Department of Biological and Pharmaceutical Engineering, Ningbo Institute of Technology, Zhejiang University, Ningbo, China.
Curr Microbiol. 2012 Dec;65(6):792-8. doi: 10.1007/s00284-012-0233-1. Epub 2012 Sep 19.
The mechanisms for the enhancement of pristinamycin production in the high-yielding recombinants of Streptomyces pristinaespiralis obtained by genome shuffling were investigated by quantitative real-time PCR (Q-PCR) and amplified fragment length polymorphism (AFLP) techniques. Q-PCR analysis showed that snaB and snbA involved, respectively, in the biosynthesis of pristinamycins II and I component had more extended high expression in the recombinant than that in the ancestor during fermentation process, indicating their expression changes might be key factors during the biosynthesis of the antibiotic. In addition, the antecedent establishment of the high self-resistance to pristinamycin, because ptr resistance gene started high-level expression ahead of the onset of the antibiotic production in the recombinant, might also lead to the increase of the antibiotics yield. AFLP analysis of these recombinants revealed genome variation of two novel genes, the homologs of AfsR regulatory gene and transposase gene, indicating these two gene variations were probably responsible for yield improvement of pristinamycin. This study provided several potential molecular clues for pristinamycin yield enhancement.
通过定量实时 PCR（Q-PCR）和扩增片段长度多态性（AFLP）技术研究了通过基因组重排获得的高产变铅青链霉菌重组体中增强普那霉素产量的机制。Q-PCR 分析表明，分别参与普那霉素 II 和 I 组分生物合成的 snaB 和 snbA 在发酵过程中重组体中的表达水平高于祖先，表明它们的表达变化可能是抗生素生物合成过程中的关键因素。此外，由于 ptr 抗性基因在抗生素产生之前就开始高水平表达，因此前体对普那霉素的高自我抗性的建立也可能导致抗生素产量的增加。对这些重组体的 AFLP 分析显示了两个新基因的基因组变异，即 AfsR 调控基因和转座酶基因的同源物，表明这两个基因变异可能是普那霉素产量提高的原因。本研究为提高普那霉素产量提供了一些潜在的分子线索。
