1. Identification and engineering of 32 membered antifungal macrolactone notonesomycins
- Author
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Shawn Hoon, Alexander Lezhava, Fernanda L. Sirota, Sharon Crasta, Frank Eisenhaber, Zann Tan, Choy Eng Nge, Swaine L. Chen, Veronica Ng, Birgit Eisenhaber, Siew Bee Ng, Mingzi M. Zhang, Huibin Zhang, Chung Yan Leong, Kia Ngee Low, Falicia Goh, Fong Tian Wong, Tian Ru Lim, Wan Lin Yeo, Yoganathan Kanagasundaram, Elena Heng, and School of Computer Science and Engineering
- Subjects
Bioengineering [Engineering] ,Antifungal ,Antifungal Agents ,Stereochemistry ,medicine.drug_class ,lcsh:QR1-502 ,Biosynthetic gene cluster ,Bioengineering ,CRISPR–Cas9 ,Polyketide ,Applied Microbiology and Biotechnology ,Streptomyces ,lcsh:Microbiology ,Natural product ,03 medical and health sciences ,chemistry.chemical_compound ,Natural Product ,Gene cluster ,medicine ,Sulfation ,Cloning, Molecular ,Streptomyces antibiotic regulatory protein ,030304 developmental biology ,0303 health sciences ,Strain (chemistry) ,biology ,030306 microbiology ,Research ,biology.organism_classification ,chemistry ,Multigene Family ,Macrolides ,Biotechnology - Abstract
Notonesomycin A is a 32-membered bioactive glycosylated macrolactone known to be produced by Streptomyces aminophilus subsp. notonesogenes 647-AV1 and S. aminophilus DSM 40186. In a high throughput antifungal screening campaign, we identified an alternative notonesomycin A producing strain, Streptomyces sp. A793, and its biosynthetic gene cluster. From this strain, we further characterized a new more potent antifungal non-sulfated analogue, named notonesomycin B. Through CRISPR-Cas9 engineering of the biosynthetic gene cluster, we were able to increase the production yield of notonesomycin B by up to 18-fold as well as generate a strain that exclusively produces this analogue. Agency for Science, Technology and Research (A*STAR) National Research Foundation (NRF) Published version This work was supported by the Agency for Science, Technology and Research (A*STAR), Singapore. WLY, EH, MMZ, FTW are supported by National Research Foundation, Singapore [NRF2013-THE001-094].
- Published
- 2020
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