Expression and characterization of a novel highly glucose-tolerant &amp;beta;-glucosidase from a soil metagenome

Lü, Jian; Du, Liqin; Wei, Yutuo; Hu, Yuanyuan; Huang, Ribo

doi:10.1093/abbs/gmt061

Cited by 50 publications

(20 citation statements)

References 36 publications

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“…yunnanensis 258 ± 5.1182 ± 3.42.2 and 1.4 for glucose and xylose, respectively842 ± 15.560~10 h at 60 °C;BacteriumPresent work S. thermophilum 204 ± 12191 ± 140.7 and 0.5 for glucose and xylose, respectively8.96020 min at 55 °CFungi[8] H. insolens 180 ± 9200 ± 110.436.46044 min at 55 °CFungi[7] H. grisea var. thermoidea 1882020.529.5507 min at 60 °CFungi[23] B. halodurans 135.0 ± 4.3131.0 ± 0.2~0.8-0.9935030 min at 50 °CBacterium[24]Uncultured bacterium1201500.3 for glucose183.950 ND Soil metagenome[5] Marinomonas sp. MWYL1~132~1450.638940~11 min at 45 °CBacterium[9]

ND not determined

a One hundred percent specific activity (control) was estimated in the absence of carbohydrates

…”

Section: Discussionmentioning

confidence: 99%

Increased enzymatic hydrolysis of sugarcane bagasse by a novel glucose- and xylose-stimulated β-glucosidase from Anoxybacillus flavithermus subsp. yunnanensis E13T

Liu

Wang

et al. 2017

BMC Biochem

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Backgroundβ-Glucosidase is claimed as a key enzyme in cellulose hydrolysis. The cellulosic fibers are usually entrapped with hemicelluloses containing xylose. So there is ongoing interest in searching for glucose- and xylose-stimulated β-glucosidases to increase the efficiency of hydrolysis of cellulosic biomass.ResultsA thermostable β-glucosidase gene (Bglp) was cloned from Anoxybacillus flavithermus subsp. yunnanensis E13T and characterized. Optimal enzyme activity was observed at 60 °C and pH 7.0. Bglp was relatively stable at 60 °C with a 10-h half-life. The kinetic parameters V max and K m for p-nitrophenyl-β-D-glucopyranoside (pNPG) were 771 ± 39 μmol/min/mg and 0.29 ± 0.01 mM, respectively. The activity of Bglp is dramatically stimulated by glucose or xylose at concentrations up to 1.4 M. After Bglp was added to Celluclast® 1.5 L, the conversion of sugarcane bagasse was 48.4 ± 0.8%, which was much higher than of Celluclast® 1.5 L alone. Furthermore, Bglp showed obvious advantages in the hydrolysis when initial concentrations of glucose and xylose are high.ConclusionsThe supplementation of BglP significantly enhanced the glucose yield from sugarcane bagasse, especially in the presence of high concentrations of glucose or xylose. Bglp should be a promising candidate for industrial applications.Electronic supplementary materialThe online version of this article (doi:10.1186/s12858-017-0079-z) contains supplementary material, which is available to authorized users.

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ND not determined

a One hundred percent specific activity (control) was estimated in the absence of carbohydrates

…”

Section: Discussionmentioning

confidence: 99%

Increased enzymatic hydrolysis of sugarcane bagasse by a novel glucose- and xylose-stimulated β-glucosidase from Anoxybacillus flavithermus subsp. yunnanensis E13T

Liu

Wang

et al. 2017

BMC Biochem

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“…For examples, β-glucosidase from Microbispora bispora was activated in the presence of 300 mM glucose [135] and that reported in T. thermosaccharolyticum DSM 571 and T. aotearoense was activated by glucose concentration < 200 and 250 mM, respectively [85,136]. Similarly, recombinant GH 1 β-glucosidase from Exiguobacterium antarcticum B7 [84], Fervidobacterium islandicum [137], Thermotoga thermarum DSM 5069T [138] and Anoxybacillus DT3-1 [83] was tolerant to high glucose concentration e.g., Ki of 0.54-15 M. On the other hand, glucose tolerant β-glucosidase with Ki of 1.0-4.8 M has been identified from different environments using metagenomic approaches [45,53,106,111,139,140].…”

