5 results about "Chalcone isomerase" patented technology

The invention discloses recombinant saccharomyces cerevisiae, a 11-seed oil fermentation product as well as a preparation method and application of the 11-seed oil fermentation product, and belongs to the technical field of synthetic biology and fermentation product preparation. According to the recombinant saccharomyces cerevisiae disclosed by the invention, a caffeic acid coenzyme A ligase 4CL coding gene, a chalcone isomerase CHI coding gene, a flavanone-3-hydroxylase F3H coding gene and a chalcone synthase CHS mutant coding gene are over-expressed, and the chalcone synthase CHS mutant is preferably Y69H / H71Y / Q161K. The preparation method of the 11-seed oil fermentation product comprises the following steps: fermenting a recombinant saccharomyces cerevisiae fermentation solution in the presence of a sophora flower bud extract and caffeic acid to obtain a first fermentation solution; fermenting the lactic acid bacteria fermentation liquor in the presence of 11 seed oil to obtain second fermentation liquor; and mixing the first fermentation liquid and the second fermentation liquid, continuing fermentation, and respectively collecting an oil phase and a water phase after fermentation is finished, so as to obtain a fermentation product. The product provided by the invention has multivitamin antioxidant activity.

The invention belongs to the technical field of biology, and discloses a saccharomyces cerevisiae engineering strain for producing naringenin, a construction method and application, the saccharomyces cerevisiae engineering strain contains tyrosine ammonia lyase gene TAL, 4-cumaric acid: coenzyme A ligase gene 4CL, chalcone synthase gene CHS, chalcone isomerase gene CHI, oligopeptide gene RIADn-RGGm and oligopeptide gene RIDD; the ARO10 gene, the FDC1 gene, the PAD1 gene and the PDC5 gene are knocked out. The engineering strain disclosed by the invention can be used for synthesizing naringenin by taking L-tyrosine as a substrate, and the engineering strain is stable in hereditary character and excellent in naringenin yield which can reach 27.45 mg / L; and the fermentation production of the product can be carried out through relatively simple method conditions, so that the method has a very wide application prospect.

The application discloses a Saccharomyces cerevisiae de novo synthesis of naringenin engineering strain and a construction method and application thereof, and the engineering strain is constructed by modifying a genome of the Saccharomyces cerevisiae CEN.PK2-1C as follows: substrate competition pathway genes are knocked out, anti-feedback inhibition genes are overexpressed, and the like, so as to improve a metabolic flux j of a precursor substance tyrosine; tyrosine deaminase, 4-coumarate:CoA-ligase, chalcone synthase and chalcone isomerase are expressed heterogeneously, so as to realize de novo synthesis of naringenin. Finally, the yield of naringenin is further improved by high-efficiency expression of Sc4CL and HaCHS at multiple copy sites.

The invention discloses chalcone isomerase capable of improving activity of chalcone synthase and application of the chalcone isomerase, compared with arabidopsis thaliana CHIL, the amino acid sequence of the chalcone isomerase has mutation H36L / M at the 36th site at the top of beta-Hairpin, or has mutation H36E at the 36th site, and meanwhile, has mutation F37T at the 37th site. The three-dimensional structure of the complex formed by the chalcone synthase and the IV type chalcone isomerase is analyzed, key amino acid residues on a contact surface when the complex is formed are identified, a catalysis and regulation mechanism of the chalcone synthase complex for exerting an enzyme activity function is disclosed, and the activity of the IV type chalcone isomerase is improved through the amino acid, located on beta-hairpin, on the IV type chalcone isomerase. According to the present invention, the mutation is performed on the naringenin, especially the H36 amino acid residue, such that the naringenin yield in the total product can be significantly improved, the naringenin content in the total product can be improved to 90% or more, and the method can be used for the field of biosynthesis and the industrial mass production of flavonoid compounds.

The invention discloses algal moss flavone synthetic route related protein as well as a coding gene and application of the algal moss flavone synthetic route related protein. The invention discloses algal moss flavone synthesis route related proteins, which are named as Tl4CL, TlCHS and TlCHI, and the amino acid sequences of the algal moss flavone synthesis route related proteins are respectively shown as SEQ ID No.2, SEQ ID No.4 and SEQ ID No.6. A test proves that the Tl4CL can efficiently catalyze p-coumaric acid to generate p-coumaryl CoA. TlCHS and TlCHI are chalcone synthetase and chalcone isomerase in algae moss respectively, p-coumaroyl CoA can be converted into naringenin under the catalysis of the two enzymes, and the yield reaches 574ng / ml through fermentation of recombinant bacteria.