Crystal structures of glycoside hydrolase family 3 β-glucosidase 1 from <i>Aspergillus aculeatus</i>

Suzuki, Kentaro; Sumitani, Jun-ichi; Nam, Young‐Woo; Nishimaki, Toru; Tani, Shuji; Wakagi, Takayoshi; Kawaguchi, Takashi; Fushinobu, Shinya

doi:10.1042/bj20130054

Cited by 81 publications

(97 citation statements)

References 58 publications

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“…The structures, which are similar to that reported for the A. aculeatus GH3 enzyme (AaG; PDB entries 4iib, 4iic, 4iid, 4iie, 4iif, 4iig and 4iih; Suzuki et al, 2013), possess the canonical GH3 three-domain structure with an active centre consistent with the recent assignment by the Brumer group (Thongpoo et al, 2013). All three proteins form dimers in the crystal.…”

Section: Introductionsupporting

confidence: 74%

“…4, the glycosylation sites are shown in a novel representation by saccharide type as recently implemented in CCP4mg (McNicholas et al, 2011). The organization of the N-glycan trees in both structures resembles that reported for AaG (Suzuki et al, 2013), with high-mannose trees of similar lengths branching from structurally conserved sites (Tables 2 and 3), with the exceptions that AfG lacks a glycan at Asn212 and that AfG and AoG have an additional GlcNAc bound to Asn543. The glycosylation sites are located in domains A and B only, and predominantly on the opposite side of the molecule with respect to the FnIII domain, with the exception of the tree that starts at Asn253 and extends past a loop of domain C near the C-terminus.…”

Section: Extensive N-glycans; Similarities To Aabg Glycansmentioning

confidence: 65%

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Three-dimensional structures of two heavily N-glycosylatedAspergillussp. family GH3 β-D-glucosidases

Agirre

Ariza

Offen

et al. 2016

Acta Crystallogr D Struct Biol

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The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. These technologies harness diverse plant degrading enzymes, classical exo-and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant -d-linked polysaccharide. A major drawback with this process is that the exo-acting cellobiohydrolases suffer from severe inhibition from their cellobiose product. -d-Glucosidases are therefore important for liberating glucose from cellobiose and thereby relieving limiting product inhibition. Here, the three-dimensional structures of two industrially important family GH3 -d-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley -d-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. Both enzymes show extensive N-glycosylation, with only a few external sites being truncated to a single GlcNAc molecule. Those glycans N-linked to the core of the structure are identified purely as highmannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. The extensive glycans pose special problems for crystallographic refinement, and new techniques and protocols were developed especially for this work. These protocols ensured that all of the d-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy 4 C 1 chair conformation and should be of general application to refinements of other crystal structures containing O-or N-glycosylation. The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal -d-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application.

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Section: Introductionsupporting

confidence: 74%

Section: Extensive N-glycans; Similarities To Aabg Glycansmentioning

confidence: 65%

Three-dimensional structures of two heavily N-glycosylatedAspergillussp. family GH3 β-D-glucosidases

Agirre

Ariza

Offen

et al. 2016

Acta Crystallogr D Struct Biol

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“…The absence of inhibition in the presence of EDTA, DTT, and 2-mercaptoethanol suggest that BxTW1 does not require metallic cations for its catalytic activity and the absence of a disulfide bond near or inside the active site. The nondependence of metal cofactors is a common feature of GH3 proteins, but there are a few solved structures displaying disulfide bonds within this group (22,23).…”

Section: Fig 2 Estimation Of Bxtw1 Molecular Mass By Sds-page (A) Andmentioning

confidence: 99%

Novel pH-Stable Glycoside Hydrolase Family 3 β-Xylosidase from Talaromyces amestolkiae: an Enzyme Displaying Regioselective Transxylosylation

Nieto-Domínguez

Eugenio

Barriuso

et al. 2015

Appl Environ Microbiol

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This paper reports on a novel ␤-xylosidase from the hemicellulolytic fungus Talaromyces amestolkiae. The expression of this enzyme, called BxTW1, could be induced by beechwood xylan and was purified as a glycoprotein from culture supernatants. We characterized the gene encoding this enzyme as an intronless gene belonging to the glycoside hydrolase gene family 3 (GH3). BxTW1 exhibited transxylosylation activity in a regioselective way. This feature would allow the synthesis of oligosaccharides or other compounds not available from natural sources, such as alkyl glycosides displaying antimicrobial or surfactant properties. Regioselective transxylosylation, an uncommon combination, makes the synthesis reproducible, which is desirable for its potential industrial application. BxTW1 showed high pH stability and Cu 2؉ tolerance. The enzyme displayed a pI of 7.6, a molecular mass around 200 kDa in its active dimeric form, and K m and V max values of 0.17 mM and 52.0 U/mg, respectively, using commercial p-nitrophenyl-␤-D-xylopyranoside as the substrate. The catalytic efficiencies for the hydrolysis of xylooligosaccharides were remarkably high, making it suitable for different applications in food and bioenergy industries. P lant biomass represents the most abundant renewable energy resource available on earth. It is composed mainly of cellulose and hemicellulose, two polysaccharides that constitute the raw material for the so-called second-generation (2G) bioethanol industry. The production of this biofuel has received special attention in recent years because it is based on the use of nonfood sources of cellulosic biomass (1). It has been pointed out that energy crops should be restricted to metal-contaminated soils in order to avoid cultivation competition against the food industry (2, 3).In order to make the production of this biofuel economically viable, many modifications have been introduced into the industrial process in recent years. Among them, the strategy of combining enzymatic hydrolysis of lignocellulose with ethanol fermentation in a single process known as simultaneous saccharification and fermentation (SSF) is a significant step forward, but reduced production costs and improved yields are still necessary (1). Most studies have been using agricultural wastes as raw materials, usually after a physicochemical pretreatment to disrupt the lignocellulose structure to enhance cellulose and hemicellulose accessibility. Nevertheless, the industrial procedure currently used to produce 2G ethanol consists of fermenting glucose, which is enzymatically released from cellulose by using Saccharomyces cerevisiae as a biocatalyst (4). To increase process yields, hemicellulose hydrolysis and pentose fermentation are extremely relevant. Within this heterogeneous group of polysaccharides, xylans are most abundant in hardwoods and grass. They are composed of a backbone of ␤-1,4-linked D-xylopyranosyl units highly substituted with arabinofuranose, glucose, glucuronic or methyl-glucuronic acid, and acetyl side groups. The enz...

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“…The crystal structure for a GH3 ␤-glucosidase from A. aculeatus has been published (PDB entry 4IIB-H) (149). AaBGL1 is a dimer in solution, and the monomer consists of three domains: a catalytic TIM barrel-like domain, an ␣/␤ sandwich domain, and an FnIII (fibronectin type III) domain.…”

Section: -5-␦-lactones (134)mentioning

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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Crystal structures of glycoside hydrolase family 3 β-glucosidase 1 from Aspergillus aculeatus

Cited by 81 publications

References 58 publications

Three-dimensional structures of two heavily N-glycosylatedAspergillussp. family GH3 β-D-glucosidases

Three-dimensional structures of two heavily N-glycosylatedAspergillussp. family GH3 β-D-glucosidases

Novel pH-Stable Glycoside Hydrolase Family 3 β-Xylosidase from Talaromyces amestolkiae: an Enzyme Displaying Regioselective Transxylosylation

Genomics Review of Holocellulose Deconstruction by Aspergilli

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