深海链霉菌Streptomyces somaliensis SCSIO ZH66 ptm生物合成基因簇中orf7功能的探究  

Function of orf7 in the ptm biosynthetic gene cluster from the deep-sea-derived Strptomyces somaliensis SCSIO ZH66

作　　者：于东琪 侯路宽 李花月 鞠建华 李文利[1,2] YU Dong-qi;HOU Lu-kuan;LI Hua-yue;JU Jian-hua;LI Wen-li(Key Laboratory of Marine Drugs,Ministry of Education,School of Medicine and Pharmacy,Ocean University of China,Qingdao 266003,China;Laboratory for Marine Drugs and Bioproducts of Qingdao Pilot National Laboratory for Marine Science and Technology,Qingdao 266237,China;CAS Key Laboratory of Marine Bio-resources Sustainable Utilization,Guangdong Key Laboratory of Marine Materia Medica,RNAM Center for Marine Microbiology,South China Sea Institute of Oceanology,Chinese Academy of Sciences,Guangzhou 510301,China)

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]青岛海洋科学与技术试点国家实验室海洋药物与生物制品功能实验室,山东青岛266237 [3]中科院南海海洋研究所中科院海洋生物资源重点实验室,广东省海洋药物重点实验室,中科院热带海洋生物资源与生态重点实验室,广东广州510301

出　　处：《中国海洋药物》2021年第3期15-21,共7页Chinese Journal of Marine Drugs

基　　金：国家自然科学基金委员会—山东联合基金重点项目(U1706206)资助。

摘　　要：目的探究深海链霉菌Streptomyces somaliensis SCSIO ZH66中编码多环特特拉姆酸大环内酰胺(polycyclic tetramate macrolactams,PTMs)类化合物somamycin A~D的生物合成基因簇ptm中orf7对该基因簇编码产物的影响。方法采用PCR-targeting策略将ptm基因簇中的orf7进行阻断,然后将野生株与突变株在相同条件下发酵后通过高效液相色谱(HPLC)分析比较代谢产物的差异。结果生物信息学分析结果表明orf7的编码产物为细胞色素P450,实验成功得到了阻断突变株Δorf7,通过发酵产物HPLC分析发现,orf7被阻断后,somamycin D的产量提高,而其14位羰基氧化物somamycin C则不再产生。结论ptm基因簇中细胞色素P450基因orf7在PTMs生物合成过程中参与了14位碳上的氧化修饰。Objective To explore the in vivo function of the putative cytochrome P450 gene orf7 in the polycyclic tetramate macrolactam biosynthetic gene cluster ptm encoding somamycins,from the deep-seaderived Streptomyces somaliensis SCSIO ZH66.Methods orf7 was inactivated by using PCR-targeting strategy.The mutant and the wild-type strains were fermented under the same conditions,and the fermentation broths were analyzed by HPLC.Results Bioinformatic analysis showed orf7 encoded a putative cytochrome P450.orf7 was knocked out successfully.HPLC analysis suggested that the disruption of orf7 led to yield increase of somamycin D,while its C14 oxide somamycin C no longer produced.Conclusion orf7 was probably involved in oxidization of the C14 hydroxyl group of somamycin D to carbonyl group.

关 键 词：深海链霉菌 多环特特拉姆酸大环内酰胺 天然产物 

分 类 号：R915[医药卫生—微生物与生化药学]