Liu Yifei, Wang Yawen, Chen Xinyue, Jin Junhua, Liu Hui, Hao Yanling, Zhang Hongxing, Xie Yuanhong
Beijing Laboratory of Food Quality and Safety, Beijing Key Laboratory of Agricultural Product Detection and Control of Spoilage Organisms and Pesticide Residue, College of Food Science and Engineering, Beijing University of Agriculture, Beijing, China.
Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China.
Front Microbiol. 2022 Apr 14;13:874789. doi: 10.3389/fmicb.2022.874789. eCollection 2022.
Plantaricin BM-1, a class IIa bacteriocin produced by BM-1, exhibits significant antibacterial activity against many gram-positive and gram-negative bacteria. However, the mechanism underlying the action of class IIa bacteriocins against gram-negative bacteria remains to be explored. This study aimed to investigate the role of the BasS/BasR two-component system (TCS) in () K12 response to plantaricin BM-1. The IC values for plantaricin BM-1 in K12, mutant ( JW4073), and mutant ( JW4074) strains were found to be 10.85, 8.94, and 7.62 mg/mL, respectively. Growth curve experiments showed that mutations in the BasS/BasR TCS led to an increase in the sensitivity of K12 to plantaricin BM-1 and that after gene complementation, the complemented mutant strain regained its original sensitivity. Proteomic analysis showed that 100 and 26 proteins were upregulated and 62 and 58 proteins were downregulated in JW4073 and JW4074, respectively. These differential proteins, which exhibited different molecular functions and participated in different molecular pathways, were mainly concentrated in the cytoplasm. More specifically, mutations in and were found to affect the synthesis and metabolism of many substances in , including many important amino acids and enzymes involved in cellular activities. In addition, 14 proteins, including 8 proteins involved in the tricarboxylic acid (TCA) cycle, were found to be downregulated in both JW4073 and JW4074. Growth curve experiments showed that the deletion of these proteins could increase the sensitivity of to plantaricin BM-1. Therefore, we speculate that TCA pathway regulation may be an important mechanism by which the BasS/BasR TCS regulates the sensitivity of to plantaricin BM-1. This finding will facilitate the determination of the mechanism underlying the action of class IIa bacteriocins against gram-negative bacteria.
植物乳杆菌素BM-1是由BM-1产生的一种IIa类细菌素,对许多革兰氏阳性菌和革兰氏阴性菌都具有显著的抗菌活性。然而,IIa类细菌素对革兰氏阴性菌的作用机制仍有待探索。本研究旨在探究BasS/BasR双组分系统(TCS)在大肠杆菌K12对植物乳杆菌素BM-1的应答中的作用。发现植物乳杆菌素BM-1对大肠杆菌K12、basS突变体(JW4073)和basR突变体(JW4074)菌株的IC值分别为10.85、8.94和7.62 mg/mL。生长曲线实验表明,BasS/BasR TCS中的突变导致大肠杆菌K12对植物乳杆菌素BM-1的敏感性增加,并且在基因互补后,互补突变株恢复了其原始敏感性。蛋白质组学分析表明,在JW4073和JW4074中分别有100和26种蛋白质上调,62和58种蛋白质下调。这些差异蛋白质具有不同的分子功能并参与不同的分子途径,主要集中在细胞质中。更具体地说,发现basS和basR中的突变会影响大肠杆菌中许多物质的合成和代谢,包括许多参与细胞活动的重要氨基酸和酶。此外,在JW4073和JW4074中均发现有14种蛋白质下调,其中包括8种参与三羧酸(TCA)循环的蛋白质。生长曲线实验表明,这些蛋白质的缺失会增加大肠杆菌对植物乳杆菌素BM-1的敏感性。因此,我们推测TCA途径调节可能是BasS/BasR TCS调节大肠杆菌对植物乳杆菌素BM-1敏感性的重要机制。这一发现将有助于确定IIa类细菌素对革兰氏阴性菌作用的机制。