CN113956988A - Two yeasts for promoting production of lactic acid and phenolic acid and application thereof in dry white wine brewing process - Google Patents
Two yeasts for promoting production of lactic acid and phenolic acid and application thereof in dry white wine brewing process Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G1/00—Preparation of wine or sparkling wine
- C12G1/02—Preparation of must from grapes; Must treatment and fermentation
- C12G1/0203—Preparation of must from grapes; Must treatment and fermentation by microbiological or enzymatic treatment
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G2200/00—Special features
- C12G2200/05—Use of particular microorganisms in the preparation of wine
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- Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
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Abstract
The invention discloses two yeasts for promoting the production of lactic acid and phenolic acid and application thereof in brewing of dry white wine. One strain is saccharomyces cerevisiae with good alcohol production rate, one strain is non-saccharomyces cerevisiae grape juice hansenula sporum with good beta-glucosidase activity through detection, and the contents of lactic acid and phenolic acid compounds of wine brewed by the two strains of yeast in a sequential inoculation fermentation mode are obviously increased. The type and content of acid are one of the most important sensory qualities of dry white wine, wherein lactic acid can blend sharp acidity taste in wine; besides affecting the taste of the wine, the phenolic acid compounds are also proved to have the effects of resisting oxidation, obesity, cancer and the like on human health. The mixed fermentation of the two strains of yeast is beneficial to brewing high-quality dry white wine with good acidity taste and high phenolic acid content, and has important significance for promoting the development of wine industry.
Description
Technical Field
The invention belongs to the field of food fermentation, and particularly relates to two yeasts for promoting the production of lactic acid and phenolic acid and application thereof in a dry white wine brewing process.
Background
The dry white wine is one of the most important global wines, wherein the acidity is an important index for evaluating the dry white wine, and balanced mouthfeel and coordinated sugar-acid ratio are often favored by consumers. The organic acid in the wine mainly comprises tartaric acid, malic acid, succinic acid, lactic acid, acetic acid, citric acid and the like, and a proper amount of organic acid can endow the wine with fresh and tasty flavor; accelerating the transformation of polysaccharide and the decomposition of pectic substance, thereby promoting the aging and clarification of the wine; inhibit the growth of harmful microorganisms and prevent the wine from putrefaction. However, if the content of the organic acid is too high, the wine body becomes hard and coarse, so that researches show that the content of the organic acid in the wine is preferably kept at 6-10 g/L. The organic acids are closely related to the yeast used in brewing, in addition to the grape material itself. Different yeasts determine the type and content of organic acids in wine, and especially influence lactic acid and succinic acid most obviously.
Phenolic acid compounds are important polyphenol compounds in wine, and are mainly p-hydroxybenzoic acid, p-hydroxycinnamic acid and derivatives thereof, wherein the p-hydroxybenzoic acid compounds mainly comprise vanillic acid, salicylic acid, gallic acid, syringic acid and the like; the derivatives of p-hydroxycinnamic acid mainly comprise p-coumaric acid, ferulic acid, caffeic acid and the like, and are widely existed in plants as secondary metabolites, and a large number of researches prove that phenolic acid substances are widely existed in daily diet of fruits, vegetables, grains and the like, and have the effects of oxidation resistance, inflammation diminishing, obesity resistance, cancer resistance and the like. The effect of yeast during wine brewing also affects the phenolic acid compound content.
In the brewing process of wine, the aims of reducing the content of malic acid and increasing the content of lactic acid are generally realized through a malic acid-lactic acid fermentation process, and methods such as a physical acid reduction method and a biological acid reduction method are researched and used for processing, wherein the biological acid reduction method using yeast is more natural and safer, so the method is particularly important for the development and utilization of biological resources. Compared with old world wine countries such as France, the research of the aspects of wine yeast screening, commercial starter research and development and the like in China is weak at present, and the utilization of a large amount of microbial resources is not sufficient. Therefore, the development and utilization of yeast resources are particularly important for the development of the wine industry, and the brewing of high-quality wine with characteristics of producing areas and varieties by using the yeast has important significance for improving the international competitiveness of the wine in China.
Disclosure of Invention
The purpose of the present invention is to provide two strains of yeast having good fermentation performance, wherein the yeast has the ability to promote the production of lactic acid and phenolic acid compounds.
The invention also aims to provide the dry white wine with excellent acidic characteristics, which has good mouthfeel and outstanding flavor.
Aiming at the purposes, the technical scheme provided by the invention is as follows:
the Saccharomyces cerevisiae (Saccharomyces cerevisiae) provided by the invention is named as YT13, is screened from a Shandong Penglai production area, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, and has the preservation address as follows: the microbial research institute of China academy of sciences, GmbH 3, GmbH, is: CGMCC NO. 22752. Unless otherwise indicated, s.cerevisiae is abbreviated herein as YT 13.
