CN112812980A - Mixed fermentation process based on Hansenula polymorpha and saccharomyces cerevisiae - Google Patents
Mixed fermentation process based on Hansenula polymorpha and saccharomyces cerevisiae Download PDFInfo
- Publication number
- CN112812980A CN112812980A CN202110189028.4A CN202110189028A CN112812980A CN 112812980 A CN112812980 A CN 112812980A CN 202110189028 A CN202110189028 A CN 202110189028A CN 112812980 A CN112812980 A CN 112812980A
- Authority
- CN
- China
- Prior art keywords
- strain
- activated
- saccharomyces cerevisiae
- fermentation
- grape juice
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
-
- 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
-
- 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
- C12G1/00—Preparation of wine or sparkling wine
- C12G1/02—Preparation of must from grapes; Must treatment and fermentation
- C12G1/04—Sulfiting the must; Desulfiting
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Abstract
The invention discloses a mixed fermentation process based on Hansenula polymorpha and saccharomyces cerevisiae of grape juice, belonging to the technical field of microorganisms. The mixed fermentation process is characterized in that hansenula polymorpha and saccharomyces cerevisiae which are contained in grape juice are subjected to mixed fermentation; the Hansenula polymorpha is Hanseniaspora uvarum strain QTX 22; the collection number of the Hanseniaspora uvarum strain QTX22 of the grape juice is CCTCC M2021083. Compared with a single-bacterium fermented product, the mixed-bacterium fermented product has more unique fragrance, can produce a wine with unique flavor, fragrance and taste, fills the consumer category and expands the consumption selection.
Description
Technical Field
The invention belongs to the technical field of beverage fermentation, and particularly relates to a mixed fermentation process based on Hansenula polymorpha and saccharomyces cerevisiae.
Background
Wine fermentation is a complex biochemical process, and Hansenula polymorpha, a grape juice, plays a very critical role in fermentation processes, such as the conversion of sugars into ethanol, carbon dioxide and other secondary metabolites, which can be thousands to tens of thousands. A large number of scientific researches show that the quality of the wine is highly dependent on the metabolic activity and fermentation behavior of Hansenula polymorpha in different grape juices, and the Hansenula polymorpha in different grape juices has important contributions to the chemical composition, the sensory characteristics, the flavor characteristics and the like of the wine. Saccharomyces cerevisiae is the strain which is widely applied in the industrial production of wine so far, has the advantages of ensuring the risk of deterioration in the fermentation process of wine, and having good fermentation power, but has the problems of single flavor characteristic, serious homogenization phenomenon and the like of wine. Therefore, in order to pursue wine style specialization and have representative, diversified and complicated aroma characteristics, brewing workers often adopt a method of mixing and fermenting saccharomyces cerevisiae and non-saccharomyces cerevisiae, particularly certain native grape juice Hansenula sporulata with strong adaptability and representativeness, so as to improve and enhance the flavor quality of wine.
Non-saccharomyces cerevisiae has become an option for improving the quality of wine. Numerous studies have shown that non-saccharomyces cerevisiae is able to produce enzymes as well as some of our desired secondary metabolites, thus improving wine aroma and flavor characteristics, and is able to control the growth of some undesirable species in wine, but has the disadvantage of insufficient fermentation power. The mixed fermentation of the non-saccharomyces cerevisiae and the saccharomyces cerevisiae can improve the aroma diversity and complexity of the wine, can make up for the problem of insufficient fermentation power of the non-saccharomyces cerevisiae, and is an effective method for improving the aroma quality of the wine.
The prior art currently reports on mixed fermentation of Hanseniaspora uvarum, a grape juice and Saccharomyces cerevisiae: the invention patent application 201911057302.1 discloses an aroma-enhancing brewing process for fresh grape brandy, which improves the aroma components and the content of fermentation ester substances of grape varieties in fermented wine base by the mixed fermentation of Hanseniaspora uvarum (Hanseniaspora uvarum) strain Yun268 with the preservation number of CCTCC NO: M2013658 and Saccharomyces cerevisiae (Saccharomyces cerevisiae).
However, the types of aroma substances generated by mixed fermentation in the prior art are relatively limited, and therefore, in order to further improve the flavor and aroma of wine, the field needs to develop more mixed fermentation processes capable of generating more aroma substances.
Disclosure of Invention
Based on the above requirements and blanks in the field, the invention provides a mixed fermentation process based on Hanseniaspora uvarum, a grape juice and Saccharomyces cerevisiae, and compared with Saccharomyces cerevisiae F33 single-strain fermentation, the mixed fermentation process has the advantages that various aroma substances which can affect the flavor of wine are remarkably improved.
The technical scheme of the invention is as follows:
a mixed fermentation process based on Hansenula polymorpha and saccharomyces cerevisiae is characterized in that the Hansenula polymorpha and saccharomyces cerevisiae are subjected to mixed fermentation;
the Hansenula polymorpha is Hanseniaspora uvarum strain QTX 22; the collection number of the Hanseniaspora uvarum strain QTX22 of the grape juice is CCTCC M2021083.
The saccharomyces cerevisiae refers to saccharomyces cerevisiae strain F33.
