CN115558611A - Schizosaccharomyces japonicus strain with high flocculation property and application thereof - Google Patents
Schizosaccharomyces japonicus strain with high flocculation property and application thereof Download PDFInfo
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- 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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- 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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- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
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Abstract
The invention belongs to the technical field of brewing engineering, and particularly relates to a high-flocculation Schizosaccharomyces japonicus strain (Schizosaccharomyces japonicum) and application thereof, wherein the Schizosaccharomyces japonicus strain is preserved in China center for type culture collection with the preservation number as follows: CCTCC NO: m20221342, preservation time: in 2022, 9 and 5 days, the strain has excellent fermentation capacity and cell aggregation capacity, can be used for brewing wine in a single strain form, can simultaneously perform flocculation and clarification of yeast cells, reduces the using amount of an adhesive material, and improves the clarity and stability of the wine in a biological clarification manner.
Description
Technical Field
The invention belongs to the technical field of brewing engineering, and particularly relates to a Schizosaccharomyces japonicus strain with high flocculation property and application thereof.
Background
The basic requirements for the clarity in the national standard organoleptic requirements of wine are clarity, gloss, no visible suspended matter. The wine is used as a colloidal solution, and after the alcoholic fermentation is finished, the macromolecular colloid which is easy to settle is separated out by developing clarification treatment process operations such as glue discharging, filtering and the like, so that the clarification 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 maintain the flavor of the wine and reduce the brewing cost of the wine, but related reports for improving the clarity of the wine by a biological clarification mode are not available at present.
Analysis of the comprehensive factors of the commercial study of wild yeasts (Wangchun et al, food and fermentation industries, 2022,48 (16): 284-290) describes the flocculation of yeasts during fermentation, mainly caused by the aggregation of cells due to the interaction of flocculants present on the surface of yeast cells with mannoproteins on the adjacent yeast cell walls. The yeast with flocculation property can be adhered to form a lump and settle in the wine fermentation process, and too high flocculation property can appear too early, so that the yeast can reduce the fermentation performance, consume incomplete sugar, prolong the fermentation time and the like caused by poor dispersibility; too low flocculation may result in slow yeast cell aggregation during the late stage of fermentation, which is detrimental to the filtration and clarification of wine. Thus, the preferred yeast having the appropriate flocculation properties facilitates the performance of wine alcohol fermentation. At present, yeast is mostly taken as a leading bacterium of alcohol fermentation in the conventional wine brewing, and after the alcohol fermentation is finished, the yeast is settled at the bottom of a fermentation tank due to flocculation and is removed by subsequent operations of tank transferring, filtering and the like. The saccharomyces cerevisiae is usually selected as the saccharomyces cerevisiae, has excellent fermentation performance but intermediate flocculation, generally has higher requirements on clarification and stability treatment of the wine, and the excessive use of the gel material in the clarification process often causes certain flavor loss while improving the clarity of the wine body.
Therefore, considering the disadvantages of Saccharomyces cerevisiae, researchers in this field try to achieve high quality wine brewing by optimizing yeast species; for example, patent publication No. CN1128796A discloses a method for brewing dry white wine, which comprises sorting, squeezing, clarifying, fermenting, filling and aging, separating, stabilizing, filtering with aging bacteria, bottling, packaging and warehousing white wine raw materials, wherein the key point is that active fission yeast replaces wine yeast adopted in the traditional process in the fermentation process. Therefore, the defects that only sugar in a fermentation substrate can be converted into alcohol, but organic acids such as malic acid and the like cannot be converted into alcohol to form high acidity and need chemical deacidification when wine yeast is used for fermentation are overcome, and the fermentation process and the deacidification process are carried out simultaneously. The percentage content (weight ratio) of the active fission yeast added during the fermentation is 5 to 10 percent, and the fermentation is carried out under the condition that the temperature is 16 to 18 ℃. The pH value of the grape juice is adjusted to 2.9-3.0 in the clarification process, which is beneficial to the activity of fission yeast. According to the technical scheme, the physical and chemical stability of the wine is mainly improved in a mode of reducing the malic acid content in the wine through biological metabolism, and the flocculation property and the influence on the clarity of active fission yeast are not mentioned.
