从清曲中分离得到的解淀粉芽孢杆菌 CH51 中家族 11 木聚糖酶基因的克隆。
Cloning of a Family 11 Xylanase Gene from Bacillus amyloliquefaciens CH51 Isolated from Cheonggukjang.
机构信息
Division of Applied Life Science (Bk21), Graduate School, Research Institute of Life Sciences, Gyeongsang National University, Jinju, Korea.
出版信息
Indian J Microbiol. 2012 Dec;52(4):695-700. doi: 10.1007/s12088-012-0260-4. Epub 2012 Mar 25.
Abstract
Bacillus amyloliquefaciens CH51, an isolate from cheonggukjang, Korean fermented soyfood, secretes several enzymes into culture medium. A gene encoding 19 kDa xylanase was cloned by PCR. Sequencing showed that the gene encoded a glycohydrolase family 11 xylanase and named xynA. xynAHis, xynA with additional codons for his-tag, was overexpressed in Escherichia coli BL21(DE3) using pET-26b(+). XynAHis was purified using HisTrap affinity column. Km and Vmax of XynAHis were 0.363 mg/ml and 701.1 μmol/min/mg, respectively with birchwood xylan as a substrate. The optimum pH and temperature were pH 4 and 25 °C, respectively. When xynA was introduced into Bacillus subtilis WB600, active XynA was secreted into culture medium.
摘要
解淀粉芽胞杆菌 CH51 是一种从韩国发酵大豆食品——清曲中分离出来的菌株，它会向培养基中分泌几种酶。通过 PCR 克隆了一个编码 19 kDa 木聚糖酶的基因。测序表明，该基因编码一个糖苷水解酶家族 11 木聚糖酶，并将其命名为 xynA。xynAHis 是在大肠杆菌 BL21(DE3)中使用 pET-26b(+)过表达的，带有用于 his-tag 的额外密码子。使用 HisTrap 亲和层析柱对 XynAHis 进行纯化。以桦木木聚糖为底物时，XynAHis 的 Km 和 Vmax 分别为 0.363 mg/ml 和 701.1 μmol/min/mg。最适 pH 和温度分别为 pH 4 和 25°C。当 xynA 被引入枯草芽孢杆菌 WB600 时，活性 XynA 被分泌到培养基中。
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