Aso Yuji, Nagao Jun-Ichi, Koga Hanako, Okuda Ken-Ichi, Kanemasa Youhei, Sashihara Toshihiro, Nakayama Jiro, Sonomoto Kenji
Laboratory of Microbial Technology, Division of Microbial Science and Technology, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
J Biosci Bioeng. 2004;98(6):429-36. doi: 10.1016/S1389-1723(05)00308-7.
Nukacin ISK-1 is a lantibiotic produced by Staphylococcus warneri ISK-1. The gene cluster of nukacin ISK-1 consists of at least nukAMTFEG, ORF1 and ORF7. In this study, we demonstrated the heterologous production of nukacin ISK-1 in Lactococcus lactis by the artificial polycistronic expression of nukAMTFEG-ORF7 under the control of the nisin-controlled expression (NICE) system. Consequently, the recombinant L. lactis showed antimicrobial activity. Mass analysis clarified the presence of nukacin ISK-1 produced in the culture supernatant. These results suggested that the recombinant L. lactis produced nukacin ISK-1 heterologously. Inactivation of nukA, -M or -T resulted in the complete loss of the nukacin ISK-1 production phenotype. This finding suggested that nukAMT are indispensably associated with the biosynthesis of nukacin ISK-1. To our knowledge, this is the first report of the heterologous production of lantibiotic using the NICE system.
Nukacin ISK-1是由沃氏葡萄球菌ISK-1产生的一种羊毛硫抗生素。Nukacin ISK-1的基因簇至少由nukAMTFEG、开放阅读框1（ORF1）和开放阅读框7（ORF7）组成。在本研究中，我们通过在乳链菌肽控制表达（NICE）系统的控制下对nukAMTFEG-ORF7进行人工多顺反子表达，证明了Nukacin ISK-1在乳酸乳球菌中的异源生产。因此，重组乳酸乳球菌表现出抗菌活性。质谱分析明确了培养上清液中产生的Nukacin ISK-1的存在。这些结果表明重组乳酸乳球菌异源生产了Nukacin ISK-1。nukA、-M或-T的失活导致Nukacin ISK-1生产表型的完全丧失。这一发现表明nukAMT与Nukacin ISK-1的生物合成不可或缺地相关。据我们所知，这是首次关于使用NICE系统异源生产羊毛硫抗生素的报道。
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