Abstract Normally, cultivation of most Streptomyces species for the purpose of mutant-screening is an arduous process, needing a large amount of material. Moreover, the quantitative analysis of corresponding metabolites is done using HPLC, which is both expensive and time-consuming, and hence not ideal for screening of large numbers of isolates. A novel two-step high-throughput screening strategy has been designed to efficiently identify high-titer virginiamycin-producing mutants of Streptomyces virginiae. The high-throughput screening strategy described here can be used to cultivate a large number of mutant S. virginiae strains in 24-well deep-well microtiter plates and rapidly evaluate a large number of strains with varying virginiamycin titers. This is done based on the cylinder-plate bioassay method using Micrococcus luteus CMCC (B) 28001 as an indicator without compromising the accuracy. After one round of microwave-mediated mutagenesis, S. virginiae strain number 74 identified from nearly 1244 isolates, was shown to have the highest titer. The titer of virginiamycin was approximately 61.26 ± 2.53 mg/L, which is nearly three times compared to the value for the parent strain. Thus, this approach holds promise in facilitating the screening of high-titer virginiamycin producing strains.

中文翻译：

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使用微量滴定板对弗吉尼亚霉素高滴度生产菌株进行高通量筛选

摘要 通常，以突变体筛选为目的的大多数链霉菌的培养是一个艰巨的过程，需要大量的材料。此外，相应代谢物的定量分析是使用 HPLC 进行的，既昂贵又耗时，因此不适合筛选大量分离物。设计了一种新的两步高通量筛选策略，以有效识别 Streptomyces virginiae 的产生高滴度维吉尼亚霉素的突变体。此处描述的高通量筛选策略可用于在 24 孔深孔微量滴定板中培养大量突变 S. virginiae 菌株，并快速评估大量具有不同维吉尼亚霉素滴度的菌株。这是基于使用藤黄微球菌 CMCC (B) 28001 作为指标的圆柱板生物测定方法完成的，而不会影响准确性。经过一轮微波介导的诱变后，从近 1244 个分离株中鉴定出的 74 号弗吉尼亚链霉菌显示出具有最高滴度。维吉尼亚霉素的滴度约为 61.26 ± 2.53 mg/L，几乎是亲本菌株值的三倍。因此，这种方法有望促进高滴度维吉尼亚霉素生产菌株的筛选。这几乎是亲本菌株值的三倍。因此，这种方法有望促进高滴度维吉尼亚霉素生产菌株的筛选。这几乎是亲本菌株值的三倍。因此，这种方法有望促进高滴度维吉尼亚霉素生产菌株的筛选。