The grape juice provided by the invention is Hanseniaspora hansenula (Hanseniasporeavum) with the name of H30, and is separated and screened from ripe red wine grape varieties, namely 'Maseran' grape fruits picked in a vineyard in the Shandong Penglai producing area. The strain is preserved in China general microbiological culture Collection center, and the preservation addresses are as follows: the microbial research institute of China academy of sciences, GmbH 3, GmbH, is: CGMCC NO. 22754. Hansenula polymorpha, succulent, is herein abbreviated H30, unless otherwise indicated.
The YT13 and H30 provided by the invention can participate in the brewing process of wine. The advantages of using YT13 and H30 in brewing wine are: during the screening process, YT13 showed good sugar consumption rate and alcohol production rate, and could complete the whole fermentation process. H30 shows excellent beta-glucosidase activity and metabolic activity. The two strains can promote the generation of phenolic acid compounds and increase the content of lactic acid and succinic acid in the wine, have important significance for brewing the wine with native characteristics, and are beneficial to promoting the industrial development of the dry white wine in the whole production area.
Biological preservation Instructions
The biological material H30 is classified and named as grape juice of Hanseniaspora uvarum, which is preserved in China general microbiological culture Collection center at 21.06.06.2021 with the address of microorganism research institute of China academy of sciences No. 3 of Beijing university Hokko No. 1 of North Chen West Lu in the Korean district, with the preservation number of CGMCC NO. 22754;
the biological material YT13 is prepared with Saccharomyces cerevisiae and deposited in China general microbiological culture Collection center in 21.06.2021 with the microbial research institute of China academy of sciences No. 3, Beijing, Chaoyang, Beicheng, Xilu No. 1, Beicheng, and the deposited number of CGMCC No. 22752.
Drawings
FIG. 1 colony morphology of strain YT13 on WL medium;
FIG. 2 colony morphology of strain H30 on WL medium;
FIG. 3 shows the variation of residual sugar content in the fermentation of Guiren grape juice with the strain YT 13;
FIG. 4 shows the content of partial organic acids in Guiren fragrant wine after fermentation with different yeasts;
FIG. 5 shows the content of some phenolic acids in the noble savoury wine after fermentation with different yeasts;
FIG. 6 content of partial organic acids in Miscanthus minimal wine after fermentation with different yeasts;
FIG. 7 shows the content of some phenolic acids in Miscanthus grape wine after fermentation with different yeasts;
in fig. 5, 6, and 7, the letters in the figures indicate that there is significant difference between the measured data for different samples, p < 0.05.
Detailed Description
For a better understanding and appreciation of the invention, reference will now be made in detail to the following examples and accompanying drawings; the examples are given solely for the purpose of illustration and are not intended to limit the scope of the invention.
The methods used in the following examples are conventional methods unless otherwise specified.
Unless otherwise indicated, the reagents used in the following examples are all of analytical grade and are commercially available from a regular channel.
Unless otherwise indicated, the quantitative tests in the following examples were set up in triplicate and the results were statistically averaged.
The media formulations referred to in the examples are as follows:
YPD liquid medium (per 1L): 10g of yeast extract powder, 20g of peptone and 20g of glucose.
Example 1 Small experiment of sequential inoculation of yeasts H30 and YT13 with fermented Guiren fragrant Dry white wine
Removing stems of mature Guirenxiang variety wine grapes, crushing and squeezing, using a 10L glass fermentation tank as a wine fermentation container, and utilizing a commercial wine yeast EC1118 single-strain fermentation, a wine yeast YT13 single-strain fermentation, sequential inoculation fermentation of H30 and the commercial wine yeast, sequential inoculation fermentation of H30 and YT13, wherein four groups are respectively arranged in parallel by 3. After being activated, the yeast is inoculated into YPD liquid culture medium and is subjected to constant temperature shaking culture at the constant temperature of 28 ℃ for 18h under the condition of 120 rpm/min. Inoculating yeast into the clarified grape juice at an inoculation amount of 1%, and fermenting at 16 deg.C. The temperature, sugar degree and specific gravity were measured and recorded every day after the start of fermentation. Stirring at proper time according to the fermentation condition. When the residual sugar content of the noble character fragrant wine is reduced to 4g/L, the supernatant is taken and filled, and the wine is sealed and stored in a refrigerator at 4 ℃.
The contents of some organic acids and phenolic acid compounds in the wine were measured by High Performance Liquid Chromatography (HPLC), and the measurement results are shown in table 1. And detecting the basic physical and chemical indexes of the wine according to GB/T15038-. After fermentation with different yeasts, the contents of partial organic acids and phenolic acids in the noble people fragrant wine are shown in fig. 4 and fig. 5.
The results of small-scale wine brewing tests show that both strains of saccharomyces cerevisiae YT13 and non-saccharomyces cerevisiae H30 can increase the content of lactic acid and succinic acid in organic acid, improve the concentration of gallic acid, chlorogenic acid, caffeic acid, vanillic acid and p-coumaric acid in phenolic acid compounds, and increase the taste of acid, so that the wine is more refreshing and delicious. The wine increasing effect of the H30+ YT13 sequentially inoculated fermentation group is more obvious.
