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

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2016年第6期17卷 455-464页

作者： Feng-ying YAN Wei XIA Xiao-xu ZHANG Sha CHEN Xin-zheng NIE Li-chun QIAN The Key Laboratory of Molecular Animal Nutrition Ministry of Education College of Animal Sciences Zhejiang University

An extracellular β-glucosidase produced by Aspergillus terreus was identified, purified, characterized and was tested for the hydrolysis of soybean isofiavone. 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 13-glucosidase showed optimal activity at pH 5.0 and 65℃ and was very stable at 50℃. 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-nitrephenyl-β-D-cjlucopyranoside （pNPG） were 1.73 mmol/L and 42.37 U/mg, respectively. The Krn 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.

关键词： β-glucosidase Aspergillus terreus Characterization Hydrolysis soybean isoflavones

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Apoptosis induction of colorectal cancer cells HTL-9 in vitro by the transformed products of soybean isoflavones by Ganoderma lucidum

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2017年第12期18卷 1101-1112页

作者： Mei-lin CUI Huan-yi YANG Guo-qing HE College of Food Science Shanxi Normal University Linfen 041004 China College of Biosystems Engineering and Food Science Zhejiang University Hangzhou 310058 China Zhejiang Provincial Key Laboratory of Food Microbiology Zhejiang University Hangzhou 310058 China

soybean isoflavones have been one of the potential preventive candidates for antitumor research in recent years. In this paper, we first studied the transformation of soybean isoflavones with the homogenized slurry of Ganoderma lucidum. The resultant transformed products（TSI） contained（703.21±4.35） mg/g of genistein, with transformed rates of 96.63% and 87.82% of daidzein and genistein, respectively, and TSI also could enrich the bioactive metabolites of G. lucidum. The antitumor effects of TSI on human colorectal cancer cell line HTL-9, human breast cancer cell line MCF-7, and human immortalized gastric epithelial cell line GES-1 were also studied. The 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide（MTT） assay showed that TSI could dramatically reduce the viability rates of HTL-9 cells and MCF-7 cells without detectable cytotoxicity on GES-1 normal cells when the TSI concentration was lower than 100 μg/ml. With 100 μg/ml of TSI, HTL-9 cells were arrested in the G1 phase, and late-apoptosis was primarily induced, accompanied with partial early-apoptosis. TSI could induce primarily earlyapoptosis by arresting cells in the G1 phase of MCF-7 cells. For HTL-9 cells, Western-blot and reverse-transcriptase polymerase chain reaction（RT-PCR） analysis showed that TSI（100 μg/ml） can up-regulate the expression of Bax, Caspase-3, Caspase-8, and cytochrome c（Cyto-c）, indicating that TSI could induce cell apoptosis mainly through the mitochondrial pathway. In addition, the expression of p53 was up-regulated, while the expression of Survivin and nuclear factor κB（NF-κB） was down-regulated. All these results showed that TSI could induce apoptosis of HTL-9 cells by the regulation of multiple apoptosis-related genes.

关键词： soybean isoflavones Ganoderma lucidum Transformation Antiproliferative activity Apoptosis

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大豆异黄酮对去卵巢大鼠肝线粒体的抗氧化作用研究

大豆异黄酮对去卵巢大鼠肝线粒体的抗氧化作用研究

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中国食品科学技术学会第五届年会暨第四届东西方食品业高层论坛

作者：蔡美琴张岚杨科峰毛洵霞马蓓蕾高蓓君上海交通大学医学院营养系

目的：探讨大豆异黄酮(soybean isoflavones,SI)对去卵巢大鼠肝线粒体抗氧化能力的影响。方法：3月龄 SD 雌性大鼠随机分为3组,每组10只,假手术对照组和模型组给予去离子水, 干预组给予大豆异黄酮提取物,灌胃。于第13W 末处死大鼠,电镜... 详细信息

目的：探讨大豆异黄酮(soybean isoflavones,SI)对去卵巢大鼠肝线粒体抗氧化能力的影响。方法：3月龄 SD 雌性大鼠随机分为3组,每组10只,假手术对照组和模型组给予去离子水, 干预组给予大豆异黄酮提取物,灌胃。于第13W 末处死大鼠,电镜观察肝线粒体形态结构变化, 氧电极法测定肝线粒体呼吸功能,并测定肝线粒体、胞浆及肝组织匀浆 MDA 的含量,肝组织匀浆 GSH-Px 活性。结果：与对照组比较,模型组和干预组大鼠肝线粒体结构和抗氧化功能均有显著下降,与模型组比较,干预组肝线粒体结构较完整,肝线粒体琥珀酸和 NADH 氧化呼吸链Ⅳ态呼吸速度(R4)显著下降,呼吸控制率(RCR)、磷氧比(P/O)和氧化磷酸化效率(OPR)有明显增加;干预组肝脏线粒体、胞浆及肝组织匀浆 MDA 含量明显下降或有下降趋势：干预组肝组织匀浆 GSH-Px 活性明显增强。结论：大豆异黄酮能够提高去卵巢大鼠肝组织的抗氧化能力。

关键词： soybean isoflavones antioxidant hepatic mitochondria MDA GSH-Px

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