Loading...
Please wait, while we are loading the content...
Similar Documents
A coupled in vitro/in vivo approach for engineering a heterologous type III PKS to enhance polyketide biosynthesis inSaccharomyces cerevisiae
| Content Provider | Scilit |
|---|---|
| Author | Vickery, Christopher R. Cardenas, Javier Bowman, Marianne E. Burkart, Michael D. Silva, Nancy A. Da Noel, Joseph P. |
| Copyright Year | 2018 |
| Description | Journal: Biotechnology & Bioengineering Polyketides are attractive compounds for uses ranging from biorenewable chemical precursors to high‐value therapeutics. In many cases, synthesis in a heterologous host is required to produce these compounds in industrially relevant quantities. The type III polyketide synthase 2‐pyrone synthase (2‐PS) from Gerbera hybrida was used for the production of triacetic acid lactone (TAL) in Saccharomyces cerevisiae. Initial in vitro characterization of 2‐PS led to the identification of active site variants with improved kinetic properties relative to wildtype. Further in vivo evaluation in S. cerevisiae suggested certain 2‐PS mutations altered enzyme stability during fermentation. In vivo experiments also revealed beneficial cysteine to serine mutations that were not initially explored due to their distance from the active site of 2‐PS, leading to the design of additional 2‐PS enzymes. While these variants showed varying catalytic efficiencies in vitro, they exhibited up to 2.5‐fold increases in TAL production when expressed in S. cerevisiae. Coupling of the 2‐PS variant [C35S,C372S] to an engineered S. cerevisiae strain led to over 10 g/L TAL at 38% of theoretical yield following fed‐batch fermentation, the highest reported to date. Our studies demonstrate the success of a coupled in vitro/in vivo approach to engineering enzymes and provide insight on cysteine‐rich enzymes and design principles toward their use in non‐native microbial hosts. |
| Related Links | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6701952/pdf |
| Ending Page | 1402 |
| Page Count | 9 |
| Starting Page | 1394 |
| e-ISSN | 10970290 |
| DOI | 10.1002/bit.26564 |
| Journal | Biotechnology & Bioengineering |
| Issue Number | 6 |
| Volume Number | 115 |
| Language | English |
| Publisher | Wiley-Blackwell |
| Publisher Date | 2018-02-19 |
| Access Restriction | Open |
| Subject Keyword | Journal: Biotechnology & Bioengineering Biotechnology and Microbiology 2‐pyrone Synthase Enzyme Engineering Saccharomyces Cerevisiae Triacetic Acid Lactone |
| Content Type | Text |
| Resource Type | Article |
