Purification and characterization of a lipopeptide produced by Bacillus thuringiensis CMB26
Purification and characterization of a lipopeptide produced by Bacillus thuringiensis CMB26
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DOI：
10.1111/j.1365-2672.2004.02356.x
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发表时间：
2004-01-01
影响因子：
4
通讯作者：
Chi, YT
中科院分区：
文献类型：
--
作者：
Kim, PI;Bai, H;Chi, YT
目的：从土壤中筛选出一株对辣椒炭疽病具有生物防治活性的拮抗菌，并对其产生的生防菌进行纯化和鉴定。方法与结果：从土壤中筛选出对辣椒炭疽病具有生物防治活性的细菌，其中苏云金芽孢杆菌（Bacillus thuringiensis）CMB 26的活性最强。由B产生的脂肽。苏云金杆菌CMB 26通过用3 N HCl调节pH 2而沉淀，并使用氯仿/甲醇（2：1，v/v）和反相HPLC进行提取。通过MALDI-TOF质谱法估计分子量为1447 Da。扫描电子显微镜和光学显微镜显示脂肽对大肠杆菌O 157：ac 88、甘蓝白色蝴蝶（Pieris raiscrucivora）幼虫和植物病原真菌具有活性。脂肽具有环状结构，氨基酸组成为l-Glu、d-Orn、l-Tyr、d-allo-Thr、d-Ala、d-Val、l-Pro和l-Ile，摩尔比为3：1：2：1：1：2：1：1。结论：从B中分离纯化了A脂肽，并对其结构进行了表征。苏云金芽孢杆菌CMB 26，并发现与脂肽丰霉素相似。该脂肽可作为生物防治剂，并具有杀菌、杀真菌和杀虫活性。研究意义和影响：与表面活性素和伊枯草菌素相比，B脂肽。苏云金杆菌CMB 26对植物病原真菌具有较强的抑菌活性。该脂肽是农业中病原体生物防治的候选者。
Aims: To isolate an antagonist for use in the biological control of phytopathogenic fungi including Colletotrichum gloeosporioides, then to purify and characterize the biocontrol agent produced by the antagonist.Methods and Results: Bacteria that exhibited antifungal activity against the causative agent pepper anthracnose were isolated from soil, with Bacillus thuringiensis CMB26 showing the strongest activity. A lipopeptide produced by B. thuringiensis CMB26 was precipitated by adjusting the pH 2 with 3 n HCl and extracted using chloroform/methanol (2 : 1, v/v) and reversed-phase HPLC. The molecular weight was estimated as 1447 Da by MALDI-TOF mass spectrometry. Scanning electron and optical microscopies showed that the lipopeptide has activity against Escherichia coli O157:ac88, larvae of the cabbage white butterfly (Pieris rapae crucivora) and phytopathogenic fungi. The lipopeptide had cyclic structure and the amino acid composition was l-Glu, d-Orn, l-Tyr, d-allo-Thr, d-Ala, d-Val, l-Pro, and l-Ile in a molar ratio of 3 : 1 : 2 : 1 : 1 : 2 : 1 : 1. The purified lipopeptide showed the same amino acid composition as fengycin, but differed slightly in fatty acid composition, in which the double bond was at carbons 13-14 (m/z 303, 316) and there was no methyl group.Conclusion: A lipopeptide was purified and characterized from B. thuringiensis CMB26 and found to be similar to the lipopeptide fengycin. This lipopeptide can function as a biocontrol agent, and exhibits fungicidal, bactericidal, and insecticidal activity.Significance and Impact of the Study: Compared with surfactin and iturin, the lipopeptide from B. thuringiensis CMB26 showed stronger antifungal activity against phytopathogenic fungi. This lipopeptide is a candidate for the biocontrol of pathogens in agriculture.