Check patentability & draft patents in minutes with Patsnap Eureka AI!

Recombinant saccharomyces cerevisiae strain for producing vitamin C, construction method and fermentation method

A technology of Saccharomyces cerevisiae and recombinant engineering strains, which is applied in the field of fermentation and production of vitamin C by Saccharomyces cerevisiae strains, and can solve the problem that the maturity and application breadth of genetic manipulation is not as good as that of Saccharomyces cerevisiae

Pending Publication Date: 2021-05-18
TIANJIN UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the maturity and breadth of its genetic manipulation is not as good as that of Saccharomyces cerevisiae

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Recombinant saccharomyces cerevisiae strain for producing vitamin C, construction method and fermentation method
  • Recombinant saccharomyces cerevisiae strain for producing vitamin C, construction method and fermentation method
  • Recombinant saccharomyces cerevisiae strain for producing vitamin C, construction method and fermentation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Construction of VC production strains

[0038] 1. Construction of coding exogenous gene elements

[0039]The present invention provides 5 genes encoding VC synthesis pathway-related enzymes derived from Arabidopsis thaliana, which are respectively the gene encoding GDP-mannose 3,5-phosphatase (GME) and the encoding GDP-L-galactose phosphorylase (GGP ), the gene encoding L-galactose 1-phosphate phosphatase (GPP), the gene encoding L-galactose dehydrogenase (L-GalDH), the gene encoding L-galactose-1,4-lactone dehydrogenase The gene of the enzyme (L-GLDH) is abbreviated as gme, vtc2, vtc4, galdh and gldh in turn. After codon optimization, each gene was paired with yeast endogenous constitutive promoter and terminator, and five exogenous gene elements were obtained by overlap extension PCR.

[0040] 2. Construction of VC production strains

[0041] The plant's VC synthesis pathway starts with glucose and includes ten reaction processes ( figure 1 ), Saccharomy...

Embodiment 2

[0056] Example 2 Optimizing Metabolic Pathways Improves VC Production

[0057] 1. Construction of galdh and gldh fusion overexpression strains

[0058] The fusion protein has the complete coding sequence of each enzyme molecule, so it has the respective catalytic functions of the enzyme molecules that make up the enzyme, using (GGGGS) 3 Flexible linker, L-GLDH and its adjacent step protein L-GalDH are fused and overexpressed to construct a recombinant strain. The fragment TDH3p-galdh-linker-gldh-ENO2t was obtained by overlap extension PCR, cloned into the PRS425 plasmid linearized with XhoI and HindIII by Gibson assembly, transformed into Escherichia coli, and positive clones were screened, and the plasmid was extracted and transformed into Saccharomyces cerevisiae Strain YLAA', the galdh and gldh fusion overexpression strain YLAAMF-1 was obtained. The whole build process is as follows Figure 4 shown.

[0059] 2. Construction of potential rate-limiting step-related gene o...

Embodiment 3

[0065]Example 3 Effect of exogenous addition of intermediate metabolites and GSH on VC production

[0066] Comparing the effects of different intermediate metabolites and GSH on VC production

[0067] In order to study which intermediate metabolites are still missing during VC synthesis, 250 mg / L L-galactose or L-galactose-1,4-lactone was added exogenously to the shake flask. In addition, glutathione (GSH) can promote thiamine / thiamine pyrophosphate (TPP) and GSH transport, increase the metabolism of the tricarboxylic acid cycle and pentose phosphate pathway, and up-regulate reactive oxygen species (ROS) detoxification proteins , at the same time, it is also an antioxidant, so this instruction also adds 200mg / L GSH to the shake flask exogenously, hoping that it can reduce the loss of VC.

[0068] Experimental material: VTC2-M

[0069] Experimental method: except that no leucine was added to the seed medium, other conditions were the same as in Example 1.

[0070] Experiment...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the field of vitamin C production by fermentation of saccharomyces cerevisiae strains, and particularly relates to a saccharomyces cerevisiae strain for producing vitamin C, a construction method and a fermentation method. Specifically, the invention provides a method for synthesizing VC by using the recombinant saccharomyces cerevisiae strain, the speed limiting step and speed limiting enzyme of a plant synthesis VC path in saccharomyces cerevisiae are explored, intracellular VC extraction and HPLC detection methods are optimized, and a contribution is made to the realization of one-step fermentation production of VC.

Description

technical field [0001] The invention relates to the field of fermentation production of vitamin C by Saccharomyces cerevisiae strains, in particular to Saccharomyces cerevisiae strains, construction methods and fermentation methods for producing vitamin C. Background technique [0002] Vitamin C (VC) is a water-soluble vitamin closely related to humans. Since the discovery of VC more than 80 years ago, people have been digging deeply into its physiological activity and biological function. Because VC has a strong reducing ability, it is an important antioxidant, free radical scavenger, and also one of the nutrients for preventing sepsis. By the 1950s, VC was considered to have anti-cancer effects, and it could participate in redox reactions to affect the growth of cancer cells and even kill cancer cells. Recently, Changxing's group at Westlake University found that VC promotes plasma cell differentiation by activating DNA demethylase TET to enhance plasma production. Mult...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N1/19C12P17/04C12R1/865
CPCC12N15/52C12N9/0006C12N9/001C12N9/1241C12N9/90C12N9/16C12P17/04C12Y101/01316C12Y103/02003C12Y207/07069C12Y501/03018C12Y301/03
Inventor 元英进周蒙雨丁明珠毕彦慧
Owner TIANJIN UNIV
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com