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Saccharomyces cerevisiae strain tolerant to vanillic acid, p-hydroxybenzoic acid and syringic acid and application of Saccharomyces cerevisiae strain

A technology of Saccharomyces cerevisiae strain and p-hydroxybenzoic acid, which is applied in the biological field, can solve the problems of inhibiting ethanol fermentation rate and lethality, and achieve good growth, ethanol fermentation performance, and enhanced tolerance

Active Publication Date: 2020-06-26
HEBEI NORMAL UNIV FOR NATTIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The object of the present invention is to provide a kind of Saccharomyces cerevisiae bacterial strain, can tolerate phenolic acid, solves the hydrolyzate vanillic acid, p-hydroxybenzoic acid and syringic acid etc. that lignocellulosic biomass obtains through different pretreatment methods to Saccharomyces cerevisiae Lethal and significantly inhibit the rate of ethanol fermentation problems

Method used

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  • Saccharomyces cerevisiae strain tolerant to vanillic acid, p-hydroxybenzoic acid and syringic acid and application of Saccharomyces cerevisiae strain
  • Saccharomyces cerevisiae strain tolerant to vanillic acid, p-hydroxybenzoic acid and syringic acid and application of Saccharomyces cerevisiae strain
  • Saccharomyces cerevisiae strain tolerant to vanillic acid, p-hydroxybenzoic acid and syringic acid and application of Saccharomyces cerevisiae strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Adaptive Evolution of Saccharomyces cerevisiae

[0038] The adaptive evolution method for obtaining the Saccharomyces cerevisiae strain PAT01 that is resistant to high concentrations of vanillic acid, p-hydroxybenzoic acid and syringic acid includes the following steps:

[0039] (1) Configuration of synthetic medium (g / L): anhydrous glucose 20, potassium dihydrogen phosphate 2, magnesium sulfate 1, ammonium sulfate 1, yeast extract 10; sterilize at 121°C for 20 minutes.

[0040] (2) The configuration of the phenolic acid inhibitor culture medium: Add a mixture of vanillic acid, p-hydroxybenzoic acid and syringic acid to the above-mentioned sterilized synthetic medium (where vanillic acid concentration is 0.80-2.50g / L, p-hydroxybenzoic acid) The concentration of benzoic acid is 0.50-1.80g / L and the concentration of syringic acid is 0.40-1.20g / L) to make phenolic acid inhibitor medium.

[0041] (3) Saccharomyces cerevisiae's adaptive evolution to mixed phenolic acid inh...

Embodiment 2

[0044] Example 2 Verification of tolerance of yeast strain PAT01 to mixed phenolic acid

[0045] (1) Comparison of the growth of the mixed phenolic acid plate: Adjust the cell density of the yeast strain PAT01 and the original yeast strain activated for 18 hours to an OD600 absorbance value of 6.0, and then dilute the cell suspension sequentially (10 -1 , 10 -2 , 10 -3 ), and pipet 5 μL of bacterial solution dropwise to different concentrations of mixed phenolic acid medium plates, and incubate at 30°C for 48 hours. Such as figure 1 As shown, the colony growth status of the yeast strain PAT01 is significantly better than the original yeast strain.

[0046] Among them, mixed phenolic acid solid medium: 1.5% (w / v) agar is added to the synthetic medium and sterilized at 121°C for 20 minutes. Vanillic acid, p-hydroxybenzoic acid and syringic acid were added to the sterilized synthetic medium to prepare mixed phenolic acid medium with different concentrations, as shown in Table 1.

[004...

Embodiment 3

[0054] Example 3 Simultaneous saccharification and ethanol fermentation using industrial corncob residue

[0055] The activated yeast strain PAT01 and the original yeast strain were respectively transferred to the industrial corn cob residue fermentation product after pre-saccharification for 12 hours at a 10% inoculum for simultaneous saccharification and ethanol fermentation. Samples were taken every 12 hours to determine the ethanol and glucose in the fermentation broth. content.

[0056] Among them: industrial corncob residue is a by-product of dilute acid treatment of corncob to produce xylose, obtained from Shandong Longli Biotechnology Co., Ltd., with a cellulose content of 56.5% (based on dry matter);

[0057] Dilute acid method treatment conditions: material-to-liquid ratio 1:10 (mass ratio), dilute sulfuric acid concentration 4%, cooking at 121°C for 1 h, solid-liquid separation, and the solid part is corncob residue.

[0058] Corncob residue pre-saccharification conditions:...

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Abstract

The invention discloses a Saccharomyces cerevisiae strain tolerant to vanillic acid, p-hydroxybenzoic acid and syringic acid and application of the Saccharomyces cerevisiae strain. The Saccharomyces cerevisiae strain is a strain PAT01 which is obtained after Saccharomyces cerevisiae is subjected to adaptive evolution by a phenolic acid inhibitor culture medium and can tolerate one or more of the vanillic acid, the p-hydroxybenzoic acid and the syringic acid with classification and nomenclature of Saccharomyces cerevisiae, the Saccharomyces cerevisiae is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 18021. The Saccharomyces cerevisiae strain can be applied to biomass conversion, and in saccharification and ethanol fermentation processes, the glucose consumption rate, the ethanol production rate and the ethanol yield of the Saccharomyces cerevisiae strain are all higher than those of an original strain.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a Saccharomyces cerevisiae strain that can tolerate vanillic acid, p-hydroxybenzoic acid and syringic acid and its application in the fermentation and production of ethanol. Background technique [0002] At present, lignocellulosic biomass has become one of the most promising alternatives to fossil resources due to its environmental friendliness and renewability, which is of great significance to the country’s security and development. But lignocellulose is a complex and dense compound composed of cellulose, hemicellulose and lignin. Its bio-resistance, which is difficult to hydrolyze, has become a major obstacle in the production of cellulosic ethanol. In order to solve this problem, it is necessary to pretreat the lignocellulosic raw materials with high temperature and high pressure and acid-base effect. However, many toxic degradation products are produced subsequently, whi...

Claims

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Application Information

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IPC IPC(8): C12N1/18C12P7/10C12R1/865
CPCC12P7/10C12R2001/865C12N1/185Y02E50/10
Inventor 顾翰琦刘晓光徐岩岩刘冉李玲李洁
Owner HEBEI NORMAL UNIV FOR NATTIES
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