摘要
目的利用海洋链霉菌Streptomyces parvus SCSIO Mla-L010中糖基转移酶的底物宽泛性对vicenistatin进行糖基定向化改造。方法采用组合生物合成技术分别将其他糖苷类天然产物生物合成途径中的糖基合成基因导入到SCSIO Mla-L010菌株中,构建糖基合成基因异源表达重组菌株;以有机溶剂萃取重组菌株发酵产物并利用现代色谱分离技术进行化合物纯化,进一步使用质谱、核磁共振光谱技术鉴定所获化合物的结构。结果将streptolydigin、grincamycin和lobophorin生物合成途径中合成d-olivose、L-rhodinose、d-kijanose和L-digitoxose糖基的基因导入到SCSIO Mla-L010菌株中,构建了3株异源糖基合成基因重组菌株,并从中分离得到4个含有不同糖基的新vicenistatin类天然产物。结论不仅丰富了糖苷类天然产物结构库,为海洋药物的研发提供化学实体,还为利用糖基转移酶的底物宽泛性对其他天然产物进行糖基定向改造提供参考和借鉴。
Objective To obtain vicenistatin analogues with different sugar moieties by applying the broad substrate specificity of glycosyl transferase in a potential vicenistatin producing strain Streptomyces parvus SCSIO Mla-L010. Methods The recombinant strains were constructed by introducing the sugar biosynthetic genes of other glycosylated natural products into the SCSIO Mla-L010 strain. The fermentation broths were extracted by solvent extraction and the targeted vicenistatin analogues were isolated and purified by a combination of chromatographic separation technologies. The structures of the purified compounds were elucidated by HRMS and intensive NMR analyses. Results The biosynthetic genes responsible for the formation of d-olivose, L-rhodinose, d-kijanose and L-digitoxose in the biosynthetic pathway of streptolydigin,grincamycin and lobophorin, respectively, were transferred into S. parvus SCSIO Mla-L010 to generate three recombinant strains. Four new vicenistatin analogues containing diverse sugar moieties were isolated from the fermentation broth of these recombinant strains. Conclusion This study not only provides chemical substances for the research and development of marine drugs, but also provides reference for the glycodiversification of other natural products by utilizing the broad substrate specificity of glycosyltransferase.
作者
梁智铖
李骏
凌春耀
许润
谭彬
黄洪波
鞠建华
李青连
LIANG Zhi-cheng;LI Jun;LING Chun-yao;XU Run;TAN Bin;HUANG Hong-bo;JU Jian-hua;LI Qing-lian(CAS Key Laboratory of Tropical Marine Bio-resources and Ecology,Guangdong Key Laboratory of Marine Materia Medica,RNAM Center for Marine Microbiology,Chinese Academy of Sceinces,South China Sea Institute of Oceanology,Guangzhou 510301,China;College of Marine Science,University of Chinese Academy of Sciences,Beijing 100049,China;Southern Marine Science and Engineering Guangdong Laboratory(Guangzhou),Guangzhou 511458,China)
出处
《中国海洋药物》
CAS
CSCD
2021年第6期1-12,共12页
Chinese Journal of Marine Drugs
基金
广东省重点领域研发计划项目(2020B1111030005)
国家自然科学基金项目(42006103)
广东省自然科学基金杰出青年项目(2021B1515020036)
广东省自然科学基金项目(2019A1515010310)资助。
