Improvement of A21978C production in Streptomyces roseosporus by reporter-guided rpsL mutation selection
Improvement of A21978C production in Streptomyces roseosporus by reporter-guided rpsL mutation selection
复制标题
通过记者引导的 rpsL 突变选择提高玫瑰孢链霉菌中 A21978C 的产量
DOI：
10.1111/j.1365-2672.2012.05302.x
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发表时间：
2012-06-01
影响因子：
4
通讯作者：
Liao, G.
中科院分区：
文献类型：
--
作者：
Wang, L.;Zhao, Y.;Liao, G.
目的：与达托霉素类似，玫瑰孢链霉菌产生的A21978C因子之一，是一种酸性环脂肽抗生素，对多种革兰氏阳性病原体具有强效活性。为了提高这种广泛使用且临床上重要的抗生素的滴度，我们采用了一种报道的rpsL突变选择系统来产生产生高水平A21978C的菌株。方法与结果：与报告基因设计相似，以dptE为过表达靶点，以neo-编码新霉素磷酸转移酶为报告基因。使用这种报告者引导的选择系统，20%的选择的耐链霉素突变体比起始菌株产生更多的A21978C。筛选系统使筛选效率提高了约10倍，在strr突变体中A21978C高产菌株的频率为1.7%。在rpsL K43N突变体中，A21978C的产量增加了约2.2倍。结论：类似于核糖体工程与报告者引导的突变体选择相结合，产生了A21978C高产菌株，其A21978C产量约为亲本菌株的两倍。研究的意义和影响：与本文所提出的策略类似，该策略结合了核糖体工程和报告者引导的突变选择的优势，可以应用于其他细菌，以提高其次生代谢物的产量。
Aims:similar to Daptomycin, one of the A21978C factors produced by Streptomyces roseosporus, is an acidic cyclic lipopeptide antibiotic with potent activity against a variety of Gram-positive pathogens. To increase the titre of this extensively used and clinically important antibiotic, we applied a reported-guided rpsL mutation selection system to generate strains producing high levels of A21978C. Methods and Results:similar to In the reporter design, dptE was chosen as the overexpressing target, and neo-encoding neomycin phosphotransferase as the reporter. Using this reporter-guided selection system, 20% of the selected, streptomycin-resistant mutants produced greater amounts of A21978C than the starting strain. The selection system increased the screening efficiency about 10-fold with a frequency of 1.7% A21978C overproducing strains among strr mutants. A21978C production was increased approximately 2.2-fold in the rpsL K43N mutant. Conclusions:similar to The combination of ribosome engineering and reporter-guided mutant selection generated an A21978C overproducing strain that produced about twice as much A21978C as the parental strain. Significance and Impact of the Study:similar to The strategies presented here, which integrated the advantages of both ribosome engineering and reporter-guided mutation selection, could be applied to other bacteria to improve their yield of secondary metabolites.