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一株类芽孢杆菌的分离及其广谱脂肽抗生素的结构解析。

Isolation of a Paenibacillus sp. strain and structural elucidation of its broad-spectrum lipopeptide antibiotic.

机构信息

Department of Food Science & Technology, Ohio State University, Columbus, Ohio, USA.

出版信息

Appl Environ Microbiol. 2012 May;78(9):3156-65. doi: 10.1128/AEM.07782-11. Epub 2012 Feb 24.

Abstract

This research was initiated to search for novel antimicrobial compounds produced by food or environmental microorganisms. A new bacterial strain, designated OSY-SE, which produces a unique and potent antimicrobial agent was isolated from soil. The isolate was identified as a Paenibacillus sp. through cultural, biochemical, and genetic analyses. An antimicrobial compound was extracted from Paenibacillus OSY-SE with acetonitrile and purified using liquid chromatography. After analyses by mass spectrometry (MS) and nuclear magnetic resonance (NMR), the antimicrobial compound was determined to be a cyclic lipopeptide consisting of a C(15) fatty acyl (FA) chain and 13 amino acids. The deduced sequence is FA-Orn-Val-Thr-Orn-Ser-Val-Lys-Ser-Ile-Pro-Val-Lys-Ile. The carboxyl-terminal Ile is connected to Thr by ester linkage. The new compound, designated paenibacterin, showed antagonistic activities against most Gram-positive and Gram-negative bacteria tested, including Listeria monocytogenes, methicillin-resistant Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella enterica serovar Typhimurium. Paenibacterin is resistant to trypsin, lipase, α-glucosidase, and lysozyme. Its antimicrobial activity was lost after digestion by pronase and polymyxin acylase. Paenibacterin is readily soluble in water and fairly stable to exposure to heat and a wide range of pH values. The new isolate and its antimicrobial agent are being investigated for usefulness in food and medical applications.

摘要

本研究旨在寻找食品或环境微生物产生的新型抗菌化合物。从土壤中分离到一株新型细菌 OSY-SE,它能产生独特而有效的抗菌剂。通过培养、生化和遗传分析,该分离株被鉴定为蜡状芽孢杆菌属。用乙腈从蜡状芽孢杆菌 OSY-SE 中提取抗菌化合物,并用液相色谱法纯化。经质谱(MS)和核磁共振(NMR)分析,该抗菌化合物被确定为一种由 C(15)脂肪酸(FA)链和 13 个氨基酸组成的环状脂肽。推断的序列为 FA-Orn-Val-Thr-Orn-Ser-Val-Lys-Ser-Ile-Pro-Val-Lys-Ile。羧基末端的 Ile 通过酯键与 Thr 连接。这种新化合物被命名为杆菌菌素,对大多数测试的革兰氏阳性和革兰氏阴性细菌表现出拮抗活性,包括单核细胞增生李斯特菌、耐甲氧西林金黄色葡萄球菌、大肠杆菌 O157:H7 和肠炎沙门氏菌血清型 Typhimurium。杆菌菌素对胰蛋白酶、脂肪酶、α-葡萄糖苷酶和溶菌酶具有抗性。经蛋白酶和多粘菌素酰化酶消化后,其抗菌活性丧失。杆菌菌素易溶于水,在暴露于热和广泛的 pH 值范围内相当稳定。正在研究新的分离株及其抗菌剂在食品和医疗应用中的用途。

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