Yarrowia guilliermondii and application thereof in lemon wine

Abstract

The invention relates to the technical field of microbial fermentation, in particular to a Jianliermondii YT-60 strain and application of the Jianliermondii YT-60 strain in lemon wine, the Jianliermondii YT-60 strain is preserved in Guangdong Microbial Culture Collection Center on April 3, 2025, the preservation number is GDMCC No: 66091, the preservation address is the 5th floor, No. 59 building, No. 100 Courtyard, Xianlie Middle Road, Guangzhou City, Guangdong Province, and the preservation number is CGMCC No: 66091. The preservation address of Guangdong Institute of Microbiology is 510070. The invention provides a novel fruit wine fermentation strain, the strain has better tolerance to pH, sugar degree, ethanol and SO2, and the total ester yield is as high as 2.60 g / L. The lemon wine fermented by combining the strain provided by the invention with saccharomyces cerevisiae has more unique flavor than the lemon wine fermented by single saccharomyces cerevisiae.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial fermentation, and specifically relates to a strain of Meyer yeast and application thereof in lemon wine. Background Art

[0002] Lemon is a highly nutritious representative of citrus fruits. Its pulp is rich in vitamin C, citric acid, flavonoids, dietary fiber and various minerals (such as potassium, calcium, magnesium, etc.). The vitamin C content can reach more than 50mg / 100g, which has significant antioxidant activity, can scavenge free radicals and enhance immunity; citric acid, as a natural organic acid, can promote the secretion of digestive juices and improve the body's acid-base balance; the unique volatile terpenes (such as limonene and linalool) and flavonoids in the peel have antibacterial, anti-inflammatory and metabolic regulating functions. In addition, the low sugar, high acid and low calorie characteristics of lemon make it an ideal choice for a healthy diet. However, fresh lemons have poor palatability when eaten directly due to their high acidity (pH 2.0-2.5), and traditional processing methods (such as juicing and drying) can easily cause the loss of active ingredients.

[0003] Lemon wine combines the nutrition of lemon with the fermentation process through bio-fermentation technology, effectively solving the pain points of strong sourness and astringency of fresh fruit and single processing form. Compared with traditional lemon products, fermented lemon wine not only retains heat-sensitive ingredients such as vitamin C and polyphenols, but also increases active substances such as free amino acids, B vitamins and short-chain fatty acids through microbial transformation. It has potential health functions such as regulating intestinal flora and promoting metabolism. In addition, the fermentation process can passivate bitter substances in lemon peel (such as limonin), improve flavor coordination, and extend the shelf life through the natural antibacterial effect of alcohol. Lemon wine is low in sugar, low in calories and naturally fermented, which meets the needs of modern consumers for functional drinks.

[0004] However, due to the high acidity of lemon, which affects the growth and reproduction of brewing yeast, the lemon wine obtained by traditional fermentation has a single flavor and poor taste. Summary of the Invention

[0005] The present invention aims to overcome the deficiencies in the prior art and provides a strain of Meyerozyma guilliermondii YT-60, which has good tolerance to pH, sugar content, ethanol, and SO2, and produces up to 2.60 g / L of total esters. Lemon wine fermented by the strain of the present invention combined with brewer's yeast has a more unique flavor than lemon wine fermented by brewer's yeast alone.

[0006] In order to achieve the above object of the invention, the specific technical solution of the present invention is:

[0007] The present invention protects a strain of Meyerozyma guilliermondii YT-60, which was deposited in the Guangdong Provincial Microbial Culture Collection Center on April 3, 2025, with a deposit number of GDMCC NO: 66091. The deposit unit address is Guangdong Institute of Microbiology, 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, and the postal code of the deposit address is: 510070.

[0008] Furthermore, the ITS sequence of Meyerozyma guilliermondii YT-60 is shown in SEQ ID NO: 1.

[0009] The present invention also protects the use of the above-mentioned Meyerozyma guilliermondii YT-60 in the preparation of fermented lemon wine.

[0010] The present invention also protects a product containing the above-mentioned Meyerozyma guilliermondii YT-60, so the product includes but is not limited to a bacterial suspension containing Meyerozyma guilliermondii YT-60, a bacterial slurry containing Meyerozyma guilliermondii YT-60, and a bacterial agent containing Meyerozyma guilliermondii YT-60.

