meijuan Xu, Yuexin Shi, Qunying Lin, Jing Wang, Shaojun Ding
Two new endo-1,4-beta-xylanases encoding genes EpXyn1 and EpXyn3 were isolated from mesophilic fungus Eupenicillium parvum 4-14. Based on analysis of catalytic domain and phylogenetic trees, the xylanases EpXYN1 (404 aa) and EpXYN3 (220 aa) belong to glycoside hydrolase (GH) family 10 and 11, respectively. Both EpXYN1 and EpXYN3 were successfully expressed in Pichia pastoris and the recombinant enzymes were characterized using beechwood xylan, birchwood xylan, or oat spelt xylan as substrates, respectively...
May 9, 2018: Applied Biochemistry and Biotechnology
Li Gao, Jianan Sun, Francesco Secundo, Xin Gao, Changhu Xue, Xiangzhao Mao
A novel chitinase gene was cloned from Streptomyces albolongus ATCC 27414, and expressed successfully in Escherichia coli BL21. The recombinant enzyme (SaChiA4) belongs to glycoside hydrolases (GH) family 18 and consists of a catalytic domain and a chitin binding domain (CBD) in its C-terminus. SaChiA4 was purified homogeneously (specific activity of 66.2 U/mg with colloidal chitin as substrate), and showed a molecular mass of approximately 47 kDa. SaChiA4 showed its optimal activity at pH 5.0 and 55 °C and exhibited remarkable pH and temperature stability...
September 30, 2018: Food Chemistry
Ruili Zhao, Rui Zhao, Yishuai Tu, Xiaoming Zhang, Liping Deng, Xiangdong Chen
A novel α-galactosidase of glycoside hydrolase family 36 was cloned from Bacillus coagulans, overexpressed in Escherichia coli, and characterized. The purified enzyme Aga-BC7050 was 85 kDa according to SDS-PAGE and 168 kDa according to gel filtration, indicating that its native structure is a dimer. With p-nitrophenyl-α-d- galactopyranoside (pNPGal) as the substrate, optimal temperature and pH were 55 °C and 6.0, respectively. At 60 °C for 30 min, it retained > 50% of its activity. It was stable at pH 5...
2018: PloS One
Anthi Karnaouri, Evangelos Topakas, Leonidas Matsakas, Ulrika Rova, Paul Christakopoulos
Non-digestible oligosaccharides (NDOs) are likely prebiotic candidates that have been related to the prevention of intestinal infections and other disorders for both humans and animals. Lignocellulosic biomass is the largest carbon source in the biosphere, therefore cello-oligosacharides (COS), especially cellobiose, are potentially the most widely available choice of NDOs. Production of COS and cellobiose with enzymes offers numerous benefits over acid-catalyzed processes, as it is milder, environmentally friendly and produces fewer by-products...
2018: Frontiers in Chemistry
Kok Jun Liew, Lily Lim, Hui Ying Woo, Kok-Gan Chan, Mohd Shahir Shamsir, Kian Mau Goh
Beta-glucosidase (BGL) is an important industrial enzyme for food, waste and biofuel processing. Jeotgalibacillus is an understudied halophilic genus, and no beta-glucosidase from this genus has been reported. A novel beta-glucosidase gene (1344 bp) from J. malaysiensis DSM 28777T was cloned and expressed in E. coli. The recombinant protein, referred to as BglD5, consists of a total 447 amino acids. BglD5 purified using a Ni-NTA column has an apparent molecular mass of 52 kDa. It achieved the highest activity at pH 7 and 65 °C...
April 30, 2018: International Journal of Biological Macromolecules
Andrew J Thompson, Richard J Spears, Yanping Zhu, Michael D L Suits, Spencer J Williams, Harry J Gilbert, Gideon J Davies
A dominant human gut microbe, the well studied symbiont Bacteroides thetaiotaomicron (Bt), is a glyco-specialist that harbors a large repertoire of genes devoted to carbohydrate processing. Despite strong similarities among them, many of the encoded enzymes have evolved distinct substrate specificities, and through the clustering of cognate genes within operons termed polysaccharide-utilization loci (PULs) enable the fulfilment of complex biological roles. Structural analyses of two glycoside hydrolase family 92 α-mannosidases, BT3130 and BT3965, together with mechanistically relevant complexes at 1...