Also, for example, patent publication No. CN108913410A discloses a method for preparing wine, comprising the steps of: (1) pretreatment of raw materials: cleaning fructus Vitis Viniferae, air drying, pulverizing to obtain fructus Vitis Viniferae mash A, pulverizing flos Lonicerae to obtain flos Lonicerae powder; (2) adding a clarifying agent: adding complex enzyme into the grape mash A, and mixing to obtain grape mash B, wherein the addition amount of the complex enzyme is 20-30mg/L; (3) adding a bacteriostatic agent: 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 is 10-20mg/L, and the honeysuckle and the SO 2 Can inhibit the generation of other mixed bacteria in the preparation process; (4) fermentation: adding yeast into the grape mash C, uniformly mixing, adding into a fermentation cylinder, performing sealed fermentation for 3-5 days, performing ultrasonic treatment for 3-6min, and filtering to obtain fermentation liquor, wherein the addition amount of the yeast is 5-10mg/L, and the ultrasonic treatment ensures that the components in the fermentation liquor are uniformly mixed, so that the dissolution rate of the pigment is improved; (5) separating: filtering the fermentation liquor by a microfiltration membrane, sterilizing to obtain the wine, wherein the microfiltration can effectively remove fine particles and bacteria in the fermentation liquor; (6) filling and storing: bottling the wine, and storing in wine cellar with humidity of 60-70% at 8-15 deg.C. The yeast is beer yeast and fission yeast, the beer yeast decomposes sugar to generate ethanol, and the fission yeast decomposes sour substances such as the sugar, malic acid and the like to generate ethanol, higher alcohol and ester; in the technical scheme, the fission yeast is mainly usedUsed for decomposing sugar and malic acid, and the flocculation property and clarification effect thereof were not evaluated.
Besides the clarity, the stability of the wine is also a crucial quality index, and phenolic substances in the wine, such as anthocyanin and tannin, belong to colloidal substances, not only directly influence the color and taste (structural sense) of the wine, but also influence the stability of the wine, which also puts higher control requirements on degumming treatment after fermentation. The related foreign research reports show that the color stability of wine can be improved by high yield of pyruvic acid or zymosan through the mixed fermentation of fission yeast and other yeast (BENITO)
(iv) the combined use of Schizosaccharomyces pombe and Lachance a thermolerans-effect on the alcoholic cell line composition. Molecules,2017,22 (5): 739), improving the stability of wine proteins and tartrates, preserving wine aroma (PORTARO L, MAIOLI F, CANUTI V, PICCHI M, LENCIONNI L, ZZNAU I, DOMIZIO P. Schizosaccharomyces japonicum/Saccharomyces cerevisiae mixed starter culture: new perspective for the immunological development of Sangiove aroma a, taste, and color. LWT,2022, 009. However, the yeast species for wine brewing disclosed at home at present have no characteristics of improving the clarity and stability of the wine, and the yeast species for wine brewing disclosed at abroad have no report on the aspect of improving the clarity of wine by applying high flocculation strains.
Disclosure of Invention
The invention provides a Schizosaccharomyces japonicus strain with high flocculation property and application thereof aiming at the defects of the prior art.
The method is realized by the following technical scheme:
the first object of the present invention is: a high flocculation fission yeast strain is provided, which is classified and named as Schizosaccharomyces japonicus (Schizosaccharomyces japonicum), the laboratory strain collection library number of which is recorded as FBKL2.9SZJ-29, and the strain collection library number is preserved in China center for type culture collection, and the preservation numbers are as follows: CCTCC NO: m20221342, preservation time: 9/5/2022.
The flocculation value range of the high flocculation property Schizosaccharomyces japonicus strain is as follows: 0.38 to 23.89 percent.
Preferably, the flocculation value range of the high flocculation property Schizosaccharomyces japonicus strain is as follows: 10.29 to 10.45 percent.
The high flocculation fission yeast strain Japanese identification characteristics are: growing on a WL culture medium, wherein bacterial colonies are light green spherical bulges with tray-shaped bottoms, and cells are in an oval or round rod shape; 5.8 the PCR product of S-ITS-RFLP analysis is 700bp, and the enzyme cutting product of Hae III is 550+150bp;26S D1/D2 area sequence analysis to obtain 586bp effective sequence, the GenBank sequence number of the strain is OP364841; the genotype of seven microsatellite loci is analyzed to be Glb-T2a-G3a-Saa-Syc-11b-12b, and 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 Schizosaccharomyces japonicus strain is a leading yeast separated and screened in the middle and later periods of natural fermentation of the crystal grapes, coexists with saccharomyces cerevisiae, and proves excellent competitiveness and tolerance of the saccharomyces japonicus strain.
