Department of Genetics, Belarusian State University, Nezavisimosty Ave., 4, 220030, Minsk, Belarus.
World J Microbiol Biotechnol. 2023 Aug 16;39(10):279. doi: 10.1007/s11274-023-03685-2.
Phenazines are heteroaromatic compounds consisting of a central pyrazine ring fused with two benzenes. Different functional groups attached to the dibenzopyrasin core cause differences in the chemical, physical, and biological properties of phenazines. Interest in these compounds has not diminished for decades. New biological activities and practical applications discovered in recent years force researchers to investigate all aspects of the synthesis, degradation, and mechanisms of action of phenazines. In this study, we have demonstrated the involvement of the coxA gene product (cytochrome c oxidase, su I) in the production of phenazines in P. chlororaphis subsp. aurantiaca. Overlap PCR was used to knock out the coxA gene and the resulting mutants were screened for their ability to grow on rich and minimal culture media and for phenazine production. The reintroduction of the full-length coxA gene into the B-162/coxA strains was used to further confirm the role of this gene product in the ability to produce phenazines. We were able to show that the product of the coxA gene is necessary for phenazine production in rich growth media. At the same time, the CoxA protein does not seem to have any effect on phenazine production in M9 minimal salt medium. We could show that knocking down even one subunit of the cytochrome c oxidase complex leads to a significant reduction (to trace concentrations) or complete suppression of phenazine antibiotic production on rich PCA medium in P. chlororaphis subsp. aurantiaca.
吩嗪是一种杂环芳香族化合物,由一个中央吡嗪环与两个苯环融合而成。连接在二苯并吡嗪核心上的不同官能团导致吩嗪的化学、物理和生物学性质存在差异。几十年来,人们对这些化合物的兴趣从未减退。近年来发现的新的生物活性和实际应用迫使研究人员研究吩嗪的合成、降解和作用机制的各个方面。在这项研究中,我们已经证明了 coxA 基因产物(细胞色素 c 氧化酶,su I)参与了 P. chlororaphis subsp. aurantiaca 中吩嗪的产生。重叠 PCR 用于敲除 coxA 基因,筛选突变体在丰富和最小培养基中的生长能力和吩嗪生产能力。将全长 coxA 基因重新引入 B-162/coxA 菌株,进一步证实了该基因产物在产生吩嗪能力中的作用。我们能够表明,coxA 基因的产物是在丰富的生长培养基中产生吩嗪所必需的。同时,CoxA 蛋白似乎对 M9 低盐培养基中吩嗪抗生素的产生没有任何影响。我们可以证明,即使敲除细胞色素 c 氧化酶复合物的一个亚基,也会导致 P. chlororaphis subsp. aurantiaca 在丰富的 PCA 培养基上产生吩嗪抗生素的产量显著降低(降至痕量浓度)或完全抑制。