A&J Science Co., Ltd., 80 Chumbok Ro, Dong Gu, Daegu 41061, Republic of Korea.
Department of Agricultural Biotechnology, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 08826, Republic of Korea.
ACS Chem Biol. 2023 Feb 17;18(2):265-272. doi: 10.1021/acschembio.2c00612. Epub 2023 Jan 24.
Micrococcin P1 and P2 are thiopeptides with a wide range of biological functions including antibacterial and antimalarial activities. We previously demonstrated optimized enzymatic sequences for the exclusive and scalable biosynthesis of micrococcin P2. Thiocillin IV is predicted to be the congener of -methylated micrococcin P2, but the exact structure has not been elucidated. In this study, we report the first scalable biosynthesis and full structural characterization of thiocillin IV, a 26-membered thiopeptide. This was achieved by generating a recombinant plasmid by inserting a gene encoding an -methyltransferase, and genes responsible for micrococcin P2 production and incorporating them into a strain. With the incorporation of precursor peptide genes and optimal culture conditions, production reached 2.4 mg/L of culture. The purified thiocillin IV structure was identified as -methylated micrococcin P2 at the 8-Thr position, and its promising biological activity toward various Gram-positive pathogens was observed. This study provides -mediated site-selective methylation and opens a biotechnological opportunity to produce selective thiopeptides.
微球菌素 P1 和 P2 是具有广泛生物学功能的噻肽类化合物，包括抗菌和抗疟活性。我们之前证明了优化的酶序列可用于微球菌素 P2 的专属性和可扩展的生物合成。硫西林 IV 被预测为 -甲基化微球菌素 P2 的同系物，但确切结构尚未阐明。在这项研究中，我们报告了第一个可扩展的生物合成和全结构表征的硫西林 IV，一种 26 元噻肽。这是通过插入一个编码 -甲基转移酶的基因并将负责微球菌素 P2 生产的基因和它们一起插入到一个菌株中来实现的。通过加入前体肽基因和优化的培养条件，产量达到了 2.4 毫克/升的培养物。纯化的硫西林 IV 结构被鉴定为在 8-Thr 位置被 -甲基化的微球菌素 P2，并且观察到它对各种革兰氏阳性病原体具有有前途的生物活性。这项研究提供了 -介导的位点选择性甲基化，并为生产选择性噻肽开辟了生物技术机会。
