Cell-based screening strategy in the search for bioactive microbial secondary metabolites

Here we present results obtained in our group from cell-based screenings of microbial secondary metabolites using intact microbial and mammalian cells having specific functions. We present summaries of the following five categories of compounds and the strategies that were used to identify them. (1) Antibacterial agents targeting cell wall peptidoglycan synthesis as well as active agents against organisms other than bacteria were identified by a combination of Bacillus subtilis and Mycoplasma as test organisms. (2) Antimetabolites, such as herbicidal agents targeting glutamine synthase, were identified using B. subtilis grown on minimal medium with or without the addition of glutamine and folate inhibitors by combination of B. subtilis and Enterococcus faecium, which have distinct folate metabolic pathways. (3) Among inhibitors of lipid metabolism, acyl-CoA synthetase inhibitors were identified using two mutants of C. lipolytica with different deletion sites in the fatty acid metabolic pathways, HMG-CoA synthase inhibitors were identified using Vero cells cultured in the presence and absence of mevalonate, and anti-atherosclerosis agents were identified using macrophages. (4) IL-6 inhibitors were detected by a combination of IL-6-dependent and independent murine hybridoma MH60 cells, and (5) Small molecules with TNF-like activities were identified using a mouse neuroblastoma cell line. Some advantages of such screening method and the significance of the identified compounds are discussed. (Contributed by Satoshi OMURA, M.J.A., Feb. 12, 2004)