CN112940885A - Combined fermentation process based on Klotch yeast and Hansenula polymorpha - Google Patents
Combined fermentation process based on Klotch yeast and Hansenula polymorpha Download PDFInfo
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Abstract
The invention discloses a combined fermentation process based on Klotch yeast and Hansenula polymorpha with grape juice, belonging to the technical field of microorganisms. The combined fermentation process based on the Kluyveromyces and the Hansenula polymorpha is characterized in that the Kluyveromyces and the Hansenula polymorpha are fermented. The fermentation product obtained by combining and fermenting the klaton yeast and the grape juice Hansenula polymorpha has more unique fragrance compared with a single-bacterium fermentation product of a substrate without amino acid, 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 combined fermentation process based on Klotch yeast and Hansenula polymorpha.
Background
Wine fermentation is a complex biochemical process, and klaton yeast plays a very critical role in the fermentation process, such as the conversion of sugars to ethanol, carbon dioxide and other secondary metabolites in the thousands. A large number of scientific studies show that the quality of the wine is highly dependent on the metabolic activity and fermentation behavior of different cladonone yeasts, and the different cladonone yeasts have important contributions to the chemical composition, the organoleptic properties, 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 make the aroma characteristics more representative, diverse and complex, brewers often adopt a method of mixing and fermenting saccharomyces cerevisiae and non-saccharomyces cerevisiae, especially some native klaton yeasts 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 related reports of fermentation by the yeast Saccharomyces cerevisiae are extremely rare at present in the field, and the combined fermentation process of the yeast Saccharomyces cerevisiae and Hanseniaspora uvarum which is a grape juice in a substrate is less frequently reported in the field.
Disclosure of Invention
Based on the above needs and blank in the art, the invention provides a combined fermentation process based on the Kluyveromyces cerevisiae and the Hanseniaspora uvarum, which has a very significant improvement on various aroma substances affecting the flavor of wine compared with the conventional fermentation of Saccharomyces cerevisiae.
The technical scheme of the invention is as follows:
a combined fermentation process based on Klotch yeast and Hansenula polymorpha with grape juice is characterized in that the Klotch yeast and the Hansenula polymorpha with grape juice are adopted for fermentation.
The yeast is Saccharomyces cerevisiae strain YC 30; the collection number of the cladonospora clavata Saccharomyces copsis crataegensis strain YC30 is CCTCC M2021089.
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 fermentation process of adding amino acid to the substrate and based on the clarithromycin comprises the following steps: the activated strain and the activated strain are inoculated to a substrate to be fermented for fermentation;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture.
The strain is selected from: saccharomyces cerevisiae strain YC30, Hanseniaspora uvarum strain QTX22, Saccharomyces cerevisiae strain F33;
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%, of a yeast extract powder of klaton, 1.0-3.0%, preferably 2%, of peptone, 1.0-5.0%, preferably 2%, of glucose, and the balance water;
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.
Preferably, the activated strain refers to: the activated yeast strain YC30 of the yeast Saccharomyces copsis crataegensis, or the activated strain QTX22 of Hanseniaspora uvarum, or the activated strain F33 of the yeast Saccharomyces cerevisiae;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the amount of inoculation of the activated strain in the substrate 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 yeast Clarithrombus and Hansenula polymorpha for grape juice fermentation are inoculated.
The yeast is Saccharomyces cerevisiae strain YC 30; the preservation number of the cladonospora clavata Saccharomyces copsis crataegensis strain YC30 is CCTCC M2021089;
preferably, the Hansenula polymorpha is Hanseniaspora uvarum strain QTX 22; the preservation number of Hanseniaspora uvarum strain QTX22 of the grape juice is CCTCC M2021083;
preferably, said one wine production method comprises: the activated strain and the activated strain are inoculated to a substrate to be fermented for fermentation.
The strain is selected from: saccharomyces cerevisiae strain YC30, Hanseniaspora uvarum strain QTX22, 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%, of a yeast extract powder of klaton, 1.0-3.0%, preferably 2%, of peptone, 1.0-5.0%, preferably 2%, of glucose, and the balance water;
preferably, the initial strain stock solution refers to a strain that is stored 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;
preferably, the activated strain refers to: the activated yeast strain YC30 of the yeast Saccharomyces copsis crataegensis, or the activated strain QTX22 of Hanseniaspora uvarum, or the activated strain F33 of the yeast Saccharomyces cerevisiae;
preferably, the activated clarithrobacter clarkii strain YC30 and the activated grape juice Hanseniaspora uvarum strain QTX22 are simultaneously inoculated with a substrate to be fermented for fermentation;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the activated yeast Saccharomyces cerevisiae strain YC30, and the activated grape juice Hanseniaspora uvarum strain QTX22 total access amount is 106-107CFU, preferably 6X 106 CFU；
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.
