CN115558611B - High flocculation property schizosaccharomyces japonica strain and application thereof - Google Patents
High flocculation property schizosaccharomyces japonica strain and application thereof Download PDFInfo
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- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
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- 229940095064 tartrate Drugs 0.000 description 1
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- 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
- C12N1/165—Yeast isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G1/00—Preparation of wine or sparkling wine
- C12G1/02—Preparation of must from grapes; Must treatment and fermentation
- C12G1/0203—Preparation of must from grapes; Must treatment and fermentation by microbiological or enzymatic treatment
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
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- 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
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Abstract
The invention belongs to the technical field of brewing engineering, and particularly relates to a high flocculation property schizosaccharomyces japonica strain (schizosaccharomyces japonica) and application thereof, which is preserved in China center for type culture collection, and the preservation number is: CCTCCNO: m20221342, the preservation time is: the strain has excellent fermentation capacity and cell aggregation capacity, is used for brewing wine in a single plant mode, can enable flocculation and clarification of yeast cells to be carried out simultaneously, reduces the dosage of gumming materials, and improves the clarity and stability of the wine in a biological clarification mode after being used for 5 days 9 of 2022.
Description
Technical Field
The invention belongs to the technical field of brewing engineering, and particularly relates to a high flocculation property schizosaccharomyces japonica strain and application thereof.
Background
The basic requirements in the national standard sensory requirements of wine regarding clarity are clear, shiny, and free of significant suspended matter. The wine is used as a colloid solution, and after the alcoholic fermentation is finished, macromolecular colloid which is easy to settle is often separated out by carrying out clarifying treatment process operations such as colloid discharging, filtering and the like, so that the clarifying degree of the wine and the stability after bottling are improved. In recent years, related researches show that excessive physical and chemical clarification treatment can cause loss of flavor substances of the wine, so that moderate clarification treatment can keep the flavor of the wine and reduce the brewing cost of the wine, but at present, no related report for improving the clarity of the wine by a biological clarification mode exists.
Comprehensive factor analysis of wild Yeast commercial research (Wang Chunxiao et al, food and fermentation industries, 2022,48 (16): 284-290) states that flocculation of yeast occurs during fermentation, mainly due to cell aggregation by interaction of flocculants present on the surface of yeast cells with mannoproteins on the cell walls of neighboring yeasts. The yeast with flocculation property can adhere to and agglomerate and settle in the fermentation process of the wine, and too high flocculation property can occur too early, so that the fermentation performance of the yeast is reduced, the sugar consumption is incomplete, the fermentation time is prolonged and the like due to poor dispersibility; too low flocculation can lead to slow flocculation of yeast cells in the late stages of fermentation, which is detrimental to filtration and clarification of wine. Therefore, preferred yeasts with suitable flocculation properties are advantageous for the alcoholic fermentation of wine. At present, the existing wine brewing mostly uses saccharomycetes as main bacteria for alcoholic fermentation, and the saccharomycetes settle at the bottom of a fermentation tank due to flocculation after the alcoholic fermentation is completed, and are removed by subsequent operations such as tank transfer, filtration and the like. The saccharomycete generally selected in the brewing process is saccharomyces cerevisiae, the fermentation performance of the saccharomyces cerevisiae is excellent, the flocculation property is centered, the requirements on the clarification and stability treatment of the wine are generally high, and the use of excessive gumming materials in the clarification process often brings certain flavor loss while improving the clarity of the wine body.
