以组合生物合成策略为导向的C-核苷类多氧霉素的研究

Research on Novel C-nucleoside Polyoxin Guided by Combinatorial Biosynthesis Strategies

基于组合生物合成策略,成功地将马来亚霉素结构中的C-核苷引入多氧霉素结构中,合成了具有C-核苷的新型多氧霉素杂合抗生素。这一过程包括通过人工设计杂合抗生素的结构,改造多氧霉素的工业菌株金产色链霉菌,并获得相应的突变菌株;接着,整合负责C-核苷合成的起始基因truD和malO,得到重组菌株PA1和PA2。通过生物活性测定和液相高分辨质谱(LC-HRMS)分析,对代谢产物进行了结构确证。此外,为了提高目标杂合抗生素的产量,增加了多氧霉素生物合成基因簇的拷贝数,获得了重组菌株PA3。重组菌株PA1和PA2能够定向产生具有C-核苷结构的假尿苷多氧霉素L(ψ-polyoxin L)和假尿苷多氧霉素K(ψ-polyoxin K)。通过基因簇加倍的策略,重组菌株PA3的ψ-polyoxin L和ψ-polyoxin K的产量均显著提高。这一新型C-核苷类多氧霉素的合成为今后相关核苷类抗生素多重组合生物的研究奠定了坚实的基础。

Based on the strategy of combinatorial biosynthesis,the paper introduces the C-nucleoside in the structure of Malayamycin into the structure of polyoxin,and obtains a new type of polyoxin hybrid antibiotic with C-nucleoside.By artificially designing the structure of hybrid antibiotics,the industrial strain Streptomyces aureogenes of polyoxin was modified and corresponding mutant strains were obtained.Then,the starting genes truD and malO responsible for C-nucleoside synthesis in the biosynthesis of Malayamycin were integrated to obtain recombinant strains PA1 and PA2.The structure of the metabolites was confirmed by biological activity measurement and liquid phase high-resolution mass spectrometry(LC-HRMS)analysis.In addition,the recombinant strain PA3 was obtained by increasing the copy number of the polyoxin biosynthesis gene cluster to increase the yield of the target heterozygous antibiotic.Recombinant strains PA1 and PA2 can selectively produce pseudouridine polyoxin L with a C-nucleoside structure(ψ-Polyoxin L),and pseudouridine polyoxin K(ψ-Polyoxin K);Gene cluster doubling in recombinant strain PA3ψ-Polyoxin L andψ-The yield of polyoxin K was significantly increased.The production of new C-nucleoside polyoxin has laid a solid foundation for the future research on the synthesis of multiple combinations of related nucleoside antibiotics.