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Recombinant distiller's yeast and use thereof in eutrit production

A technology of Saccharomyces cerevisiae and xylitol, applied in the field of microbial engineering

Active Publication Date: 2008-12-17
SHANDONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The corn cob acid hydrolysis solution contains toxic substances such as furfural, phenol and other anti-nutritional factors, and it is difficult for common Saccharomyces cerevisiae to grow well in it. Therefore, a starting strain that can adapt to the corn cob acid hydrolysis solution and reproduce rapidly in it is obtained especially important

Method used

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  • Recombinant distiller's yeast and use thereof in eutrit production
  • Recombinant distiller's yeast and use thereof in eutrit production
  • Recombinant distiller's yeast and use thereof in eutrit production

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Embodiment 1

[0034] A Saccharomyces cerevisiae (Saccharomyces cerevisiae) GZ-5-T2, which has been preserved in the General Microorganism Center of China Committee for the Collection of Microorganisms on July 10, 2008, and its preservation number is CGMCC No.2581. The rDNA repeat region of the Saccharomyces cerevisiae GZ-5-T2 chromosome has multiple xylose reductase genes XYL1 derived from Pichia spp. The homologous integration of the rDNA region of the chromosome of the industrial production strain GZ-5-T2 was realized. At the same time, according to the requirements of stability, the rDNA sequence of Saccharomyces cerevisiae was analyzed, and the length of the three regions spanning 17S, 5.8S and 26S on the selected rDNA was A 2.2 kb fragment served as the integration site.

Embodiment 2

[0036] The construction method of Saccharomyces cerevisiae GZ-5-T2:

[0037] (1) Recombinant plasmid pYMIC-xyl1 was linearized at the 6453bp enzyme cutting site with restriction endonuclease HpaI.

[0038] (2) The Saccharomyces cerevisiae industrial strain GZ-5 was transformed by the lithium acetate transformation method, and the recombinant strain GZ-5-T2 was obtained by screening with YEPD plates added with copper ions.

[0039]Wherein, the PGK promoter (PGK promoter) and PGK terminator (PGK terminator) in the recombinant plasmid vector pYMIC-xyl1 in step (1) are cloned in the yeast chromosome DNA, and the promoter and the terminator are inserted and cloned in the trunk of Pichia Xylose reductase gene XYL1 of yeast chromosomal DNA;

[0040] Wherein the copper ion resistance gene (CUP I) in the recombinant plasmid vector pYMIC-xyl1 in the step (1) is cloned in the chromosomal DNA of Saccharomyces cerevisiae as a screening marker.

[0041] Wherein the rDNA sequence in the re...

Embodiment 3

[0044] The application of Saccharomyces cerevisiae GZ-5-T2 in the production of xylitol, the specific steps are as follows:

[0045] (1) Use 0.1wt% hydrochloric acid together with 0.01w% ferric chloride promoter to completely convert the hemicellulose in the corncob powder into xylose, arabinose, and glucose to obtain sugar liquid;

[0046] (2) decolouring, electrodialysis deacidification and ion exchange resin removal of impurities with the sugar solution obtained in step (1) to obtain a fermentation substrate; after electrodialysis deacidification, the acid concentration in the acid water reaches 0.04wt%, and reuse can save nearly 20% acid dosage;

[0047] (3) adding mass-volume ratio to the fermented substrate prepared in step (2) is 0.5% yeast powder, and mass-volume ratio is 0.5% peptone, and inoculate Saccharomyces cerevisiae GZ-5-T2 by volume percentage 8% inoculum , fermented for 60 hours at a temperature of 26°C, a pH of 4.6, and an aeration rate of 0.1vvm to obtain ...

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Abstract

The invention relates to a recombinant Saccharomyces cerevisiae and application thereof in the production of xylitol, which pertains to the technical field of microbial engineering. A Saccharomyces cerevisiae GZ-5-T2 is preserved in the Common Microorganisms Center of China Committee for Culture Collection of Microorganisms on July 10, 2008, with the strain accession number of CGMCC No.2581. The strain can be applied to the production of the xylitol. The Saccharomyces cerevisiae preserved by the invention reduces the stable integration and high efficient expression of enzyme genes XYL1 in a chromosome rDNA area by xylose on the basis of keeping the original growth feature, thus obtaining the property of utilizing the xylose in the acidolysis solution of corncob to produce the xylitol.

Description

technical field [0001] The invention relates to a recombinant saccharomyces cerevisiae and its application in producing xylitol, which belongs to the technical field of microbial engineering. Background technique [0002] Xylitol is a pentavalent alcohol, one of the intermediate products of xylose metabolism, and its chemical formula is C 5 h 12 o 5 , the shape is white crystal. As an important functional polysaccharide alcohol, xylitol has been widely recognized for its good physiological functions on the human body, mainly in: caries prevention, no increase in blood sugar level, bifidus factor effect, obesity relief and liver function improvement. The sweetness of xylitol is comparable to that of sucrose, and its sweetness is higher at low temperature, but it does not increase blood sugar, so it is a good substitute for sucrose. Due to its special physiological functions, xylitol is widely used in industries such as food and medicine, and the international demand is in...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12P19/02C12R1/865
Inventor 鲍晓明程少博刘怀伟肖林
Owner SHANDONG UNIV
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