According to the invention, the combination fermentation is carried out by simultaneously inoculating the Saccharomyces cerevisiae and Hanseniaspora uvarum which are Hanseniaspora uvarum in grape juice in a substrate, and compared with the single use of commercial Saccharomyces cerevisiae fermentation, the yield of a plurality of fragrant substances in the generated dozens of fragrant substances is improved, for example, 2-ethylhexanol is improved by 83%, 1-pentanol is improved by 11 times, ethyl lactate is improved by 1.25 times, acetaldehyde is improved by 82%, diethyl succinate is improved by 6.3 times, 2-methylbutyric acid is improved by 14.9 times, hexanol is improved by 97%, hexanoic acid is improved by 30 times, and 1, 3-propylene glycol monoethyl ether is improved by 2.3 times; ethyl myristate was increased by 1.5 times, phenethylacetate by 3.56 times, ethyl palmitate by 28%, 4-vinyl-2-methoxyphenol by 71%, citronellol by 2.19 times, ethyl hexanoate by 84%, octanoic acid by 9.9 times, and glacial acetic acid by 1.03 times; the combined fermentation process of the present invention also produces aroma that cannot be produced by fermentation using commercial saccharomyces cerevisiae alone, such as: ethyl 3-phenylpropionate, trans-3-hexen-1-ol, isovaleraldehyde, 1-decanol, (2S-cis) -tetrahydro-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, 3-hydroxy-2-butanone, 2, 3-butanedione, butyric acid, 3-methyl-1-pentanol, isoamyloctanoate, 3-methyl-3-buten-1-ol, ethyl isobutyrate. 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 strain YC30 of the yeast Saccharomyces capsisacratae used in the experimental examples is a new strain screened by the applicant's laboratory and has the following deposit information:
naming: YC30
And (4) classification name: clarithromycin yeast
The name of Latin is: saccharomyces copesis crataegensis
The preservation number is as follows: CCTCC NO: M2021089
The preservation organization: china center for type culture Collection; the address of the depository: wuhan university of Wuhan, China
The preservation date is as follows: 1 month 15 days 2021;
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; the address of the depository: wuhan university of Wuhan, China
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, fermentation Process for substrate addition of amino acids according to the invention
The present group of embodiments provides a combined fermentation process based on yeast klaton and hansenula polymorpha. All embodiments of this group share the following common features: fermentation was performed using Klotch yeast and Hansenula polymorpha.
Those skilled in the art can appreciate that any fermentation and production activities using Klotch yeast and Hansenula polymorpha will fall within the scope of the present invention, in accordance with the teachings of the present invention. Target products of fermentation include, but are not limited to: wine, dairy products, flour products, etc.
Including but not limited to yogurt, fermented milk, milk drinks, and the like; such dough products include, but are not limited to: bread, cakes, steamed stuffed buns, steamed bread, steamed rolls and the like.
In a preferred embodiment, the yeast species kluyveromyces is kluyveromyces clavuligerus strain YC 30; the collection number of the cladonospora clavata Saccharomyces copsis crataegensis strain YC30 is CCTCC M2021089.
In other embodiments, 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 further embodiment, the fermentation process of a substrate supplemented with amino acids and based on klaton yeast comprises the following steps: the activated strain and the activated strain are inoculated to a substrate to be fermented for fermentation;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
in a preferred embodiment, the strain is selected from the group consisting of: saccharomyces cerevisiae strain YC30, Hanseniaspora uvarum strain QTX22, Saccharomyces cerevisiae strain F33;
in addition to the demonstration of the combined fermentation effect of the Saccharomyces cerevisiae strain and the Hanseniaspora uvarum strain of Saccharomyces cerevisiae and the single-strain fermentation effect of the commercial Saccharomyces cerevisiae strain, compared with the single-strain fermentation of the commercial Saccharomyces cerevisiae strain, the combined fermentation of the Saccharomyces cerevisiae strain and the Hanseniaspora uvarum strain of Saccharomyces cerevisiae of the invention can achieve higher yield of aroma substances, so that the skilled person can select other commercial strains to combine with the Saccharomyces cerevisiae strain and the Hanseniaspora uvarum strain of Saccharomyces cerevisiae, and can also select various commercial strains on the market, such as Saccharomyces cerevisiae V, Saccharomyces cerevisiae, VL1 strains to inoculate amino acids in fermentation medium, such as VL 52 and VL 52, it is contemplated that technical effects similar to those of the present invention may be achieved.
