Improving acetyl-CoA biosynthesis in Saccharomyces cerevisiae via the overexpression of pantothenate kinase and PDH bypass

• Main Authors: Liu, Wenshan, Zhang, Bo, Jiang, Rongrong
• Other Authors: School of Chemical and Biomedical Engineering
• Format: Article
• Language: English
• Published: 2018
• Subjects: Pantothenate Kinase; DRNTU::Engineering::Bioengineering; Acetyl-CoA
• Online Access: https://hdl.handle.net/10356/87852; http://hdl.handle.net/10220/46887
• Institution: Nanyang Technological University

Background: Acetyl-CoA is an important precursor in Saccharomyces cerevisiae. Various approaches have been adopted to improve its cytosolic level previously with the emphasis on engineering the “acetyl-” part of acetyl-CoA. To the best of our knowledge, there have been no reports on engineering the “-CoA” part so far. Results: In this study, we had tried to engineer S. cerevisiae from both the “-CoA” part via pantothenate kinase overexpression [PanK from S. cerevisiae, the rate-limiting enzyme for CoA synthesis] and the “acetyl-“part through PDH bypass introduction (ALD6 from S. cerevisiae and SeAcs L641P from Salmonella enteric). A naringenin-producing reporter strain had been constructed to reflect cytosolic acetyl-CoA level as acetyl-CoA is the precursor of naringenin. It was found that PanK overexpression or PDH bypass introduction alone only led to a twofold or 6.74-fold increase in naringenin titer, but the combination of both (strain CENFPAA01) had resulted in 24.4-fold increase as compared to the control (strain CENF09) in the presence of 0.5 mM substrate p-coumaric acid. The supplement of PanK substrate pantothenate resulted in another 19% increase in naringenin production. Conclusions: To greatly enhance acetyl-CoA level in yeast cytosol, it is feasible to engineer both the “acetyl-” part and the “-CoA” part simultaneously. Insufficient CoA supply might aggravate acetyl-CoA shortage and cause low yield of target product.