海洋芽孢杆菌B-9987中macrolactin生物合成基因簇分析及反式酰基转移酶高表达  

Characterization of macrolactin gene cluster from Bacillus marinus B-9987 and overexpression of trans-acyl transferase

作　　者：刘扬[1] 郑华[1] 姚婷婷[1] 田黎[2,3] 李文利[1] 

机构地区：[1]中国海洋大学海洋药物教育部重点实验室医药学院,山东青岛266003 [2]国家海洋局第一海洋研究所,山东青岛266061 [3]青岛科技大学化工学院,山东青岛266042

出　　处：《中国海洋药物》2014年第3期69-75,共7页Chinese Journal of Marine Drugs

基　　金：国家自然科学基金项目(31070072;31171201);教育部新世纪优秀人才支持计划项目(NCET-09-0717)资助

摘　　要：目的对海洋芽孢杆菌B-9987中macrolactin生物合成基因簇及其关键基因进行分析、鉴定及功能研究。方法通过生物信息学手段对基因簇的基因组成和功能结构域进行了分析;构建了用于酰基转移酶基因高表达的大肠杆菌—芽孢杆菌穿梭载体,采用电击转化方法导入B-9987之中进行高表达。结果 Macrolactins是由位于同1个操纵子的9个基因bmmA-I所编码的反式酰基转移酶聚酮合酶体系组装而成,反式酰基转移酶(trans-acyltransferase,trans-AT)BmmA的高表达使macrolactin A的产量提高了约0.6倍。结论 Macrolactin生物合成基因簇在具有生物防治活性的芽孢杆菌中普遍存在,且高度保守;反式酰基转移酶的高表达能够增加macrolactin A的产量。Objective To identify and characterize the macrolactin gene cluster from Bacillus rnarinus I3- 9987. Methods By elaborate bioinformatics analysis, the gene organization and functional domain composition of the macrolactin gene cluster were predicted; for overexpression of the trans-acyl transferase （trans-AT） gene, an E. colffBacillus shuttle vector was constructed and introduced into B. marinus 13- 9987 by electroporation. Results Macrolactins were assembled via a trans-AT polyketide synthase sys- tem, which was encoded by 9 genes, brnrnA-I, located in an operon; overexpression of trans-AT BrnrnA led to the improved macrolactin A production by about 0. 6-fold. Conclusion Macrolactin biosyn- thetic gene clusters are highly conserved among biocontrol Bacillus strains; overexpression of trans-acyl transferase BmmA is able to increase macrolactin A production.

关 键 词：海洋芽孢杆菌 Macrolactin 生物合成基因簇 反式酰基转移酶聚酮合酶 

分 类 号：R915[医药卫生—微生物与生化药学]