Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Saccharomyces cerevisiae gene engineering bacterium capable of utilizing xylose and glucose jointly as well as construction method and application of saccharomyces cerevisiae gene engineering bacterium

A technology of genetically engineered bacteria and Saccharomyces cerevisiae is applied in the field of using xylose saccharomyces cerevisiae strains and its construction, which can solve problems such as imbalance, carbon utilization of engineered bacteria and reduction of ethanol production, and achieve the effect of simple operation.

Active Publication Date: 2017-01-04
NANJING UNIV OF TECH
View PDF2 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The imbalance of cofactors leads to the secretion of xylitol, which reduces the carbon utilization and ethanol production of engineered bacteria

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
  • Saccharomyces cerevisiae gene engineering bacterium capable of utilizing xylose and glucose jointly as well as construction method and application of saccharomyces cerevisiae gene engineering bacterium
  • Saccharomyces cerevisiae gene engineering bacterium capable of utilizing xylose and glucose jointly as well as construction method and application of saccharomyces cerevisiae gene engineering bacterium
  • Saccharomyces cerevisiae gene engineering bacterium capable of utilizing xylose and glucose jointly as well as construction method and application of saccharomyces cerevisiae gene engineering bacterium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Using the xylose-utilizing Saccharomyces cerevisiae strain BY4741 (MATa; ura3; his3; leu2; met15) as the starting strain, the expression vector constitutive high-copy plasmid pYX212 was used to overexpress XR and XDH with TEF1p as the promoter to obtain the recombinant strain XR-XDH The Saccharomyces cerevisiae strain BY4741 (MATa; ura3; his3; leu2; met15) containing the empty plasmid is the control bacteria CON; the source strain Candida tropicalis 121 of XR and XDH is also used as the control bacteria (hereinafter abbreviated as Ctp strain).

[0052] Use the above three strains to do glucose aerobic fermentation experiments:

[0053] The seed medium is a synthetic medium YPD, and the fermentation medium is a YPD medium with an initial sugar concentration of 100 g / L. Pick an appropriate amount from a single colony in a 30% glycerol tube stored at -80°C with an inoculation loop or a pipette tip and inoculate it into 5ml of YPD liquid medium, culture at 30°C and 200rpm f...

Embodiment 2

[0060] Carry out glucose, xylose mixed sugar fermentation experiment to three bacterial strains among the embodiment 1. Seed culture method is the same as embodiment 1. The fermentation medium is 20g / L glucose, 40g / L glucose, 10g / L yeast powder, and 20g / L peptone. The detection method is the same as in Example 1.

[0061] Experimental results such as Figure 4 as shown, Figure 4 Saccharomyces cerevisiae control strain CON, Candida tropicalis Ctp, Saccharomyces cerevisiae strain XR-XDH overexpressing XR and XDH with TEF1 as promoter, glucose and xylose mixed sugar fermentation experiments.

[0062] The results show that the Ctp strain can consume xylose in 3 days, but the CON and XR-XDH strains can hardly utilize xylose, and the bacterial growth and ethanol production are limited to the glucose utilization stage. After glucose is exhausted, they turn to use ethanol Used as a carbon source to sustain growth.

Embodiment 3

[0064] Xylulokinase XK and transaldolase TAL derived from Candida tropicalis were further overexpressed in the XR-XDH strain to obtain strains RH-TAL and RH-XK for glucose fermentation. The cultivation and detection methods are the same as in Example 1.

[0065] Experimental results such as Figure 5 as shown, Figure 5 The strain XR-XDH of XR and XDH with TEF1 as the promoter, the strain RH-TAL that further overexpresses the transaldolase derived from Candida tropicalis, and the strain XK that further overexpresses the xylulokinase derived from Candida tropicalis Glucose fermentation experiment of the strain RH-XK.

[0066] The results showed that overexpression of xylulokinase XK, a key enzyme in the non-oxidative part of the PPP pathway, could significantly increase the glucose consumption rate (RH-XK 28h vs. RH 48h), and the cell growth was significantly accelerated, but the ethanol production was reduced. Analysis The reason may be that overexpression of XK increases t...

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 microorganisms and molecular biology and in particular relates to a strain, a construction method thereof and fermentation application thereof. By taking a high-copy free type plasmid pYX212 as a carrier, xylose reductase (XR) and xylitol dehydrogenase (XDH) from candida pseudotropicalis are over-expressed in a saccharomyces cerevisiae strain, and the enzyme activity of the XDH is regulated through saccharomyces cerevisiae endogenous promoters TEF1 and PGK1, so that a recombinant saccharomyces cerevisiae strain capable of improving the consumption speed of xylose in joint utilization of the xylose and glucose can be obtained. According to the strain, the construction method thereof and the fermentation application thereof, molecular manipulation is simple and the joint utilization of the xylose and the glucose can be primarily realized without the need of a site-directed mutation or long-period evolution process.

Description

technical field [0001] The invention relates to the fields of microorganisms and molecular biology, in particular to a strain of Saccharomyces cerevisiae utilizing xylose and its construction method and application. Background technique [0002] Currently, ethanol is produced primarily from starch or sucrose from some crops such as corn, sugar cane and sugar beets. Bioethanol production from agricultural waste and other low-cost carbohydrate feedstocks, including corn stover, bagasse, wheat straw, non-recyclable paper, and switchgrass, has received widespread attention due to economic and environmental considerations. Lignocellulosic biomass is mainly composed of cellulose, hemicellulose, pectin and lignin, and the main sugar component is glucose, but contains 20% pentose sugars, such as D-xylose and L-arabinose. [0003] The production of bioethanol from lignocellulosic feedstocks has gained much attention in recent years because this feedstock is plentiful and inexpensive...

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): C12N1/19C12N15/81C12P7/06C12R1/865
CPCC12N9/0006C12N15/81C12P7/06C12Y101/01009C12Y101/01307C12N2800/102Y02E50/10
Inventor 应汉杰邹亚男施欣驰陈勇
Owner NANJING UNIV OF TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com