Purification and Characterization of a Glucose-tolerant<i>β</i>-Glucosidase from<i>Aspergillus niger</i>CCRC 31494

Yan, Tsong‐Rong; Lin, Chun-Lieh

doi:10.1271/bbb.61.965

Cited by 119 publications

(24 citation statements)

References 13 publications

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“…Of the five cellooligosaccharides tested, As BG1 demonstrated much higher activities, approximately twofold, towards cellotriose to cellohexaose than to cellobiose. The results are contrary to the β-glucosidases derived from Aspergillus oryzae and Aspergillus niger with increased activities on cellobiose [27, 39]. The catalytic efficiencies of As BG1 towards p NPG and cellobiose were 215 and 9 s −1 mM −1 , respectively, which are much higher than the β-glucosidases from Humicola insolens and Trichoderma reesei [18, 33], but lower than Aa β–gly [17].…”

Section: Discussionmentioning

confidence: 88%

A highly glucose-tolerant GH1 β-glucosidase with greater conversion rate of soybean isoflavones in monogastric animals

Cao

Zhang

Shi

et al. 2018

Journal of Industrial Microbiology and Biotechnology

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In the feed industry, β-glucosidase has been widely used in the conversion of inactive and bounded soybean isoflavones into active aglycones. However, the conversion is frequently inhibited by the high concentration of intestinal glucose in monogastric animals. In this study, a GH1 β-glucosidase (AsBG1) with high specific activity, thermostability and glucose tolerance (IC50 = 800 mM) was identified. It showed great glucose tolerance against substrates with hydrophobic aryl ligands (such as pNPG and soy isoflavones). Using soybean meal as the substrate, AsBG1 exhibited higher hydrolysis efficiency than the GH3 counterpart Bgl3A with or without the presence of glucose in the reaction system. Furthermore, it is the first time to find that the endogenous β-glucosidase of soybean meal, mostly belonging to GH3, plays a role in the hydrolysis of soybean isoflavones and is highly sensitive to glucose. These findings lead to a conclusion that the GH1 rather than GH3 β-glucosidase has prosperous application advantages in the conversion of soybean isoflavones in the feed industry.Electronic supplementary materialThe online version of this article (10.1007/s10295-018-2040-6) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 88%

A highly glucose-tolerant GH1 β-glucosidase with greater conversion rate of soybean isoflavones in monogastric animals

Cao

Zhang

Shi

et al. 2018

Journal of Industrial Microbiology and Biotechnology

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“…The optimum pH (6.0) of the purified enzyme is similar to that of Flavoba cterium (18), and higher than that of Aspergillus ornatus (4.6) (19) and A. niger CCRC 31494 (5.0) (20). The enzyme had a wide pH range of 3.0-9.0 for its activity.…”

Section: Discussionmentioning

confidence: 79%

Purification and characterization of an extracellular β-glucosidase with high transglucosylation activity and stability from Aspergillus niger No. 5.1

Xie

Gao

Chen

2004

Appl Biochem Biotechnol

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An extracellular beta-glucosidase was extracted from the culture filtrate of Aspergillus niger No. 5.1 and purified to homogeneity by using ammonium sulfate precipitation, Chitopearl-DEAE chromatography, and Sephadex G-100 chromatography. The specific activity of the enzyme was enriched 6.33-fold, with a recovery of 11.67%. The enzyme was a monomer and the molecular mass was 67.5 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis and 66.5 kDa by gel-filtration chromatography. The enzyme had optimum activity at pH 6.0 and 60 degrees C and was stable over the pH range of 3.0-9.0. It showed specificity of hydrolysis for p-nitrophenyl-beta-D-glucoside and cellobiose. The Km and Vmax values of the enzyme for cellobiose and salicin were 5.34 mM, 2.57 micromol/(mL.s), and 3.09 mM, 1.34 micromol/(mL.s), respectively. Both amino acid composition and N-terminal amino acid sequence of the enzyme were determined, which provides useful information for cloning of this enzyme.

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“…To date, several glucose-tolerant BGLs have been identified in insects (Uchima et al, 2011), fungi (Saha and Bothast, 1996; Yan and Lin, 1997; Riou et al, 1998; Decker et al, 2001; Zanoelo et al, 2004; Souza et al, 2014), bacteria (Pérez-Pons et al, 1995), and metagenomes (Fang et al, 2010; Biver et al, 2014). Recently, we have identified a glucose-tolerant BGL (Td2F2) in a wood compost metagenomic library (Uchiyama et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

Glucose-tolerant Î²-glucosidase retrieved from a Kusaya gravy metagenome

2015

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β-glucosidases (BGLs) hydrolyze cello-oligosaccharides to glucose and play a crucial role in the enzymatic saccharification of cellulosic biomass. Despite their significance for the production of glucose, most identified BGLs are commonly inhibited by low (∼mM) concentrations of glucose. Therefore, BGLs that are insensitive to glucose inhibition have great biotechnological merit. We applied a metagenomic approach to screen for such rare glucose-tolerant BGLs. A metagenomic library was created in Escherichia coli (∼10,000 colonies) and grown on LB agar plates containing 5-bromo-4-chloro-3-indolyl-β-D-glucoside, yielding 828 positive (blue) colonies. These were then arrayed in 96-well plates, grown in LB, and secondarily screened for activity in the presence of 10% (w/v) glucose. Seven glucose-tolerant clones were identified, each of which contained a single bgl gene. The genes were classified into two groups, differing by two nucleotides. The deduced amino acid sequences of these genes were identical (452 aa) and found to belong to the glycosyl hydrolase family 1. The recombinant protein (Ks5A7) was overproduced in E. coli as a C-terminal 6 × His-tagged protein and purified to apparent homogeneity. The molecular mass of the purified Ks5A7 was determined to be 54 kDa by SDS-PAGE, and 160 kDa by gel filtration analysis. The enzyme was optimally active at 45°C and pH 5.0–6.5 and retained full or 1.5–2-fold enhanced activity in the presence of 0.1–0.5 M glucose. It had a low KM (78 μM with p-nitrophenyl β-D-glucoside; 0.36 mM with cellobiose) and high Vmax (91 μmol min-1 mg-1 with p-nitrophenyl β-D-glucoside; 155 μmol min-1 mg-1 with cellobiose) among known glucose-tolerant BGLs and was free from substrate (0.1 M cellobiose) inhibition. The efficient use of Ks5A7 in conjunction with Trichoderma reesei cellulases in enzymatic saccharification of alkaline-treated rice straw was demonstrated by increased production of glucose.

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Purification and Characterization of a Glucose-tolerantβ-Glucosidase fromAspergillus nigerCCRC 31494

Cited by 119 publications

References 13 publications

A highly glucose-tolerant GH1 β-glucosidase with greater conversion rate of soybean isoflavones in monogastric animals

A highly glucose-tolerant GH1 β-glucosidase with greater conversion rate of soybean isoflavones in monogastric animals

Purification and characterization of an extracellular β-glucosidase with high transglucosylation activity and stability from Aspergillus niger No. 5.1

Glucose-tolerant Î²-glucosidase retrieved from a Kusaya gravy metagenome

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