Method for Regulating and Controlling Heterologous Synthetic Flavonoid Compound and Use Thereof
OwnerYong Wang, Jianhua Li +1inventors
InventorsYong Wang, Jianhua Li, Dongni Ji
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Abstract
Provided is a method for synthesizing a flavonoid compound. The method comprises providing a recombinant prokaryotic cell, wherein, in the prokaryotic cell, the transmembrane protein rhodanese Ygap of Escherichia coli is up-regulated or a target gene or target gene combination selected from the following groups is down-regulated: pyrB, accC, accB, purC, glyA, tktA, fabB, leuD, leuC, glpC, folK and leuA. Also provided are a prokaryotic cell for synthesizing a flavonoid compound and the use thereof, and the use of a kit and a regulation and control reagent. The present disclosure achieves significant improvement in the yield of the flavonoid compound.
Claims18
A method for synthesizing a flavonoid compound, wherein the method comprises:
- (1) providing a recombinant prokaryotic cell, comprising exogenous genes encoding the following enzymes:
- phenylalanine ammonia lyase, 4-coumaryl-CoA ligase, chalcone synthase, chalcone isomerase and flavone synthase I;
- and, in the prokaryotic cell, the transmembrane protein rhodanese Ygap of Escherichia coli is up-regulated or a target gene or target gene combination selected from the following group is down-regulated:
- pyrB, accC, accB, purC, glyA, tktA, fabB, leuD, leuC, glpC, folk, leuA;
- (2) using the formula (I) as a substrate to synthesize flavonoids with the prokaryotic cells of claim 1, wherein the flavonoid is chrysin compound;
- wherein, R comprises H or OH.
A prokaryotic cell for synthesizing flavonoid compound, wherein, the prokaryotic cell comprises exogenous genes encoding the following enzymes:
- phenylalanine ammonia lyase, 4-coumaryl-CoA ligase, chalcone synthase, chalcone isomerase and flavone synthase I;
- and, in the prokaryotic cell, the transmembrane protein rhodanese Ygap of Escherichia coli is up-regulated or a target gene or target gene combination selected from the following groups is down-regulated:
- pyrB, accC, accB, purC, glyA, tktA, fabB, leuD, leuC, glpC, folk, leuA;
- the flavonoid compound is chrysin compound.
(canceled)
A kit for constructing host cells for synthesizing flavonoid compounds, wherein the kit comprises:
- (a) a construct expressing phenylalanine ammonia lyase, 4-coumaryl-CoA ligase, chalcone synthase, chalcone isomerase and flavone synthase I;
- (b) a construct expressing transmembrane protein rhodanese Ygap of Escherichia coli;
- or a construct expressing a down-regulator, wherein the down-regulator down-regulates the target gene or target gene combination selected from the following group:
- pyrB, accC, accB, purC, glyA, tktA, fabB, leuD, leuC, glpC, folk, leuA;
- optionally, the kit further comprises a construct expressing genes encoding flavone 6-hydroxylase and cytochrome P450 oxidoreductase.
(canceled)
Dependencies are inferred from the claim text.