World Journal of Microbiology and Biotechnology ( IF 4.0 ) Pub Date : 2021-09-14 , DOI: 10.1007/s11274-021-03135-x Amrutha Bindu 1 , N. Lakshmidevi 1
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The focus of the present study was to characterize antimicrobial peptide produced by potential probiotic cultures of Enterococcus durans DB-1aa (MCC4243), Lactiplantibacillus plantarum Cu2-PM7 (MCC4246) and Limosilactobacillus fermentum Cu3-PM8 (MCC4233) against Staphylococus aureus MTCC 96 and Escherichia coli MTCC118. The growth kinetic assay revealed 24 h of incubation to be optimum for bacteriocin production. The partially purified compound of all the three selected cultures after ion-exchange chromatography was found to be thermoresistant and stable under a wide range of pH. The compound was sensitive to proteinase-K, but resistant to trypsin, α-amylase and lipase. Comparatively, bacteriocins from L. fermentum Cu3-PM8 and L. plantarum Cu2-PM7 showed higher stability under studied parameter, hence was taken up for further investigation. The apparent molecular weight of bacteriocin from L. fermentum MCC4233 and L. plantarum MCC4246 was found to be 3.5 kDa. Further, plantaricin gene from MCC4246 was characterized in silico. The translated partial amino acid sequence of the plnA gene in MCC4246 displayed 48 amino acids showing 100 % similarity with plantaricin A of Lactobacillus plantarum (WP_0036419). The sequence revealed 7 β sheets, 6 α sheets, 6 predicted coils and 9 predicted turns. The predicted properties of the peptide included an isoelectric point of 10.82 and a hydrophobicity of 48.6 %. The molecular approach of using Geneious Prime software and protein prediction data base for characterization of bacteriocin is novel and predicts “KSSAYSLQMGATAIKQVKKLFKKWGW” to be a peptide responsible for antimicrobial activity. The study provides information about a broad spectrum bacteriocin in native probiotic culture and paves a way towards its application in functional foods as a biopreservative agent.
Graphic abstract
中文翻译：
用于表征来自潜在益生菌培养物的抗金黄色葡萄球菌和大肠杆菌的抗菌肽的体外和计算机方法
本研究的重点是表征由耐久肠球菌DB-1aa (MCC4243)、植物乳杆菌Cu2-PM7 (MCC4246) 和发酵乳杆菌Cu3-PM8 (MCC4233)的潜在益生菌培养物产生的抗金黄色葡萄球菌和 MTCC 的抗菌肽大肠杆菌MTCC118。生长动力学分析表明，24 小时的孵育最适合细菌素的产生。离子交换层析后所有三种选定培养物的部分纯化化合物均具有耐热性，并且在很宽的 pH 范围内稳定。该化合物对蛋白酶 K 敏感，但对胰蛋白酶、α-淀粉酶和脂肪酶具有抗性。相比之下，细菌素来自发酵乳杆菌Cu3-PM8 和植物乳杆菌Cu2-PM7 在研究参数下表现出更高的稳定性，因此被用于进一步研究。发现来自发酵乳杆菌MCC4233和植物乳杆菌MCC4246的细菌素的表观分子量为 3.5 kDa。此外，来自 MCC4246 的植物素基因在计算机上进行了表征。MCC4246 中pln A 基因的翻译部分氨基酸序列显示 48 个氨基酸，与植物乳杆菌的植物蛋白 A 具有 100% 的相似性(WP_0036419)。该序列揭示了 7 个 β 片层、6 个 α 片层、6 个预测的线圈和 9 个预测的匝数。肽的预测特性包括 10.82 的等电点和 48.6% 的疏水性。使用 Geneious Prime 软件和蛋白质预测数据库表征细菌素的分子方法是新颖的，并预测“KSSAYSLQMGATAIKQVKKLFKKWGW”是一种负责抗菌活性的肽。该研究提供了有关天然益生菌培养物中广谱细菌素的信息，并为其在功能性食品中作为生物防腐剂的应用铺平了道路。
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