Example 2 sequential inoculation of yeasts H30 and YT13 in a bench test for fermenting Miscanthus Dry white wine
The method comprises the steps of removing stems, crushing and squeezing mature Musson brewing grapes, taking a 10L glass fermentation tank as a wine fermentation container, and utilizing a commercial saccharomyces cerevisiae EC1118 single-strain fermentation strain, a saccharomyces cerevisiae YT13 single-strain fermentation strain, a commercial saccharomyces cerevisiae sequential inoculation fermentation strain of H30 and H30 and YT13 sequential inoculation fermentation strain, wherein 3 strains are arranged in parallel in total. After the yeast is activated, the yeast is inoculated into YPD liquid culture medium and is cultured for 18h at the constant temperature of 28 ℃ by shaking. Inoculating 1% of the single yeast liquid into the clarified grape juice, sequentially inoculating H30, respectively inoculating commercial Saccharomyces cerevisiae and YT13 on the 4 th day of fermentation, and fermenting at 16 deg.C. The temperature and sugar levels were measured and recorded daily after the start of fermentation. Stirring at proper time according to the fermentation condition. When the residual sugar content of the Miscanthus sinensis wine is reduced to 4g/L, taking the supernatant, filling, sealing and storing in a refrigerator at 4 ℃.
The contents of some organic acids and phenolic acid compounds in the wine were determined by high performance liquid chromatography, and the results are shown in table 2. And detecting the basic physical and chemical indexes of the wine according to GB/T15038-. The contents of partial organic acids and phenolic acids in Miscanthus sinensis wine after fermentation with different yeasts are shown in FIGS. 6 and 7.
The results of small-scale brewing experiments show that both saccharomyces cerevisiae YT13 and non-saccharomyces cerevisiae H30 can increase the content of lactic acid in organic acid and increase the concentration of gallic acid, chlorogenic acid, caffeic acid, vanillic acid and p-coumaric acid in phenolic acid compounds. The wine adding effect of sequential inoculation fermentation groups is more obvious.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.
Claims (5)
1. A Hansenula polymorpha strain is characterized by having a preservation number of: CGMCC NO. 22754.
2. The saccharomyces cerevisiae is characterized by comprising the following preservation numbers: CGMCC NO. 22752.
3. A method for sequential inoculation of yeast, which comprises inoculating Hansenula polymorpha with the grape juice of claim 1 at a ratio of 1-3% into clarified grape juice, inoculating Saccharomyces cerevisiae with the grape juice of claim 2 at the 4 th day of fermentation at a ratio of 1-3%.
4. Use of two yeasts according to claims 1 and 2 for the production of dry white wine, wherein the dry white wine enriched with lactic acid and phenolic acid compounds is obtained by single strain/sequential mixing inoculation of the two yeasts for the fermentation of the raw white wine fruit material.
5. Dry white wine brewed with the yeast according to claims 1 and 2, comprising the steps of:
(1) taking out the strain preserved in glycerol at-80 deg.C, thawing, returning to room temperature, inoculating to YPD liquid culture medium at a ratio of 8-12% (v/v), culturing in a constant temperature shaking incubator until the strain grows to 10%7Obtaining a single bacterium liquid after the concentration is more than CFU/mL;
(2) picking up Guirenxiang grapes, selecting seeds, removing stems, crushing, squeezing, introducing grape self-flowing juice into a constant-temperature fermentation tank, and adding a clarifying agent to obtain clarified grape juice;
(3) inoculating the clarified white grape juice with yeast of a monosperm solution by the inoculation method according to claim 3;
(4) and controlling the temperature in the fermentation process at 16-18 ℃ until the residual sugar content is lower than 4g/L, and adding potassium metabisulfite to terminate the fermentation.
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CN106916758A (en) * | 2017-05-08 | 2017-07-04 | 中国农业大学 | A kind of Hansenula yeast and its application in wine production |
CN110923080A (en) * | 2019-10-30 | 2020-03-27 | 镇江瑞德酒业有限公司 | Flavor enhancing brewing process for fresh grape brandy |
CN112812980A (en) * | 2021-02-19 | 2021-05-18 | 宁夏农产品质量标准与检测技术研究所(宁夏农产品质量监测中心) | Mixed fermentation process based on Hansenula polymorpha and saccharomyces cerevisiae |
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CN106916758A (en) * | 2017-05-08 | 2017-07-04 | 中国农业大学 | A kind of Hansenula yeast and its application in wine production |
CN110923080A (en) * | 2019-10-30 | 2020-03-27 | 镇江瑞德酒业有限公司 | Flavor enhancing brewing process for fresh grape brandy |
CN112812980A (en) * | 2021-02-19 | 2021-05-18 | 宁夏农产品质量标准与检测技术研究所(宁夏农产品质量监测中心) | Mixed fermentation process based on Hansenula polymorpha and saccharomyces cerevisiae |
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