May 1, 2018: Acta Crystallographica. Section D, Structural Biology
Monika Schmoll
Trichoderma reesei (syn. Hypocrea jecorina ) is the model organism for industrial production of plant cell wall degradating enzymes. The integration of light and nutrient signals for adaptation of enzyme production in T. reesei emerged as an important regulatory mechanism to be tackled for strain improvement. Gene regulation specific for cellulase inducing conditions is different in light and darkness with substantial regulation by photoreceptors. Genes regulated by light are clustered in the genome, with several of the clusters overlapping with CAZyme clusters...
2018: Fungal Biology and Biotechnology
Jenny Arnling Bååth, Antonio Martínez-Abad, Jennie Berglund, Johan Larsbrink, Francisco Vilaplana, Lisbeth Olsson
Background: Galactoglucomannan (GGM) is the most abundant hemicellulose in softwood, and consists of a backbone of mannose and glucose units, decorated with galactose and acetyl moieties. GGM can be hydrolyzed into fermentable sugars, or used as a polymer in films, gels, and food additives. Endo -β-mannanases, which can be found in the glycoside hydrolase families 5 and 26, specifically cleave the mannan backbone of GGM into shorter oligosaccharides. Information on the activity and specificity of different mannanases on complex and acetylated substrates is still lacking...
2018: Biotechnology for Biofuels
Suman Yadav, Suresh Kumar Dubey
The aim of this work was to study cellulose degradation and whole genome sequence of Paenibacillus lautus BHU3 isolate. The 16S rRNA gene sequence analysis revealed genetic relatedness (99%) of Iso 7 with Paenibacillus lautus, Iso 8 with Paenibacillus lactis, and Iso 9 with Bacillus amyloliquefaciens. Clear zone formation followed by CMCase and FPase assays exhibited cellulolytic potential in the order: P. lautus > P. lactis > B. amyloliquefaciens. The most potent isolate, Paenibacillus lautus strain BHU3 was subjected to whole genome analysis with reference to the genomic basis of cellulose degradation...
April 19, 2018: Bioresource Technology
Ali Rohman, Niels van Oosterwijk, Ni Nyoman Tri Puspaningsih, Bauke W Dijkstra
Complete degradation of the xylan backbone of hemicellulosic plant cell walls requires the synergistic action of endo-xylanases and β-1,4-xylosidases. While endo-xylanases produce xylooligosaccharides from xylan, β-1,4-xylosidases degrade the xylooligosaccharides into xylose monomers. The glycoside hydrolase family 43 β-1,4-xylosidase from Geobacillus thermoleovorans IT-08 is a promising, heat stable catalyst for the saccharification of hemicellulosic material into simple fermentable sugars, but it is competitively inhibited by its products arabinose and xylose...
2018: PloS One
Jose Sergio Hleap, Christian Blouin
The Glycoside Hydrolase Family 13 (GH13) is both evolutionarily diverse and relevant to many industrial applications. Its members hydrolyze starch into smaller carbohydrates and members of the family have been bioengineered to improve catalytic function under industrial environments. We introduce a framework to analyze the response to selection of GH13 protein structures given some phylogenetic and simulated dynamic information. We find that the TIM-barrel (a conserved protein fold consisting of eight α-helices and eight parallel β-strands that alternate along the peptide backbone, common to all amylases) is not selectable since it is under purifying selection...
2018: PloS One
Yuan Yuan, Xiaohua Jin, Juan Liu, Xing Zhao, Junhui Zhou, Xin Wang, Deyi Wang, Changjiangsheng Lai, Wei Xu, Jingwen Huang, Liangping Zha, Dahui Liu, Xiao Ma, Li Wang, Menyan Zhou, Zhi Jiang, Hubiao Meng, Huasheng Peng, Yuting Liang, Ruiqiang Li, Chao Jiang, Yuyang Zhao, Tiegui Nan, Yan Jin, Zhilai Zhan, Jian Yang, Wenkai Jiang, Luqi Huang
We present the 1.06 Gb sequenced genome of Gastrodia elata, an obligate mycoheterotrophic plant, which contains 18,969 protein-coding genes. Many genes conserved in other plant species have been deleted from the G. elata genome, including most of those for photosynthesis. Additional evidence of the influence of genome plasticity in the adaptation of this mycoheterotrophic lifestyle is evident in the large number of gene families that are expanded in G. elata, including glycoside hydrolases and urease that likely facilitate the digestion of hyphae are expanded, as are genes associated with strigolactone signaling, and ATPases that may contribute to the atypical energy metabolism...
April 24, 2018: Nature Communications
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