The mixed fermentation process based on Hansenula polymorpha and saccharomyces cerevisiae comprises the following steps: the activated strain and the activated strain are inoculated into a substrate for secondary fermentation for fermentation.
Preferably, said strain refers to: hanseniaspora uvarum strain QTX22, Hanseniaspora uvarum, or Saccharomyces cerevisiae strain F33;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
preferably, the culture temperature is 24-30 ℃, preferably 28 ℃; the culture time is 20-30h, preferably 24h, the rotation speed is 120-250rpm, preferably 150 rpm;
preferably, the inoculation refers to inoculating the initial strain stock solution into the culture medium according to the inoculation amount of 2-4%, preferably 3% in volume ratio;
preferably, the medium is a YPD medium; preferably, the YPD medium comprises the following components in mass-to-volume ratio: 0.5-3.5%, preferably 1%, yeast extract, 1.0-3.0%, preferably 2%, peptone, 1.0-5.0%, preferably 2%, glucose, and water in balance;
preferably, the initial strain stock solution refers to the strain stock in 15-35% by volume, preferably 25% glycerol/YPD medium;
more preferably, the activation of the strain is performed 1-3 times, preferably 2 times.
The activated strain refers to: the activated grape juice contains Hanseniaspora uvarum Hanseniaspora strain QTX22, or the activated saccharomyces cerevisiae strain F33;
preferably, the fermentation of the activated substrate into the secondary fermentation refers to: firstly, inoculating an activated Hanseniaspora uvarum strain QTX22 of grape juice to a substrate for fermentation for 0-96h, preferably 48h, and then inoculating an activated saccharomyces cerevisiae strain F33 for continuous fermentation;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the activated grape juice has Hanseniaspora uvarum strain QTX22, and the total inoculum size of the activated Saccharomyces cerevisiae strain F33 is 106-107CFU, preferably 6X 106CFU；
Preferably, the fermentation is terminated when the substrate weight loss does not change for 3 consecutive days;
preferably, the substrate is grape juice.
A production method of wine is characterized in that grape juice is used as a substrate, and hansenula polymorpha and saccharomyces cerevisiae which are used as grape juice are subjected to mixed fermentation;
the Hansenula polymorpha is Hanseniaspora uvarum strain QTX 22; the collection number of the Hanseniaspora uvarum strain QTX22 of the grape juice is CCTCC M2021083.
The saccharomyces cerevisiae refers to saccharomyces cerevisiae strain F33.
The production method of the wine comprises the following steps: the activated strain and the activated strain are inoculated into a substrate for secondary fermentation for fermentation.
Preferably, said strain refers to: hanseniaspora uvarum strain QTX22, Hanseniaspora uvarum, or Saccharomyces cerevisiae strain F33;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
preferably, the culture temperature is 24-30 ℃, preferably 28 ℃; the culture time is 20-30h, preferably 24h, the rotation speed is 120-250rpm, preferably 150 rpm;
preferably, the inoculation refers to inoculating the initial strain preservation solution into the culture medium according to the inoculation amount of 3% by volume;
preferably, the medium is a YPD medium; preferably, the YPD medium comprises the following components in mass-to-volume ratio: 0.5-3.5%, preferably 1%, yeast extract, 1.0-3.0%, preferably 2%, peptone, 1.0-5.0%, preferably 2%, glucose, and water in balance;
preferably, the initial strain stock solution refers to the strain stock in 15-35% by volume, preferably 25% glycerol/YPD medium;
more preferably, the activation of the strain is performed 1-3 times, preferably 2 times.
The activated strain refers to: the activated grape juice contains Hanseniaspora uvarum Hanseniaspora strain QTX22, or the activated saccharomyces cerevisiae strain F33;
preferably, the fermentation of the activated substrate into the secondary fermentation refers to: firstly, inoculating an activated Hanseniaspora uvarum strain QTX22 of grape juice to a substrate for fermentation for 0-96h, preferably 48h, and then inoculating an activated saccharomyces cerevisiae strain F33 for continuous fermentation;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the activated grape juice has Hanseniaspora uvarum strain QTX22 of Hanseniaspora, and the total inoculated amount of the activated Saccharomyces cerevisiae strain F33 is 106-107CFU, preferably 6X 106CFU；
Preferably, the fermentation is terminated when the substrate weight loss is not changed for 3 consecutive days.
The wine production method further comprises the following steps: preparing grape juice;
the preparation of the grape juice comprises the following steps: squeezing grape fruit grains at low temperature;
preferably, the harvested grape fruit particles are pressed with ice by an air bag press;
preferably, pressing is performed while adding sulfur dioxide or K2S2O5And, pectinase;
preferably, sulfur dioxide or K2S2O5The addition amount of (B) is 30-100mg/L, preferably 50 mg/L; the addition amount of pectinase is 10-30mg/L, preferably 20 mg/L;
preferably, the grape fruit pieces are uniformly sized fruit pieces;
preferably, the grape fruit particles are grape fruit particles collected when the temperature of the mature grapes continuously drops below 24 hours to 8 ℃.
The wine is characterized by being produced by the wine production method.
