Biosynthetic engineering of the antifungal, anti-MRSA auroramycin

• Published in: Microbial cell factories Vol. 19; no. 1; p. 3
• Main Authors: Yeo, Wan Lin, Heng, Elena, Tan, Lee Ling, Lim, Yi Wee, Ching, Kuan Chieh, Tsai, De-Juin, Jhang, Yi Wun, Lauderdale, Tsai-Ling, Shia, Kak-Shan, Zhao, Huimin, Ang, Ee Lui, Zhang, Mingzi M, Lim, Yee Hwee, Wong, Fong T
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 06.01.2020; BioMed Central; BMC
• Subjects: Acylation; Analogs; Annotations; Anti-Bacterial Agents - biosynthesis; Anti-Bacterial Agents - chemistry; Anti-Bacterial Agents - pharmacology; Antibiotics; Antifungal activity; Antifungal agents; Antifungal Agents - chemistry; Antifungal Agents - pharmacology; Biological activity; Biosynthesis; Biosynthetic Pathways - genetics; Combinatorial analysis; Combinatorial biosynthesis; CRISPR; CRISPR-Cas Systems; Disaccharides; Drug resistance; Engineering; Fungicides; Gene Editing - methods; Genes; Genome editing; Genomes; Genomics; Glycosylation; Hydroxylation; Metabolic Engineering - methods; Methicillin-Resistant Staphylococcus aureus - drug effects; Methods; Methylation; Microbial Sensitivity Tests; Microbiological synthesis; Natural products; Polyene macrolactam; Polyenes - chemistry; Staphylococcus aureus; Streptomyces; Streptomyces - genetics; Streptomyces - metabolism; Streptomycetes; Sugar; Testing; China; Combinatorial biosynthesis; Streptomyces; Polyene macrolactam
• ISSN: 1475-2859; 1475-2859
• DOI: 10.1186/s12934-019-1274-y

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.