Purification, Characterization, Cloning, and Expression of a Novel Xyloglucan-specific Glycosidase, Oligoxyloglucan Reducing End-specific Cellobiohydrolase

Yaoi, Katsuro; Mitsuishi, Yasushi

doi:10.1074/jbc.m208443200

Cited by 74 publications

(80 citation statements)

References 30 publications

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“…1) to generate the oligosaccharides XXXG, XLXG, XXLG and XLLG, which differ in their degree of side-chain galactosylation (Figs 6 and 7). This is the most common cleavage pattern observed for GH74 endo-xyloglucanases [38,41,[48][49][50], although certain members are known to cleave between branched backbone glucosyl units [42,43,51,52].…”

Section: Bond Cleavage Specificity and Mode Of Action Of Cjgh74mentioning

confidence: 99%

Functional and structural characterization of a potent GH74 endo‐xyloglucanase from the soil saprophyte Cellvibrio japonicus unravels the first step of xyloglucan degradation

et al. 2016

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The heteropolysaccharide xyloglucan (XyG) comprises up to one‐quarter of the total carbohydrate content of terrestrial plant cell walls and, as such, represents a significant reservoir in the global carbon cycle. The complex composition of XyG requires a consortium of backbone‐cleaving endo‐xyloglucanases and side‐chain cleaving exo‐glycosidases for complete saccharification. The biochemical basis for XyG utilization by the model Gram‐negative soil saprophytic bacterium Cellvibrio japonicus is incompletely understood, despite the recent characterization of associated side‐chain cleaving exo‐glycosidases. We present a detailed functional and structural characterization of a multimodular enzyme encoded by gene locus CJA_2477. The CJA_2477 gene product comprises an N‐terminal glycoside hydrolase family 74 (GH74) endo‐xyloglucanase module in train with two carbohydrate‐binding modules (CBMs) from families 10 and 2 (CBM10 and CBM2). The GH74 catalytic domain generates Glc4‐based xylogluco‐oligosaccharide (XyGO) substrates for downstream enzymes through an endo‐dissociative mode of action. X‐ray crystallography of the GH74 module, alone and in complex with XyGO products spanning the entire active site, revealed a broad substrate‐binding cleft specifically adapted to XyG recognition, which is composed of two seven‐bladed propeller domains characteristic of the GH74 family. The appended CBM10 and CBM2 members notably did not bind XyG, nor other soluble polysaccharides, and instead were specific cellulose‐binding modules. Taken together, these data shed light on the first step of xyloglucan utilization by C. japonicus and expand the repertoire of GHs and CBMs for selective biomass analysis and utilization. Database Structural data have been deposited in the RCSB protein database under the Protein Data Bank codes: http://www.rcsb.org/pdb/search/structidSearch.do?structureId=5FKR, http://www.rcsb.org/pdb/search/structidSearch.do?structureId=5FKS, http://www.rcsb.org/pdb/search/structidSearch.do?structureId=5FKT and http://www.rcsb.org/pdb/search/structidSearch.do?structureId=5FKQ.

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Section: Bond Cleavage Specificity and Mode Of Action Of Cjgh74mentioning

confidence: 99%

Functional and structural characterization of a potent GH74 endo‐xyloglucanase from the soil saprophyte Cellvibrio japonicus unravels the first step of xyloglucan degradation

et al. 2016

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“…Two GH74 enzymes have been shown to be reducing-end-specific cellobiohydrolases (EC 3.2.1.150) (195,196) releasing XG, LG, or FG from xyloglucan [X, G, L, and (197). The exo-acting activity is believed to be due to a loop insertion closing off the positive subsites (198).…”

