Substrate specificity of glycoside hydrolase family 1 β-glucosidase AtBGlu42 from Arabidopsis thaliana and its molecular mechanism

2021 ◽  
Author(s):  
Shu Horikoshi ◽  
Wataru Saburi ◽  
Jian Yu ◽  
Hideyuki Matsuura ◽  
James R Ketudat Cairns ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Glycoside Hydrolase ◽  
Sequence Similarity ◽  
Glycoside Hydrolase Family ◽  
Its Sequence ◽  
Cell Wall Metabolism ◽  
Bioactive Substances ◽  
Chain Length Specificity ◽  
Glycoside Hydrolase Family 1 ◽  
Hydrolase Family

ABSTRACT Plants possess many glycoside hydrolase family 1 (GH1) β-glucosidases, which physiologically function in cell wall metabolism and activation of bioactive substances, but most remain uncharacterized. One GH1 isoenzyme AtBGlu42 in Arabidopsis thaliana has been identified to hydrolyze scopolin using the gene deficient plants, but no enzymatic properties were obtained. Its sequence similarity to another functionally characterized enzyme Os1BGlu4 in rice suggests that AtBGlu42 also acts on oligosaccharides. Here, we show that the recombinant AtBGlu42 possesses high kcat/Km not only on scopolin, but also on various β-glucosides, cellooligosaccharides, and laminarioligosaccharides. Of the cellooligosaccharides, cellotriose was the most preferred. The crystal structure, determined at 1.7 Å resolution, suggests that Arg342 gives unfavorable binding to cellooligosaccharides at subsite +3. The mutants R342Y and R342A showed the highest preference on cellotetraose or cellopentaose with increased affinities at subsite +3, indicating that the residues at this position have an important role for chain length specificity.

Functional genomic analysis of Arabidopsis thaliana glycoside hydrolase family 1

2004 ◽  
Vol 55 (3) ◽  
pp. 343-367 ◽  
Author(s):  
Zhiwei Xu ◽  
Luis Escamilla-Trevi�o ◽  
Lihui Zeng ◽  
Mallikarjun Lalgondar ◽  
David Bevan ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Glycoside Hydrolase ◽  
Genomic Analysis ◽  
Glycoside Hydrolase Family ◽  
Functional Genomic ◽  
Functional Genomic Analysis ◽  
Glycoside Hydrolase Family 1 ◽  
Hydrolase Family

scholarly journals Analysis of the diversity of the glycoside hydrolase family 130 in mammal gut microbiomes reveals a novel mannoside-phosphorylase function

2020 ◽  
Vol 6 (10) ◽  
Author(s):  
Ao Li ◽  
Elisabeth Laville ◽  
Laurence Tarquis ◽  
Vincent Lombard ◽  
David Ropartz ◽  
...  
Keyword(s):  
Glycoside Hydrolase ◽  
Sequence Similarity ◽  
Gut Bacteria ◽  
Glycoside Hydrolase Family ◽  
Sequence Alignments ◽  
Multiple Sequence ◽  
Content Type ◽  
Multiple Sequence Alignments ◽  
Hydrolase Family

Mannoside phosphorylases are involved in the intracellular metabolization of mannooligosaccharides, and are also useful enzymes for the in vitro synthesis of oligosaccharides. They are found in glycoside hydrolase family GH130. Here we report on an analysis of 6308 GH130 sequences, including 4714 from the human, bovine, porcine and murine microbiomes. Using sequence similarity networks, we divided the diversity of sequences into 15 mostly isofunctional meta-nodes; of these, 9 contained no experimentally characterized member. By examining the multiple sequence alignments in each meta-node, we predicted the determinants of the phosphorolytic mechanism and linkage specificity. We thus hypothesized that eight uncharacterized meta-nodes would be phosphorylases. These sequences are characterized by the absence of signal peptides and of the catalytic base. Those sequences with the conserved E/K, E/R and Y/R pairs of residues involved in substrate binding would target β-1,2-, β-1,3- and β-1,4-linked mannosyl residues, respectively. These predictions were tested by characterizing members of three of the uncharacterized meta-nodes from gut bacteria. We discovered the first known β-1,4-mannosyl-glucuronic acid phosphorylase, which targets a motif of the Shigella lipopolysaccharide O-antigen. This work uncovers a reliable strategy for the discovery of novel mannoside-phosphorylases, reveals possible interactions between gut bacteria, and identifies a biotechnological tool for the synthesis of antigenic oligosaccharides.

scholarly journals Functional metagenomics reveals novel β-galactosidases not predictable from gene sequences

2016 ◽  
Author(s):  
Jiujun Cheng ◽  
Tatyana Romantsov ◽  
Katja Engel ◽  
Andrew C. Doxey ◽  
David R. Rose ◽  
...  
Keyword(s):  
Glycoside Hydrolase ◽  
Sinorhizobium Meliloti ◽  
Biochemical Analysis ◽  
Sequence Similarity ◽  
Metagenomic Library ◽  
Glycoside Hydrolases ◽  
Glycoside Hydrolase Family ◽  
Galactosidase Activity ◽  
Functional Metagenomics ◽  
Hydrolase Family

AbstractA soil metagenomic library carried in pJC8 (an IncP cosmid) was used for functional complementation for β-galactosidase activity in bothα-Proteobacteria (Sinorhizobium meliloti)andγ-Proteobacteria (Escherichia coli).Oneβ-galactosidase, encoded by overlapping clones selected in both hosts, was identified as a member of glycoside hydrolase family 2. ORFs obviously encoding possible β-galactosidases were not identified in 19 other clones that were only able to complementS. meliloti.Based on low sequence similarity to known glycoside hydrolases but not β-galactosidases, three ORFs were examined further. Biochemical analysis confirmed that all encodedβ-galactosidase activity. Bioinformatic and structural modeling implied that Lac161_ORF10 protein represented a novel enzyme family with a five-bladed propeller glycoside hydrolase domain.

scholarly journals Protein-carbohydrate Interactions Leading to Hydrolysis and Transglycosylation in Plant Glycoside Hydrolase Family 1 Enzymes

2012 ◽  
Vol 59 (2) ◽  
pp. 51-62 ◽  
Author(s):  
James R. Ketudat Cairns ◽  
Salila Pengthaisong ◽  
Sukanya Luang ◽  
Sompong Sansenya ◽  
Anupong Tankrathok ◽  
...  
Keyword(s):  
Glycoside Hydrolase ◽  
Glycoside Hydrolase Family ◽  
Glycoside Hydrolase Family 1 ◽  
Hydrolase Family
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