Thus, considering the disadvantages of Saccharomyces cerevisiae, researchers in the field have attempted to achieve brewing of high quality wine by optimizing the type of yeasts; the brewing method of dry white wine is disclosed in the patent with the publication number of CN1128796A, which comprises the steps of sorting, squeezing, clarifying, fermenting, fully filling and ageing raw materials of white grape, separating, stabilizing, chen Jun filtering, bottling, packaging and warehousing, and is characterized in that active schizosaccharomyces is used for replacing wine yeast adopted in the traditional process in the fermentation process. Thus, the defect that the acidity is high and chemical deacidification is needed because only sugar in a fermentation substrate can be converted into alcohol but organic acid such as malic acid cannot be converted into alcohol when wine yeast is adopted for fermentation is overcome, and the fermentation process and the deacidification process are carried out simultaneously. The active schizosaccharomyces is added in the fermentation process, the percentage content (weight ratio) is 5-10%, and the fermentation is carried out under the condition that the temperature is 16-18 ℃. The pH value of the grape juice is regulated to 2.9-3.0 in the clarification process, which is beneficial to the activity of schizosaccharomyces. The improvement of the physical and chemical stability of the wine is mainly realized by reducing the malic acid content in the wine through biological metabolism, and the flocculation property and the influence on the clarity of active schizosaccharomyces are not mentioned.
As another example, patent publication No. CN108913410a discloses a method for preparing wine, comprising the steps of: raw material pretreatment: washing grape, air drying, pulverizing to obtain grape mash A, and pulverizing flos Lonicerae to obtain flos Lonicerae powder; (2) addition of a clarifying agent: adding complex enzyme into the grape mash A, and uniformly mixing to obtain grape mash B, wherein the addition amount of the complex enzyme is 20-30mg/L; (3) addition of bacteriostat: adding flos Lonicerae powder and SO into grape mash B 2 Mixing to obtain grape mash C, SO 2 The addition amount of the honeysuckle and the SO is 10-20mg/L 2 Can inhibit the generation of other miscellaneous bacteria in the preparation process; (4) fermentation: adding saccharomycetes into the grape mash C, uniformly mixing, adding the grape mash C into a fermentation cylinder, performing sealed fermentation for 3-5 days, performing ultrasonic treatment for 3-6min, filtering to obtain fermentation liquor, wherein the adding amount of the saccharomycetes is 5-10mg/L, and the ultrasonic treatment ensures that the components in the fermentation liquor are more uniformly mixed, so that the dissolution rate of pigment is improved; (5) separation: filtering the fermentation liquor by a microfiltration membrane, and sterilizing to obtain wine, wherein the microfiltration can effectively remove fine particles and bacteria in the fermentation liquor; and (6) filling and storing: bottling the wine, and storing in cellar with humidity of 60-70% at 8-15deg.C. The microzyme is beer microzyme and schizosaccharomyces, wherein the beer microzyme decomposes sugar to produce ethanol, and the schizosaccharomyces breaks down sour substances such as sugar, malic acid and the like to produce ethanol, higher alcohol and ester; in the technical scheme, the schizosaccharomyces is mainly used for decomposing sugar and malic acid, and flocculation property and clarification effect of the schizosaccharomyces are not evaluated.
In addition to clarity, wine stability is also criticalThe required quality index is that phenolic substances such as anthocyanin and tannin in the wine belong to colloid substances, and not only directly influence the color and the taste (structural sense) of the wine, but also influence the stability of the wine, so that higher control requirements are also provided for the post-fermentation gumming treatment. Relevant foreign research reports indicate that schizosaccharomyces can improve the color stability of wine through high-yield pyruvic acid or zymosan through mixed fermentation with other saccharomycetes (BENITOF, bento S.the combined use of Schizosaccharomyces pombe and Lachancea thermotolerans-effect on the anthocyanin wine composition. Molecules,2017,22 (5): 739), improving the stability of wine proteins and tartrate, retaining wine aroma (PORTRO L, MAIOLI F, CANUTI V, PICCHI M, LENCIONI L, MANAZZU I, DOMIZIO P.Schizosaccharomyces japonica/Saccharomyces cerevisiae mixed starter cultures: new perspectives for the improvement of Sangiovese aroma, paste, and color stability.LWT,2022, 156:113009). However, none of the yeast types for wine brewing disclosed in China has the characteristics of improving the clarity and the stability of the wine, and none of the yeast types for wine brewing disclosed in foreign countries has been reported on the aspect of improving the clarity of wine by applying high flocculation strains.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a high flocculation property schizosaccharomyces japonica strain and application thereof.