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%, of a yeast extract powder of klaton, 1.0-3.0%, preferably 2%, of peptone, 1.0-5.0%, preferably 2%, of glucose, and the balance water;
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.
Preferably, the activated strain refers to: the activated yeast strain YC30 of the yeast Saccharomyces copsis crataegensis, or the activated strain QTX22 of Hanseniaspora uvarum, or the activated strain F33 of the yeast Saccharomyces cerevisiae;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the amount of inoculation of the activated strain in the substrate 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.
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: inoculating yeast Clarithromycin into grape juice as substrate, and fermenting.
In a preferred embodiment, the yeast species kluyveromyces is kluyveromyces clavuligerus strain YC 30; the collection number of the cladonospora clavata Saccharomyces copsis crataegensis strain YC30 is CCTCC M2021089.
In other embodiments, 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 other embodiments, the method of wine production comprises: adding amino acid, activated strain into grape juice, inoculating the activated strain into a substrate for secondary fermentation, and fermenting;
in some embodiments, the strain is selected from the group consisting of: saccharomyces cerevisiae strain YC30, Hanseniaspora uvarum strain QTX22, 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%, of a yeast extract powder of klaton, 1.0-3.0%, preferably 2%, of peptone, 1.0-5.0%, preferably 2%, of glucose, and the balance water;
preferably, the initial strain stock solution refers to a strain that is stored 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;
preferably, the activated strain refers to: the activated yeast strain of the Clarithromy coprisscrataegensis YC30, or the activated yeast strain of Saccharomyces cerevisiae F33, or the activated strain of Hanseniaspora uvarum QTX 22;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the total inoculum size of activated strain YC30 of the yeast Saccharomyces cerevisiae and strain QTX22 of Hanseniaspora uvarum, after activation, is 106-107CFU, preferably 6X 106 CFU；
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, 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 ℃. 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: ethyl 3-phenylpropionate, trans-3-hexen-1-ol, isovaleraldehyde, 1-decanol, (2S-cis) -tetrahydro-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, 3-hydroxy-2-butanone, 2, 3-butanedione, butyric acid, 3-methyl-1-pentanol, isoamyloctanoate, 3-methyl-3-buten-1-ol, ethyl isobutyrate.
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, the strain Saccharomyces cerevisiae YC30 isolated and selected according to the invention, and Hanseniaspora uvarum strain QTX22, Hanseniaspora uvarum.
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). The grape vines are planted in 2013, small-canopy-frame cultivation is adopted, the plant row spacing is 1.0m multiplied by 2.0m, the grape vines are harvested in 2017, the grape vines are not harvested after being mature, the grape vines are harvested when the temperature is reduced to be below 24 hours-8 ℃, small fruit grains are removed, and ice grape fruits with the same size are randomly selected and squeezed at low temperature. To pairThe harvested ice grape is pressed with ice by air bag press, and sulfur dioxide (50mg/L K) is added2S2O5) 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
3 strains: saccharomyces cerevisiae F33 strain, Saccharomyces cerevisiae strain YC30 strain, and Hanseniaspora uvarum strain QTX22 strain of Hanseniaspora uvarum, Saccharomyces cerevisiae, all were stored in 25% by volume glycerol/YPD medium prior to use. YPD medium was 1% of yeast extract powder of Kluyveromyces crassipes, 2% of peptone and 2% of glucose. Respectively inoculating the strain with 3% of volume ratio into a 50mL triangular flask containing 40mL YPD medium and a 250mL triangular flask containing 150mL YPD 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 40mL YPD medium and a 250mL triangular flask containing 150mL YPD medium, and repeating the above culture to complete the 2 nd passage activation, thus obtaining the activated bacterial liquid. Inoculating activated Saccharomyces cerevisiae strain YC30 and activated Hanseniaspora uvarum strain QTX22 into collected grape juice, wherein the total inoculation amount of the strains 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 ℃.
The yeast Saccharomyces cerevisiae strain YC30 and the Hanseniaspora uvarum strain QTX22 of Hanseniaspora uvarum, grape juice are fermented in a combined manner for 3 times of parallel fermentation treatment, and S. the single strain F33 of the yeast is fermented for 3 times of parallel fermentation treatment.
