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Saccharomyces cerevisiae and application thereof in food fermentation

A technology for Saccharomyces cerevisiae and fermented food, which is applied in the directions of fermentation, preparation of alcoholic beverages, enzymes, etc., can solve the problems of long metabolic pathway of β-phenylethanol, low yield and many branches, etc., and achieves good alcohol production characteristics and catalytic effect. The effect of power enhancement, high alcohol production capacity

Inactive Publication Date: 2018-04-20
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] From the above we can see that the metabolic pathway of β-phenylethanol is too long, there are too many branches, and there are many inhibitory effects, and the final yield is not high

Method used

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  • Saccharomyces cerevisiae and application thereof in food fermentation
  • Saccharomyces cerevisiae and application thereof in food fermentation
  • Saccharomyces cerevisiae and application thereof in food fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Example 1: Screening and Sequence Alignment of High-Producing β-Phenylalcohol Yeast Strains

[0038] The Saccharomyces cerevisiae strain of the present invention is screened out from rice wine fermentation mash, and the process flow of rice wine fermentation mash made by adding wine medicine is shown in figure 1 , the alcohol content can reach 13°, and the phenylethanol content is about 120mg / L. The rice wine fermented mash has good alcohol characteristics and is resistant to phenylethanol. The rice wine mash was extracted, diluted step by step, coated with YPD medium, purified by streaking, and the microorganisms in the rice wine fermentation mash were screened. Finally, we obtained a high-yielding and phenylethanol-tolerant Saccharomyces cerevisiae 11-1. It was deposited in the China Center for Type Culture Collection on September 11, 2017. The deposit address is Wuhan University, Wuhan, China, and the deposit number is CCTCCNO:M2017488.

[0039] We used high-fidelit...

Embodiment 2

[0041] Embodiment 2: the enzymatic properties of prephenate dehydratase PHA2P

[0042] Compared with the model yeast strain S288c-pha2 gene, it was found that using the Saccharomyces cerevisiae 11-1 genome as a template, natural mutant genes (I161K, L239P, Q250H) were amplified by PCR. The prokaryotic expression of the body protein was purified by affinity chromatography with HisTrapTMHP under non-denaturing conditions, and the electrophoretic pure target protein was successfully obtained, and the size was consistent with the expectation.

[0043] Enzyme activity assay reaction system: 0.4 ml assay system

[0044]

[0045] Add water to make up the volume, incubate at 45°C for 5 minutes, add 800ul 1.5mol / L NaOH to terminate the reaction, centrifuge for 5 minutes to remove protein precipitates, transfer 200ul to a 96-well microplate reader, measure the light absorption value at 320nm, and use benzene Pyruvate was used as a standard curve to calculate the reaction enzyme acti...

Embodiment 3

[0055] Embodiment 3: the application of high-yielding β-phenylethanol yeast in yellow rice wine

[0056] The model yeast strain S288c and Saccharomyces cerevisiae 11-1 were respectively used to make distiller's mother, and then used for rice wine fermentation, and detected by liquid chromatography, the detection method of phenylethanol (using waters X bridge C18 column, mobile phase methanol: water = 1: 1, flow rate 0.8ml / min, column temperature 30°C).

[0057] (1) Production of distiller's yeast: 50mLYPD shake flask was inoculated with yeast strain, cultured at 30°C and 200r / min for 24h. Add 1L of water and 0.05kg of wheat koji to 1kg of steamed rice (moisture content 70%), stir evenly, keep warm at 60°C for 4h, inoculate yeast liquid at 5% after cooling, and incubate at 30°C and 200r / min for 16h;

[0058] (2) Steam cooked rice (moisture content 70%), add equal weight of clear water, 2% wheat koji, 5% distiller's mother, and stir evenly.

[0059] (3) Fermentation and stirri...

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Abstract

Belonging to the technical field of industrial microbiology, the invention discloses a saccharomyces cerevisiae and application thereof in food fermentation. The saccharomyces cerevisiae strain 11-1 is preserved in China Center for Type Culture Collection on September 11, 2017 at Wuhan University, Wuhan, China, and the preservation number is CCTCC NO:M 2017488. Sense mutation occurs to the key gene pha2 in the path of synthesizing beta-phenylethyl alcohol from the strain, and the catalytic ability of mutated prephenate dehydratase PHA2P is strengthened by 22%. The invention also finds that thesaccharomyces cerevisiae pha2p suffers from the feedback inhibition of L-phenylalanine, when the strain is used for fermenting yellow rice wine, the alcoholic strength can reach 13 degrees, and the yield of beta-phenylethyl alcohol can reach 120mg / L.

Description

technical field [0001] The invention relates to a strain of Saccharomyces cerevisiae and its application in fermented food, belonging to the technical field of industrial microorganisms. Background technique [0002] β-Phenethyl alcohol is an aromatic alcohol with rose flavor, which naturally exists in essential oils of plants such as jasmine and rose, and is also widely used as an important flavor and fragrance component in cosmetics, tobacco and daily chemical products. As a secondary metabolite of microorganisms, β-phenylethanol has a high content in fermented products such as rice wine, cooking wine, brewed vinegar, soy sauce, and white wine. At the same time, as a characteristic flavor substance in fermented products, it can improve the flavor and overall quality of fermented products . [0003] Phenylethyl alcohol is synthesized during the decarboxylation of phenylpyruvate, a precursor of phenylalanine. For Saccharomyces cerevisiae, the starting material for the synt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12P7/22C12G3/02C12J1/04A23L27/50C12R1/865
CPCA23L27/50C12G3/02C12J1/04C12N9/88C12P7/22C12Y402/01051
Inventor 毛健刘双平白梅
Owner JIANGNAN UNIV
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