|
|||||
|
|
| Lactobacillus reuteri PYR8亚油酸异构酶基因在大肠杆菌中的表达与活性检测 | |
| 引用本文： | 张艳禾,张兰威,胡森,步志高.Lactobacillus reuteri PYR8亚油酸异构酶基因在大肠杆菌中的表达与活性检测[J].农业生物技术学报,2007,15(1):147-152. |
| 作者姓名： | 张艳禾 张兰威 胡森 步志高 |
| 作者单位： | 1. 东北农业大学食品科学学院,哈尔滨,150030 2. 哈尔滨工业大学食品科学与工程学院 3. 中国农业科学院哈尔滨兽医研究所,哈尔滨,150001 |
| 基金项目： | 黑龙江省科学技术重大项目;科技部农业科技成果转化基金 |
| 摘 要： | 以Lactobacillus reuteri PYR8菌株基因组为模板,利用PCR方法扩增出亚油酸异构酶(linoleate isomerase,LI)基因,亚克隆到pET30a中构建原核表达载体pET30a-LI,并转化入大肠杆菌(Escherichia coli)BL21(DE3)中。其重组菌株在37℃条件下,经1.0mmol/L IPTG诱导表达重组LI蛋白,该蛋白以包涵体形式存在,表达量占菌体总蛋白的39.2%。采用Ni2 -NTA柱上复性法获得有活性的纯化重组亚油酸异构酶,其比活力5.12U/mg,是天然亚油酸异构酶比活力的2.5倍。 |
| 关 键 词： | 亚油酸异构酶 大肠杆菌 活性检测 |
| 文章编号： | 1006-1304(2007)01-0147-06 |
| 修稿时间： | 2006年2月20日 |
Expression of Linoleate Isomerase Gene from Lactobacillus reuteri PYR8 in Escherichia coli and Assay of Its Bioactivity |
|
| ZHANG Yan-he,ZHANG Lan-wei,HU Sen,BU Zhi-gao.Expression of Linoleate Isomerase Gene from Lactobacillus reuteri PYR8 in Escherichia coli and Assay of Its Bioactivity[J].Journal of Agricultural Biotechnology,2007,15(1):147-152. | |
| Authors: | ZHANG Yan-he ZHANG Lan-wei HU Sen BU Zhi-gao |
| Abstract: | Linoleate isomerase(LI ) gene was amplified by PCR from chromosome of Lactobacillus reuteri PYR8,and was inserted into pET30a to construct the prokaryotic expression vector pET30a-LI.The recombinant LI protein was highly expressed at yield about 39.2% of total protein in Escherichia coli BL21(DE3)strain by 1.0 mmol/L IPTG induced and was only present in the inclusion body. And purified soluble linoleate isomerase was gained by refolding of denatured LI protein bound to Ni2 -NTA, and the purified recombinant linoleate isomerase possessed biological activity(5.12 U/mg),which was 2.5 times of natural linoleate isomerase. |
| Keywords: | Lactobacillus reuteri PYR8 |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |