CN108060091B - Aroma-producing yeast and application thereof in Chinese wolfberry fruit wine - Google Patents
Aroma-producing yeast and application thereof in Chinese wolfberry fruit wine Download PDFInfo
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Abstract
An aroma-producing yeast GF-60 is a pure aroma-producing yeast GF-60 obtained by culturing, separating, screening and purifying the materials obtained in a medlar plantation or a medlar fruit wine production place. The aroma-producing yeast provided by the invention is preserved in China general microbiological culture Collection center on 29 th 09.29 th 2017, and the preservation number is CGMCC No. 14774. The invention also applies the aroma-producing yeast GF-60 strain to the brewing of the medlar fruit wine. The aroma-producing yeast provided by the invention is suitable for brewing the medlar fruit wine by using most varieties of medlar, mainly aims at the problems of thin wine body and unobvious aroma characteristics of the medlar fruit wine caused by raw materials and processes in the fermentation process of the medlar fruit wine, supplements the aroma-producing yeast in time at the initial stage of fermentation, improves the aroma of the medlar fruit wine and improves the product quality. The strain makes up the defects of thin wine body and insufficient aroma of commercial aroma-producing yeast fermented Chinese wolfberry fruit wine, and improves the quality of the wine body.
Description
The technical field is as follows:
the invention relates to the technical field of aroma-producing yeast and wine brewing, in particular to aroma-producing yeast and application of the aroma-producing yeast in Chinese wolfberry fruit wine.
Background art:
the brewing of the fermented medlar wine is a complex microbiological process, the metabolite of the microorganism directly influences the color, taste and aroma of the wine, and the effective components in medlar are reserved. Among the factors influencing the quality of the fruit wine, the interaction of the factors of the variety of the medlar, the fermentation process, the fermentation equipment and the fermentation microorganisms finally determines the quality of the medlar wine. In industrial production, once the variety, fermentation process and fermentation equipment of the medlar are determined, the high-quality fermented medlar wine is obtained, and the selected yeast becomes important.
The high-quality medlar fruit wine depends on aroma and taste to a great extent, the main source of the aroma is the medlar and the aroma generated in the fermentation process, but compared with other fruit wine raw materials, the fruit aroma of the raw materials used by the medlar fruit wine is not prominent, and the surface of the medlar fruit needs to be cleaned before the medlar fruit wine is brewed, aroma-producing yeast on the surface of the medlar fruit is lost, and the deficiency is not compensated by post-process treatment, so that the quality of the medlar fruit wine is influenced by the problems of thin wine body, unobvious aroma characteristics, unobvious style and the like of the produced medlar fruit wine.
The invention content is as follows:
in view of the above, there is a need for aroma-producing yeast for fermentation of lycium barbarum.
Also provides a method for brewing the medlar fruit wine by adopting aroma-producing yeast.
The aroma-producing yeast provided by the invention is preserved in China general microbiological culture Collection center (CGMCC) in 29.09.2017 with the preservation number of CGMCC (China general microbiological culture Collection center) and is suitable for brewing medlar fruit wine.
The aroma-producing yeast CGMCC No.14774 is Hanseniaspora uvarum (Hanseniaspora uvarum), the aroma-producing yeast CGMCC No.14774 is an aroma-producing yeast strain GF-60, the colony color of the strain on a WL culture medium is dark green, the colony shape is flat, the surface is smooth and opaque, the strain is in a butter shape, and the strain is a lemon-shaped colony under a microscope.
A method for brewing fructus Lycii wine with aroma-producing yeast CGMCC No.14774 comprises the following steps:
collecting Chinese wolfberry fruits, and preparing the Chinese wolfberry fruits into Chinese wolfberry fruit juice;
putting the medlar juice into a medlar juice fermentation tank, and adding pectinase and sulfur dioxide;
inoculating, performing aroma-producing fermentation, and inoculating aroma-producing yeast CGMCC No.14774 into fermented mash in the medlar juice fermentation tank;
step four, alcoholic fermentation, namely fermenting the inoculated aroma-producing yeast CGMCC No.14774 medlar fermentation mash for 0-4 days, and then carrying out alcoholic fermentation;
and step five, clarifying and filtering, compounding bentonite and gelatin, adding the mixture into a wolfberry juice fermentation tank, pouring the mixture into the wolfberry juice fermentation tank while stirring the mixture uniformly, clarifying and placing the mixture for 15 to 16 days at the temperature of between 18 and 22 ℃, filtering, sterilizing and bottling the mixture to obtain the finished wolfberry fruit wine.
Step one, the dry wolfberry fruit is selected, the fresh and ripe fruits and rotten fruits are removed, and the wolfberry fruit is sprayed and washed with purified water, drained and added with 10 times of purified water to be rehydrated and extracted to prepare wolfberry fruit juice; or selecting fresh fructus Lycii, removing fresh and rotten fructus Lycii, spray rinsing, draining, adding 8 times of purified water, and squeezing to obtain fructus Lycii juice.
In the second step, the addition amount of the pectinase added into the medlar juice fermentation tank is 30-50g of pectinase added into each t of medlar juice, and 40-60ppm of 6% sulfur dioxide is added into the medlar juice fermentation tank.
And (3) adjusting components before inoculation, adding white granulated sugar and citric acid into the medlar juice fermentation tank to adjust the components of medlar fermentation mash so that the sugar content reaches 150g/L, and the pH value is 3.5-4.0.
In the third step, the required aroma-producing yeast GF-60 is activated in advance according to the time required by the strain to reach the peak of logarithmic phase according to the growth curve of the aroma-producing yeast CGMCC No.14774, and then the strain is inoculated into the fermented mash of medlar.
And in the fourth step, performing alcohol fermentation, namely inoculating aroma-producing yeast CGMCC No.14774 into the medlar juice fermentation tank, fermenting for 0-4 days, and adding activated AC commercial aroma-producing yeast according to 200g/t to start the alcohol fermentation.
And (3) carrying out secondary sugar supplement after the alcoholic fermentation, adding 70g/L of white granulated sugar again after the alcoholic fermentation is started for 48 hours, adding 100g/t of fermentation aid, monitoring the specific gravity change, and separating wine lees after the specific gravity is reduced to 0.996 and the sugar content is less than or equal to 4 g/L.
And in the fifth step, clarifying and filtering, namely using bentonite and gelatin to compound and glue, pouring the mixture in sequence while pouring the mixture evenly, clarifying and placing the mixture for 15 to 16 days at the temperature of between 18 and 22 ℃, and filtering the mixture by using a 0.45-micron microporous filter membrane.
The aroma-producing yeast provided by the invention is suitable for brewing the medlar fruit wine by using most varieties of medlar, mainly aims at the problems of thin wine body and unobvious aroma characteristics of the medlar fruit wine caused by raw materials and processes in the fermentation process of the medlar fruit wine, supplements the aroma-producing yeast in time at the initial stage of fermentation, improves the aroma of the medlar fruit wine and improves the product quality. The strain makes up the defects of thin wine body and insufficient aroma of commercial aroma-producing yeast fermented Chinese wolfberry fruit wine, and improves the quality of the wine body.
According to the invention, the aroma-producing yeast resource special for the medlar wine is excavated, and the yeast which can embody the special flavor and taste of medlar raw materials and has good aroma-producing performance is screened out through the deep systematic research on the fermentation characteristic of medlar yeast, so that a good strain with strong adaptability can be provided for the production of high-quality medlar fruit wine. The strain provided by the invention makes a great contribution to the sensory quality of the medlar fruit wine, and the aroma-producing yeast strain has the capability of producing a plurality of extracellular hydrolases, and the enzymes can accelerate the clarification and filtration of the wine, and are beneficial to the extraction of fruit juice pigment, the release of aromatic substances and the stability of the wine body. According to the invention, the proper aroma-producing yeast GF-60 is added in the brewing process of the fruit wine to make up for the defects that the fruit aroma of raw materials used by the medlar fruit wine is not prominent, the surface of the medlar fruit needs to be cleaned before the medlar fruit wine is brewed, the aroma-producing yeast on the surface of the medlar fruit is lost, the produced medlar fruit wine is thin in body, unobvious in aroma characteristics, unobvious in style and the like, and the quality of the medlar fruit wine is improved.
Description of the drawings:
FIG. 1 shows the microscopic morphology of aroma-producing yeast GF-60.
FIG. 2 shows the colony morphology of the aroma-producing yeast GF-60 medium.
The specific implementation mode is as follows:
the aroma-producing yeast provided by the invention is suitable for brewing medlar fruit wine, is preserved in China general microbiological culture Collection center in 29 th 09.2017, and has the preservation addresses as follows: the collection number of No.1 Xilu of Beijing, Chaoyang district is CGMCC No. 14774.
The aroma-producing yeast CGMCC No.14774 is Hanseniaspora uvarum (Hanseniaspora uvarum), the aroma-producing yeast CGMCC No.14774 is an aroma-producing yeast strain GF-60, the colony color of the strain on a WL culture medium is dark green, the colony shape is flat, the surface is smooth and opaque, the strain is in a butter shape, and the strain is a lemon-shaped colony under a microscope. The cell morphology of the fungus column is shown in figure 1, and the colony morphology of the aroma-producing yeast GF-60 culture medium is shown in figure 2.
The aroma-producing yeast strain GF-60 is obtained by obtaining materials in relevant places of medlar plantation and medlar fruit wine production, culturing, separating and screening pure aroma-producing yeast strain GF-60, carrying out physiological and biochemical experiments, identifying the gene sequence of yeast to determine the species of the yeast, simultaneously carrying out fermentation performance experiments on the yeast, selecting pure aroma-producing yeast with good fermentation performance and applying the pure aroma-producing yeast strain GF-60 to the brewing of medlar fruit wine.
The main experimental materials and equipment for GF-60 test of aroma-producing yeast are as follows:
first, experimental material
Second, experimental facilities
The invention provides a better implementation mode for brewing medlar fruit wine by adopting aroma-producing yeast GF-60.
Separation and purification: separating yeast from the orchard soil, the surface of fresh wolfberry fruit, the naturally fermented wolfberry liquid and wolfberry distiller's grains, purifying and preserving. Simultaneously carrying out three-stage screening on the separated saccharomycetes;
physiological and biochemical identification: identifying the attribution of the yeast according to different colony forms and colors of the yeast grown on a WL culture medium;
fermentation performance study: respectively adding alcohol, glucose, sulfurous acid and the like into an YDP liquid culture medium with a Du's tube to form a certain concentration, inoculating a certain amount of aroma-producing yeast, recording the fermentation time of the aroma-producing yeast, and evaluating the alcohol resistance, the sugar resistance and the sulfur dioxide resistance of the aroma-producing yeast; inoculating GF-60 to a BiGGY culture medium, culturing and observing the color of the culture medium to qualitatively detect the hydrogen sulfide production; selecting a strain with better fermentation performance to determine a growth curve;
leaching: selecting ripe dry fruits of Ningxia Zhongning fructus Lycii, selecting and removing fresh fruits and rotten fruits, spraying and washing with purified water, draining, adding 10 times of purified water, and extracting with water again;
pretreatment: putting the medlar juice into a tank, and adding 30-50g/t of pectinase and 40-60ppm of 6% sulfur dioxide;
component adjustment: adding white sugar and citric acid to adjust fermented liquor components of fructus Lycii until sugar content reaches 150g/L and pH value is 3.5-4.0;
fragrance production and fermentation: inoculating the aroma-producing yeast GF-60 which is required to be activated in advance according to the time required by the GF-60 growth curve strain to reach the peak of logarithmic phase into the fermented mash of the medlar;
alcohol fermentation: inoculating aroma-producing yeast for 2-6 days, adding activated AC commercial aroma-producing yeast at 200g/t to start alcohol fermentation;
and (3) secondary sugar supplement: after the alcohol fermentation is started for 48 hours, adding 70g/L of white granulated sugar again and adding 100g/t of fermentation aid, monitoring the change of specific gravity, and separating wine lees after the specific gravity is reduced to 0.996 and the sugar content is less than or equal to 4 g/L;
clarifying and filtering: adding bentonite and gelatin, pouring in sequence while stirring, standing at 18-22 deg.C for 15 days, and filtering with 0.45 μm microporous membrane;
comparing the quality evaluation and physical and chemical indexes with the medlar fruit wine fermented by commercial yeast AC single strain;
the strain is obtained from a Ningxia Zhongning wolfberry plantation and Ningqi No. seven wolfberry natural fermentation liquor through cultivation, separation, three-stage screening and purification.
The strain is subjected to a hydrogen sulfide production experiment, and is colorless (the lighter the color is, the less hydrogen sulfide is) after being cultured on a BIGGY culture medium for 60 hours; the tolerance test shows that: the yeast grows vigorously by fermenting for 48 hours in various environments with the sulfur dioxide concentration below 250mg/l, the sugar concentration below 300g/l and the temperature below 35 ℃.
The growth curve of aroma-producing yeast GF-60 was determined, which after 14 hours of fermentation the yeast proliferated in large amounts and reached a maximum at 22 hours.
Through the identification of ITS (internal transcribed spacer) discriminators of aroma-producing yeast GF-60 bacteria, the strain belongs to the Hansenula polymorpha with grape juice, and the identification sequence result is as follows:
TTGTTGCTCGAGTTCTTGTTTAGATCTTTTACAATAATGTGTATCTTTACTGAAGATGT GCGCTTAATTGCGCTGCTTCTTTAGAGTGTCGCAGTGAAAGTAGTCTTGCTTGAATCTC AGTCAACGCTACACACATTGGAGTTTTTTTACTTTAATTTAATTCTTTCTGCTTTGAAT CGAAAGGTTCAAGGCAAAAAACAAACACAAACAATTTTATTTTATTATAATTTTTTAAA CTAAACCAAAATTCCTAACGGAAATTTTAAAATAATTTAAAACTTTCAACAACGGATCT CTTGGTTCTCGCATCGATGAAGAACGTAGCGAATTGCGATAAGTAATGTGAATTGCAGA TACTCGTGAATCATTGAATTTTTGAACGCACATTGCGCCCTTGAGCATTCTCAGGGGCA TGCCTGTTTGAGCGTCATTTCCTTCTCAAAAGATAATTTATTATTTTTTGGTTGTGGGC GATACTCAGGGTTAGCTTGAAATTGGAGACTGTTTCAGTCTTTTTTAATTCAACACTTA GCTTCTTTGGAGACGCTGTTCTCGCTGTGATGTATTTATGGATTTATTCGTTTTACTTT ACAAGGGAAATGGTAACGTACCTTAGGCAAAGGGTTGCTTTTAATATTCATCAAGTTTG ACCTCAAATCAGGTAGGATTACCCGCTGAACTTAAGCATATCAATAAGCGGAGGAA.
the aroma-producing yeast GF-60 and the commercial aroma-producing yeast AC are inoculated simultaneously, and the inoculation amount is 3 times of the inoculation amount of the AC yeast. Compared with the medlar fruit wine without adding aroma-producing yeast GF-60, the medlar fruit wine has 7 kinds of substances, 2 kinds of lipid substances and enriched aroma.
According to the invention, the aroma-producing yeast resource special for the medlar wine is excavated, and the yeast which can embody the special flavor and taste of medlar raw materials and has good aroma-producing performance is screened out through the deep systematic research on the fermentation characteristic of medlar yeast, so that a good strain with strong adaptability can be provided for the production of high-quality medlar fruit wine. The strain provided by the invention makes a great contribution to the sensory quality of the medlar fruit wine, and the aroma-producing yeast strain has the capability of producing a plurality of extracellular hydrolases, and the enzymes can accelerate the clarification and filtration of the wine, and are beneficial to the extraction of fruit juice pigment, the release of aromatic substances and the stability of the wine body. According to the invention, the proper aroma-producing yeast GF-60 is added in the brewing process of the fruit wine to make up for the defects that the fruit aroma of raw materials used by the medlar fruit wine is not prominent, the surface of the medlar fruit needs to be cleaned before the medlar fruit wine is brewed, the aroma-producing yeast on the surface of the medlar fruit is lost, the produced medlar fruit wine is thin in body, unobvious in aroma characteristics, unobvious in style and the like, and the quality of the medlar fruit wine is improved.
Claims (7)
1. An aroma-producing yeast CGMCC No.14774, which is characterized in that: the aroma-producing yeast strain has the preservation number of CGMCC No.14774 and is preserved in the China general microbiological culture Collection center on 29 th 09.2017;
the aroma-producing yeast CGMCC No.14774 is Hanseniaspora uvarum (Hanseniaspora uvarum), the aroma-producing yeast CGMCC No.14774 is an aroma-producing yeast strain GF-60, the colony color of the strain on a WL culture medium is dark green, the colony shape is flat, the surface is smooth and opaque, the strain is in a butter shape, and the strain is a lemon-shaped colony under a microscope.
