生物加工过程2011，Vol.9Issue(3)：6-10,5.DOI:10.3969/j.issn.1672-3678.2011.03.002
异戊二烯合成酶(IspS)在大肠杆菌中的表达及其产异戊二烯的研究
Expression of isoprene synthase in Escherichia coli for isoprene production
摘要
Abstract
The Populus alba isoprene synthase (IspS) was optimized to the preferred codon usage of Escherichia coli, cloned into vector pACYCDuet-1 and then heterologously expressed in E. coli BL21 ( DE3).The recombinant protein was purified by Ni2+ affinity chromatography and the enzyme activity was determined in vitro. Isoprene production by the recombinant strain was evaluated by shake-flask fermentation.The results showed that the isoprene synthase was efficiently expressed in E. coli. After purified by Ni2+columns, SDS-PAGE analysis revealed a special band corresponding to the fusion protein. The activity determination confirmed that the enzyme could catalyze the formation of isoprene. Under shake-tlask conditions, the isoprene productivity of the engineered strain was 60 μg/L.关键词
银白杨/异戊二烯合成酶/亲和层析Key words
Populus alba/ isoprene synthase/ affinity chromatography分类
生物科学引用本文复制引用
苏思正,刘建忠,杨建明,刘炜,王乾,咸漠..异戊二烯合成酶(IspS)在大肠杆菌中的表达及其产异戊二烯的研究[J].生物加工过程,2011,9(3):6-10,5.基金项目
中国科学院知识创新工程重要方向项目(KGCXZ-YW-801) （KGCXZ-YW-801）
