Improving CsOAC Activity in Saccharomyces cerevisiae for Directed Production of Olivetolic Acid through Rational Design.

Autor: Spitzer S; Technical Biochemistry Laboratory, Faculty of Biochemical and Chemical Engineering, TU Dortmund University, Emil-Figge-Strasse 66, 44227, Dortmund, Germany., Aras M; Technical Biochemistry Laboratory, Faculty of Biochemical and Chemical Engineering, TU Dortmund University, Emil-Figge-Strasse 66, 44227, Dortmund, Germany., Kayser O; Technical Biochemistry Laboratory, Faculty of Biochemical and Chemical Engineering, TU Dortmund University, Emil-Figge-Strasse 66, 44227, Dortmund, Germany.
Jazyk: angličtina
Zdroj: Chembiochem : a European journal of chemical biology [Chembiochem] 2024 Oct 17, pp. e202400651. Date of Electronic Publication: 2024 Oct 17.
DOI: 10.1002/cbic.202400651
Abstrakt: Olivetolic acid (OA) is an essential precursor in the cannabinoid biosynthesis. It is produced through a unique interaction between the two proteins, olivetol synthase (CsOLS) and olivetolic acid cyclase (CsOAC). When the OA biosynthesis is transferred to Saccharomyces cerevisiae, olivetol (OL) is produced as a side product, even with a high enhancement of copy number of CsOAC. In order to increase the OA titer while decreasing the OL titer in S. cerevisiae, rational design was applied to CsOAC using in silico approaches such as protein-ligand docking to find potential protein variants. In vivo screening and also testing different approaches for both proteins was applied to identify the best performing variants of CsOAC. Four variants were identified that gave the desired properties. The best CsOAC variant, G82 A/L92Y, resulted in a 1.7-fold increase in OA production and a shift in the ratio between the two products towards OA.
(© 2024 Wiley-VCH GmbH.)
Databáze: MEDLINE
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