Engelke G, Gutowski-Eckel Z, Kiesau P, Siegers K, Hammelmann M, Entian K D
Institut für Mikrobiologie, Johann Wolfgang Goethe-Universität Frankfurt, Federal Republic of Germany.
Appl Environ Microbiol. 1994 Mar;60(3):814-25. doi: 10.1128/aem.60.3.814-825.1994.
The biosynthetic genes of the nisin-producing strain Lactococcus lactis 6F3 are organized in an operon-like structure starting with the structural gene nisA followed by the genes nisB, nisT, and nisC, which are probably involved in chemical modification and secretion of the prepeptide (G. Engelke, Z. Gutowski-Eckel, M. Hammelmann, and K.-D. Entian, Appl. Environ. Microbiol. 58:3730-3743, 1992). Subcloning of an adjacent 5-kb downstream region revealed additional genes involved in nisin biosynthesis. The gene nisI, which encodes a lipoprotein, causes increased immunity after its transformation into nisin-sensitive L. lactis MG1614. It is followed by the gene nisP, coding for a subtilisin-like serine protease possibly involved in processing of the secreted leader peptide. Adjacent to the 3' end of nisP the genes nisR and nisK were identified, coding for a regulatory protein and a histidine kinase, showing marked similarities to members of the OmpR/EnvZ-like subgroup of two-component regulatory systems. The deduced amino acid sequences of nisR and nisK exhibit marked similarities to SpaR and SpaK, which were recently identified as the response regulator and the corresponding histidine kinase of subtilin biosynthesis. By using antibodies directed against the nisin prepeptide and the NisB protein, respectively, we could show that nisin biosynthesis is regulated by the expression of its structural and biosynthetic genes. Prenisin expression starts in the exponential growth phase and precedes that of the NisB protein by approximately 30 min. Both proteins are expressed to a maximum in the stationary growth phase.
产乳链菌肽的乳酸乳球菌6F3的生物合成基因以操纵子样结构组织，起始于结构基因nisA，随后是基因nisB、nisT和nisC，它们可能参与前肽的化学修饰和分泌（G. Engelke、Z. Gutowski-Eckel、M. Hammelmann和K.-D. Entian，《应用与环境微生物学》58:3730 - 3743，1992年）。对相邻的5 kb下游区域进行亚克隆揭示了参与乳链菌肽生物合成的其他基因。编码脂蛋白的基因nisI转化到对乳链菌肽敏感的乳酸乳球菌MG1614后可增强免疫力。接着是基因nisP，其编码一种可能参与分泌前导肽加工的枯草杆菌蛋白酶样丝氨酸蛋白酶。在nisP的3'端相邻位置鉴定出基因nisR和nisK，它们分别编码一种调节蛋白和一种组氨酸激酶，与双组分调节系统的OmpR/EnvZ样亚组成员有显著相似性。nisR和nisK推导的氨基酸序列与最近被鉴定为枯草菌素生物合成的应答调节因子和相应组氨酸激酶的SpaR和SpaK有显著相似性。通过分别使用针对乳链菌肽前肽和NisB蛋白的抗体，我们能够表明乳链菌肽的生物合成受其结构和生物合成基因表达的调控。前乳链菌肽的表达在指数生长期开始，比NisB蛋白的表达提前约30分钟。两种蛋白在稳定生长期都表达至最大值。