The invention adopts Hanseniaspora uvarum which is a grape juice Hanseniaspora uvarum and saccharomyces cerevisiae to carry out mixed fermentation, and the output of most aroma substances in dozens of aroma substances generated is improved, for example, isoamyl acetate is improved by 158 percent, methyl acetate is improved by about 61 percent, methyl acetate is improved by 180 percent, caprylic acid is improved by about 46 percent, n-capric acid is improved by 167 percent, 9-decenoic acid is improved by about 54 percent, 2-methylbutyric acid is improved by 42 percent, 2-oxocaprylic acid is improved by 37 percent, dimethyl silanediol is improved by about 2 times, acetaldehyde is improved by 38 percent, n-hexanal is improved by about 138 percent, vinyl ether is improved by 22 percent, ethyl laurate is improved by 68 percent, ethyl heptanoate is improved by 21 percent, ethyl hexanoate is improved by 34 percent, ethyl butyrate is improved by 73 percent, ethyl myristate is improved by 1 time, ethyl 3-phenylpropionate is improved by 39 percent, the trans-4-ethyl decenoate is improved by 18 percent, the 9-ethyl hexadecenoate is improved by 50 percent, the diisobutyl phthalate is improved by 65 percent, the 2,2, 4-trimethyl pentanediol isobutyl ester is improved by 36 percent, the 2,2, 4-trimethyl-1, 3-pentanediol monoisobutyrate is improved by 71 percent, and the vinyl acetate is improved by 43 percent; meanwhile, aroma substances which cannot be produced by single-strain fermentation of saccharomyces cerevisiae are also produced, such as: isobutyl acetate, phenethylacetate, lauric acid, 2-benzylpropionic acid, 3-methyl-1-pentanol, 2-ethylhexanol, propylene glycol, (Z) -3-methyl-2-hepten-1-ol, 2-tridecanone, isoamyl decanoate, m-xylene, alpha-terpineol, levo-beta-pinene, 2, 4-di-tert-butylphenol. The production of the aroma substances or the increase of the yield of the aroma substances can exert certain influence on the flavor and aroma of the beverage obtained by fermentation, so that the mixed fermentation product presents more unique aroma compared with a single-bacterium fermentation product, a wine beverage with unique flavor, aroma and taste can be produced, the consumer class is enriched, and the consumption selection is expanded.
Detailed Description
The following detailed description of the present invention will be made with reference to specific examples, but the scope of the present invention is not limited thereto.
Sources and documentations of biological materials
The Hanseniaspora uvarum strain QTX22, a novel strain selected in the laboratory of the applicant, was used in the experimental examples of grape juice with the following deposit information:
naming: QTX22
And (4) classification name: hansenula polymorpha of grape juice
The name of Latin is: hanseniaspora uvarum
The preservation number is as follows: CCTCC NO: m2021083
The preservation organization: china center for type culture Collection
And (4) storage address: wuhan university school of eight-channel 299 # Wuhan university in Wuchang district of Wuhan city, Hubei province (first attached to small opposite surface of Wuhan university), Wuhan university collection center
The preservation date is as follows: 1 month 15 days 2021;
saccharomyces cerevisiae F33 was a commercial strain purchased from Lafford (Laffort) France.
The grape variety used was a wital ice grape, purchased from Ningxia Bagges drunk intersomatic wine village, Inc.
Group 1 example, Mixed fermentation Process of the invention
The embodiment of the group provides a mixed fermentation process based on Hansenula polymorpha and saccharomyces cerevisiae. All embodiments of this group share the following common features: carrying out mixed fermentation on Hansenula polymorpha and saccharomyces cerevisiae; the Hansenula polymorpha is Hanseniaspora uvarum strain QTX 22; the collection number of the Hanseniaspora uvarum strain QTX22 of the grape juice is CCTCC M2021083.
Those skilled in the art can produce and use Hanseniaspora uvarum strain QTX22 of Hanseniaspora uvarum, which is a strain of grape juice, in combination with Saccharomyces cerevisiae, and the invention also falls within the scope of the present invention. Target products of fermentation include, but are not limited to: wine drink, fermented milk, bread, etc.
In a specific embodiment, the Saccharomyces cerevisiae is Saccharomyces cerevisiae strain F33.
According to the teaching of the present invention, other commercial strains can be selected by those skilled in the art for mixed fermentation, and besides F33, there are many commercial strains on the market, such as Saccharomyces cerevisiae V1116, Saccharomyces cerevisiae VL1, Saccharomyces cerevisiae X16, etc., which can be used for mixed fermentation with Hanseniaspora uvarum strain QTX22 of the grape juice of the present invention to obtain similar technical effects as the present invention.
In some embodiments, the mixed fermentation process based on Hansenula polymorpha and Saccharomyces cerevisiae comprises the following steps: the activated strain and the activated strain are inoculated into a substrate for secondary fermentation for fermentation.
Preferably, said strain refers to: hanseniaspora uvarum strain QTX22, Hanseniaspora uvarum, or Saccharomyces cerevisiae strain F33;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
preferably, the culture temperature is 24-30 ℃, preferably 28 ℃; the culture time is 20-30h, preferably 24h, the rotation speed is 120-250rpm, preferably 150 rpm;
preferably, the inoculation refers to inoculating the initial strain stock solution into the culture medium according to the inoculation amount of 2-4%, preferably 3% in volume ratio;
preferably, the medium is a YPD medium; preferably, the YPD medium comprises the following components in mass-to-volume ratio: 0.5-3.5%, preferably 1%, yeast extract, 1.0-3.0%, preferably 2%, peptone, 1.0-5.0%, preferably 2%, glucose, and water in balance;
preferably, the initial strain stock solution refers to the strain stock in 15-35% by volume, preferably 25% glycerol/YPD medium;
more preferably, the activation of the strain is performed 1-3 times, preferably 2 times.
