棘白菌素B高产菌株Aspergillus delacroxii转化体系的构建与优化  

Establishment and Optimization of Transformation System for an Echinocandin B Overproduction Strain of Aspergillus delacroxii

作　　者：闵涛玲[1] 高苹[1] 熊磊[1] 陈昌发[1] 胡海峰[1] MIN Taoling;GAO Ping;XIONG Lei;CHEN Changfa;HU Haifeng(State Key Lab.of New Drug and Pharmaceutical Process,Shanghai Institute of Pharmaceutical Industry,China State Institute of Pharmaceutical Industry,Shanghai 201203)

机构地区：[1]中国医药工业研究总院上海医药工业研究院创新药物与制药工艺国家重点实验室

出　　处：《中国医药工业杂志》2019年第9期981-986,共6页Chinese Journal of Pharmaceuticals

基　　金：上海市科委“创新行动计划”基础重点项目(08JC1418400);上海市科技支撑项目(12431901102);上海市科技人才计划(11XD1423700)

摘　　要：Aspergillus delacroxii SIPI-W15是棘白菌素B的高产菌株,目前尚缺少相关遗传操作信息,极大地限制了其代谢机制和分子机制的研究。本研究考察了Asp. delacroxii SIPI-W15 转化过程中的主要影响因素,建立了高效的遗传转化体系: Agrobacterium tumefaciens AGL-1作为介导菌株、新鲜孢子浓度1×10^6cuf/ml、农杆菌浓度OD600 0.7、24 ℃共转化3d。在最佳转化体系下,转化效率可达96个转化子(以10^6个分生孢子计),外源基因可以稳定遗传。基于上述遗传转化技术,获得棘白菌素B产量提高22%的转化子(产量达到2468mg/L),为进一步研究棘白菌素B 生物合成的功能基因和提高其产量提供了有力手段。The genetic transformation of Aspergillus delacroxii SIPI-W15 with high echinocandin B productivity is not reported, which greatly limits the molecular mechanism and the metabolomics development of Asp. delacroxii SIPI-W15. The optimum genetic transformation of Asp. delacroxii SIPI-W15 mediated by Agr. tumefaciens was determined with Agr. tumefaciens AGL-1 as the mediated strain, the fresh Asp. delacroxii SIPI-W15 spores concentration of 10^6 cuf/ml suspension, the Agr. tumefaciens AGL-1 concentration of OD600 0.7, and the cocultivation time of 3 days at 24 ℃. Under such optimal transformation conditions, the transformation frequency was 96 transformants per plate using 10^6 conidia. Based on the genetic transformation, one mutant strain that high-yield of echinoncandin B was screened, with a productivity of 2 468 mg/L. AMT was applied in Asp. delacroxii and this transformation system will provide a powerful tool for the further study on function genes of echinocandin B biosynthesis.

关 键 词：棘白菌素B ASPERGILLUS delacroxii 农杆菌介导转化 

分 类 号：R978.5[医药卫生—药品] Q784[医药卫生—药学]