Engineering Saccharomyces cerevisiae with the deletion of endogenous glucosidases for the production of flavonoid glucosides
نویسندگان
چکیده
BACKGROUND Glycosylation of flavonoids is a promising approach to improve the pharmacokinetic properties and biological activities of flavonoids. Recently, many efforts such as enzymatic biocatalysis and the engineered Escherichia coli biotransformation have increased the production of flavonoid glucosides. However, the low yield of flavonoid glucosides can not meet the increasing demand for human medical and dietary needs. Saccharomyces cerevisiae is a generally regarded as safe (GRAS) organism that has several attractive characteristics as a metabolic engineering platform for the production of flavonoid glucosides. However, endogenous glucosidases of S. cerevisiae as a whole-cell biocatalyst reversibly hydrolyse the glucosidic bond and hinder the biosynthesis of the desired products. In this study, a model flavonoid, scutellarein, was used to exploit how to enhance the production of flavonoid glucosides in the engineered S. cerevisiae. RESULTS To produce flavonoid glucosides, three flavonoid glucosyltransferases (SbGTs) from Scutellaria baicalensis Georgi were successfully expressed in E. coli, and their biochemical characterizations were identified. In addition, to synthesize the flavonoid glucosides in whole-cell S. cerevisiae, SbGT34 was selected for constructing the engineering yeast. Three glucosidase genes (EXG1, SPR1, YIR007W) were knocked out using homologous integration, and the EXG1 gene was determined to be the decisive gene of S. cerevisiae in the process of hydrolysing flavonoid glucosides. To further enhance the potential glycosylation activity of S. cerevisiae, two genes encoding phosphoglucomutase and UTP-glucose-1-phosphate uridylyltransferase involved in the synthetic system of uridine diphosphate glucose were over-expressed in S. cerevisiae. Consequently, approximately 4.8 g (1.2 g/L) of scutellarein 7-O-glucoside (S7G) was produced in 4 L of medium after 54 h of incubation in a 10-L fermenter while being supplied with ~3.5 g of scutellarein. CONCLUSIONS The engineered yeast harbouring SbGT with a deletion of glucosidases produced more flavonoid glucosides than strains without a deletion of glucosidases. This platform without glucosidase activity could be used to modify a wide range of valued plant secondary metabolites and to explore of their biological functions using whole-cell S. cerevisiae as a biocatalyst.
منابع مشابه
Statistical Optimization of Conditions for Maximize Production of Mannan by Saccharomyces Cerevisiae Using Response Surface Methodology
In view of the increase in Saccharomyces cerevisiae mannan content, the culture condition for S.cerevisiae were optimized in this study. The influence of culture condition such as original pH, inoculum size, and temperature on mannan production were evaluated using Response surface methodology. The mathematical model was established by the quadratic rotary combination design. with the order of ...
متن کاملKinetics Studies Impact of Initial pH and Addition of Yeast Saccharomyces cerevisiae on Biogas Production from Tofu Wastewater in Indonesia
The purpose of this work was to study the effect of initial pH and yeast Saccharomyces Cerevisiae on biogas production from tofu wastewater (TW). The initial pH was varied in ranging of 5 – 9 in substrate without yeast (T5-T9) and with yeast (TY5-TY9). The results showed that optimum initial pH was 8. The maximum biogas was resulted in T8 (275 mL) and TY8 (421 mL). Yeast addition increased tota...
متن کاملProduction of Single Cell Protein from Sugarcane Bagasse by Saccharomyces cerevisiae in Tray Bioreactor
In this study, solid state fermentation (SSF) was carried out to produce single cell protein (SCP) from sugarcane bagasse using Saccharomyces cerevisiae. The SSF experiment were performed in a tray bioreactor. The influence of several parameters including extraction buffer, initial moisture content of substrate, fermentation time, relative humidity in bioreactor, the bioreactor temperature and ...
متن کاملUtilizing an endogenous pathway for 1-butanol production in Saccharomyces cerevisiae.
Microbial production of higher alcohols from renewable feedstock has attracted intensive attention thanks to its potential as a source for next-generation gasoline substitutes. Here we report the discovery, characterization and engineering of an endogenous 1-butanol pathway in Saccharomyces cerevisiae. Upon introduction of a single gene deletion adh1Δ, S. cerevisiae was able to accumulate more ...
متن کاملStatistical Optimization of Conditions for Maximize Production of Mannan by Saccharomyces Cerevisiae Using Response Surface Methodology
In view of the increase in Saccharomyces cerevisiae mannan content, the culture condition for S.cerevisiae were optimized in this study. The influence of culture condition such as original pH, inoculum size, and temperature on mannan production were evaluated using Response surface methodology. The mathematical model was established by the quadratic rotary combination design. with the order of ...
متن کامل