2. A method for brewing medlar wine by aroma-producing yeast CGMCC No.14774 as defined in claim 1, which is characterized in that: the method for brewing the medlar fruit wine by adopting aroma-producing yeast CGMCC No.14774 comprises the following steps:
collecting Chinese wolfberry fruits, and preparing the Chinese wolfberry fruits into Chinese wolfberry fruit juice;
putting the medlar juice into a medlar juice fermentation tank, and adding pectinase and sulfur dioxide;
inoculating, performing aroma-producing fermentation, and inoculating aroma-producing yeast CGMCC No.14774 into fermented mash in the medlar juice fermentation tank;
step four, alcoholic fermentation, namely fermenting the inoculated aroma-producing yeast CGMCC No.14774 medlar fermentation mash for 0-4 days, and then carrying out alcoholic fermentation;
step five, clarifying and filtering, namely compounding bentonite and gelatin, adding the mixture into a medlar juice fermentation tank, pouring the mixture into the medlar juice fermentation tank while stirring the mixture uniformly, then clarifying and placing the mixture for 15 to 16 days at the temperature of between 18 and 22 ℃, then filtering, sterilizing and bottling the mixture to obtain a finished medlar fruit wine product;
selecting dry fructus Lycii, removing fresh and rotten fructus Lycii, spraying purified water, draining, adding 10 times of purified water, and extracting to obtain fructus Lycii juice; or selecting fructus Lycii from fresh fructus Lycii, removing fresh and ripe fruit and rotten fruit, spray rinsing, draining, adding 8 times of purified water, and squeezing to obtain fructus Lycii juice;
in the second step, the addition amount of the pectinase added into the medlar juice fermentation tank is 30-50g of pectinase added into each t of medlar juice, and 40-60ppm of 6% sulfur dioxide is added into the medlar juice fermentation tank.
3. The method for brewing medlar wine by adopting aroma-producing yeast CGMCC No.14774 as claimed in claim 2, which is characterized in that: and (3) adjusting components before inoculation, adding white granulated sugar and citric acid into the medlar juice fermentation tank to adjust the components of medlar fermentation mash so that the sugar content reaches 150g/L, and the pH value is 3.5-4.0.
4. The method for brewing medlar wine by adopting aroma-producing yeast CGMCC No.14774 as claimed in claim 2, which is characterized in that: in the third step, the aroma-producing yeast GF-60 required by the activation is advanced according to the time required by the strain of the aroma-producing yeast CGMCC No.14774 growth curve to reach the peak of the logarithmic phase, and then the strain is inoculated into the fermented mash of the medlar.
5. The method for brewing medlar wine by adopting aroma-producing yeast CGMCC No.14774 as claimed in claim 2, which is characterized in that: and in the fourth step, performing alcohol fermentation, namely inoculating aroma-producing yeast CGMCC No.14774 into the medlar juice fermentation tank, fermenting for 0-4 days, and adding activated AC commercial aroma-producing yeast according to 200g/t to start the alcohol fermentation.
6. The method for brewing medlar wine by adopting aroma-producing yeast CGMCC No.14774 as claimed in claim 2, which is characterized in that: and (3) carrying out secondary sugar supplement after the alcoholic fermentation, adding 70g/L of white granulated sugar again after the alcoholic fermentation is started for 48 hours, adding 100g/t of fermentation aid, monitoring the specific gravity change, and separating wine lees after the specific gravity is reduced to 0.996 and the sugar content is less than or equal to 4 g/L.
7. A method for brewing medlar wine by adopting aroma-producing yeast CGMCC No.14774 as claimed in any one of claims 2 to 6, wherein: and step five, clarifying and filtering, namely using bentonite and gelatin to compound and glue, pouring the mixture in sequence while pouring the mixture evenly, clarifying and placing the mixture for 15 to 16 days at the temperature of between 18 and 22 ℃, and filtering the mixture by using a 0.45-micron microporous filter membrane.
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| CN113150902B (en) * | 2021-03-24 | 2023-07-18 | 西北农林科技大学 | Fermentation process for adding amino acid into substrate and based on Hansenula polymorpha of grape juice |
| CN114276943B (en) * | 2022-01-27 | 2023-08-15 | 宁夏红枸杞产业有限公司 | Saccharomyces cerevisiae, microbial preparation and medlar wine and preparation method thereof |
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