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
Measuring the content of the aroma substances by taking the fermentation products obtained by each parallel fermentation treatment in the part 3 as different samples, and measuring each sample in parallel for 3 times respectively to obtain an average value. 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(CAMO ASA, 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 aroma content of grape juice fermentation product
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. The meanings of the respective symbols in the header are listed below:
sc + Hu: the average value of the aroma substance content of the fermentation products obtained by 3 times of parallel fermentation treatments of combined fermentation of a Kluyveromyces saccharolytica capsicacratae gene strain YC30 and a Hanseniaspora uvarum strain QTX22 of Hanseniaspora uvarum, which are used as the fermentation liquid, is respectively determined;
f33: and S, respectively measuring the average value of the content of the aroma substances of fermentation products obtained by performing 3 times of parallel fermentation treatment on the single-strain fermentation of the cerevisiae F33.
It is known in the field of fermentation that many and complex factors affect fermentation, and the fermented product components 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-bacterium fermentation is higher than amino acid addition fermentation on certain aroma components can also occur, which is a normal phenomenon in the field.
Claims (10)
1. A combined fermentation process based on Klotch yeast and Hansenula polymorpha with grape juice is characterized in that the Klotch yeast and the Hansenula polymorpha with grape juice are adopted for fermentation.
2. The combined fermentation process based on yeast Hansenula polymorpha with Alternaria uvarum, according to claim 2, wherein the yeast is Saccharomyces cerevisiae strain YC 30; the collection number of the cladonospora clavata Saccharomyces copsis crataegensis strain YC30 is CCTCC M2021089.
3. The combined fermentation process based on Hansenula crassa and Hansenula polymorpha, as claimed in claim 1, wherein the Hansenula polymorpha refers to Hanseniaspora uvarum strain QTX 22; the grape juice has Hanseniaspora uvarum Hanseniaspora strain QTX22 with the preservation number of CCTCC M2021083.
4. A substrate amino acid addition and klaton yeast based fermentation process according to any one of claims 1-3, comprising the steps of: the activated strain and the activated strain are inoculated to a substrate to be fermented for fermentation;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture.
5. A substrate amino acid addition and Clarithromycin based fermentation process as claimed in claim 4, wherein the strain is selected from the group consisting of: saccharomyces cerevisiae strain YC30, Hanseniaspora uvarum strain QTX22, Saccharomyces cerevisiae strain F33;
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%, of a yeast extract powder of klaton, 1.0-3.0%, preferably 2%, of peptone, 1.0-5.0%, preferably 2%, of glucose, and the balance water;
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;
preferably, the activated strain refers to: the activated yeast strain YC30 of the yeast Saccharomyces copsis crataegensis, or the activated strain QTX22 of Hanseniaspora uvarum, or the activated strain F33 of the yeast Saccharomyces cerevisiae;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferablyThe amount of the activated strain inoculated into the substrate 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.
6. A production method of wine is characterized in that grape juice is used as a substrate, and yeast Clarithrombus and Hansenula polymorpha for grape juice fermentation are inoculated.
7. A wine production process according to claim 6 wherein the yeast species Clarithromium species is Saccharomyces cerevisiae strain YC 30; the preservation number of the cladonospora clavata Saccharomyces copsis crataegensis strain YC30 is CCTCC M2021089;
preferably, the Hansenula polymorpha is Hanseniaspora uvarum strain QTX 22; the preservation number of the Hanseniaspora uvarum strain QTX22 of the grape juice is CCTCC M2021083;
preferably, said one wine production method comprises: the activated strain and the activated strain are inoculated to a substrate to be fermented for fermentation.
8. A wine production process according to claim 7, wherein said strain is selected from the group consisting of: saccharomyces cerevisiae strain YC30, Hanseniaspora uvarum strain QTX22, 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%, of a yeast extract powder of klaton, 1.0-3.0%, preferably 2%, of peptone, 1.0-5.0%, preferably 2%, of glucose, and the balance water;
preferably, the initial strain stock solution refers to a strain that is stored 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;
preferably, the activated strain refers to: the activated yeast strain YC30 of the yeast Saccharomyces copsis crataegensis, or the activated strain QTX22 of Hanseniaspora uvarum, or the activated strain F33 of the yeast Saccharomyces cerevisiae;
preferably, the activated clarithrobacter clarkii strain YC30 and the activated grape juice Hanseniaspora uvarum strain QTX22 are simultaneously inoculated with a substrate to be fermented for fermentation;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the total inoculum size of activated strain YC30 of the yeast Saccharomyces cerevisiae and strain QTX22 of Hanseniaspora uvarum, after activation, 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 6 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 6 to 9.
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| 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 |
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| 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 |
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