In other embodiments, the activated strain refers to: the activated grape juice contains Hanseniaspora uvarum Hanseniaspora strain QTX22, or the activated saccharomyces cerevisiae strain F33;
preferably, the fermentation of the activated substrate into the secondary fermentation refers to: firstly, inoculating an activated Hanseniaspora uvarum strain QTX22 of grape juice to a substrate for fermentation for 0-96h, preferably 48h, and then inoculating an activated saccharomyces cerevisiae strain F33 for continuous fermentation;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the activated glucanHanseniaspora uvarum strain QTX22, and the total inoculated amount of the activated Saccharomyces cerevisiae strain F33 was 106-107CFU, preferably 6X 106CFU；
Preferably, the fermentation is terminated when the substrate weight loss does not change for 3 consecutive days;
preferably, the substrate is grape juice.
Group 2 example, Process for the production of wine according to the invention
The present group of embodiments provides a method of producing wine. The present group of embodiments all have the following common features: taking grape juice as a substrate, and carrying out mixed fermentation on the grape juice with Hansenula polymorpha and saccharomyces cerevisiae; the Hansenula polymorpha is Hanseniaspora uvarum strain QTX 22; the collection number of the Hanseniaspora uvarum strain QTX22 of the grape juice is CCTCC M2021083.
In a specific embodiment, the Saccharomyces cerevisiae is Saccharomyces cerevisiae strain F33.
In other specific embodiments, the wine production method comprises: the activated strain and the activated strain are inoculated into a substrate for secondary fermentation for fermentation.
Preferably, said strain refers to: hanseniaspora uvarum strain QTX22, Hanseniaspora uvarum, or Saccharomyces cerevisiae strain F33;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
preferably, the culture temperature is 24-30 ℃, preferably 28 ℃; the culture time is 20-30h, preferably 24h, the rotation speed is 120-250rpm, preferably 150 rpm;
preferably, the inoculation refers to inoculating the initial strain stock solution into the culture medium according to the inoculation amount of 2-4%, preferably 3% in volume ratio;
preferably, the medium is a YPD medium; preferably, the YPD medium comprises the following components in mass-to-volume ratio: 0.5-3.5%, preferably 1%, yeast extract, 1.0-3.0%, preferably 2%, peptone, 1.0-5.0%, preferably 2%, glucose, and water in balance;
preferably, the initial strain stock solution refers to the strain stock in 15-35% by volume, preferably 25% glycerol/YPD medium;
more preferably, the activation of the strain is performed 1-3 times, preferably 2 times.
In specific embodiments, the activated strain refers to: the activated grape juice contains Hanseniaspora uvarum Hanseniaspora strain QTX22, or the activated saccharomyces cerevisiae strain F33;
preferably, the fermentation of the activated substrate into the secondary fermentation refers to: firstly, inoculating an activated Hanseniaspora uvarum strain QTX22 of grape juice to a substrate for fermentation for 0-96h, preferably 48h, and then inoculating an activated saccharomyces cerevisiae strain F33 for continuous fermentation;
when the time for fermenting by inoculating the Hanseniaspora uvarum strain QTX22 of the activated grape juice to the substrate is 0h, the Hanseniaspora uvarum strain QTX22 of the activated grape juice and the activated saccharomyces cerevisiae strain F33 can be simultaneously inoculated to the substrate for fermenting until the substrate weight loss is not changed any more for 3 days, and the fermentation is stopped.
Preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the activated grape juice has Hanseniaspora uvarum strain QTX22 of Hanseniaspora, and the total inoculated amount of the activated Saccharomyces cerevisiae strain F33 is 106-107CFU, preferably 6X 106CFU；
Preferably, the fermentation is terminated when the substrate weight loss is not changed for 3 consecutive days.
In a further embodiment, the method of wine production further comprises: preparing grape juice;
the preparation of the grape juice comprises the following steps: squeezing grape fruit grains at low temperature;
preferably, the harvested grape fruit particles are pressed with ice by an air bag press;
preferably, pressing is performed while adding sulfur dioxide or K2S2O5And, pectinase;
preferably, the addition amount of sulfur dioxide or K2S2O5 is 30-100mg/L, preferably 50 mg/L; the addition amount of pectinase is 10-30mg/L, preferably 20 mg/L;
preferably, the grape fruit pieces are uniformly sized fruit pieces;
preferably, the grape fruit particles are grape fruit particles collected when the temperature of the mature grapes continuously drops below 24 hours to 8 ℃. Group 3 examples, wine of the invention
The present group of embodiments provides a wine. All embodiments of this group share the following common features: produced by the wine production method of any one of the group 2 examples.
