脂环酸芽孢杆菌Alicyclobacillus sp. A15木聚糖酶基因xynA15的克隆表达及性质研究  

Cloning,Expression and Characterization Studies on Xylanase Gene xynA15 from Alicyclobacillus sp. A15

作　　者：王建设[1,2] 柏映国[2] 尹俊[1] 姚斌[2] 

机构地区：[1]内蒙古农业大学生命科学学院,呼和浩特010018 [2]中国农业科学院饲料研究所,农业部饲料生物技术重点开放实验室,北京100081

出　　处：《中国农业科技导报》2010年第6期114-119,共6页Journal of Agricultural Science and Technology

基　　金：国家转基因生物新品种培育重大专项(2008ZX003-002);国家肉鸡产业技术体系(nycytx-42-G2-05)资助

摘　　要：从云南保山市的一眼温泉中分离到一株嗜热嗜酸菌,脂环酸芽孢杆菌Alicyclobacillus sp. A15。通过同源克隆和TAIL-PCR方法,从该菌株中克隆得到一个木聚糖酶基因xynA15,全长990 bp,编码329个氨基酸和一个终止密码子。将xynA15基因克隆到原核表达载体pET-22b（＋）上,并转化至BL21（DE3）,经过IPTG诱导,其表达产物具有木聚糖酶的活性。对其酶学性质研究发现,XynA15的最适温度为50℃,最适pH为7.0,不仅在较广的温度范围（35℃-60℃）内具有较高的酶活,而且在50℃具有较好的热稳定性。Alicyclobacillus sp.A15 was isolated from the outflow of a hot spring in Baoshan City,Yunnan Province,China.xynA15 was cloned from the strain through homogolous cloning and TAIL-PCR,which consists of 990 bp and encodes 329 amino acids and one stop codon.Then it was inserted into the prokaryotic expression vector pET-22b（＋） and transformed into Escherichia coli BL21（DE3）.Xylanase activity was detected in the cultured supernantant after induced by isopropyl-β-d-1-thiogalactopyranoside（IPTG）.The optimal temperature and pH was 50℃ and 7.0,respectively for XynA15 activity.XynA15 contains higher activity at 35-60℃ and better thermal stability at 50℃.

关 键 词：ALICYCLOBACILLUS sp. A15 木聚糖酶 酶学性质 原核表达 

分 类 号：Q786[生物学—分子生物学]