Shemediuk A L, Dolia B S, Ochi K, Fedorenko V O, Ostash B O
Ivan Franko National University of Lviv, 79005 Lviv, Ukraine.
Hiroshima Institute of Technology, 731-5193 Hiroshima, Japan.
Cytol Genet. 2022;56(1):31-36. doi: 10.3103/S009545272201011X. Epub 2022 Feb 11.
The J1074 strain remains one of the most popular platforms for the discovery of new natural compounds due to the expression of biosynthetic gene clusters (BGCs) from the microorganisms of the class. Different methods were tested to provide a maximal expression of heterologous BGCs in this strain. However, there is still no description of the properties of spontaneous J1074 mutants in the gene encoding a ribosomal protein S12. The interest in such mutations in actinobacteria is due to the fact that they provide a considerable increase in the antibiotic activity. In this work, we describe the isolation and characterization of the KO-1297 strain, which contains a spontaneous missense mutation in the gene leading to a Lys88Glu substitution in the protein S12. As compared with the initial strain, this mutant exhibits an increased resistance to streptomycin and higher antibiotic productivity. The KO-1297 strain and genetically engineered mutant K88E are not identical in their ability to produce antibiotics. KO-1297 also exhibits a certain level of instability of mutation. The genomes of KO-1297 and its revertant contain the mutations that can cause phenotypic differences between these strains (as well as between them and SAM2 and K88E strains).
由于该菌株能够表达来自该类微生物的生物合成基因簇(BGC),J1074菌株仍然是发现新天然化合物最常用的平台之一。为了使该菌株中异源BGC实现最大程度的表达,人们测试了不同的方法。然而,目前仍没有关于核糖体蛋白S12编码基因中J1074自发突变体特性的描述。对放线菌中此类突变感兴趣的原因在于,它们能使抗生素活性显著提高。在这项研究中,我们描述了KO - 1297菌株的分离和特性,该菌株在该基因中存在一个自发的错义突变,导致蛋白S12中的赖氨酸88被谷氨酸取代。与原始菌株相比,该突变体对链霉素的抗性增强,抗生素产量更高。KO - 1297菌株和基因工程突变体K88E在产生抗生素的能力方面并不相同。KO - 1297还表现出一定程度的突变不稳定性。KO - 1297及其回复菌株的基因组中存在一些突变,这些突变可能导致这些菌株之间(以及它们与SAM2和K88E菌株之间)出现表型差异。