Section: Glucose Sensitivity and Tolerancementioning

confidence: 99%

Microbial β-Glucosidases: Screening, Characterization, Cloning and Applications

Ahmed¹,

Aslam²,

Ashraf³

et al. 2017

JAEM

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Cellulose is the most abundant biomaterial in the biosphere and the major component of plant biomass.Cellulase is an enzymatic system required for conversion of renewable cellulose biomass into free sugar for subsequent use in different applications. Cellulase system mainly consists of three individual enzymes namely: endoglucanase, exoglucanase and β-glucosidases. β-Glucosidases are ubiquitous enzymes found in all living organisms with great biological significance. β-Glucosidases have also tremendous biotechnological applications such as biofuel production, beverage industry, food industry, cassava detoxification and oligosaccharides synthesis. Microbial β-glucosidases are preferred for industrial uses because of robust activity and novel properties exhibited by them. This review aims at describing the various biochemical methods used for screening and evaluating β-glucosidases activity from microbial sources. Subsequently, it generally highlights techniques used for purification of β-glucosidases. It then elaborates various biochemical and molecular properties of this valuable enzyme such as pH and temperature optima, glucose tolerance, substrate specificity, molecular weight, and multiplicity. Furthermore, it describes molecular cloning and expression of bacterial, fungal and metagenomic β-glucosidases. Finally, it highlights the potential biotechnological applications of β-glucosidases.

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“…Cocktails have generally been based on commercial enzymes isolated from different microorganisms, which may have different optimal growth conditions. Recently, the isolation of novel enzymes, including thermostable enzymes from metagenomic libraries, has been extensively studied, leading to the isolation of many enzymes with special characteristics such as being cold- (Wierzbicka-Wos et al, 2013 ) or heat-active (Schroder et al, 2014 ), and ionic liquid- (Ilmberger et al, 2012 ) or product-tolerant (Lu et al, 2013 ). Microorganisms in the same metagenome generally live together for a relatively long time, and should have similar tolerance for environmental conditions, such as temperature, pH, and oxidation-reduction potentials, which may result in isolation of enzymes with similar optimal conditions.…”

Section: Discussionmentioning

confidence: 99%

Synergistic function of four novel thermostable glycoside hydrolases from a long-term enriched thermophilic methanogenic digester

et al. 2015

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In biofuel production from lignocellulose, low thermostability and product inhibition strongly restrict the enzyme activities and production process. Application of multiple thermostable glycoside hydrolases, forming an enzyme “cocktail”, can result in a synergistic action and therefore improve production efficiency and reduce operational costs. Therefore, increasing enzyme thermostabilities and compatibility are important for the biofuel industry. In this study, we reported the screening, cloning and biochemical characterization of four novel thermostable lignocellulose hydrolases from a metagenomic library of a long-term dry thermophilic methanogenic digester community, which were highly compatible with optimal conditions and specific activities. The optimal temperatures of the four enzymes, β-xylosidase, xylanase, β-glucosidase, and cellulase ranged from 60 to 75°C, and over 80% residual activities were observed after 2 h incubation at 50°C. Mixtures of these hydrolases retained high residual synergistic activities after incubation with cellulose, xylan, and steam-exploded corncob at 50°C for 72 h. In addition, about 55% dry weight of steam-exploded corncob was hydrolyzed to glucose and xylose by the synergistic action of the four enzymes at 50°C for 48 h. This work suggested that since different enzymes from a same ecosystem could be more compatible, screening enzymes from a long-term enriching community could be a favorable strategy.

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Expression and characterization of a novel highly glucose-tolerant β-glucosidase from a soil metagenome

Cited by 50 publications

References 36 publications

Increased enzymatic hydrolysis of sugarcane bagasse by a novel glucose- and xylose-stimulated β-glucosidase from Anoxybacillus flavithermus subsp. yunnanensis E13T

Increased enzymatic hydrolysis of sugarcane bagasse by a novel glucose- and xylose-stimulated β-glucosidase from Anoxybacillus flavithermus subsp. yunnanensis E13T

Microbial β-Glucosidases: Screening, Characterization, Cloning and Applications

Synergistic function of four novel thermostable glycoside hydrolases from a long-term enriched thermophilic methanogenic digester

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