摘要
目的对海洋芽孢杆菌B-9987中difficidins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difficidins基因簇的基因组成和功能结构域进行了分析;结合其结构特征及聚酮化合物的生物合成原理,初步推导其生物合成途径;构建了烯酰辅酶A水合酶基因difO双交换阻断突变株,对其功能进行初步验证并对基因簇进行确认。结果从B-9987发酵液中分离纯化得到化合物oxydifficidin,鉴定了B-9987中该化合物的生物合成基因簇,其是由位于同一个操纵子的15个基因difA-O所编码的反式酰基转移酶聚酮合酶体系组装而成;difO基因阻断后,oxydifficidin不再产生。结论 difficidins生物合成基因簇的鉴定为后续研究其生物合成途径及相关基因的功能奠定了基础。
Objective To identify and characterize the difficidin biosynthetic gene cluster from Bacillus methylotrophicus B-9987.Methods By bioinformatics analysis,the gene organization and functional domain composition of the difficidin gene cluster were predicted.The biosynthetic pathway was deduced by combining the structural characteristics of difficidins and the biosynthesis principle of polyketides.To identify the gene cluster,the difOdouble crossover mutant was constructed followed by HPLC analysis of fermentation and to probe its function.Results Oxydifficidin was obtained from B-9987 fermentation broth.The difficidin gene cluster of B-9987 was identified,which was encoded by 15 genes,difA-O,located in an operon and assembled via a trans-AT polyketide synthase system.Difficidins were no longer produced in the difOgene deletion mutant.Conclusion The characterization of difficidin gene cluster fromBacillus methylotrophicus B-9987 set the stage for elucidation of the biosynthetic molecular mechanism and further combinatorial biosynthesis of difficidins.
作者
高宇佳
张君
李花月
李文利
GAO Yu-jia;ZHANG Jun;LI Hua-yue;LI Wen-li(Key Laboratory of Marine Drugs,Ministry of Education,School of Medicine and Pharmacy,Ocean University of China,Qingdao 266003,China)
出处
《中国海洋药物》
CAS
CSCD
2019年第5期1-8,共8页
Chinese Journal of Marine Drugs
基金
国家自然科学基金项目（31570032）资助
