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一株云南传统发酵酸乳中产细菌素乳酸菌的分离鉴定及其生物学特性研究

A novel bacteriocin against Staphylococcus aureus from Lactobacillus paracasei isolated from Yunnan traditional fermented yogurt: Purification, antibacterial characterization, and antibiofilm activity.

机构信息

Faculty of Life Science and Technology, Kunming University of Science and Technology, Yunnan Kunming 650500, China.

State Key Laboratory of Plateau Ecology and Agriculture and Qinghai Academy of Animal and Veterinary Science, Qinghai University, Qinghai Xining 810000, China.

出版信息

J Dairy Sci. 2022 Mar;105(3):2094-2107. doi: 10.3168/jds.2021-21126.

Abstract

Staphylococcus aureus and its biofilm have emerged as a significant threat to the safety of dairy products. In recent years, lactic acid bacteria (LAB) bacteriocins have been widely acknowledged as the potential natural antibacterial substance in food biopreservation due to their excellent antibacterial effects. However, few LAB bacteriocins with antibacterial and antibiofilm activity against S. aureus have been reported in dairy products. In the present study, a novel bacteriocin LSX01 of Lactobacillus paracasei LS-6 isolated from a traditional fermented yogurt produced in Yunnan, China, was purified and characterized extensively. The LSX01 possessed a molecular weight of 967.49 Da and an AA sequence of LDQAGISYT. The minimum inhibitory concentration of LSX01 against S. aureus_45 was 16.90 μg/mL, which was close to or lower than the previously reported bacteriocins. The LSX01 exhibited an extensive antimicrobial spectrum against both gram-positive and gram-negative bacteria. Moreover, LSX01 exhibited excellent tolerance to heat and acid-base treatments, and sensitivity to the proteolytic enzymes, such as pepsin and proteinase K. Furthermore, the treatment of S. aureus_45 planktonic cells with LSX01 significantly reduced their metabolic activity and disrupted the cell membrane integrity. Scan electron microscopy results demonstrated that LSX01 induced cytoplasmic content leakage and cell deformation. Additionally, biofilm formation of S. aureus_45 was also significantly inhibited by LSX01. Overall, the results suggested that the novel LAB bacteriocin LSX01 possessed antibacterial activity and antibiofilm activity against S. aureus and, hence, could have potential for improving safety of dairy products.

摘要

金黄色葡萄球菌及其生物膜已成为乳制品安全的重大威胁。近年来,由于具有优异的抗菌效果,乳酸菌(LAB)细菌素已被广泛认为是食品生物保鲜的潜在天然抗菌物质。然而,在乳制品中报道的具有抗金黄色葡萄球菌活性和抗生物膜活性的 LAB 细菌素很少。在本研究中,从中国云南传统发酵酸奶中分离到的副干酪乳杆菌 LS-6 的新型细菌素 LSX01 被广泛纯化和表征。LSX01 的分子量为 967.49 Da,AA 序列为 LDQAGISYT。LSX01 对金黄色葡萄球菌_45 的最小抑菌浓度为 16.90 μg/mL,接近或低于先前报道的细菌素。LSX01 对革兰氏阳性和革兰氏阴性细菌均具有广泛的抗菌谱。此外,LSX01 对热和酸碱处理具有出色的耐受性,并且对胃蛋白酶和蛋白酶 K 等蛋白水解酶敏感。此外,LSX01 处理金黄色葡萄球菌_45 浮游细胞可显著降低其代谢活性并破坏细胞膜完整性。扫描电子显微镜结果表明,LSX01 诱导细胞质内容物泄漏和细胞变形。此外,LSX01 还显著抑制了金黄色葡萄球菌_45 的生物膜形成。总体而言,结果表明新型 LAB 细菌素 LSX01 对金黄色葡萄球菌具有抗菌活性和抗生物膜活性,因此可能有助于提高乳制品的安全性。

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