1. Biosynthetic engineering of the antifungal, anti-MRSA auroramycin.
- Author
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Yeo WL, Heng E, Tan LL, Lim YW, Ching KC, Tsai DJ, Jhang YW, Lauderdale TL, Shia KS, Zhao H, Ang EL, Zhang MM, Lim YH, and Wong FT
- Subjects
- Anti-Bacterial Agents chemistry, Anti-Bacterial Agents pharmacology, Antifungal Agents pharmacology, CRISPR-Cas Systems, Gene Editing methods, Methicillin-Resistant Staphylococcus aureus drug effects, Microbial Sensitivity Tests, Polyenes chemistry, Streptomyces metabolism, Anti-Bacterial Agents biosynthesis, Antifungal Agents chemistry, Biosynthetic Pathways genetics, Metabolic Engineering methods, Streptomyces genetics
- Abstract
Using an established CRISPR-Cas mediated genome editing technique for streptomycetes, we explored the combinatorial biosynthesis potential of the auroramycin biosynthetic gene cluster in Streptomyces roseosporous. Auroramycin is a potent anti-MRSA polyene macrolactam. In addition, auroramycin has antifungal activities, which is unique among structurally similar polyene macrolactams, such as incednine and silvalactam. In this work, we employed different engineering strategies to target glycosylation and acylation biosynthetic machineries within its recently elucidated biosynthetic pathway. Auroramycin analogs with variations in C-, N- methylation, hydroxylation and extender units incorporation were produced and characterized. By comparing the bioactivity profiles of five of these analogs, we determined that unique disaccharide motif of auroramycin is essential for its antimicrobial bioactivity. We further demonstrated that C-methylation of the 3, 5-epi-lemonose unit, which is unique among structurally similar polyene macrolactams, is key to its antifungal activity.
- Published
- 2020
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