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Grape juice yeast strain with low higher alcohol yield and strong malic acid degradation and application of grape juice yeast strain

A grape juice yeast and yeast strain technology, applied in the fields of bioengineering and genetic engineering, can solve problems such as fermentation obstruction, wine disease, delay, etc., achieve broad market prospects, shorten the fermentation cycle, and eliminate the effects of influence

Active Publication Date: 2020-05-12
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, after the alcoholic fermentation is over, the higher alcohol content, lower pH value and residual sugar content of the wine body will inhibit the normal metabolism of lactic acid bacteria and hinder the fermentation, and the bacteriophages in the wine body will also cause malolactic fermentation. Delayed or inhibited, the fermentation of spoilage bacteria produces off-flavors and off-flavors, leading to wine diseases and reducing the flavor quality of wines

Method used

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  • Grape juice yeast strain with low higher alcohol yield and strong malic acid degradation and application of grape juice yeast strain
  • Grape juice yeast strain with low higher alcohol yield and strong malic acid degradation and application of grape juice yeast strain
  • Grape juice yeast strain with low higher alcohol yield and strong malic acid degradation and application of grape juice yeast strain

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Construction of grape juice yeast overexpressing malate permease and malolactase

[0046] (1) Construction of recombinant plasmid Yep-KPm1S

[0047] The construction process of the recombinant plasmid Yep-Pm1 is as follows: figure 1 shown;

[0048] Using the plasmid pPGK1 as a template, PGK-F (SEQ ID NO:5) and PGK-R (SEQ ID NO:6) as primers, PCR amplifies the PGK1 gene fragment (SEQ ID NO:4), PCR reaction conditions: 95°C 5min; 94°C for 40s, 56°C for 1min, 72°C for 108s, 30 cycles; 72°C for 10min. Plasmid Yep532 and PGK1 gene fragments were digested with restriction endonucleases BamHI and SalI, and the two were ligated to construct plasmid Yep-P; using the genome of Schizosaccharomyces pombe CICC1757 strain as a template, mae1-F (SEQ ID NO: 7) and mae1-R (SEQ ID NO: 8) as primers, PCR amplified fragment mae1 (SEQ ID NO: 1), PCR reaction conditions: 95°C for 5min; 94°C for 40s, 56°C for 1min, 72°C for 108s, 30 cycles; 10 min at 72°C. The fragment was ligated with t...

Embodiment 2

[0054] Fermentation Experiment of Grape Juice Yeast Strain with Low Production of Higher Alcohol

[0055] (1) Wine fermentation experiments of recombinant strains and starting strains

[0056] ①Fermentation process roadmap:

[0057] Grape raw materials; screening, cleaning, drying, destemming; crushing; sugar adjustment, acid adjustment; adding sulfurous acid, sterilization; inoculation; pre-fermentation; skin residue separation; determination indicators.

[0058] ②Process conditions: sugar content: 20.45Brix; acidity: pH 3.5; SO 2 Dosage: 80mg / L, let stand at 4°C for 12h; liquid volume: 190mL grape juice in a 250mL triangular bottle; inoculation volume: 1×10 8 CFU / mL; fermentation temperature and time: 25°C, 5 days; wine steaming conditions: 100mL fermentation broth, add 100mL water, steam 100mL wine sample.

[0059] Carry out wine fermentation experiment to grape juice yeast starting strain CICC1465 and the bacterial strain WY-m1S of selection embodiment according to abov...

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Abstract

The invention provides a grape juice yeast strain with low higher alcohol yield and strong malic acid degradation. According to the grape juice yeast strain, under the condition that other fermentation properties are not influenced and after wine is fermented for 5 days, the contents of isobutanol, isoamyl alcohol and phenethyl alcohol are 28.18 mg / L, 171.76 mg / L and 13.60 mg / L separately, and arereduced by 20.28%, 14.77% and 11.26% separately compared with those of an original strain; the total content of main higher alcohols (normal propyl alcohol, isobutanol, isoamyl alcohol and phenethylalcohol) is reduced by 12.97%; the content of malic acid after fermentation is reduced to 1.13 g / L; the fermentation period is greatly shortened; the influences of lactic acid bacteria fermentation ina common fermentation process are overcome, and flavor incompatibility caused by relatively high content of higher alcohol is also overcome; and the grape juice yeast strain has important meanings onreducing consumption of the wine brewing industry and improving the production technology level of the wine industry in China.

Description

Technical field: [0001] The invention belongs to the technical field of bioengineering and genetic engineering, and relates to the breeding and application of industrial microorganisms, in particular to a grape juice yeast strain with low yield of higher alcohol and strong malic acid degradation and its application in wine preparation. Background technique: [0002] Wine is a low-alcohol fruit wine made from fresh grapes or grape juice through full or partial fermentation. It is the product of the harmonious coexistence between man and nature. Cultivate suitable grape varieties, combined with unique technology, to brew wines with different flavors and characteristics. Wine flavor is a comprehensive reflection of the complementary balance of various flavor substances, and it is an important indicator to measure the quality of wine. Higher alcohol is the main by-product of wine fermentation and the main component of wine taste. A proper amount of higher alcohol cooperates wit...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12N15/81C12G1/022C12R1/85
CPCC12N9/88C12N9/0006C12N15/81C12G1/0203C12Y401/01C12Y101/0104C12G2200/11
Inventor 张翠英李彤李凭肖冬光郭学武林良才陈叶福于爱群
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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