Asano Yuri, Saito Shun, Ujie Yukiko, Iwata Chisato, Yaguchi Takashi, Arai Midori A
Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, Kanagawa 223-8522, Japan.
Medical Mycology Research Center, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8673, Japan.
ACS Omega. 2024 Oct 10;9(42):43129-43137. doi: 10.1021/acsomega.4c07124. eCollection 2024 Oct 22.
The activation of secondary metabolism plays a pivotal role in the discovery of novel natural products. We recently developed a coculture method involving actinomycetes and mouse macrophage-like cells to stimulate the production of bioactive compounds. A black koji mold, IFM 61405, markedly enhanced the production of (3,8)-8-hydroxy-3-carboxy-2-methylenenonanoic acid (), (3,8)-8-hydroxy-3-carboxy-2-methylenenonanoic acid (), and (3)-9-hydroxy-3-carboxy-2-methylenenonanoic acid () when coincubated with J774.1 mouse macrophage cells. The production of and increased by at least 3.5-fold and 2.7-fold, respectively, compared to monoculture after 7 days. A mechanistic investigation revealed that a protease from strain IFM 61405 plays a key role in enhancing the production of and . This enhancement was not replicated in IFM 59706, a nonkoji mold, despite the presence of biosynthetic genes for and in IFM 59706. Furthermore, the addition of protease inhibitors suppressed the production of and , suggesting that proteins secreted from animal cells, likely degraded by proteases secreted by strain IFM 61405, serve as precursors for and . The results show that the strategy of coculturing koji mold with animal cells has the potential to enhance the production of natural products.
次级代谢的激活在新型天然产物的发现中起着关键作用。我们最近开发了一种共培养方法，涉及放线菌和小鼠巨噬细胞样细胞，以刺激生物活性化合物的产生。一种黑曲霉IFM 61405，与J774.1小鼠巨噬细胞共孵育时，显著提高了(3,8)-8-羟基-3-羧基-2-亚甲基壬酸、(3,8)-8-羟基-3-羧基-2-亚甲基壬酸和(3)-9-羟基-3-羧基-2-亚甲基壬酸的产量。与7天后的单培养相比，和的产量分别增加了至少3.5倍和2.7倍。机理研究表明，来自菌株IFM 61405的一种蛋白酶在提高和的产量中起关键作用。尽管IFM 59706中存在和的生物合成基因，但在非曲霉IFM 59706中并未重复这种提高。此外，添加蛋白酶抑制剂会抑制和的产生，这表明动物细胞分泌的蛋白质，可能被菌株IFM 61405分泌的蛋白酶降解，可作为和的前体。结果表明，将曲霉与动物细胞共培养的策略有可能提高天然产物的产量。