The method is realized by the following technical scheme:
the first object of the invention is: there is provided a high flocculation property schizosaccharomyces japonica strain classified and named as schizosaccharomyces japonica (schizosaccharomyces japonica), the laboratory strain deposit accession number is FBKL2.9SZJ-29, deposited in the China center for type culture Collection, deposit number is: cctccc NO: m20221342, the preservation time is: 2022, 9 and 5.
Flocculation value range of the high flocculation property schizosaccharomyces japonica strain: 0.38 to 23.89 percent.
Preferably, the flocculation value of the high flocculation property schizosaccharomyces japonica strain ranges: 10.29% -10.45%.
The identification characteristics of the high flocculation property schizosaccharomyces japonica strain are as follows: growing on a WL culture medium, wherein the colony is a light green spherical bulge with a tray-shaped bottom, and the cells are elliptical or round rod-shaped; 5.8S-ITS-RFLP analysis of 700bp PCR product and 550+150bp HaeIII cleavage product; 26S D1/D2 region sequence analysis to obtain 586bp effective sequence, the GenBank serial number of the strain is OP364841; the genotype of the seven microsatellite loci is Glb-T2a-G3a-Saa-Syc-11b-12b, specifically, the microsatellite locus GA1 amplification product is 190bp, the microsatellite locus TG2 amplification product is 280bp, the microsatellite locus CG3 amplification product is 280bp, the microsatellite locus SaGAA1 amplification product is 110bp, the microsatellite locus SyGAA2 amplification product is 110+80bp, the microsatellite locus C11 amplification product is 170bp and the microsatellite locus C12 amplification product is 190bp.
The high flocculation property schizosaccharomyces japonica strain is a dominant saccharomycete for separating and screening during the middle and later periods of natural fermentation of the crystal grapes, and coexists with saccharomyces cerevisiae, so that excellent competitiveness and tolerance are proved.
The second object of the invention is: provides an application of a high flocculation property schizosaccharomyces japonica strain in wine brewing.
In particular, the use of the high flocculation property schizosaccharomyces japonica strain in the alcoholic fermentation process to improve the stability and clarity of wine in a way that flocculation and clarification of yeast cells are carried out simultaneously.
In particular to the application of the high flocculation schizosaccharomyces japonica strain in improving the clarity and stability of wine bodies during the fermentation and gumming of the wine.
The method for obtaining the high flocculation schizosaccharomyces japonica strain comprises the following steps: mature crystal grape grains are obtained, crushed and placed in a sterile fermentation bottle, closed fermentation is carried out at 28 ℃, the fermentation progress is judged by monitoring the carbon dioxide release amount, and sampling is carried out at the later stage of fermentation, and the mature crystal grape grains are obtained by adopting a culture separation technology.
The adding amount of the broken mash of the crystalline grapes in the fermentation bottle is 800mL/L, and the following needs to be stated: the natural fermentation in the invention is limited aerobic fermentation, namely, the fermentation bottle does not carry out vacuumizing or inert gas filling treatment, so that the fermentation bottle contains a small amount of oxygen, but the oxygen is consumed until no oxygen exists in the early stage of fermentation, and the closed fermentation only allows carbon dioxide to be discharged and does not allow other gases to enter, so that the middle and later stages of fermentation belong to anaerobic fermentation.
The application of the high flocculation property schizosaccharomyces japonica strain in wine brewing comprises the following steps:
1) Crushing grapes; 2) Performing enzymolysis; 3) Inoculating high flocculation property schizosaccharomyces japonica strains into crushed and enzymatically hydrolyzed grape mash to carry out alcoholic fermentation; 4) Filtering, centrifuging and discharging glue.
The inoculation amount of the high flocculation schizosaccharomyces japonica strain is 10 6 cells/mL。
The beneficial effects are that:
1. the schizosaccharomyces japonica strain provided by the invention is a novel strain, and the strain separation sieve is selected from middle and later periods of natural fermentation of the crystal grapes, and has the characteristic of high flocculation property.
