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)

Vol. 11 1 6197

Subject Categories

Biochemistry and Molecular Biology

Biocatalysis and Enzyme Technology

Other Industrial Biotechnology

DOI

10.1038/s41467-020-19984-3

PubMed

33273470

More information

Latest update

12/18/2020