文献类型：期刊文献

中文题名：短芽孢杆菌耐碱性木聚糖酶(xylB)的分子生物学研究

英文题名：Study on Xylanase(xylB) Gene from Bacillus brevi

作者：胡春霞[1,2];陆平[1];李卫芬[1];许梓荣[1]

机构：[1]浙江大学教育部动物分子营养重点实验室,浙江杭州310029;[2]绍兴文理学院元培学院生命科学系,浙江绍兴312000

年份：2009

卷号：28

期号：1

起止页码：86

中文期刊名：食品与生物技术学报

外文期刊名：Journal of Food Science and Biotechnology

收录：CSTPCD、、Scopus、CSCD2011_2012、北大核心2008、北大核心、CSCD

基金：浙江省重大科技专项(2006C12086)

语种：中文

中文关键词：木聚糖酶;克隆;表达;大肠杆菌;短芽孢杆菌

外文关键词：alkaline xylanase, gene cloning and expression, sequence analysis, Bacillus brevi, Escherichia coli

中文摘要：根据已发表的环状芽孢杆菌(Bacillus circulan)和枯草芽孢杆菌(Bacillus subtilis)木聚糖酶基因序列设计引物,首次扩增出短芽孢杆菌(Bacillus brevis)中的-β1,4-内切木聚糖酶(以下简称木聚糖酶,E.C.3.2.1.8)基因片段。序列分析表明,该基因与已登录的木聚糖酶基因AF490979.1和AF490980.1分别有97%和96%的同源性,与其它芽孢杆菌属的同源性也较高。将此基因片段插入表达载体pET-30a(+)构建重组质粒,转化大肠杆菌E.coliBL21(DE3)。重组基因工程菌破碎后进行SDS-PAGE电泳检测,结果表明,IPTG诱导后,木聚糖酶基因在大肠杆菌的胞内获得高效表达,且酶活力最高可达26.14 U/mL。重组木聚糖酶最适温度为50℃,最适pH为9.0。

外文摘要：In this study, a pairs of primers were designed according to the published nucleotide sequences of putative xylanase genes of Bacillus circulan and Bacillus subtilis. With the specific primers, a target fragment of the β-1,4-endo-xylanase gene was firstly amplified from Bacillus brevi L8. Sequence analyses showed the homology of the cloned gene to AF490979. land AF490980.1 were 97% and 96%, respectively. The recombinant expression plasmid was constructed and transformed into E. coli BL21 （DE3）. SDS-PAGE demonstrated that the β-1, 4- endo-xylanase protein gene were expressed extracellularly in E. coli BL21 （DE3） successfully. A maximum activity of 26.14 U/mL achieved gotten from cellular extract of pET-30a-xylB[BL21] induced by IPTG. The optimum pH and temperature of the purified enzyme were 9.0 and 50℃,respectively.