Nikandrova Arina A, Petriakova Anna D, Izzi Anton R, Petrosyan Garegin A, Tashlitsky Vadim N, Alferova Vera A, Panova Tatiana V, Khrenova Maria G, Biryukov Mikhail V, Zakalyukina Yuliya V, Zvereva Maria I, Lukianov Dmitrii A, Sergiev Petr V
Center for Molecular and Cellular Biology, 121205 Moscow, Russia.
Department of Biology, Lomonosov Moscow State University, 119991 Moscow, Russia.
Int J Mol Sci. 2024 Dec 12;25(24):13328. doi: 10.3390/ijms252413328.
Antibiotic resistance has been and remains a major problem in our society. The main solution to this problem is to search and study the mechanisms of antibiotic action. Many groups of secondary metabolites, including antimicrobial ones, are produced by the phylum. The actinobacterial strains isolated from habitats that have not been well studied are of great interest. Due to high resource competition, antibiotics are now considered a 'trump card in the game of life' due to their presence in natural substrates with limited nutrients. Potentially, strains isolated from such habitats can be producers of novel or poorly studied antibiotics. In the current research, we identified the strain sp. AP22 from the soils of the Akhshatyrsky Gorge, which is capable of producing pentalenolactone. This study describes the phenotypic and morphological characteristics of sp. AP22 and its biological activity. Pentalenolactone is a known inhibitor of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an important enzyme involved in glycolysis. We identified a previously unknown mutation in the gene encoding glyceraldehyde-3-phosphate dehydrogenase that confers resistance to this antibiotic compound. This antibiotic is not used in clinical practice, so its application as a selectable marker will not lead to the creation of pathogens resistant to clinically relevant antibiotics. In this case, the selectable marker is based on a genetic construct containing the glyceraldehyde-3-phosphate dehydrogenase gene with a resistance mutation. The use of this selectable marker can be applied to various genetic and molecular techniques, such as cloning and transformation. This can help to facilitate genetic and molecular biology studies of strains resistant to standard selectable markers such as kanamycin or ampicillin.
抗生素耐药性一直是且仍然是我们社会中的一个主要问题。解决这个问题的主要办法是寻找和研究抗生素作用机制。包括抗菌性次级代谢产物在内的许多类次级代谢产物是由该门产生的。从尚未得到充分研究的栖息地分离出的放线菌菌株备受关注。由于资源竞争激烈,抗生素因其存在于营养有限的天然底物中,现在被视为“生命游戏中的王牌”。潜在地,从这些栖息地分离出的菌株可能是新型或研究较少的抗生素的生产者。在当前的研究中,我们从阿克沙特尔斯基峡谷的土壤中鉴定出了能够产生戊烯内酯的菌株sp. AP22。本研究描述了sp. AP22的表型和形态特征及其生物活性。戊烯内酯是甘油醛 - 3 - 磷酸脱氢酶(GAPDH)的已知抑制剂,GAPDH是参与糖酵解的一种重要酶。我们在编码甘油醛 - 3 - 磷酸脱氢酶的基因中鉴定出一个先前未知的突变,该突变赋予了对这种抗生素化合物的抗性。这种抗生素不在临床实践中使用,因此将其用作选择标记不会导致产生对临床相关抗生素耐药的病原体。在这种情况下,选择标记基于一个包含具有抗性突变的甘油醛 - 3 - 磷酸脱氢酶基因的遗传构建体。这种选择标记的使用可应用于各种遗传和分子技术,如克隆和转化。这有助于促进对诸如卡那霉素或氨苄青霉素等标准选择标记耐药的菌株的遗传和分子生物学研究。