The second object of the present invention is: provides an application of a Schizosaccharomyces japonicus strain with high flocculation in wine brewing.
Specifically, the high flocculation fission yeast strain of Japan is applied to the improvement of the stability and the clarity of the wine in a way that the flocculation and the clarification of the yeast cells are simultaneously carried out in the alcohol fermentation process.
In particular, the application of the high flocculation fission yeast strain in improving the clarity and stability of wine during fermentation and degumming of wine.
The method for obtaining the high-flocculation fission yeast strains comprises the following steps: obtaining mature crystal grape fruit grains, crushing, placing in an aseptic fermentation bottle, performing closed fermentation at 28 ℃, judging the fermentation process by monitoring the release amount of carbon dioxide, and sampling in the middle and later stages of fermentation by adopting a culturable separation technology.
The addition amount of the broken grape mash in the fermentation bottle is 800mL/L, and the following items are required: the natural fermentation in the invention is limited aerobic fermentation, namely, because the fermentation bottle is not vacuumized or filled with inert gas, the fermentation bottle contains a small amount of oxygen, but the oxygen is consumed until the fermentation bottle is anaerobic in the early stage, and the closed fermentation only allows carbon dioxide to be discharged and does not allow other gases to enter, so the fermentation bottle belongs to anaerobic fermentation in the middle and later stages.
The application of the high-flocculation Schizosaccharomyces japonicus strain in wine brewing comprises the following steps:
1) Crushing grapes; 2) Carrying out enzymolysis; 3) Inoculating a high-flocculation Schizosaccharomyces japonicus strain into the crushed and enzymolyzed grape mash to perform alcoholic fermentation; 4) Filtering, centrifuging and gluing.
The inoculation amount of the high flocculation fission yeast strain in Japan is 10 6 cells/mL。
Has the advantages that:
1. the Schizosaccharomyces japonicus strain provided by the invention is a new strain, and the strain separation sieve is selected from the middle and later stages 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 content of reducing sugar in the obtained wine is about 2.93g/L.
4. The strain of the invention has scientific alcohol production capacity; the alcohol content of the obtained wine was about 11.3% vol.
5. The strain of the invention has scientific organic acid metabolism capability, and the obtained wine has the tartaric acid content of about 3.66g/L, the malic acid content of about 2.21g/L and the citric acid content of about 0.50g/L.
6. The strain is utilized to brew the wine, and the obtained wine has moderate contents of glycerin and total phenols; the content of glycerin in the wine is about 5.89g/L, and the content of total phenols is 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 has excellent cell aggregation capacity, can be well aggregated in fermentation liquor and wine in the alcohol fermentation process, is beneficial to combining macromolecular colloidal substances and removing the macromolecular colloidal substances through filtration, reduces the use amount of lower gel materials such as bentonite and the like, and improves the clarity and the stability of the wine.
9. The strain is separated from the middle and later stages of natural fermentation, is a dominant bacterium, has excellent ethanol tolerance and capability of competing with saccharomyces cerevisiae, and has the concentration (10) of living cells higher than or equal to that of the saccharomyces cerevisiae under the condition that various yeasts die in the middle and later stages of natural fermentation 6 ～10 8 cfu/mL)。
Drawings
FIG. 1: morphological characteristics of the highly flocculating Schizosaccharomyces japonicus of the present invention on WL medium;
FIG. 2: microscopically the high flocculation fission yeast Japanese cell morphology;
FIG. 3: the yeast cell aggregation state in the wine after the alcoholic fermentation is finished; wherein, A: high flocculating fission yeast of Japan; b: schizosaccharomyces japonicus having medium flocculation property; c: low flocculation fission yeast japan; d: and (4) medium flocculation saccharomyces cerevisiae.
Detailed Description
The following is a detailed description of the embodiments of the present invention, but the present invention is not limited to these embodiments, and any modifications or substitutions in the basic spirit of the embodiments are included in the scope of the present invention as claimed in the claims.
