Cloning of a Family 11 Xylanase Gene from Bacillus amyloliquefaciens CH51 Isolated from Cheonggukjang

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. K-m and V-max of XynAHis were 0.363 mg/ml and 701.1 mu mol/min/mg, respectively with birchwood xylan as a substrate. The optimum pH and temperature were pH 4 and 25 A degrees C, respectively. When xynA was introduced into Bacillus subtilis WB600, active XynA was secreted into culture medium.