Institute of Medical Microbiology, Immunology and Parasitology, University of Bonn, Bonn, Germany.
PLoS One. 2011;6(7):e22389. doi: 10.1371/journal.pone.0022389. Epub 2011 Jul 21.
Lantibiotics are small peptide antibiotics that contain the characteristic thioether amino acids lanthionine and methyllanthionine. As ribosomally synthesized peptides, lantibiotics possess biosynthetic gene clusters which contain the structural gene (lanA) as well as the other genes which are involved in lantibiotic modification (lanM, lanB, lanC, lanP), regulation (lanR, lanK), export (lanT(P)) and immunity (lanEFG). The lantibiotic mersacidin is produced by Bacillus sp. HIL Y-85,54728, which is not naturally competent.
METHODOLOGY/PRINCIPAL FINDINGS: The aim of these studies was to test if the production of mersacidin could be transferred to a naturally competent Bacillus strain employing genomic DNA of the producer strain. Bacillus amyloliquefaciens FZB42 was chosen for these experiments because it already harbors the mersacidin immunity genes. After transfer of the biosynthetic part of the gene cluster by competence transformation, production of active mersacidin was obtained from a plasmid in trans. Furthermore, comparison of several DNA sequences and biochemical testing of B. amyloliquefaciens FZB42 and B. sp. HIL Y-85,54728 showed that the producer strain of mersacidin is a member of the species B. amyloliquefaciens.
CONCLUSIONS/SIGNIFICANCE: The lantibiotic mersacidin can be produced in B. amyloliquefaciens FZB42, which is closely related to the wild type producer strain of mersacidin. The new mersacidin producer strain enables us to use the full potential of the biosynthetic gene cluster for genetic manipulation and downstream modification approaches.
类杆菌肽是一种含有硫醚氨基酸即赖氨酰和甲硫氨酰的小肽抗生素。作为核糖体合成的肽,类杆菌肽具有生物合成基因簇,其中包含结构基因(lanA)以及参与类杆菌肽修饰(lanM、lanB、lanC、lanP)、调控(lanR、lanK)、输出(lanT(P))和免疫(lanEFG)的其他基因。杆菌肽 mersacidin 由 Bacillus sp. HIL Y-85,54728 产生,该菌自然没有感受态。
方法/主要发现:这些研究的目的是检验是否可以通过转化感受态的方法将 mersacidin 的生产转移到自然感受态的芽孢杆菌菌株,采用生产菌株的基因组 DNA。选择地衣芽孢杆菌 FZB42 进行这些实验,因为它已经拥有 mersacidin 免疫基因。在通过感受态转化转移生物合成基因簇的合成部分后,在转座质粒中获得了活性 mersacidin 的生产。此外,对几种 DNA 序列的比较和对地衣芽孢杆菌 FZB42 和 Bacillus sp. HIL Y-85,54728 的生化测试表明,mersacidin 的生产菌株是地衣芽孢杆菌的一个成员。
结论/意义:杆菌肽 mersacidin 可在与 mersacidin 野生型生产菌株密切相关的地衣芽孢杆菌 FZB42 中产生。新型 mersacidin 生产菌株使我们能够充分利用生物合成基因簇进行遗传操作和下游修饰方法。