Pathway engineering in yeast for synthesizing the complex polyketide bikaverin
Journal article, 2020
Fungal polyketides display remarkable structural diversity and bioactivity, and therefore the biosynthesis and engineering of this large class of molecules is therapeutically significant. Here, we successfully recode, construct and characterize the biosynthetic pathway of bikaverin, a tetracyclic polyketide with antibiotic, antifungal and anticancer properties, in S. cerevisiae. We use a green fluorescent protein (GFP) mapping strategy to identify the low expression of Bik1 (polyketide synthase) as a major bottleneck step in the pathway, and a promoter exchange strategy is used to increase expression of Bik1 and bikaverin titer. Then, we use an enzyme-fusion strategy to directly couple the monooxygenase (Bik2) and methyltransferase (Bik3) to efficiently channel intermediates between modifying enzymes, leading to an improved titer of bikaverin at 202.75 mg/L with flask fermentation (273-fold higher than the initial titer). This study demonstrates that the biosynthesis of complex fungal polyketides can be established and efficiently engineered in S. cerevisiae, highlighting the potential for natural product synthesis and large-scale fermentation in yeast.
Author
Meng Zhao
NYU Langone Health
Tianjin University
Yu Zhao
NYU Langone Health
Mingdong Yao
Tianjin University
Hala Iqbal
NYU Langone Health
Qi Hu
Tianjin University
Hong Liu
Tianjin University
Bin Qiao
Tianjin University
Chun Li
Tianjin University
Christine A.S. Skovbjerg
Technical University of Denmark (DTU)
Jens Christian Froslev Nielsen
Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology
Jens B Nielsen
Technical University of Denmark (DTU)
Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology
Rasmus John Normand Frandsen
Technical University of Denmark (DTU)
Yingjin Yuan
Tianjin University
Jef D. Boeke
NYU Langone Health
New York University
Nature Communications
2041-1723 (ISSN) 20411723 (eISSN)
Vol. 11 1 6197Subject Categories
Biochemistry and Molecular Biology
Biocatalysis and Enzyme Technology
Other Industrial Biotechnology
DOI
10.1038/s41467-020-19984-3
PubMed
33273470