Functional identification of plant secondary metabolite biosynthesis gene in Saccharomyces cerevisiae by cell feeding / 中草药

ABSTRACT

Objective: To screen the most efficient cell feeding way and provide guidance for functional verification of genes in Saccharomyces cerevisiae. Methods: Firstly, the recombinant industrial S. cerevisiae strains containing PsCPR and PsP6H genes were constructed. Then four different cell feeding methods were used. At the end, the downstream product dihydrosanguinarine was detected via UPLC Q-TOF MS/MS. Results: The best method of cell feeding was that the recombinant industrial strain induced by galactose for 31 h and then incubated in the TE solution buffer (pH = 8.0) for 24 h. Conclusion: Different cell feeding ways have different effects on the downstream products of recombinant industrial S. cerevisiae. The optimized cell feeding method shows stability and good repeatability.