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Recombinant saccharomyces cerevisiae as well as construction method and application thereof

A technology for Saccharomyces cerevisiae and a construction method, applied in the field of genetic engineering, can solve the problems of low utilization rate of yeast cells, high cost of naringenin and the like, and achieve the effects of low cost, good economic benefit and few by-products

Active Publication Date: 2017-07-07
TIANJIN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows yeast or fungi (Coriolus) produce eight different types of compounds called quaternary neurotoxicity factor B1(QNB1) through their ability to use certain sugars like glucose instead of ribulosesulfonates. By combining these sugar molecules together they create an enzymatic reaction system where some specific substances are produced while others remain unchanged. These reactions help protect plants against harmful insect attacks without causing any damage during harvesting processes. Overall this innovation makes it possible to efficiently make various valuable medicines such as citrullum berberide and selenazedoxan at very affordable prices.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the efficiency or effectiveness of producing flavonols (flavones) through biosynthesis methods like Microbial Synthase Ferrooxygenases (MFO).

Method used

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  • Recombinant saccharomyces cerevisiae as well as construction method and application thereof
  • Recombinant saccharomyces cerevisiae as well as construction method and application thereof
  • Recombinant saccharomyces cerevisiae as well as construction method and application thereof

Examples

Experimental program
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Embodiment 1

[0045] Embodiment 1, the construction of the recombinant Saccharomyces cerevisiae strain that produces naringenin

[0046] 1. Acquisition of exogenous functional gene elements

[0047] The exogenous genes are the key enzymes TAL, 4CL, CHS and CHI genes for the synthesis of naringenin: the source of the TAL gene is Rhodosporidium toruloides (R. toruloides), referred to as Sc_TAL. The source of the 4CL gene is parsley (petroselinum crispum), referred to as Sc_4CL. Gene sources of CHS include Arabidopsis thaliana, grape (Vitis vinifera), Polygonum cuspidatum, marine bacteria (Rhodopirellula baltica), soybean (Glycine max), etc., which are abbreviated as Sc_CHS1, Sc_CHS2, Sc_CHS3, Sc_CHS4, Sc_CHS7, the gene sources of CHI include soybean 1 (Glycine max), soybean 2 (Glycine max), tangerine (Citrus unshiu), Arabidopsis thaliana (Arabidopsis thaliana), corn (Zea mays), peanut (Arachis hypogaea), etc., abbreviated in order Sc_CHI1, Sc_CHI2, Sc_CHI4, Sc_CHI5, Sc_CHI6, Sc_CHI7. The s...

Embodiment 2

[0094] Example 2, Expression of exogenous gene N8DT in recombinant Saccharomyces cerevisiae to synthesize 8-dimethylprenylnaringenin

[0095] 1. Acquisition of exogenous functional gene element N8DT

[0096] The exogenous gene is the key enzyme N8DT for 8-dimethylprenylnaringenin: the selected one is naringenin prenyltransferase (N8DT) from Sophora flavescens, the specific gene The sequence is shown in SEQ ID No:14. The above-mentioned genes are artificially synthesized after Saccharomyces cerevisiae codon optimization.

[0097] 2. Construction of a recombinant Saccharomyces cerevisiae strain producing 8-prenylnaringenin

[0098] The obtained exogenous N8DT gene was digested with BsaI and connected to the module library pRS425K-ENO2t-PDC2p-GPM2t plasmid (available for free at http: / / synbioml.org / ) using T4 ligase, and the recombinant The plasmid was transformed into the SyBE_Sc02050031 chassis strain, using Sc-LEU solid plate (synthetic yeast nitrogen source YNB 6.7g / L, glu...

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Abstract

The invention relates to the technical field of genetic engineering and discloses a recombinant saccharomyces cerevisiae strain as well as a construction method and application thereof. The recombinant saccharomyces cerevisiae realizes production of 8-dimethyl isopentenyl naringenin with tyrosine as a substrate, in the recombinant saccharomyces cerevisiae strain, rhodosporidium toruloides is sourced from a TAL gene, parsley is sourced from the TAL gene, arabidopsis is sourced from a CHS gene, sweet mandarin is sourced from a CHI gene and sophora flavescens is sourced from an N8DT gene combination, thereby having the effect of producing the 8-dimethyl isopentenyl naringenin with high yield, and experiments show that the yield of the 8-dimethyl isopentenyl naringenin can reach 36.7mu g/L. The recombinant saccharomyces cerevisiae strain is utilized for producing the 8-dimethyl isopentenyl naringenin, cost is low, byproducts are fewer and pollution is low compared with plant extraction and chemical synthesis, a feasible method is provided for production of the 8-dimethyl isopentenyl naringenin and related products thereof, and a foundation is laid for biosynthesis of icaritin and icariin.

Description

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Claims

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Application Information

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Owner TIANJIN UNIV
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