2. The strain of the invention can be used for brewing wine in a single strain mode without being combined with saccharomyces cerevisiae.
3. The strain of the invention has excellent fermentation capacity, the fermentation time is about 15 days, and the reducing sugar content in the obtained wine is about 2.93g/L.
4. The strain of the invention has scientific alcohol production capacity; the alcoholicity of the wine obtained is about 11.3% vol.
5. The strain provided by the invention has scientific organic acid metabolism capability, the tartaric acid content in the obtained wine is about 3.66g/L, the malic acid content is about 2.21g/L, and the citric acid content is about 0.50g/L.
6. The strain of the invention is utilized for brewing wine, and the content of glycerin and total phenol in the obtained wine is moderate; the glycerol content of the wine was about 5.89g/L and the total phenol content was about 1.35g/L.
7. The strain can improve the clarity of the wine in a biological clarification mode, thereby saving the clarification cost and improving the clarification effect.
8. The strain provided by the invention has excellent cell aggregation capability, can well aggregate in fermentation liquor and wine in the alcoholic fermentation process, is favorable for combining macromolecular colloid substances and removing the macromolecular colloid substances through filtration, reduces the use amount of colloid materials such as bentonite and the like, and improves the clarity and stability of the wine.
9. The strain of the invention is separated from the middle and later period of natural fermentation, is dominant bacteria, has excellent ethanol tolerance and capability of competing with saccharomyces cerevisiae, and has a living cell concentration higher than or equal to that of the saccharomyces cerevisiae (10 6 ～10 8 cfu/mL)。
Drawings
Fig. 1: morphological characteristics of the high flocculation merozoite Japanese of the invention on WL medium;
fig. 2: the high flocculation property schizosaccharomyces japonica cell morphology of the invention under a microscope;
fig. 3: the aggregation state of yeast cells in the wine after the alcoholic fermentation is finished; wherein A: high flocculation property schizosaccharomyces japonica; b: middle flocculation schizosaccharomyces japonica; c: low flocculation schizosaccharomyces japonica; d: saccharomyces cerevisiae, medium flocculation property.
Detailed Description
The following detailed description of the invention is provided in further detail, but the invention is not limited to these embodiments, any modifications or substitutions in the basic spirit of the present examples, which still fall within the scope of the invention as claimed.
Example 1
A method for obtaining high flocculation property schizosaccharomyces japonica strain, which comprises the following steps:
(1) Mature crystal grape fruit grains are obtained, crushed and placed in a sterile fermentation bottle, closed fermentation is carried out at 28 ℃, and the addition amount of the crushed mash of the crystal grape in the fermentation bottle is 800mL/L, and the following is required: the fermentation bottle is not vacuumized or filled with inert gas, so that the fermentation bottle contains a small amount of oxygen, but the oxygen is consumed until no oxygen exists in the early stage of fermentation, and the sealed fermentation only allows carbon dioxide to be discharged and does not allow other gases to enter, so that the middle and later stages of fermentation belong to anaerobic fermentation;
(2) Judging the fermentation process by monitoring the carbon dioxide release amount, sampling at different stages of fermentation, separating 63 strains of schizosaccharomyces japonica by adopting a culturable separation technology in the natural fermentation process of the crystal grapes in Guizhou province, and detecting the flocculation value of the schizosaccharomyces japonica by a citrate method; mixing schizosaccharomyces japonica with yeast 10 6 Inoculating the cells/mL into liquid YPD, culturing to a stationary phase middle period, centrifuging at 12000rpm for 5min, and collecting thalli; washing the thalli for 2 times by using deflocculating buffer solution, removing supernatant, washing the thalli for 2 times by using sterile water, suspending the thalli in the flocculation buffer solution, and detecting an OD600 value (marked as A) by using an enzyme-labeling instrument; culturing the rest suspension at 28deg.C for 2 hr, shaking, standing for 30min, and detecting OD600 value (denoted as B) with enzyme-labeled instrument; calculating flocculation value of each strain of yeast according to the flocculation value Flo=B/A×100%; the pH=3.0 of the deflocculating buffer solution consists of 50mmol/L sodium citrate and 5mmol/L EDTA; the pH of the flocculation buffer=2.2, consisting of 50mmol/L sodium citrate, 20mmol/L CaCl 2 0.1mol/L citric acid;