The wine of the invention produces the following aroma substances which cannot be produced by the wine fermented by single bacteria: isobutyl acetate, phenethylacetate, lauric acid, 2-benzylpropionic acid, 3-methyl-1-pentanol, 2-ethylhexanol, propylene glycol, (Z) -3-methyl-2-hepten-1-ol, 2-tridecanone, isoamyl decanoate, m-xylene, α -terpineol, l- β -pinene, 2, 4-di-t-butylphenol, while being much higher in the content of the following aroma substances than in single-fermented wines: isoamyl acetate, methyl acetate, octanoic acid, n-decanoic acid, 9-decenoic acid, 2-methylbutyric acid, 2-oxooctanoic acid, dimethylsilanediol, acetaldehyde, n-hexanal, vinyl ethyl ether, ethyl laurate, ethyl heptanoate, ethyl hexanoate, ethyl butyrate, ethyl myristate, ethyl 3-phenylpropionate, ethyl trans-4-decenoate, ethyl 9-hexadecenoic acid, diisobutyl phthalate, isobutyl 2,2, 4-trimethylpentanediol, isobutyl 2,2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, vinyl acetate.
Experimental example, mixed bacteria fermentation process and fermentation data of the invention
1. Bacterial strains
The strains used in this experiment were: commercial s.cerevisiae F33 and the isolated and screened grape juice of Hanseniaspora uvarum strain QTX22 of the present invention.
2. Grape juice
The Weidai ice grape raw material is planted in Yuquan Yingning county, Yinchuan city, Ningxia Bagges Zuius boundary wine village GmbH (E106.02 degree, N38.24 degree). Grapevine plantingIn 2013, small shed frames are adopted for cultivation, the row spacing of the plants is 1.0m multiplied by 2.0m, the test is carried out in 2017, the grapes are not picked after being ripe, the grapes are picked when the temperature is continuously reduced to below 24 hours to 8 ℃, small fruit grains are removed, and the ice grape fruits with the same size are randomly selected and pressed at low temperature. Pressing harvested ice grape with ice by air bag press while adding sulfur dioxide (50mg/L K)2S2O5) And 20mg/L of pectinase (more than or equal to 500U/mg), inhibits bacteria and improves the juice yield. The squeezed grape juice has sugar content of 432g/dm3Acidity of 4.65g/dm3(tartaric acid) pH 4.21.
3. Fermentation operation
2 strains: saccharomyces cerevisiae F33 strain, Hanseniaspora uvarum strain QTX22, Hanseniaspora uvarum, were all stored in 25% by volume glycerol/YPD medium prior to use. YPD medium is 1% yeast extract powder, 2% peptone and 2% glucose. Respectively inoculating the strain with 3% of volume ratio into a 50mL triangular flask containing 40mLYPD culture medium and a 250mL triangular flask containing 150mLYPD culture medium, culturing at 28 deg.C and 150rpm for 24h to obtain the 1 st activated bacterial liquid, respectively inoculating the strain with 3% of volume ratio into a 50mL triangular flask containing 40mLYPD culture medium and a 250mL triangular flask containing 150mLYPD culture medium, and repeating the above culture to complete the 2 nd passage activation, thus obtaining the activated bacterial liquid. Inoculating activated Hanseniaspora uvarum strain QTX22 into collected grape juice, inoculating into S.cerevisiae F33 strain at an inoculation ratio of QTX22/F33 of 2:1 after 48 hours, wherein the total inoculation amount of the strain is controlled at 6 × 106CFU, using S.cerevisiae F33 pure fermented grape juice as blank control, fermenting at 18 + -2 deg.C, and stopping fermentation when grape juice weight loss is not changed for three consecutive days. All wine samples were centrifuged at 7500rpm for 8 minutes and the supernatant was stored at 4 ℃.
4. Method for quantifying aroma substance
Headspace-solid phase microextraction-gas mass spectrometry (HS-SPME-GC/MS) was used. An 8mL sample of wine was accurately weighed into a headspace bottle containing 1.5g NaCl, while 394.08. mu.g/L4-methyl-1-pentanol (internal standard) was capped and sealed. Inserting CAR/DVB/PDMS extraction fiber, adsorbing at 45 deg.C for 30min, desorbing at 250 deg.C for 3 min, and performing GC-MS analysis. A chromatographic column: InertCap WAX polar chromatography column (60m × 0.25mm, 0.25 μm); the temperature rising procedure is as follows: maintaining at 40 deg.C for 5min, heating to 120 deg.C at 3 deg.C/min, heating to 230 deg.C at 8 deg.C/min, and maintaining for 10 min; the flow rate of the carrier gas (He) was 0.8mL/min, and was not split. Electron bombardment ion source; electron energy 70 eV; the transmission line temperature was 275 ℃; the ion source temperature is 230 ℃; the activation voltage is 1.5V; the filament flow is 0.25 mA; the mass scanning range m/z is 33-450. Compound quantitative analysis was performed using external standard quantitation method.
5. Data analysis method
All samples were averaged in 3 replicates respectively. Single-factor analysis of variance (ANOVA) and Duncan's multi-range test (P <0.05) were performed using SPSS 22.0for Windows (SPSS inc., Chicago, IL, US); unscamblebler 9.7(CAMOASA, Norway) was analyzed by partial least squares.