Example 1
A method for obtaining a high flocculation fission yeast strain comprises the following steps:
(1) Obtaining mature crystal grape fruit particles, crushing the crystal grape fruit particles, putting the crushed crystal grape fruit particles into a sterile fermentation bottle, and performing closed fermentation at 28 ℃, wherein the addition amount of the crushed crystal grape mash in the fermentation bottle is 800mL/L, and the following items are required: because the fermentation bottle is not vacuumized or filled with inert gas for treatment, a small amount of oxygen is contained in the fermentation bottle, but the oxygen is consumed until the fermentation is anaerobic in the early stage, and the closed fermentation only allows carbon dioxide to be discharged and does not allow other gases to enter, so the fermentation is anaerobic in the middle and later stages;
(2) Judging the fermentation process by monitoring the release amount of carbon dioxide, sampling at different stages of fermentation, separating 63 Schizosaccharomyces japonicum strains 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 japonicum strains by a citrate method; schizosaccharomyces japonicus was cultured at 10 6 Inoculating cells/mL into liquid YPD to culture to stationary phase middle stage, centrifuging at 12000rpm for 5min, and collecting thallus; washing the thallus with deflocculating buffer solution for 2 times, removing supernatant, washing with sterile water for 2 times, suspending the thallus in the deflocculating buffer solution, and detecting OD600 value (marked as A) by using an enzyme-labeling instrument; culturing the rest suspension at 28 deg.C for 2 hr, shaking, standing for 30min, and detecting OD600 value (labeled as B) with enzyme-labeling instrument; the flocculation value of each yeast was calculated as flocculation value Flo = B/a × 100%; the deflocculating buffer has a pH =3.0 and consists of 50mmol/L sodium citrate, 5mmol/L EDTA; the flocculation buffer has pH =2.2 and is composed of 50mmol/L sodium citrate and 20mmol/L CaCl 2 0.1mol/L citric acid;
(3) The flocculation property of 63 strains of Schizosaccharomyces japonicus is determined according to the flocculation value and is divided into three levels, namely high level, middle level and low level: the flocculation value is high flocculation property within the range of 0-30%, the flocculation value is medium flocculation property within the range of 30-70%, and the flocculation value is low flocculation property within the range of 70-100%. Through detection, 61 high flocculation fission yeast, 1 flocculation fission yeast among the strains and 1 low flocculation fission yeast are obtained;
seven microsatellite loci are the first creation of the inventor, so that the genotype characteristic of the schizosaccharomyces japonicus with high flocculation can be characterized in the horizontal molecular characteristic of the strain 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 elliptic or round rod-shaped (figure 2);
2) Molecular characteristics: 5.8S-ITS-RFLP analysis PCR product is 700bp, hae III enzyme cutting product is 550+150bp;26S D1/D2 area sequence analysis to obtain 586bp effective sequence, the GenBank sequence number of the strain is OP364841; the genotype of seven microsatellite loci is analyzed to be Glb-T2a-G3a-Saa-Syc-11b-12b, and 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
The application of a high flocculation fission yeast strain in wine brewing; the high flocculation fission yeast strain is classified and named as Schizosaccharomyces japonicus (Schizosaccharomyces japonicum), the laboratory strain collection library number of which is recorded as FBKL2.9SZJ-29, and the strain collection library is preserved in China center for type culture collection with the preservation number of: CCTCC NO: m20221342, preservation time: 9/month 5/2022; its flocculation value is about 10.37 + -0.08%;
the wine brewing method comprises the following steps:
1) Crushing grape, adding 60mg/L SO 2 (ii) a 2) Adding 16mg/L pectinase into the crushed grape material obtained in the step 1) for treatment for 3h to obtain grape mash; 3) At 10 6 Inoculating a high-flocculation Schizosaccharomyces japonicus strain into the grape mash at the cell/mL concentration, controlling the alcoholic fermentation temperature at 28 ℃, and monitoring the fermentation process by adopting a weight loss method; 4) Filtering with sterile gauze after fermentation, centrifuging at 8000rpm for 5min, standing at 4 deg.C for 7d, and analyzing basic components of wine; 5) Treating the filtrate with a sizing material, and storing at 4 ℃;
basic component analysis indexes of the wine: reducing sugar (feilin reagent method), total acid (acid-base titration method), pH, total phenol (furilin phenol method), glycerol (enzymatic assay kit), alcoholicity (alcometer method), organic acid (high performance liquid chromatography) and volatile acid (GB/T15038 assay method);
