Pathway engineering in yeast for synthesizing the complex polyketide bikaverin
Artikel i vetenskaplig tidskrift, 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.
Författare
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
Danmarks Tekniske Universitet (DTU)
Jens Christian Froslev Nielsen
Chalmers, Biologi och bioteknik, Systembiologi
Jens B Nielsen
Danmarks Tekniske Universitet (DTU)
Chalmers, Biologi och bioteknik, Systembiologi
Rasmus John Normand Frandsen
Danmarks Tekniske Universitet (DTU)
Yingjin Yuan
Tianjin University
Jef D. Boeke
NYU Langone Health
New York University
Nature Communications
2041-1723 (ISSN) 20411723 (eISSN)
Vol. 11 1 6197Ämneskategorier
Biokemi och molekylärbiologi
Biokatalys och enzymteknik
Annan industriell bioteknik
DOI
10.1038/s41467-020-19984-3
PubMed
33273470