(3) Determining flocculation property of 63 strains of schizosaccharomyces japonica according to flocculation value, wherein the flocculation property is divided into three levels of high, medium and low: the flocculation value is high in the range of 0-30%, the flocculation value is medium in the range of 30-70%, and the flocculation value is low in the range of 70-100%. Through detection, 61 high flocculation merozoites japonica, 1 flocculation merozoites japonica in 1 and 1 low flocculation merozoites japonica are obtained in total;
the identification characteristics of the high flocculation property schizosaccharomyces japonica strain are the first creation of the inventor, so that the genotype characteristics of the schizosaccharomyces japonica strain with high flocculation property can characterize the strain level molecular characteristics in the application process:
1) Morphological characteristics: growing on WL culture medium, wherein the colony is light green spherical bulge with tray-shaped bottom (figure 1), and the cell is elliptical or round rod-shaped (figure 2);
2) Molecular characteristics: 5.8S-ITS-RFLP analysis of 700bp PCR product and 550+150bp HaeIII cleavage product; 26S D1/D2 region sequence analysis to obtain 586bp effective sequence, the GenBank serial number of the strain is OP364841; the genotype of the seven microsatellite loci is Glb-T2a-G3a-Saa-Syc-11b-12b, specifically, the microsatellite locus GA1 amplification product is 190bp, the microsatellite locus TG2 amplification product is 280bp, the microsatellite locus CG3 amplification product is 280bp, the microsatellite locus SaGAA1 amplification product is 110bp, the microsatellite locus SyGAA2 amplification product is 110+80bp, the microsatellite locus C11 amplification product is 170bp and the microsatellite locus C12 amplification product is 190bp.
Example 2
Use of high flocculation property Schizosaccharomyces japonicus strain in wine brewing; the high flocculation property schizosaccharomyces japonica strain is classified and named as schizosaccharomyces japonica (schizosaccharomyces japonica), and the laboratory strain collection number is recorded as FBKL2.9SZJ-29 and is stored in China center for type culture collection, and the storage number is: cctccc NO: m20221342, the preservation time is: 2022, 9, 5; the flocculation value is about 10.37 plus or minus 0.08 percent;
the wine brewing method comprises the following steps:
1) Crushing grape, adding 60mg/L SO 2 The method comprises the steps of carrying out a first treatment on the surface of the 2) Adding 16mg/L pectase into the crushed grape material obtained in the step 1) for treatment for 3 hours to obtain grape mash; 3) At 10 6 Inoculating high flocculation schizosaccharomyces japonica strain into grape mash at a concentration of cells/mL, controlling the alcoholic fermentation temperature at 28 ℃, and monitoring the fermentation process by adopting a weightlessness method; 4) Filtering with sterile gauze after fermentation, centrifuging at 8000rpm for 5min, standing at 4deg.C for 7d, and analyzing basic components of wine; 5) Treating the filtrate with the adhesive material, and preserving at 4deg.C;
basic component analysis index of grape wine: reducing sugar (filin reagent method), total acid (acid-base titration method), pH, total phenol (Fu Lin Fenfa), glycerol (enzyme method detection kit), alcoholicity (alcohol meter method), organic acid (high performance liquid chromatography) and volatile acid (GB/T15038 detection method);
in this example, a comparative example was also set, and wine was brewed using low flocculation property (flocculation value 77.11.+ -. 0.46%), medium flocculation property (flocculation value 64.60.+ -. 0.17%) and commercial Saccharomyces cerevisiae (flocculation value 55.14.+ -. 2.46%) and wild-type preferred Saccharomyces cerevisiae (flocculation value 55.27.+ -. 1.11%) separated from example 1, respectively, in the form of single strain inoculation, and the basic characteristics of the obtained wine are shown in Table 1:
TABLE 1