The final statistical interpretation gives the following Table 1, where the data are in μ g/L, meaning: the content of the aroma substances in each liter of wine.
TABLE 1
The aroma threshold value in the above table 1 refers to the lower limit value of the lowest concentration at which a human can smell the substance, and the aroma description and the threshold value are subject to the relevant literature reports that the aroma description and the threshold value of the aroma substance are recorded, and the aroma substance without the aroma description and the threshold value in the table is the literature that the relevant literature about the aroma description and the threshold value of the substance is not searched. In the table, "F33" refers to data obtained by single fermentation using a commercial strain, Saccharomyces cerevisiae F33, and "F33 _ QTX 22" refers to data obtained by mixed fermentation using a commercial strain, Saccharomyces cerevisiae F33 and Hanseniaspora uvarum strain QTX22, a Hanseniaspora uvarum.
As is well known in the field of fermentation, factors influencing fermentation are numerous and complex, and the components of a fermented product can change due to the change of factors such as raw material batches, raw material components, fermentation conditions, temperature, time and the like, so that the condition that single-strain fermentation is higher than mixed-strain fermentation on certain aroma components can occur, which is a normal phenomenon in the field.
Claims (10)
1. A mixed fermentation process based on Hansenula polymorpha and saccharomyces cerevisiae is characterized in that the Hansenula polymorpha and saccharomyces cerevisiae are subjected to mixed fermentation;
the Hansenula polymorpha is Hanseniaspora uvarum strain QTX 22; the collection number of the Hanseniaspora uvarum strain QTX22 of the grape juice is CCTCC M2021083.
2. The mixed fermentation process based on Hansenula polymorpha and Saccharomyces cerevisiae of claim 1, wherein the Saccharomyces cerevisiae is Saccharomyces cerevisiae strain F33.
3. The mixed fermentation process based on Hansenula polymorpha and Saccharomyces cerevisiae, as claimed in claim 1, comprises the following steps: the activated strain and the activated strain are inoculated into a substrate for secondary fermentation for fermentation.
Preferably, said strain refers to: hanseniaspora uvarum strain QTX22, Hanseniaspora uvarum, or Saccharomyces cerevisiae strain F33;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
preferably, the culture temperature is 24-30 ℃, preferably 28 ℃; the culture time is 20-30h, preferably 24h, the rotation speed is 120-250rpm, preferably 150 rpm;
preferably, the inoculation refers to inoculating the initial strain stock solution into the culture medium according to the inoculation amount of 2-4%, preferably 3% in volume ratio;
preferably, the medium is a YPD medium; preferably, the YPD medium comprises the following components in mass-to-volume ratio: 0.5-3.5%, preferably 1%, yeast extract, 1.0-3.0%, preferably 2%, peptone, 1.0-5.0%, preferably 2%, glucose, and water in balance;
preferably, the initial strain stock solution refers to the strain stock in 15-35% by volume, preferably 25% glycerol/YPD medium;
more preferably, the activation of the strain is performed 1-3 times, preferably 2 times.
4. The mixed fermentation process based on Hansenula polymorpha and Saccharomyces cerevisiae of grape juice as claimed in claim 3, wherein the activated strains refer to: the activated grape juice contains Hanseniaspora uvarum Hanseniaspora strain QTX22, or the activated saccharomyces cerevisiae strain F33;
preferably, the fermentation of the activated substrate into the secondary fermentation refers to: firstly, inoculating an activated Hanseniaspora uvarum strain QTX22 of grape juice to a substrate for fermentation for 0-96h, preferably 48h, and then inoculating an activated saccharomyces cerevisiae strain F33 for continuous fermentation;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the activated grape juice has Hanseniaspora uvarum strain QTX22, and the total inoculum size of the activated Saccharomyces cerevisiae strain F33 is 106-107CFU, preferably 6X 106CFU；
Preferably, the fermentation is terminated when the substrate weight loss does not change for 3 consecutive days;
preferably, the substrate is grape juice.
5. A production method of wine is characterized in that grape juice is used as a substrate, and hansenula polymorpha and saccharomyces cerevisiae which are used as grape juice are subjected to mixed fermentation;
the Hansenula polymorpha is Hanseniaspora uvarum strain QTX 22; the collection number of the Hanseniaspora uvarum strain QTX22 of the grape juice is CCTCC M2021083.
6. A wine production method according to claim 5, characterised in that the Saccharomyces cerevisiae is Saccharomyces cerevisiae strain F33.
7. A wine production process according to claim 5 or 6, comprising: the activated strain and the activated strain are inoculated into a substrate for secondary fermentation for fermentation.
Preferably, said strain refers to: hanseniaspora uvarum strain QTX22, Hanseniaspora uvarum, or Saccharomyces cerevisiae strain F33;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
preferably, the culture temperature is 24-30 ℃, preferably 28 ℃; the culture time is 20-30h, preferably 24h, the rotation speed is 120-250rpm, preferably 150 rpm;
preferably, the inoculation refers to inoculating the initial strain preservation solution into the culture medium according to the inoculation amount of 3% by volume;
preferably, the medium is a YPD medium; preferably, the YPD medium comprises the following components in mass-to-volume ratio: 0.5-3.5%, preferably 1%, yeast extract, 1.0-3.0%, preferably 2%, peptone, 1.0-5.0%, preferably 2%, glucose, and water in balance;
preferably, the initial strain stock solution refers to the strain stock in 15-35% by volume, preferably 25% glycerol/YPD medium;
more preferably, the activation of the strain is performed 1-3 times, preferably 2 times.
