Department of Biochemical and Chemical Engineering, TU Dortmund University, Emil-Figge-Straße 66, 44227 Dortmund, Germany.
Biomolecules. 2021 Jan 30;11(2):193. doi: 10.3390/biom11020193.
Over the past decade, the one strain many compounds (OSMAC) approach has been established for the activation of biosynthetic gene clusters (BGCs), which mainly encode the enzymes of secondary metabolite (SM) biosynthesis pathways. These BGCs were successfully activated by altering various culture conditions, such as aeration rate, temperature, and nutrient composition. Here, we determined the biosynthetic potential of 43 bacteria using the genome mining tool antiSMASH. Based on the number of BGCs, biological safety, availability of deposited cultures, and literature coverage, we selected five promising candidates: DSM7, DSM2259, HKI727, DSM44719, and DSM40499. The bacteria were cultivated under a broad range of OSMAC conditions (nutrient-rich media, minimal media, nutrient-limited media, addition of organic solvents, addition of biotic additives, and type of culture vessel) to fully assess the biosynthetic potential. In particular, we investigated so far scarcely applied OSMAC conditions to enhance the diversity of SMs. We detected the four predicted compounds bacillibactin, desferrioxamine B, myxochelin A, and surfactin. In total, 590 novel mass features were detected in a broad range of investigated OSMAC conditions, which outnumber the predicted gene clusters for all investigated bacteria by far. Interestingly, we detected mass features of the bioactive compounds cyclo-(Tyr-Pro) and nocardamin in extracts of DSM7 and DSM2259. Both compounds were so far not reported for these strains, indicating that our broad OSMAC screening approach was successful. Remarkably, the infrequently applied OSMAC conditions in defined medium with and without nutrient limitation were demonstrated to be very effective for BGC activation and for SM discovery.
在过去的十年中，人们已经建立了一种单一菌株多种化合物（OSMAC）的方法，用于激活生物合成基因簇（BGCs），这些基因簇主要编码次生代谢物（SM）生物合成途径的酶。这些 BGC 通过改变各种培养条件（例如通气率、温度和营养成分）成功地被激活。在这里，我们使用基因组挖掘工具 antiSMASH 确定了 43 种细菌的生物合成潜力。基于 BGC 的数量、生物安全性、已存培养物的可用性以及文献报道，我们选择了五个有前途的候选菌株：DSM7、DSM2259、HKI727、DSM44719 和 DSM40499。这些细菌在广泛的 OSMAC 条件下（富含营养的培养基、最小培养基、营养限制培养基、添加有机溶剂、添加生物添加剂和培养容器类型）进行培养，以充分评估其生物合成潜力。特别是，我们研究了迄今为止应用较少的 OSMAC 条件，以增强 SM 多样性。我们检测到了四种预测化合物 bacillibactin、desferrioxamine B、myxochelin A 和 surfactin。在广泛的 OSMAC 条件下，总共检测到 590 种新的质量特征，远远超过了所有被研究细菌的预测基因簇。有趣的是，我们在 DSM7 和 DSM2259 的提取物中检测到了生物活性化合物 cyclo-(Tyr-Pro) 和 nocardamin 的质量特征。到目前为止，这些化合物在这两个菌株中都没有报道过，这表明我们广泛的 OSMAC 筛选方法是成功的。值得注意的是，在限定培养基中添加和不添加营养物质的不常应用的 OSMAC 条件被证明对 BGC 激活和 SM 发现非常有效。