this example was carried out while setting comparative examples, and wine brewing was carried out in the form of single inoculation using low flocculation property (flocculation value of 77.11. + -. 0.46%) of Schizosaccharomyces japonicus, medium flocculation property (flocculation value of 64.60. + -. 0.17%) of Schizosaccharomyces japonicus, commercial Saccharomyces cerevisiae (flocculation value of 55.14. + -. 2.46%) and wild preferred Saccharomyces cerevisiae (flocculation value of 55.27. + -. 1.11%) isolated from example 1, respectively, and the basic characteristics of the obtained wine are shown in Table 1:
TABLE 1
As can be seen from Table 1: a strain of high flocculation fission yeast Schizosaccharomyces japonicus shows more excellent fermentation capability (lower reducing sugar and shorter fermentation time after the end of fermentation), similar alcohol production capability, weaker organic acid metabolism capability and glycerol production capability, and partial phenol loss caused by high flocculation compared with the medium flocculation and low flocculation fission yeast Schizosaccharomyces japonicus. A strain of highly flocculating Schizosaccharomyces japonicus 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 embodies slightly stronger tartaric acid metabolism) and glycerol production capacity compared with 2 strains of flocculating Saccharomyces cerevisiae, and phenol loss caused by flocculation is similar to that of commercial Saccharomyces cerevisiae and higher than that of wild preferred Saccharomyces cerevisiae. Therefore, the basic wine brewing characteristics of a high flocculation fission yeast strain are closer to the wine brewing characteristics of saccharomyces cerevisiae compared with other fission yeasts, and the description shows that: the high flocculation fission yeast Schizosaccharomyces japonicus single strain embodies the basic fermentation capability of wine brewing, and can perform wine fermentation in a single strain mode.
Example 3
The application of a high flocculation fission yeast schizosaccharomyces japonicus in improving the stability of wine; the high flocculation fission yeast strain (Schizosaccharomyces japonicum) has the laboratory strain collection number of FBKL2.9SZJ-29, is preserved in China center for type culture collection, and has the preservation number: CCTCC NO: m20221342, preservation time: 9/5/2022; its flocculation value is about 10.37 + -0.08%;
according to the wine brewing method of the embodiment 2, after the fermentation is finished, the aggregation state of the yeast cells in the raw wine, the absorbance of the raw wine after centrifugation and the gel test are carried out, and the specific scheme is as follows:
(1) Analysis of aggregation status 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 x) manner, wherein as shown in figure 3, a high-flocculation fission yeast strain shows a very obvious large aggregation state, a middle-flocculation fission yeast strain shows partial aggregation, and a low-flocculation fission yeast strain shows a non-aggregation state basically; the large aggregation state of the fermented yeast cells is beneficial to the sedimentation and removal of the yeast cells;
(2) Absorbance analysis of the wine base after centrifugation: after the alcoholic fermentation is finished, centrifuging the wine base (12000rpm, 5 min), and detecting the absorbance value of the wine base under 680nm by using an enzyme-linked immunosorbent assay (ELIAS. RTM.) as shown in Table 2;
(3) Glue-dropping experiment of wine: after the grape wine base after the alcoholic fermentation is finished is kept still at 4 ℃ for 7d, bentonite with different concentrations is used for glue adding experiments, the bentonite is added and then shaken evenly and kept still at 4 ℃ for 24h, an enzyme-labeling instrument is used for detecting the absorbance of supernatant, and the corresponding bentonite concentration when the absorbance value is minimum is used as the glue adding concentration of the grape wine base; as shown in table 2;
this example was carried out while setting comparative examples, and wine brewing was carried out in the form of single inoculation using low flocculation property (flocculation value of 77.11. + -. 0.46%), medium flocculation property (flocculation value of 64.60. + -. 0.17%) Schizosaccharomyces japonicus, commercial Saccharomyces cerevisiae (flocculation value of 55.14. + -. 2.46%), and wild preferred Saccharomyces cerevisiae (flocculation value of 55.27. + -. 1.11%) isolated from example 1, respectively;
TABLE 2
Brewing wine with fructus Vitis Viniferae as raw material, inoculating with single strain before alcoholic fermentation, with inoculation amount of 10 6 cells/mL, after the alcoholic fermentation, observing the aggregation state of yeast cells in the raw wine, and knowing that the high flocculation fission yeast Saccharomyces cerevisiae is very obviously aggregated (figure 3A), the absorbance of the raw wine at 680nm after centrifugation is lowest, and the middle flocculation fission yeast and the low flocculation fission yeast are partially aggregated (figure 3B) or not aggregated (figure 3C), and the absorbance of the raw wine after centrifugation is higher than that of the raw wine obtained by the high flocculation fission yeast fermentation; whereas, using commercial s.cerevisiae and wild preferred s.cerevisiae as controls, the s.cerevisiae did not aggregate in the wine base after the end of alcoholic fermentation (FIG. 3D), and thus was difficult to remove.