8. A wine production process according to claim 7, wherein the activated strain is: the activated grape juice contains Hanseniaspora uvarum Hanseniaspora strain QTX22, or the activated saccharomyces cerevisiae strain F33;
preferably, the fermentation of the activated substrate into the secondary fermentation refers to: firstly, inoculating an activated Hanseniaspora uvarum strain QTX22 of grape juice to a substrate for fermentation for 0-96h, preferably 48h, and then inoculating an activated saccharomyces cerevisiae strain F33 for continuous fermentation;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the activated grape juice has Hanseniaspora uvarum strain QTX22 of Hanseniaspora, and the total inoculated amount of the activated Saccharomyces cerevisiae strain F33 is 106-107CFU, preferably 6X 106CFU；
Preferably, the fermentation is terminated when the substrate weight loss is not changed for 3 consecutive days.
9. A wine production process according to any one of claims 5 to 8, further comprising: preparing grape juice;
the preparation of the grape juice comprises the following steps: squeezing grape fruit grains at low temperature;
preferably, the harvested grape fruit particles are pressed with ice by an air bag press;
preferably, pressing is performed while adding sulfur dioxide or K2S2O5And, pectinase;
preferably, sulfur dioxide or K2S2O5The addition amount of (B) is 30-100mg/L, preferably 50 mg/L; the addition amount of pectinase is 10-30mg/L, preferably 20 mg/L;
preferably, the grape fruit pieces are uniformly sized fruit pieces;
preferably, the grape fruit particles are grape fruit particles collected when the temperature of the mature grapes continuously drops below 24 hours to 8 ℃.
10. Wine produced by the wine production method of any one of claims 5 to 9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110189028.4A CN112812980B (en) | 2021-02-19 | 2021-02-19 | Mixed fermentation process based on Hansenula polymorpha and Saccharomyces cerevisiae |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110189028.4A CN112812980B (en) | 2021-02-19 | 2021-02-19 | Mixed fermentation process based on Hansenula polymorpha and Saccharomyces cerevisiae |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112812980A true CN112812980A (en) | 2021-05-18 |
| CN112812980B CN112812980B (en) | 2023-08-01 |
Family
ID=75865604
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110189028.4A Active CN112812980B (en) | 2021-02-19 | 2021-02-19 | Mixed fermentation process based on Hansenula polymorpha and Saccharomyces cerevisiae |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112812980B (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112940885A (en) * | 2021-03-26 | 2021-06-11 | 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) | Combined fermentation process based on Klotch yeast and Hansenula polymorpha |
| CN113331412A (en) * | 2021-07-14 | 2021-09-03 | 山西农业大学山西功能食品研究院 | Method for fermenting enzyme of jujubes |
| CN113416658A (en) * | 2021-05-28 | 2021-09-21 | 湖南省农产品加工研究所 | Hansenula sordida and application thereof in yellow peach fruit wine |
| CN113773975A (en) * | 2021-10-26 | 2021-12-10 | 中国农业大学 | Two aroma-producing yeasts and application thereof in dry white wine brewing process |
| CN113897297A (en) * | 2021-09-30 | 2022-01-07 | 甘肃农业大学 | Beta-glucosidase yeast with high enzyme activity under low pH condition and application thereof |
| CN113956988A (en) * | 2021-10-26 | 2022-01-21 | 中国农业大学 | Two yeasts for promoting production of lactic acid and phenolic acid and application thereof in dry white wine brewing process |
| CN114032185A (en) * | 2021-08-24 | 2022-02-11 | 甘肃农业大学 | Hansenula polymorpha culture medium and culture method for grape juice |
| CN114451450A (en) * | 2022-02-24 | 2022-05-10 | 兰州大学 | Hansenula polymorpha capable of antagonizing botrytis cinerea and application thereof |
| CN114854609A (en) * | 2022-05-13 | 2022-08-05 | 江南大学 | Hansenula polymorpha capable of increasing fermentation aroma of red date lactobacillus beverage and application of Hansenula polymorpha |
| CN117165459A (en) * | 2023-10-31 | 2023-12-05 | 华熙生物科技股份有限公司 | Hansenula polymorpha of grape juice and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN110951631A (en) * | 2019-12-31 | 2020-04-03 | 江南大学 | Hansenula polymorpha capable of producing geraniol and fermentation method thereof |
-
2021
- 2021-02-19 CN CN202110189028.4A patent/CN112812980B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN110951631A (en) * | 2019-12-31 | 2020-04-03 | 江南大学 | Hansenula polymorpha capable of producing geraniol and fermentation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 卑诺: "《葡萄酒科学与工艺》", 28 February 1992 * |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112940885A (en) * | 2021-03-26 | 2021-06-11 | 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) | Combined fermentation process based on Klotch yeast and Hansenula polymorpha |
