School of Pharmacy, Walailak University, Nakhon Si Thammarat 80161, Thailand.
Drug and Cosmetics Excellence Center, Walailak University, Nakhon Si Thammarat 80161, Thailand.
Int J Mol Sci. 2023 Jul 27;24(15):12016. doi: 10.3390/ijms241512016.
sp. SPR20 produced potentially antibacterial substances against methicillin-resistant (MRSA). The synthesis of these substances is controlled by their biosynthetic gene clusters. Several mutagenesis methods are used to overcome the restriction of gene regulations when genetic information is absent. Atmospheric and room temperature plasma (ARTP) is a powerful technique to initiate random mutagenesis for microbial strain improvement. This study utilized an argon-based ARTP to conduct the mutations on SPR20. The positive mutants of 40% occurred. The M27 mutant exhibited an increase in anti-MRSA activity when compared to the wild-type strain, with the MIC values of 250-500 and 500 μg/mL, respectively. M27 had genetic stability because it exhibited constant activity throughout fifteen generations. This mutant had similar morphology and antibiotic susceptibility to the wild type. Comparative proteomic analysis identified some specific proteins that were upregulated in M27. These proteins were involved in the metabolism of amino acids, cell structure and movement, and catalytic enzymes. These might result in the enhancement of the anti-MRSA activity of the ARTP-treated SPR20 mutant. This study supports the ARTP technology designed to increase the production of valuable antibacterial agents.
sp. SPR20 产生了针对耐甲氧西林金黄色葡萄球菌 (MRSA) 的潜在抗菌物质。这些物质的合成受其生物合成基因簇控制。当遗传信息缺失时，几种诱变方法被用于克服基因调控的限制。大气压室温等离子体 (ARTP) 是一种强大的技术，可以引发随机突变，从而改善微生物菌株。本研究利用基于氩气的 ARTP 对 SPR20 进行了突变。阳性突变体的发生率为 40%。与野生型菌株相比，M27 突变株的抗-MRSA 活性增加，MIC 值分别为 250-500 和 500μg/mL。M27 具有遗传稳定性，因为它在十五代中表现出恒定的活性。该突变体的形态和抗生素敏感性与野生型相似。比较蛋白质组学分析鉴定出 M27 中一些上调的特定蛋白质。这些蛋白质参与氨基酸代谢、细胞结构和运动以及催化酶。这可能导致 ARTP 处理的 SPR20 突变体抗-MRSA 活性的增强。本研究支持旨在增加有价值抗菌剂产量的 ARTP 技术。
