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Cloning and expression of β-glucosidases from Bifidobacterium lactis AD011

机译：乳酸双歧杆菌AD011中β-葡萄糖苷酶的克隆与表达

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摘要

For the conversion of β-glycosides, 3 genes (BLA_0039, BLA_0141, and BLA_0893) annotated as β-glucosidase (E.C 3.2.1.21) from the genome of Bifidobacterium lactis AD011 were cloned and expressed both in Escherichia coli DH5α and Bifidobacterium bifidum BGN4 using previously established bifidobacterial expression system under the control of 16S rRNA promoter. Deduced amino acid sequence analysis revealed that BLA_0141 is highly similar to β-d-glucosidase from Bifidobacterium breve clb and is included in glycosyl hydrolase family 1 (GHF1) while BLA_0893 belongs to glycosyl hydrolase family 3 (GHF3). Recombinant BLA_0893 showed β-glucosidase activity and converted ginsenoside Rb1 to Rd. Recombinant BLA_0141 showed broad range of activities including β-glucosidase, β-galactosidase, β-xylosidase, cellobiosidase, and α-arabinopyranosidase and converted ginsenoside Rb1 and Rb2 to Rd. They also hydrolized cellobiose into glucose. The recombinant enzyme activities of both BLA_0141 and BLA_0893 were stable around pH 4.5–7.5 and below 50°C.
机译：为了转化β-糖苷，克隆了来自乳酸双歧杆菌AD011基因组的注释为β-葡萄糖苷酶（EC 3.2.1.21）的3个基因（BLA_0039，BLA_0141和BLA_0893），并在大肠杆菌DH5α和双歧双歧杆菌BGN4中进行了表达。以前在16S rRNA启动子的控制下建立的双歧杆菌表达系统。推导的氨基酸序列分析表明，BLA_0141与短双歧杆菌clb的β-d-葡萄糖苷酶高度相似，并包含在糖基水解酶家族1（GHF1）中，而BLA_0893属于糖基水解酶家族3（GHF3）。重组BLA_0893显示出β-葡糖苷酶活性并将人参皂苷Rb1转化为Rd。重组BLA_0141表现出广泛的活性，包括β-葡萄糖苷酶，β-半乳糖苷酶，β-木糖苷酶，纤维二糖苷酶和α-阿拉伯吡喃糖苷酶，并将人参皂苷Rb1和Rb2转化为Rd。他们还将纤维二糖水解为葡萄糖。 BLA_0141和BLA_0893的重组酶活性在pH 4.5–7.5和低于50°C的温度下均稳定。

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  • 来源
    《Food Science and Biotechnology》 |2012年第3期|p.731-738|共8页
  • 作者

    Jin Yong Kim; Yan Wang; Su Ji Park; Geun Eog Ji; Myeong Soo Park;

  • 作者单位

    Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University, Seoul, 151-742, Korea;

    Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University, Seoul, 151-742, Korea;

    Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University, Seoul, 151-742, Korea;

    Research Institute, BIFIDO Co., Ltd., Hongcheon, Gangwon, 250-804, Korea;

    Research Institute, BIFIDO Co., Ltd., Hongcheon, Gangwon, 250-804, Korea;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Bifidobacterium; β-glucosidase; cloning; expression;

    机译：双歧杆菌;β-葡萄糖苷酶;克隆;表达;

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