摘要
对菌株Y-2维生素K_2合成途径中的6个主要酶基因分别进行克隆和表达,以探索不同酶基因对MK-7产量的影响。参照《分子克隆实验指南》完成对基因重组质粒的构建,并对spizizen转化法进行改进,实现重组质粒对菌株Y-2的转化。结果表明,分别过量表达维生素K_2合成途径中的6个基因均能提高MK-7的产量。需要指出的是,过表达基因hep S对MK-7产量的影响最大,其中在摇瓶培养组的产量提高了38%,静置培养组的产量提高了48%。其余几个基因过表达后对MK-7产量的影响则根据培养方式的不同而各有差异。
This paper focused on understanding the effects of different enzyme genes on MK-7 production by cloning and expressingthe genes of 6 enzymes in the biosynthetic pathway of vitamin K_2 with strain Y-2. Firstly,the recombinant plasmids with single gene of the 6genes were constructed according to the description of Guide to Molecular Cloning Experiment. Subsequently,the recombinant plasmids weresuccessfully transformed into the strain Y-2 via the improved ＂spizizen＂ transformation method. Results showed that respective overexpressionof the 6 genes involved in vitamin K_2 biosynthetic pathway all was beneficial to raise MK-7 production. It should be noted that overexpressionof gene hep S had the greatest effects on MK-7 production among these 6 genes,which resulted in an increase by 38% in shaking group and by48% in static group. However,the effects of other genes on MK-7 production varied depending on the culture mode.
出处
《生物技术通报》
CAS
CSCD
北大核心
2016年第11期248-254,共7页
Biotechnology Bulletin
基金
江苏省自然科学基金-青年基金项目(BK20150149)
关键词
解淀粉芽孢杆菌
甲萘醌-7
合成途径
催化酶
Bacillus amyloliquefaciens
menaquinone-7
biosynthetic pathway
catalyzing enzyme
