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首页> 外文期刊>ACS Omega >Development of a Robust and Quantitative High-Throughput Screening Method for Antibiotic Production in Bacterial Libraries
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Development of a Robust and Quantitative High-Throughput Screening Method for Antibiotic Production in Bacterial Libraries

机译：细菌库中抗生素生产的鲁棒定量高通量筛选方法的建立

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Over the past 30 years, there has been a dramatic rise in the number of infections caused by multidrug-resistant bacteria, which have proliferated due to the misuse and overuse of antibiotics. Over this same time period, however, there has also been a decline in the number of antibiotics with novel mechanisms of action coming to market. Therefore, there is a growing need for an increase in the speed at which new antibiotics are discovered and developed. Natural products produced by bacteria have been and continue to be a robust source of novel antibiotics; however, new and complementary methods for screening large bacterial libraries for novel antibiotic production are needed due to the current agar methods being limited in scope, time consuming, and prone to error. Herein, we describe a rapid, robust, and quantitative high-throughput liquid culture screening method for antibiotic production by bacteria. This method has the ability to screen both mono- and coculture mixtures of bacteria in vitro and be adapted to other phenotypic natural product analyses. Over 260 bacterial species were screened in monoculture, and 38 and 34% were found to produce antibiotics capable of inhibition of Staphylococcus aureus or Escherichia coli, respectively, with 8 and 4% being classified as strong producers (≥30% growth inhibition), respectively. Bacteria found to not produce antibiotics in monoculture were also screened in coculture using an adaptation of this method. Of the more than 270 cocultures screened, 14 and 30% were found to produce antibiotics capable of inhibition of S. aureus or E. coli, respectively. Of those bacteria found to produce antibiotics in monoculture, 43 bacteria were subjected to 16S rRNA sequencing and found to be majority Pseudomonas (37%), Serratia (19%), and Bacillus (14%) bacteria, but two novel producers, Herbaspirillum and Kluyvera, were also found.
机译：在过去的30年中，由多重耐药性细菌引起的感染数量急剧增加，这些细菌由于滥用和过度使用抗生素而激增。然而，在同一时期，具有新颖作用机制的抗生素数量也出现了下降。因此，越来越需要提高发现和开发新抗生素的速度。由细菌产生的天然产物已经并且将继续是新型抗生素的强大来源。然而，由于目前的琼脂方法的范围，耗时且容易出错，因此需要用于筛选大型细菌文库以生产新型抗生素的新的补充方法。本文中，我们描述了一种快速，健壮和定量的高通量液体培养筛选方法，用于细菌生产抗生素。该方法具有体外筛选细菌的单培养混合物和共培养混合物的能力，并适用于其他表型天然产物分析。在单一培养中筛选出超过260种细菌，发现38和34％的抗生素分别能够抑制金黄色葡萄球菌或大肠杆菌，其中8％和4％被归为强力生产者（生长抑制率≥30％）。 。使用此方法的一种适应方法，还可以在共培养物中筛选发现在单培养中不产生抗生素的细菌。在筛选的270多个共培养物中，发现14％和30％分别产生能够抑制金黄色葡萄球菌或大肠杆菌的抗生素。在单一培养中发现可产生抗生素的细菌中，有43种细菌经过了16S rRNA测序，发现多数为假单胞菌（37％），沙雷氏菌（19％）和芽孢杆菌（14％），但有两种新型生产者，即草螺旋藻和还发现了克鲁维拉。

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