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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 analyze, identify and study the macrolactin biosynthetic gene cluster and its key genes from Bacillus subtilis B-9987. Methods The gene cluster and functional domains of the gene cluster were analyzed by bioinformatics methods. The E. coli-Bacillus shuttle vector for high expression of acyltransferase gene was constructed and introduced into B-9987 by electroporation High expression. Results Macrolactins were assembled by trans-acyltransferase (trans-AT) BmmA, a trans acyltransferase polyketide synthase encoded by 9 genes bmmA-I of the same operon. High expression increased the yield of macrolactin A about 0.6-fold. Conclusion Macrolactin biosynthetic gene cluster is prevalent in Bacillus that has biological control activity and is highly conserved. The high expression of trans acyltransferase can increase the yield of macrolactin A.