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Process for producing alcohol by co-fermentation of glucose and xylose

A technology of co-fermentation of glucose and co-fermentation, which is applied in the field of co-fermentation of glucose and xylose to produce alcohol, can solve the problems of insufficient stability of recombinant strains in alcohol production, and the problem of integration area is not considered, and achieve stable properties of strains, simple fermentation process and high expression level high effect

Inactive Publication Date: 2007-05-23
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are works to achieve high-level stable expression by integrating xylose metabolism genes in the yeast rDNA region with high copies, but the specific integration region is not considered. Therefore, the recombinant strain used for ethanol production still has the defect of insufficient stability. Used after acclimatization in medium with xylose as the only carbon source

Method used

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  • Process for producing alcohol by co-fermentation of glucose and xylose
  • Process for producing alcohol by co-fermentation of glucose and xylose
  • Process for producing alcohol by co-fermentation of glucose and xylose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 The construction method of the recombinant Saccharomyces cerevisiae NAN127 of the present invention:

[0037] (1) Construct the recombinant plasmid pYMIK-127 with the yeast universal single-copy integration vector YIp5 as the basic backbone (Figure 2).

[0038] (2) Linearize the recombinant plasmid pYMIK-127 at the 5232bp restriction site with restriction endonuclease HpaI.

[0039] (3) The industrial Saccharomyces cerevisiae strain NAN27 was transformed by the universal lithium acetate transformation method, and the recombinant strain NAN127 was obtained by screening with the YEPD plate added G418.

[0040] Among them, the PGK promoter and terminator (PGK promoter and terminator) in the recombinant plasmid vector pYMIK-127 in step (1) are cloned into the yeast chromosomal DNA, and inserted between the promoter and the terminator into the Pichia stipitis chromosomal DNA The xylitol dehydrogenase gene XYL2 of, the entire fragment is connected to the HindIII (32bp) ...

Embodiment 2

[0048] Example 2 Using recombinant engineering bacteria to simultaneously ferment glucose and xylose to produce alcohol, the method is as follows

[0049] (1) Selection of strains: select Saccharomyces cerevisiae NAN127, and the preservation number is CGMCC No.1848;

[0050] (2) Slant culture: Connect 1-2 loops of the above-mentioned recombinant Saccharomyces cerevisiae strain NAN127 to a YEPD medium slant containing 2% agar (w / v), and cultivate at 30°C for 24 hours;

[0051] (3) Seed culture: Connect NAN127 to 50mL YEPD liquid medium in the step 1~2 step (2), 30℃, 150~180r / min shaking culture for 24 hours;

[0052] (4) Expanded culture: Transfer the NAN127 in step (3) to 500mL YEPD liquid medium at a volume percentage of 10% of the inoculum, and culture at 30°C with a shaker of 150~180r / min for 24 hours;

[0053] (5) Alcohol fermentation: Transfer the NAN127 in step (4) to the fermenter for fermentation at a volume percentage of 10% inoculum. The fermentation conditions are at 30...

Embodiment 3

[0057] Example 3: Using recombinant engineering bacteria to simultaneously ferment glucose and xylose to produce alcohol, the method is as follows

[0058] (1) Selection of strains: select Saccharomyces cerevisiae NAN127, and the preservation number is CGMCC No.1848;

[0059] (2) Slant culture: Connect 1-2 loops of the above-mentioned recombinant Saccharomyces cerevisiae strain NAN127 to a YEPD medium slant containing 2% agar (w / v), and cultivate at 26°C for 36 hours;

[0060] (3) Seed culture: Connect NAN127 to 50mL YEPD liquid medium in step 1 to 2 in step (2), culture at 26°C, 150~180r / min shaking for 36 hours;

[0061] (4) Expanded culture: Transfer the NAN127 in step (3) to 500mLYEPD liquid medium at 5% of the inoculum amount by volume percentage, and culture for 36 hours at 26°C with a shaker of 150-180r / min;

[0062] (5) Alcohol fermentation: According to the volume percentage of 5% of the inoculum, transfer the NAN127 in step (4) to the fermenter for fermentation, the ferm...

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Abstract

The invention discloses a method for producing alcohol by co-fermenting glucose with recombinant Saccharomyces cerevisiae strains for industrial production. The invention comprises the steps of: (1) selecting recombinant Saccharomyces cerevisiae NAN127 (CGMCC No.1848), (2) slant-culturing, (3) culturing for seed, (4) enlargement-culturing, (5) fermenting, and (6) distilling to obtain alcohol. The method can utilize glucose and xylose in the medium at the same time, has the advantages of high utilization of xylose, high alcohol converting rate, and broad prospect in production alcohol with lignocellulose.

Description

Technical field [0001] The invention relates to a method for producing alcohol, in particular to a method for producing alcohol by co-fermentation of glucose and xylose. Background technique [0002] Petroleum is a non-renewable energy material, and the modern industrialized social development model based on this will be difficult to sustain. my country is an oil importer. In order to promote stable economic development and ensure national security, it is necessary to find renewable alternative energy sources. Alcohol is a clean and renewable liquid fuel. Many countries are already using gasoline with a certain proportion of alcohol-gasoline alcohol. This new fuel not only slows down the rate of oil consumption, but also reduces automobile exhaust pollution, and has great potential for application and development. my country has also begun to promote the use of gasoline alcohol since 2001. Currently, gasoline alcohol accounts for 20% of the total gasoline fuel consumption, and it...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/06C12N1/19C12R1/865
CPCY02E50/17Y02E50/10
Inventor 鲍晓明
Owner SHANDONG UNIV
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