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Culture medium suitable for improvement of tolerance of glucose and xylose co-fermentation saccharomyces cerevisiae for multiple kinds of pretreatment inhibitors and application

A technology of Saccharomyces cerevisiae and pretreatment, applied in the field of microorganisms

Inactive Publication Date: 2018-09-07
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After searching, there are few reports about the high-efficiency medium suitable for improving the tolerance of Saccharomyces cerevisiae for co-fermentation of glucose and xylose to various pretreatment inhibitors at home and abroad.

Method used

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  • Culture medium suitable for improvement of tolerance of glucose and xylose co-fermentation saccharomyces cerevisiae for multiple kinds of pretreatment inhibitors and application
  • Culture medium suitable for improvement of tolerance of glucose and xylose co-fermentation saccharomyces cerevisiae for multiple kinds of pretreatment inhibitors and application
  • Culture medium suitable for improvement of tolerance of glucose and xylose co-fermentation saccharomyces cerevisiae for multiple kinds of pretreatment inhibitors and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Preparation of Screening Medium Containing Various Pretreatment Inhibitors

[0030] Specific steps are as follows:

[0031] (1) Pretreatment of corn stalk with dilute acid to prepare corn stalk pretreatment solution: prepare a dilute sulfuric acid solution with a sulfuric acid mass fraction of 1.0%, as a cooking solution for dilute acid pretreatment, mix corn stalk (pulverized to 20-80 mesh) with The cooking liquid is mixed evenly in a 1.5L small steel tank at a material-liquid ratio of 1:10. After sealing, the small tank is placed in the cooking pot. At the upper end of the small pot in the middle, seal the cooking pot and set the required reaction temperature to 170°C and start heating, and keep the reaction for 25 minutes after reaching the temperature. Immediately after the reaction, the canister was taken out and put into cold water so that the temperature of the canister was lowered to room temperature. The pretreatment liquid in the small tank is sep...

Embodiment 2

[0034] Example 2: Effect of medium containing dilute acid and AHP pretreatment hydrolyzate inhibitor on yeast growth

[0035] Specific steps are as follows:

[0036] (1) Dilution coating experiment: The industrial strain of Saccharomyces cerevisiae (deposit number: CGMCC No. 11331), which has undergone metabolic engineering and can efficiently co-metabolize glucose and xylose, was used as the starting strain for this experiment, and 5 mL of YPD was added to it. (YPD medium formula: yeast powder 10g / L, peptone 20g / L, glucose 20g / L.), culture overnight, take 1ml of bacterial suspension, centrifuge at 8000r / min, remove the supernatant, wash and suspend with 1mL sterile water Bacteria, centrifuge, remove supernatant, adjust OD 600 = about 1.0, diluted 1000 times, take 100 μL and apply it to YPX plate, (YPX + dilute acid pretreatment) plate and (YPX + AHP) plate, cultivate at 30 ° C for 3 to 4 days, observe the growth of the strain, take pictures, the results show ( like figure ...

Embodiment 3

[0038] Example 3: Using ARTP mutagenesis technology to improve the ability of Saccharomyces cerevisiae to tolerate AHP pretreatment corn stover hydrolyzate inhibitor

[0039] Specific steps are as follows:

[0040] (1) Activation of the starting strain: Inoculate the starting strain on the YPD plate, inoculate it upside down at 30°C for 2 to 3 days, then inoculate it into the YPD liquid, shake it for 12h to 24 hours, and transfer the bacterial liquid to fresh YPD to control the bacteria. Concentration OD 600 = 0.1, incubate at 30°C with shaking for several hours, and control the final concentration of bacteria OD 600 = within 1.0, as the ARTP mutagenesis starting strain.

[0041] (2) ARTP mutagenesis: take 1 mL of bacterial solution, suspend it in a 1.5 mL EP tube, centrifuge at 8000 r / min for 2 min, and discard the supernatant. After washing twice with normal saline, diluted to make the bacterial concentration at 10 6 ~10 7 Between the bacterial suspension, take 10 μL of...

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Abstract

The invention discloses a culture medium suitable for improvement of tolerance of glucose and xylose co-fermentation saccharomyces cerevisiae for multiple kinds of pretreatment inhibitors. The culturemedium takes a lignocellulose pretreatment solution containing multiple kinds of inhibitor components as a matrix, and yeast powder, peptone and xylose which are suitable for growth of xylose metabolism bacterial strains are added to the culture medium. The culture medium is suitable for improvement of tolerance of glucose and xylose co-fermentation saccharomyces cerevisiae for multiple kinds ofpretreatment inhibitors and beneficial to screening of glucose and xylose co-fermentation saccharomyces cerevisiae with improved tolerance for the pretreatment inhibitors. Compared with an existing screening culture medium only by adding one kind of inhibitor component, the culture medium has the advantage that the screening efficiency of high-tolerance saccharomyces cerevisiae can be greatly improved.

Description

technical field [0001] The invention relates to a screening medium and application thereof, in particular to a medium and application suitable for improving the tolerance of Saccharomyces cerevisiae for co-fermentation of glucose and xylose to various pretreatment inhibitors, belonging to the technical field of microorganisms. Background technique [0002] Crop straws are discarded and burned at will, causing waste of resources and one of the important reasons for the formation of haze weather in recent years. Converting lignocellulose in straw into clean energy such as fuel ethanol has broad application prospects. Lignocellulosic raw materials are mainly composed of cellulose, hemicellulose and lignin, and the structure of each component is stable and complex. The use of lignocellulose to produce fuel ethanol and other chemicals mainly includes pretreatment of raw materials, enzymatic hydrolysis of cellulose and microbial fermentation of sugar [Sun Y, Cheng J.Hydrolysis of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/18C12N1/36C12R1/865
CPCC12N1/18C12N1/36
Inventor 徐丽丽鲍晓明李洪兴刘同军
Owner QILU UNIV OF TECH
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