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CLC number: Q55

On-line Access: 2016-06-03

Received: 2015-12-15

Revision Accepted: 2016-05-05

Crosschecked: 2016-05-12

Cited: 1

Clicked: 1459

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Feng-ying Yan

http://orcid.org/0000-0002-6884-7094

Li-chun Qian

http://orcid.org/0000-0001-8699-1885

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Journal of Zhejiang University SCIENCE B 2016 Vol.17 No.6 P.455-464

10.1631/jzus.B1500317


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


Author(s):  Feng-ying Yan, Wei Xia, Xiao-xu Zhang, Sha Chen, Xin-zheng Nie, Li-chun Qian

Affiliation(s):  The Key Laboratory of Molecular Animal Nutrition, Ministry of Education, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China

Corresponding email(s):   lcqian@zju.edu.cn

Key Words:  β, -glucosidase, Aspergillus terreus, Characterization, Hydrolysis, Soybean isoflavones


Feng-ying Yan, Wei Xia, Xiao-xu Zhang, Sha Chen, Xin-zheng Nie, Li-chun Qian. Characterization of β-glucosidase from Aspergillus terreus and its application in the hydrolysis of soybean isoflavones[J]. Journal of Zhejiang University Science B, 2016, 17(6): 455-464.

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author="Feng-ying Yan, Wei Xia, Xiao-xu Zhang, Sha Chen, Xin-zheng Nie, Li-chun Qian",
journal="Journal of Zhejiang University Science B",
volume="17",
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pages="455-464",
year="2016",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1500317"
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%0 Journal Article
%T Characterization of β-glucosidase from Aspergillus terreus and its application in the hydrolysis of soybean isoflavones
%A Feng-ying Yan
%A Wei Xia
%A Xiao-xu Zhang
%A Sha Chen
%A Xin-zheng Nie
%A Li-chun Qian
%J Journal of Zhejiang University SCIENCE B
%V 17
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%P 455-464
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%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1500317

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T1 - Characterization of β-glucosidase from Aspergillus terreus and its application in the hydrolysis of soybean isoflavones
A1 - Feng-ying Yan
A1 - Wei Xia
A1 - Xiao-xu Zhang
A1 - Sha Chen
A1 - Xin-zheng Nie
A1 - Li-chun Qian
J0 - Journal of Zhejiang University Science B
VL - 17
IS - 6
SP - 455
EP - 464
%@ 1673-1581
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PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1500317


Abstract: 
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-of-flight/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 Km (apparent Michaelis-Menten constant) and Vmax (maximal reaction velocity) for p-nitrophenyl-β;-D-glucopyranoside (pNPG) were 1.73 mmol/L and 42.37 U/mg, respectively. The Km and Vmax 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.

土曲霉来源的β-葡萄糖苷酶的酶学特性及其对大豆异黄酮的水解

目的:分离纯化土曲霉来源的β-葡萄糖苷酶(At-Bgl),探究其酶学特性及其对大豆异黄酮的水解效果。
创新点:成功将At-Bgl分离纯化出来,并首次将分离纯化后的At-Bgl应用于水解大豆异黄酮糖苷。
方法:利用超滤、透析、阴离子交换柱层析和聚丙烯酰氨凝胶电泳(SDS-PAGE)等手段分离纯化土曲霉来源的β-葡萄糖苷酶(表1),并用解析电离串联飞行时间质谱技术(MALDI-TOF/TOF MS)鉴定蛋白条带。以对硝基苯基β-D-葡萄糖苷(pNPG)为底物进行酶学特性研究;以pNPG和纤维二糖为底物,进行酶动力学参数研究 (表2);以胃蛋白酶和胰蛋白酶模拟动物胃肠道酸性环境,进行酸耐受性研究。通过高效液相色谱(HPLC)检测At-Bgl对大豆异黄酮糖苷的水解效果(表3)。
结论:At-Bgl属糖苷水解酶第三家族(GH3),分子量约为120 kDa(图1),最适酶解条件为pH 5.0和65 °C,具有良好的热稳定性和pH稳定性(图2),且胃蛋白酶和胰蛋白酶耐受性强(图3)。At-Bgl可将大豆异黄酮糖苷高效转化为异黄酮苷元 (图4)。综上所述,At-Bgl在增强动物胃肠道对大豆异黄酮的水解方面具有重要应用价值。

关键词:β-葡萄糖苷酶;土曲霉;酶学特性;水解;大豆异黄酮

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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[39]List of electronic supplementary materials

[40]Fig. S1 Morphology of A. terrues culture mycelium

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