Functional Characterization of the Glycoside Hydrolase Encoding GeneOsBE1during Chloroplast Development inOryza sativa

2014 ◽  
Vol 40 (12) ◽  
pp. 2090 ◽  
Author(s):  
Xing-Chun WANG ◽  
Min WANG ◽  
Zhi-Juan JI ◽  
Zhao CHEN ◽  
Wen-Zhen LIU ◽  
...  
Keyword(s):  
Glycoside Hydrolase ◽  
Chloroplast Development ◽  
Functional Characterization

scholarly journals Biochemical and Functional Characterization of Glycoside Hydrolase Family 16 Genes in Aedes aegypti Larvae: Identification of the Major Digestive β-1,3-Glucanase

2019 ◽  
Vol 10 ◽  
Author(s):  
Raquel Santos Souza ◽  
Maiara do Valle Faria Gama ◽  
Renata Schama ◽  
José Bento Pereira Lima ◽  
Hector Manuel Diaz-Albiter ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Glycoside Hydrolase ◽  
Functional Characterization ◽  
Glycoside Hydrolase Family ◽  
Hydrolase Family

Origin, evolution and functional characterization of the land plant glycoside hydrolase subfamily GH5_11

2019 ◽  
Vol 138 ◽  
pp. 205-218 ◽  
Author(s):  
Rujia Chen ◽  
Youli Yao ◽  
Huimin Fang ◽  
Enying Zhang ◽  
Pengcheng Li ◽  
...  
Keyword(s):  
Glycoside Hydrolase ◽  
Functional Characterization ◽  
Land Plant

scholarly journals High-Throughput Recovery and Characterization of Metagenome-Derived Glycoside Hydrolase-Containing Clones as a Resource for Biocatalyst Development

mSystems ◽  
2019 ◽  
Vol 4 (4) ◽  
Author(s):  
Zachary Armstrong ◽  
Feng Liu ◽  
Sam Kheirandish ◽  
Hong-Ming Chen ◽  
Keith Mewis ◽  
...  
Keyword(s):  
High Throughput ◽  
Glycoside Hydrolase ◽  
Plant Biomass ◽  
Functional Characterization ◽  
Environmental Dna ◽  
Functional Screening ◽  
Biomass Degradation ◽  
Genomic Context ◽  
Or Gene

ABSTRACT Functional metagenomics is a powerful tool for both the discovery and development of biocatalysts. This study presents the high-throughput functional screening of 22 large-insert fosmid libraries containing over 300,000 clones sourced from natural and engineered ecosystems, characterization of active clones, and a demonstration of the utility of recovered genes or gene cassettes in the development of novel biocatalysts. Screening was performed in a 384-well-plate format with the fluorogenic substrate 4-methylumbelliferyl cellobioside, which releases a fluorescent molecule when cleaved by β-glucosidases or cellulases. The resulting set of 164 active clones was subsequently interrogated for substrate preference, reaction mechanism, thermal stability, and optimal pH. The environmental DNA harbored within each active clone was sequenced, and functional annotation revealed a cornucopia of carbohydrate-degrading enzymes. Evaluation of genomic-context information revealed both synteny and polymer-targeting loci within a number of sequenced clones. The utility of these fosmids was then demonstrated by identifying clones encoding activity on an unnatural glycoside (4-methylumbelliferyl 6-azido-6-deoxy-β-d-galactoside) and transforming one of the identified enzymes into a glycosynthase capable of forming taggable disaccharides. IMPORTANCE The generation of new biocatalysts for plant biomass degradation and glycan synthesis has typically relied on the characterization and investigation of one or a few enzymes at a time. By coupling functional metagenomic screening and high-throughput functional characterization, we can progress beyond the current scale of catalyst discovery and provide rapid annotation of catalyst function. By functionally screening environmental DNA from many diverse sources, we have generated a suite of active glycoside hydrolase-containing clones and demonstrated their reaction parameters. We then demonstrated the utility of this collection through the generation of a new catalyst for the formation of azido-modified glycans. Further interrogation of this collection of clones will expand our biocatalytic toolbox, with potential application to biomass deconstruction and synthesis of glycans.

scholarly journals Structural and Functional Characterization of a Ruminal β-Glycosidase Defines a Novel Subfamily of Glycoside Hydrolase Family 3 with Permuted Domain Topology

2016 ◽  
Vol 291 (46) ◽  
pp. 24200-24214 ◽  
Author(s):  
Mercedes Ramírez-Escudero ◽  
Mercedes V. del Pozo ◽  
Julia Marín-Navarro ◽  
Beatriz González ◽  
Peter N. Golyshin ◽  
...  
Keyword(s):  
Glycoside Hydrolase ◽  
Functional Characterization ◽  
Glycoside Hydrolase Family ◽  
Hydrolase Family

Structural and functional characterization of a glycoside hydrolase family 3 β-N-acetylglucosaminidase from Paenibacillus sp. str. FPU-7

2019 ◽  
Vol 166 (6) ◽  
pp. 503-515
Author(s):  
Takafumi Itoh ◽  
Tomomitsu Araki ◽  
Tomohiro Nishiyama ◽  
Takao Hibi ◽  
Hisashi Kimoto
Keyword(s):  
Crystal Structure ◽  
Glycoside Hydrolase ◽  
Catalytic Mechanism ◽  
Functional Characterization ◽  
Structural Features ◽  
Expression Cloning ◽  
Glycoside Hydrolase Family ◽  
Paenibacillus Sp ◽  
Hydrolase Family

Abstract Chitin, a β-1,4-linked homopolysaccharide of N-acetyl-d-glucosamine (GlcNAc), is one of the most abundant biopolymers on Earth. Paenibacillus sp. str. FPU-7 produces several different chitinases and converts chitin into N,N′-diacetylchitobiose ((GlcNAc)2) in the culture medium. However, the mechanism by which the Paenibacillus species imports (GlcNAc)2 into the cytoplasm and divides it into the monomer GlcNAc remains unclear. The gene encoding Paenibacillus β-N-acetyl-d-glucosaminidase (PsNagA) was identified in the Paenibacillus sp. str. FPU-7 genome using an expression cloning system. The deduced amino acid sequence of PsNagA suggests that the enzyme is a part of the glycoside hydrolase family 3 (GH3). Recombinant PsNagA was successfully overexpressed in Escherichia coli and purified to homogeneity. As assessed by gel permeation chromatography, the enzyme exists as a 57-kDa monomer. PsNagA specifically hydrolyses chitin oligosaccharides, (GlcNAc)2–4, 4-nitrophenyl N-acetyl β-d-glucosamine (pNP-GlcNAc) and pNP-(GlcNAc)2–6, but has no detectable activity against 4-nitrophenyl β-d-glucose, 4-nitrophenyl β-d-galactosamine and colloidal chitin. In this study, we present a 1.9 Å crystal structure of PsNagA bound to GlcNAc. The crystal structure reveals structural features related to substrate recognition and the catalytic mechanism of PsNagA. This is the first study on the structural and functional characterization of a GH3 β-N-acetyl-d-glucosaminidase from Paenibacillus sp.

Functional characterization of ObgC in ribosome biogenesis during chloroplast development

2012 ◽  
Vol 71 (1) ◽  
pp. 122-134 ◽  
Author(s):  
Woo Young Bang ◽  
Ji Chen ◽  
In Sil Jeong ◽  
Sam Woong Kim ◽  
Chul Wook Kim ◽  
...  
Keyword(s):  
Ribosome Biogenesis ◽  
Chloroplast Development ◽  
Functional Characterization
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