Molecular cloning and characterization of a novel β-glucosidase with high hydrolyzing ability for soybean isoflavone glycosides and glucose-tolerance from soil metagenomic library

Li, Gang; Jiang, Yang; Fan, Xiaoran; Liu, Yuhuan

doi:10.1016/j.biortech.2012.07.083

Cited by 89 publications

(56 citation statements)

References 34 publications

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“…2a), which was slightly higher than the previous reports (Li et al, 2012;2013;Fujita et al, 2015;Kaur and Chadha, 2015;Zhao et al, 2015). In addition, the enzyme showed more than 97% relative activity at 75 °C, and showed no significant difference from the activity at 65 °C (P>0.05), while other fungal counterparts were generally inactivated at 70 °C (Li et al, 2013;Fujita et al, 2015;Kaur and Chadha, 2015;Zhao et al, 2015).…”

Section: Optimal Ph and Temperature And Stabilitycontrasting

confidence: 54%

“…A control reaction of extract samples without enzyme was set up in the same manner. The determination of isoflavone in hydrolyzed samples was determined by high performance liquid chromatography (HPLC) as previously described (Li et al, 2012). HPLC analyses were performed using a Waters HP1100 HPLC system (Milford, MA, USA) equipped with a Diamonsil C18 column (5 μm, 250 mm×4.6 mm; Dima Co., Ltd., Orlando, FL, USA), and six types of high purity soybean isoflavones, daidzin, glycitin, genistin, daidzein, glycitein, and genistein, were used as standards.…”

Section: Isoflavone Extraction From Soybean Flour and Enzymatic Hydromentioning

confidence: 99%

“…2c), maintaining 90.0% and 65.1% of the initial activity after incubation at 50 and 60 °C for 1 h, respectively, though it decreased significantly after incubation at 50, 60, or 70 °C for 5 min (P<0.05). These superior properties make this purified enzyme more advantageous for industrial processes, since most applications of β-glucosidase require high temperatures (50 °C or above) (Li et al, 2012).…”

Section: Optimal Ph and Temperature And Stabilitymentioning

confidence: 99%

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Characterization of β-glucosidase from Aspergillus terreus and its application in the hydrolysis of soybean isoflavones

Yan

Xia

Zhang

et al. 2016

J. Zhejiang Univ. Sci. B

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An extracellular β-glucosidase produced by Aspergillus terreus was identified, purified, characterized and was tested for the hydrolysis of soybean isoflavone. Matrix-assisted laser desorption/ionization with tandem time-offlight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS) revealed the protein to be a member of the glycosyl hydrolase family 3 with an apparent molecular mass of about 120 kDa. The purified β-glucosidase showed optimal activity at pH 5.0 and 65 °C and was very stable at 50 °C. Moreover, the enzyme exhibited good stability over pH 3.0-8.0 and possessed high tolerance towards pepsin and trypsin. The kinetic parameters K m (apparent MichaelisMenten constant) and V max (maximal reaction velocity) for p-nitrophenyl-β-D-glucopyranoside (pNPG) were 1.73 mmol/L and 42.37 U/mg, respectively. The K m and V max for cellobiose were 4.11 mmol/L and 5.7 U/mg, respectively. The enzyme efficiently converted isoflavone glycosides to aglycones, with a hydrolysis rate of 95.8% for daidzin, 86.7% for genistin, and 72.1% for glycitin. Meanwhile, the productivities were 1.14 mmol/(L·h) for daidzein, 0.72 mmol/(L·h) for genistein, and 0.19 mmol/(L·h) for glycitein. This is the first report on the application of A. terreus β-glucosidase for converting isoflavone glycosides to their aglycones in soybean products.

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Section: Optimal Ph and Temperature And Stabilitycontrasting

confidence: 54%

Section: Isoflavone Extraction From Soybean Flour and Enzymatic Hydromentioning

confidence: 99%

Section: Optimal Ph and Temperature And Stabilitymentioning

confidence: 99%

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Characterization of β-glucosidase from Aspergillus terreus and its application in the hydrolysis of soybean isoflavones

Yan

Xia

Zhang

et al. 2016

J. Zhejiang Univ. Sci. B

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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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“…More than 99% of the microorganisms cannot be cultured using conventional methods, which leaves a great deal of industry-potential β-glucosidases unmined [18]. The metagenomic approach has been successfully employed in the isolation and identification of novel β-glucosidases from various samples [19][20][21][22]. Nevertheless, the hydrolysis activity of these β-glucosidases for polydatin has not been investigated until now.…”

Section: Introductionmentioning

confidence: 99%

Characterization of a Metagenome-Derived β-Glucosidase and Its Application in Conversion of Polydatin to Resveratrol

Mai

Zhang

2016

Catalysts

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Abstract:For the beneficial pharmacological properties of resveratrol, there is increasingly interest in enzymatic conversion of polydatin to resveratrol. The metagenomic technique provides an effective strategy for mining novel polydatin-hydrolysis enzymes from uncultured microorganisms. In this study, a metagenomic library of mangrove soil was constructed and a novel β-glucosidase gene MlBgl was isolated. The deduced amino acid sequences of MlBgl showed the highest identity of 64% with predicted β-glucosidase in the GenBank database. The gene was cloned and overexpressed in Escherichia coli BL21(DE3). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) assay demonstrated the purified recombinant β-glucosidase r-MlBgl with a molecular weight approximately of 71 kDa. The optimal pH and temperature of purified recombinant r-MlBgl were 7.0 and 40˝C, respectively. r-MlBgl could hydrolyze polydatin effectively. The k cat and k cat /K m values for polydatin were 989 s´1 and 1476 mM´1¨s´1, respectively. These properties suggest that -r-MlBgl has potential application in the enzymatic conversion of polydatin to resveratrol for further study.

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Molecular cloning and characterization of a novel β-glucosidase with high hydrolyzing ability for soybean isoflavone glycosides and glucose-tolerance from soil metagenomic library

Cited by 89 publications

References 34 publications

Characterization of β-glucosidase from Aspergillus terreus and its application in the hydrolysis of soybean isoflavones

Characterization of β-glucosidase from Aspergillus terreus and its application in the hydrolysis of soybean isoflavones

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

Characterization of a Metagenome-Derived β-Glucosidase and Its Application in Conversion of Polydatin to Resveratrol

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