As can be seen from table 1: the high flocculation property schizosaccharomyces japonica is superior to the medium flocculation property and low flocculation property schizosaccharomyces japonica in fermentation capacity (lower reducing sugar and shorter fermentation time after fermentation), similar alcohol production capacity, weaker organic acid metabolism capacity and glycerol production capacity and partial phenol loss caused by high flocculation property. The high flocculation property schizosaccharomyces japonica exhibits similar fermentation capacity (lower reducing sugar, slightly longer fermentation time), alcohol production capacity (similar to wild preferred saccharomyces cerevisiae, slightly weaker than commercial saccharomyces cerevisiae), organic acid metabolism capacity (commercial saccharomyces cerevisiae shows slightly stronger tartaric acid metabolism capacity) and glycerol production capacity compared with the flocculation property saccharomyces cerevisiae in 2 strains, and the phenol loss caused by flocculation property is similar to that of the commercial saccharomyces cerevisiae and higher than that of the wild preferred saccharomyces cerevisiae. Thus, the basic characteristics of wine brewing by a strain of highly flocculating schizosaccharomyces cerevisiae, compared to other schizosaccharomyces japonica, are closer to those of wine brewing by saccharomyces cerevisiae, demonstrating: the high flocculation property schizosaccharomyces japonica single strain of the invention shows basic fermentation capacity of wine brewing, and can perform wine fermentation in a single strain mode.
Example 3
Application of high flocculation schizosaccharomyces japonica in improving stability of wine; the high flocculation property schizosaccharomyces japonica strain (schizosaccharomyces japonica) is recorded as FBKL2.9SZJ-29 in a laboratory strain collection number, and is deposited in China center for type culture collection, and the deposit number is: cctccc NO: m20221342, the preservation time is: 2022, 9, 5; the flocculation value is about 10.37 plus or minus 0.08 percent;
according to the method for brewing wine of example 2, after fermentation is completed, the aggregation state of yeast cells in the raw wine, the absorbance of the raw wine after centrifugation and the development of a gumming test are analyzed, and the specific scheme is as follows:
(1) Analysis of aggregation state of yeast cells in wine base: after the alcoholic fermentation is finished, shaking up the fermentation liquor, taking the fermentation liquor to observe the aggregation state of yeast cells in a microscopic (400×), as shown in fig. 3, a high-flocculation-property schizosaccharomyces japonica is in a very obvious large aggregation state, medium-flocculation-property schizosaccharomyces japonica cells are partially aggregated, and low-flocculation-property schizosaccharomyces japonica and medium-flocculation-property saccharomyces cerevisiae are in a non-aggregation state basically; the large amount of aggregation state after fermentation of the yeast cells is beneficial to sedimentation and removal of the yeast cells;
(2) Absorbance analysis of the wine base after centrifugation: after the alcoholic fermentation, the base wine was centrifuged (12000 rpm,5 min), and the absorbance value of the base wine at 680nm was measured by using a microplate reader, as shown in Table 2;
(3) And (3) a wine gluing experiment: after the alcoholic fermentation is finished, the grape wine is kept stand at 4 ℃ for 7d, then the bentonite with different concentrations is used for the glue-removing experiment, the bentonite is added, and then the mixture is shaken uniformly and kept stand at 4 ℃ for 24h, the absorbance of the supernatant is detected by adopting an enzyme-labeled instrument, and the bentonite concentration corresponding to the minimum absorbance value is used as the glue-removing concentration of the grape wine; as shown in table 2;
in this example, a comparative example was also set up, and wine brewing was carried out as single inoculation using Schizosaccharomyces japonica having low flocculation property (flocculation value 77.11.+ -. 0.46%), medium flocculation property (flocculation value 64.60.+ -. 0.17%), commercial Saccharomyces cerevisiae (flocculation value 55.14.+ -. 2.46%), and wild-type preferred Saccharomyces cerevisiae (flocculation value 55.27.+ -. 1.11%) separated from example 1;
TABLE 2
Brewing wine with grape as raw material, inoculating single strain with inoculation amount of 10 before alcoholic fermentation 6 cell/mL, observing the fermentation in the raw wine after the alcoholic fermentation is finishedThe aggregation state of parent cells shows that the aggregation of high flocculation property schizosaccharomyces japonica is obvious (figure 3A), the absorbance of the raw wine after centrifugation is the lowest at 680nm, and the absorbance of the raw wine after centrifugation is higher than that of the raw wine obtained by fermenting the high flocculation property schizosaccharomyces japonica after the partial aggregation of medium flocculation property and low flocculation property schizosaccharomyces japonica (figure 3B) or no aggregation (figure 3C); in contrast, using commercial and wild-preferred Saccharomyces cerevisiae as a control, saccharomyces cerevisiae did not aggregate in the base wine after the end of the alcoholic fermentation (FIG. 3D), and was therefore difficult to remove.
