In vitro and in vivo characterization of three Cellvibrio japonicus glycoside hydrolase family 5 members reveals potent xyloglucan backbone-cleaving functions

Biotechnology for Biofuels
Mohamed A AttiaHarry Brumer

Abstract

Xyloglucan (XyG) is a ubiquitous and fundamental polysaccharide of plant cell walls. Due to its structural complexity, XyG requires a combination of backbone-cleaving and sidechain-debranching enzymes for complete deconstruction into its component monosaccharides. The soil saprophyte Cellvibrio japonicus has emerged as a genetically tractable model system to study biomass saccharification, in part due to its innate capacity to utilize a wide range of plant polysaccharides for growth. Whereas the downstream debranching enzymes of the xyloglucan utilization system of C. japonicus have been functionally characterized, the requisite backbone-cleaving endo-xyloglucanases were unresolved. Combined bioinformatic and transcriptomic analyses implicated three glycoside hydrolase family 5 subfamily 4 (GH5_4) members, with distinct modular organization, as potential keystone endo-xyloglucanases in C. japonicus. Detailed biochemical and enzymatic characterization of the GH5_4 modules of all three recombinant proteins confirmed particularly high specificities for the XyG polysaccharide versus a panel of other cell wall glycans, including mixed-linkage beta-glucan and cellulose. Moreover, product analysis demonstrated that all three enzymes g...Continue Reading

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Citations

Jul 22, 2019·The Journal of Biological Chemistry·Gregory ArnalHarry Brumer
Aug 20, 2019·Applied and Environmental Microbiology·Guillaume DéjeanHarry Brumer
Nov 2, 2019·Microbiology Resource Announcements·Cecelia A GarciaJeffrey G Gardner
Oct 14, 2020·Chembiochem : a European Journal of Chemical Biology·Elias K ZegeyeAaron T Wright
Feb 8, 2021·Current Opinion in Biotechnology·Evan GlasgowBrian G Fox
Jan 28, 2019·Journal of Molecular Biology·Evan M GlasgowBrian G Fox
May 15, 2021·ACS Chemical Biology·Namrata JainHarry Brumer

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Datasets Mentioned

BETA
ACE84905.1
ACE83841.1
ACE86198.1
GSE109594

Methods Mentioned

BETA
PCR
RNAseq
lipidation
X-ray

Software Mentioned

Coot
lalign
ClustalW
Chromeleon
Buccaneer
Phaser
MolProbity
Privateer
Sigmaplot
AIMLESS

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