Section: Hemicellulasesmentioning

confidence: 99%

Genomics Review of Holocellulose Deconstruction by Aspergilli

Segato

Damásio

Lucas

et al. 2014

Microbiol Mol Biol Rev

115

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SUMMARY Biomass is constructed of dense recalcitrant polymeric materials: proteins, lignin, and holocellulose, a fraction constituting fibrous cellulose wrapped in hemicellulose-pectin. Bacteria and fungi are abundant in soil and forest floors, actively recycling biomass mainly by extracting sugars from holocellulose degradation. Here we review the genome-wide contents of seven Aspergillus species and unravel hundreds of gene models encoding holocellulose-degrading enzymes. Numerous apparent gene duplications followed functional evolution, grouping similar genes into smaller coherent functional families according to specialized structural features, domain organization, biochemical activity, and genus genome distribution. Aspergilli contain about 37 cellulase gene models, clustered in two mechanistic categories: 27 hydrolyze and 10 oxidize glycosidic bonds. Within the oxidative enzymes, we found two cellobiose dehydrogenases that produce oxygen radicals utilized by eight lytic polysaccharide monooxygenases that oxidize glycosidic linkages, breaking crystalline cellulose chains and making them accessible to hydrolytic enzymes. Among the hydrolases, six cellobiohydrolases with a tunnel-like structural fold embrace single crystalline cellulose chains and cooperate at nonreducing or reducing end termini, splitting off cellobiose. Five endoglucanases group into four structural families and interact randomly and internally with cellulose through an open cleft catalytic domain, and finally, seven extracellular β-glucosidases cleave cellobiose and related oligomers into glucose. Aspergilli contain, on average, 30 hemicellulase and 7 accessory gene models, distributed among 9 distinct functional categories: the backbone-attacking enzymes xylanase, mannosidase, arabinase, and xyloglucanase, the short-side-chain-removing enzymes xylan α-1,2-glucuronidase, arabinofuranosidase, and xylosidase, and the accessory enzymes acetyl xylan and feruloyl esterases.

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“…For example, isoprimeverose-producing oligoxyloglucan hydrolase (IPase) (EC 3.2.1.120), which cleaves xyloglucan oligosaccharide at the nonreducing end to produce an isoprimeverose [␣-D-Xylp-(136)-␤-D-Glcp-] residue, is a useful tool for analyzing xyloglucan and for preparing various xyloglucan oligosaccharide structures (9,(11)(12)(13)(14)(15)33). An oligoxyloglucan reducing-endspecific cellobiohydrolase (OXG-RCBH) (EC 3.2.1.150), which is a unique exoglucanase that cleaves xyloglucan oligosaccharides at their reducing ends (30), and a xyloglucan-specific endo-␤-1,4-glucanase (xyloglucanase [XEG]) (29) from Geotrichum sp. strain M128, which we recently screened and cloned, have also been used for analyzing xyloglucan structure (11,12).…”

mentioning

confidence: 99%

Cloning and Characterization of Two Xyloglucanases from Paenibacillus sp. Strain KM21

Yaoi

Nakai

Kameda

et al. 2005

Appl Environ Microbiol

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Two xyloglucan-specific endo-␤-1,4-glucanases (xyloglucanases [XEGs]), XEG5 and XEG74, with molecular masses of 40 kDa and 105 kDa, respectively, were isolated from the gram-positive bacterium Paenibacillus sp. strain KM21, which degrades tamarind seed xyloglucan. The genes encoding these XEGs were cloned and sequenced. Based on their amino acid sequences, the catalytic domains of XEG5 and XEG74 were classified in the glycoside hydrolase families 5 and 74, respectively. XEG5 is the first xyloglucanase belonging to glycoside hydrolase family 5. XEG5 lacks a carbohydrate-binding module, while XEG74 has an X2 module and a family 3 type carbohydrate-binding module at its C terminus. The two XEGs were expressed in Escherichia coli, and recombinant forms of the enzymes were purified and characterized. Both XEGs had endoglucanase active only toward xyloglucan and not toward Avicel, carboxymethylcellulose, barley ␤-1,3/1,4-glucan, or xylan. XEG5 is a typical endo-type enzyme that randomly cleaves the xyloglucan main chain, while XEG74 has dual endo-and exo-mode activities or processive endo-mode activity. XEG5 digested the xyloglucan oligosaccharide XXXG XXXG to produce XXXG, whereas XEG74 digestion of XXXGXXXG resulted in XXX, XXXG, and GXXXG, suggesting that this enzyme cleaves the glycosidic bond of unbranched Glc residues. Analyses using various oligosaccharide structures revealed that unique structures of xyloglucan oligosaccharides can be prepared with XEG74.

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Purification, Characterization, Cloning, and Expression of a Novel Xyloglucan-specific Glycosidase, Oligoxyloglucan Reducing End-specific Cellobiohydrolase

Cited by 74 publications

References 30 publications

Functional and structural characterization of a potent GH74 endo‐xyloglucanase from the soil saprophyte Cellvibrio japonicus unravels the first step of xyloglucan degradation

Functional and structural characterization of a potent GH74 endo‐xyloglucanase from the soil saprophyte Cellvibrio japonicus unravels the first step of xyloglucan degradation

Genomics Review of Holocellulose Deconstruction by Aspergilli

Cloning and Characterization of Two Xyloglucanases from Paenibacillus sp. Strain KM21

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