As can be seen from table 2: the bentonite consumption of the wine base brewed by the high-flocculation-property schizosaccharomyces japonica in the colloid-making link is the lowest, and the bentonite consumption is reduced by 21.43 percent and 31.25 percent respectively compared with that of medium-flocculation-property commercial saccharomyces cerevisiae and medium-flocculation-property wild preferred saccharomyces cerevisiae, and the bentonite consumption is reduced by 8.33 percent and 38.89 percent respectively compared with that of medium-flocculation-property schizosaccharomyces japonica and low-flocculation-property schizosaccharomyces japonica; and the development of the sizing according to the sizing concentration shown in Table 2 shows that the absorbance of the obtained wine has no obvious difference except that the wild preferred Saccharomyces cerevisiae is higher, so that the high flocculation property Schizosaccharomyces japonica of the invention needs minimum sizing materials for fermenting the wine and has excellent clarity.
As can be seen from table 2: at the end of fermentation, the total phenol content in the high-flocculation-property schizosaccharomyces japonica and medium-flocculation-property commercial saccharomyces cerevisiae fermented brewed wine is relatively lower than that of the wine fermented by other strains, and after 4-month standing at the temperature of 4 ℃, the total phenol content in the high-flocculation-property schizosaccharomyces japonica and medium-flocculation-property commercial saccharomyces cerevisiae fermented wine is found to be slightly increased, while the total phenol content of other strains is reduced, so that the pigment and tannin stability of the high-flocculation-property schizosaccharomyces japonica fermented wine in the ageing process is reflected.
Claims (6)
1. A schizosaccharomyces japonica strain with high flocculation property is characterized by being classified and named as schizosaccharomyces japonicaSchizosaccharomyces japonicus) The laboratory strain collection number is recorded as FBKL2.9SZJ-29, and is stored in China center for type culture Collection, and the collection number is: cctccc NO: m20221342, the preservation time is: 2022, 9 and 5.
2. Use of a high flocculation property schizosaccharomyces japonica strain according to claim 1 in wine brewing.
3. The use according to claim 2, wherein the application method comprises the steps of:
1) Crushing grapes;
2) Performing enzymolysis;
3) Inoculating high flocculation property schizosaccharomyces japonica strains into crushed and enzymatic hydrolyzed grape mash, and carrying out alcoholic fermentation;
4) After fermentation, filtering, centrifuging, standing and gumming.
4. The use according to claim 3, wherein the high flocculation property schizosaccharomyces japonica strain is inoculated in an amount of 10 6 cells/mL。
5. The use according to claim 2, wherein flocculation of the yeast cells in wine brewing is performed simultaneously with clarification.
6. Use of the high flocculation property schizosaccharomyces japonica strain according to claim 1 for improving clarity and stability of wine during fermentation and gumming of wine.
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