| CN113416658B (en) * | 2021-05-28 | 2022-04-12 | 湖南省农产品加工研究所 | Hansenula sordida and application thereof in yellow peach fruit wine |
| CN113416658A (en) * | 2021-05-28 | 2021-09-21 | 湖南省农产品加工研究所 | Hansenula sordida and application thereof in yellow peach fruit wine |
| CN113331412A (en) * | 2021-07-14 | 2021-09-03 | 山西农业大学山西功能食品研究院 | Method for fermenting enzyme of jujubes |
| CN114032185A (en) * | 2021-08-24 | 2022-02-11 | 甘肃农业大学 | Hansenula polymorpha culture medium and culture method for grape juice |
| CN113897297A (en) * | 2021-09-30 | 2022-01-07 | 甘肃农业大学 | Beta-glucosidase yeast with high enzyme activity under low pH condition and application thereof |
| CN113897297B (en) * | 2021-09-30 | 2023-08-08 | 甘肃农业大学 | Beta-glucosidase yeast with high enzyme activity under low pH condition and application thereof |
| CN113773975A (en) * | 2021-10-26 | 2021-12-10 | 中国农业大学 | Two aroma-producing yeasts and application thereof in dry white wine brewing process |
| CN113956988A (en) * | 2021-10-26 | 2022-01-21 | 中国农业大学 | Two yeasts for promoting production of lactic acid and phenolic acid and application thereof in dry white wine brewing process |
| CN114451450A (en) * | 2022-02-24 | 2022-05-10 | 兰州大学 | Hansenula polymorpha capable of antagonizing botrytis cinerea and application thereof |
| CN114854609A (en) * | 2022-05-13 | 2022-08-05 | 江南大学 | Hansenula polymorpha capable of increasing fermentation aroma of red date lactobacillus beverage and application of Hansenula polymorpha |
| CN114854609B (en) * | 2022-05-13 | 2023-08-25 | 江南大学 | Hansenula polymorpha serving as grape juice capable of increasing fermentation aroma of red date lactobacillus beverage and application of Hansenula polymorpha |
| CN117165459A (en) * | 2023-10-31 | 2023-12-05 | 华熙生物科技股份有限公司 | Hansenula polymorpha of grape juice and application thereof |
| CN117165459B (en) * | 2023-10-31 | 2024-02-23 | 华熙生物科技股份有限公司 | Hansenula polymorpha of grape juice and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112812980B (en) | 2023-08-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112812980B (en) | Mixed fermentation process based on Hansenula polymorpha and Saccharomyces cerevisiae | |
| CN108239608B (en) | Torulaspora delbrueckii and application thereof in wine brewing | |
| CN109097291B (en) | Composite fermentation agent and application thereof in brewing cabernet sauvignon anhydration wine | |
| CN112746029B (en) | Hansenula polymorpha strain QTX22 for producing aroma substances in grape juice and application of strain | |
| CN109182156B (en) | Saccharomyces cerevisiae suitable for brewing red-core pitaya wine and application thereof | |
| CN108179119B (en) | Fermentation process for improving aroma quality of ice wine by using non-saccharomyces cerevisiae | |
| CN112795448B (en) | Mixed bacteria fermentation process based on lactobacillus plantarum and saccharomyces cerevisiae | |
| CN112812914A (en) | Mixed fermentation process based on pichia kluyveri and saccharomyces cerevisiae | |
| Ezeronye | Nutrient utilization profile of Saccharomyces cerevisiae from palm wine in tropical fruit fermentation | |
| CN112812915B (en) | Mixed fermentation process based on candida lycopi and saccharomyces cerevisiae | |
| CN112694981B (en) | Candida zemplina strain YC32 of Candida zemplina and application thereof | |
| CN113150897B (en) | Mixed fermentation process based on Mei Ji Yeast and Saccharomyces cerevisiae | |
| CN112725107B (en) | Mixed fermentation process based on Klatong yeast and Saccharomyces cerevisiae | |
| CN113773975A (en) | Two aroma-producing yeasts and application thereof in dry white wine brewing process | |
| CN113150898B (en) | Mixed fermentation process based on yeast Naganishia albida and saccharomyces cerevisiae | |
| CN109913379B (en) | High-esterase-activity Meiji yeast and application thereof in fermentation of mead | |
| CN112980705B (en) | Combined fermentation process based on klatomyces, hansenula polymorpha and saccharomyces cerevisiae | |
| CN112940884B (en) | Mixed bacteria fermentation process based on Curvibasidium pallidicorallinum | |
| CN112852569B (en) | Mixed fermentation process based on rhodosporidium rhodochrous and saccharomyces cerevisiae | |
| CN113150899B (en) | Mixed fermentation process based on yeast Papiliotrema laurentii and saccharomyces cerevisiae | |
| CN113150896B (en) | Mixed fermentation process based on cryptococcus gracilis and saccharomyces cerevisiae | |
| CN113150902B (en) | Fermentation process for adding amino acid into substrate and based on Hansenula polymorpha of grape juice | |
| CN113061540B (en) | Fermentation process based on klaton yeast with amino acid added into substrate | |
| CN112940885A (en) | Combined fermentation process based on Klotch yeast and Hansenula polymorpha | |
| CN112625924B (en) | Cryptococcus albidus Naganishia albica strain YC22 for high yield of aroma substances and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |