核糖体工程技术提高荧光假单胞菌Pf-5菌株中次级代谢产物产量
Improving the Production of Active Secondary Metabolites in Pseudomonas Pf-5 by Using Ribosome Engineering Technology
利用核糖体工程技术，来提高荧光假单胞Pf-5菌株活性次级代谢产物的产量。在高浓度的利福平抗性筛选下，分离得60株到利福平抗性突变株，通过对真菌抑菌活性比较化，以及通过色谱分析比较发现，其中突变株R55相比原始菌，其多个抑菌活性产物大幅提高。经色谱、质谱测定表明，其中三种物质已知,有一种物质未知。对突变菌株R55的 DNA 序列进行测序分析，发现在编码RNA聚合酶β亚基的rpoB基因出现点突变，第1570 位的胞嘧啶(C) 突变为腺嘌呤( A)，相应的氨基酸残基由组氨酸突变为天冬酰胺。
In this study, we used the Ribosome engineering technology to improve the yield of secondary metabolites produced by Pseudomonas fluorescens Pf-5 strain. In a high concentrations of rifampicin, 60 spontaneous rifampicin resistant mutants have been isolated, by comparison of antibacterial activity against fungi, and combined with the chromatographic analysis. Among plenty of mutants, the R55 displayed the greatest increase in the yield of several antibiotics than that in parent strain. By HPLC and 2D-MS/MS demonstrate that three of metbolites is known, and one is unknown. The DNA sequencing and analysis show that R55 mutant existed one point mutations in the rpoB gene which encoding β subunit of RNA polymerase. The mutations occurred at nucleotide pair 1570, which resulted in the mutation of Histidine (523) to Asparagine.
谢亚丽、胡胜标、刘正强
生物工程学微生物学分子生物学
核糖体工程荧光假单胞利福平次级代谢产物
Ribosome engineering technologyPseudomonas fluorescensrifampicinsecondary metabolites
谢亚丽,胡胜标,刘正强.核糖体工程技术提高荧光假单胞菌Pf-5菌株中次级代谢产物产量[EB/OL].(2016-01-08)[2025-08-10].http://www.paper.edu.cn/releasepaper/content/201601-171.点此复制
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