Construction and application of a "superplasmid" for enhanced production of antibiotics.
Construction and application of a "superplasmid" for enhanced production of antibiotics.
复制标题
用于增强抗生素生产的“超级质粒”的构建和应用。
DOI：
10.1007/s00253-019-10283-6
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发表时间：
2020
影响因子：
5
通讯作者：
Jing He
中科院分区：
文献类型：
--
作者：
Qin Liu;Qin Lin;Xinying Li;Muhammad Ali;Jing He
超过三分之二的已知抗生素是由链霉菌属的放线菌产生的。不幸的是，链霉菌天然抗生素的生产速度非常慢，因此不能满足工业需求。在本研究中，一个由PermE*启动子控制的超质粒促进链霉菌产生抗生素，该超质粒包括全球调控因子SafsR、环腺苷受体蛋白(CRP)、带点突变的RNA聚合酶β亚基(RpoB)和乙酰辅酶A羧基酶基因(AccA2BE)，然后通过连接将这些元件转移到天蓝色链霉菌M145、变色链霉菌TRM45540、吸水链霉菌XM201和吸水链霉菌CC29253中产生接合子。核磁共振、高效液相和液质联用分析表明，该超质粒导致放线菌素(101.90%)和十一烷基灵芝菌素(181.60%)INS产量过高。天蓝色M145：：pLQ003，雷帕霉素(110%)，潮霉素A(163.4%)INS。吸湿剂ATCC29253：：pLQ003；放线菌素D(11.78%)Ins。突变株TRM45540：：pLQ003，也与格尔达霉素INS的下调有关。吸湿菌XM201，但我们发现突变株中有突变株OFS。插入调控因子的吸湿菌XM201出现了野生型菌株所没有的几个峰。本工作的结果表明，链霉菌中的调控网络并不统一，我们构建的超质粒可能导致了不同链霉菌的过量生产和下调。
More than two-third of known antibiotics are produced by actinomycetes of the genusStreptomyces. Unfortunately, the production rate fromStreptomycesnatural antibiotic is extremely slow and thus cannot satisfy industrial demand. In this study, the production of antibiotics byStreptomycesis enhanced by a “superplasmid” which including global regulatory factorsafsR, cyclic adenosine receptor protein (CRP), RNA polymerase beta subunits (rpoB) with point mutation and acetyl coenzyme A carboxylase gene (accA2BE), these elements are controlled by the PermE* promoter and then transfer intoStreptomyces coelicolorM145, Streptomyces mutabilisTRM45540, Streptomyces hygroscopicusXM201, andStreptomyces hygroscopicusATCC29253 by conjugation to generate exconjugants. NMR, HPLC, and LC–MS analyses revealed that the superplasmid led to the overproduction of actinorhodin (101.90%), undecylprodigiosin (181.60%) inS. coelicolorM145:: pLQ003, of rapamycin (110%), hygrocin A (163.4%) inS. hygroscopicusATCC29253:: pLQ003, and of actinomycin D (11.78%) inS. mutabilisTRM45540:: pLQ003, and also to the downregulation of geldanamycin inS. hygroscopicusXM201, but we found that mutant strains in mutant strains ofS. hygroscopicusXM201 with regulatory factors inserted showed several peaks that were not found in wild-type strains. The results of the present work indicated that the regulator net working inStreptomyceswas not uniform, the superplasmid we constructed possibly caused this overproduction and downregulation in differentStreptomyces.