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多粘芽孢杆菌E681中新型羊毛硫抗生素多粘菌素兰生物合成基因簇的鉴定及其产物的表征

Identification of the biosynthesis gene cluster for the novel lantibiotic paenilan from Paenibacillus polymyxa E681 and characterization of its product.

作者信息

Park J-E, Kim H-R, Park S-Y, Choi S-K, Park S-H

机构信息

Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Yuseong-gu, Daejeon, Korea.

Department of Biosystems and Bioengineering, KRIBB School of Biotechnology, Korea University of Science and Technology (UST), Yuseong-gu, Daejeon, Korea.

出版信息

J Appl Microbiol. 2017 Nov;123(5):1133-1147. doi: 10.1111/jam.13580. Epub 2017 Oct 13.

Abstract

AIMS

To analyse the function of a putative lantibiotic gene cluster of Paenibacillus polymyxa E681 and to characterize its product, paenilan.

METHODS AND RESULTS

Comparative analysis of the lantibiotic gene cluster of E681 revealed that the cluster, consisting of 11 open reading frames, is involved in the biosynthesis of a class I lantibiotic. The pnlA gene encoding the prepeptide PnlA was identified and P. polymyxa strain EPT14 producing only paenilan was constructed by knockout of the other five antibiotic biosynthetic gene clusters of E681. Paenilan was purified from EPT14 culture by solvent partitioning, ODS silica gel chromatography and reversed-phase preparative HPLC. The molecular mass (2510·10 Da) and structure of paenilan analysed by Nanoelectrospray ionization mass spectrometry (MS) and MS/MS showed that paenilan is a novel class I lantibiotic. Paenilan exhibited antimicrobial activity against Gram-positive bacteria such as Bacillus cereus, Micrococcus luteus and Paenibacillus durus. The paenilan gene is well-conserved in different Paenibacillus sp. isolated from globally distant places.

CONCLUSIONS

The lantibiotic gene cluster of P. polymyxa E681 was analysed and its product, a novel and useful lantibiotic named paenilan that inhibits the growth of some Gram-positive bacteria, was characterized.

SIGNIFICANCE AND IMPACT OF THE STUDY

Paenibacillus species are a good source of new lantibiotics, and the conservation of the paenilan gene among Paenibacillus sp. implies paenilan has an important function(s) for their survival.

摘要

目的

分析多粘芽孢杆菌E681中一个假定的羊毛硫抗生素基因簇的功能,并对其产物多粘菌素进行表征。

方法与结果

对E681的羊毛硫抗生素基因簇进行比较分析,发现该基因簇由11个开放阅读框组成,参与I类羊毛硫抗生素的生物合成。鉴定了编码前肽PnlA的pnlA基因,并通过敲除E681的其他五个抗生素生物合成基因簇构建了仅产生多粘菌素的多粘芽孢杆菌菌株EPT14。通过溶剂分配、ODS硅胶柱色谱和反相制备型高效液相色谱从EPT14培养物中纯化多粘菌素。通过纳米电喷雾电离质谱(MS)和MS/MS分析的多粘菌素的分子量(2510·10 Da)和结构表明,多粘菌素是一种新型的I类羊毛硫抗生素。多粘菌素对革兰氏阳性菌如蜡样芽孢杆菌、藤黄微球菌和耐久芽孢杆菌具有抗菌活性。多粘菌素基因在从全球不同地方分离的不同芽孢杆菌属中保守性良好。

结论

分析了多粘芽孢杆菌E681的羊毛硫抗生素基因簇,并对其产物——一种名为多粘菌素的新型且有用的抑制某些革兰氏阳性菌生长的羊毛硫抗生素进行了表征。

研究的意义和影响

芽孢杆菌属是新羊毛硫抗生素的良好来源,多粘菌素基因在芽孢杆菌属中的保守性意味着多粘菌素对它们的生存具有重要功能。

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