Division of Applied Life Science (Bk21), Graduate School, Research Institute of Life Sciences, Gyeongsang National University, Jinju, Korea.
Indian J Microbiol. 2012 Dec;52(4):695-700. doi: 10.1007/s12088-012-0260-4. Epub 2012 Mar 25.
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.
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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