As can be seen from table 2: the raw wine brewed by a strain of high flocculation fission yeast Japanese has the lowest consumption of bentonite in the glue adding process, and is respectively reduced by 21.43 percent and 31.25 percent compared with middle flocculation commercial saccharomyces cerevisiae and middle flocculation wild preferred saccharomyces cerevisiae, and is respectively reduced by 8.33 percent and 38.89 percent compared with middle flocculation fission yeast Japanese and low flocculation fission yeast Japanese; and the gel-adding is carried out according to the gel-adding concentration shown in the table 2, the absorbance of the obtained wine is higher except that the wild preferred saccharomyces cerevisiae, and other differences are not obvious, which shows that the gel-adding material required by the high flocculation schizosaccharomyces japan fermentation wine is least, and the high flocculation schizosaccharomyces japan fermentation wine has excellent clarity.
As can be seen from Table 2: at the end of fermentation, the total phenol content of the high-flocculation fission yeast and the middle-flocculation commercial saccharomyces cerevisiae fermented and brewed wine is relatively lower than that of wine fermented and brewed by other strains, after the wine is placed at 4 ℃ for 4 months, the total phenol content of the high-flocculation fission yeast and the middle-flocculation commercial saccharomyces cerevisiae fermented and brewed wine is slightly increased, and the total phenol content of other strains is reduced, so that the stability of pigment and tannin in the high-flocculation fission yeast fermented wine during the aging process is reflected.
Claims (10)
1. A highly flocculating Schizosaccharomyces japonicus strain, which is classified and named Schizosaccharomyces japonicus (Schizosaccharomyces japonica), has a laboratory strain collection number of FBKL2.9SZJ-29, is deposited in China center for type culture collection, and has a deposition number of: CCTCC NO: m20221342, preservation time: 9/5/2022.
2. The highly flocculating schizosaccharomyces japonicus strain of claim 1, wherein the highly flocculating flocculation value ranges from: 0.38 to 23.89 percent.
3. The highly flocculating fission yeast japan according to claim 1 or 2, wherein the highly flocculating fission yeast strain has a flocculation value range of: 10.29 to 10.45 percent.
4. The highly flocculating schizosaccharomyces japan strain of yeast according to claim 1, wherein the highly flocculating schizosaccharomyces japan strain is characterized by: growing on a WL culture medium, wherein the bacterial colony is a light green spherical bulge with a tray-shaped bottom, and the cell is in an oval or round rod shape; 5.8S-ITS-RFLP analysis PCR product is 700bp, hae III enzyme cutting product is 550+150bp;26SD1/D2 area sequence analysis obtains 586bp effective sequence, the GenBank sequence number of the strain is OP364841; the genotype of seven microsatellite loci is analyzed to be Glb-T2a-G3a-Saa-Syc-11b-12b, and 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.
5. The highly flocculating schizosaccharomyces japan strain according to claim 1, wherein the highly flocculating schizosaccharomyces japan strain is a dominant yeast isolated and selected in the middle and later stages of the natural fermentation of grapevine.
6. Use of a highly flocculating Schizosaccharomyces japonicus strain according to any one of claims 1 to 5 for brewing wine.
7. Use of a highly flocculating Schizosaccharomyces japonicus strain according to any one of claims 1 to 5 for simultaneous flocculation and clarification of yeast cells in alcoholic fermentation.
8. Use of a highly flocculating Schizosaccharomyces japonicus strain according to any one of claims 1 to 5 to improve the clarity and stability of the wine during fermentation, degumming of the wine.
9. The use of a highly flocculating Schizosaccharomyces japonicus strain according to claim 6 in wine brewing, comprising the steps of:
1) Crushing grapes;
2) Carrying out enzymolysis;
3) Inoculating a high-flocculation Schizosaccharomyces japonicus strain into the crushed and enzymolyzed grape mash, and carrying out alcoholic fermentation;
4) After fermentation, filtering, centrifuging, standing and adding glue.
10. The use of a highly flocculating Schizosaccharomyces japonicus strain as claimed in claim 6 in wine brewing, wherein the amount of inoculation of the highly flocculating Schizosaccharomyces japonicus strain is 10 6 cells/mL。
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