[0011] The present invention also protects a fermented lemon wine, in which the above-mentioned Meyerozyma guilliermondii YT-60 or the product containing Meyerozyma guilliermondii YT-60 is used in the preparation process.

[0012] The present invention also protects a method for preparing fermented lemon wine, which comprises the following steps:

[0013] 1) Fresh lemons are washed, seeded, peeled, and juiced, and then purified water and lemon peel without white pulp are added. Potassium metabisulfite is then added for sterilization, followed by pectinase enzymolysis, and the sugar concentration is adjusted;

[0014] 2) preparing a bacterial suspension containing Meyerozyma guilliermondii YT-60; then adding the bacterial suspension and Angel Wine Yeast SY to the lemon juice treated in step 1) for static fermentation to obtain lemon wine;

[0015] 3) filtering the lemon wine in step 2) through a filter screen and then filling it into a sterile glass bottle, sterilizing it, aging it, and collecting the supernatant to obtain a finished lemon wine.

[0016] As a preferred embodiment, in step 1) of the method for preparing fermented lemon wine, the amount of purified water added is 1-3 times the volume of fresh lemon juice; the amount of lemon peel without white pulp added is 1 / 9-1 / 10 the volume of fresh lemon juice; the amount of potassium metabisulfite added is 0.08-0.1 g / L; the amount of pectinase added is 0.2-0.4 g / L; enzymatic hydrolysis is carried out under constant temperature conditions, the enzymatic hydrolysis temperature is 45-55° C., the hydrolysis time is 2-3 hours, and the sugar concentration is adjusted to 23 Brix with white sugar.

[0017] As a preferred embodiment, in step 2) of the method for preparing fermented lemon wine, the preparation steps of the bacterial suspension containing Meyerozyma guilliermondii YT-60 are as follows:

[0018] The seed liquid was inoculated into YPD liquid medium to obtain culture solution; the culture solution was then fermented to obtain fermentation stock solution. At this time, the strain concentration was 2×10 8 CFU / mL-6×10 8 CFU / mL;

[0019] The fermentation stock solution was centrifuged at 8000 rpm and 4° C. for 8 min, the supernatant was removed, and the solution was washed three times with sterile saline and finally resuspended with an equal volume of saline to obtain a bacterial suspension.

[0020] As a preferred embodiment, in step 2) of the fermented lemon wine preparation method, the amount of the bacterial suspension containing Meyerozyma guilliermondii YT-60 added is 3% of the volume of the treated lemon juice; the amount of Angel Wine Yeast SY added is 0.2-0.8 g / L; the static fermentation temperature is 20-25°C (specifically 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, etc.), and the fermentation time is 10-12 days.

[0021] As a preferred embodiment, in step 3) of the method for preparing fermented lemon wine, the sterilization temperature is 80° C. and the time is 20 minutes; the aging temperature is 4° C. and the time is 1 month.

[0022] The present invention also protects fermented lemon wine that can be prepared by adopting the above combination of method steps.

[0023] Furthermore, fermented lemon wine contains 20 volatile compounds, with a high content of ethyl acetate. The geraniol contained in it gives the fermented lemon wine a tropical fruit and grassy flavor; the glycerol contained in it can make the wine body more rounded, and the ethyl palmitate contained in it gives the lemon wine a unique creamy aroma.

[0024] Compared with the existing technology, the beneficial effects of the present invention are:

[0025] (1) The present invention provides a strain of Meyerozyma guilliermondii YT-60 that can be used for fermentation. The strain has the characteristics of high ester production, strong acid resistance, SO2 resistance, sugar resistance, and strong alcohol resistance.

[0026] (2) The strain can be used for fruit wine fermentation and can supplement the strain library for fruit wine fermentation. The lemon wine fermented by the strain of the present invention has a more unique flavor than that fermented by a single strain of Saccharomyces cerevisiae.

[0027] (3) The lemon wine obtained by fermentation of the present invention is obtained by liquid fermentation, which effectively improves the sensory organs and has the advantages of high efficiency and low cost. The fermented lemon wine has a unique flavor. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. It should be understood that the following drawings only represent some embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0029] Figure 1 1 is a microscopic morphology diagram of some strains of the present invention;

[0030] Figure 2 Schematic diagram of color development of some strains of the present invention on an ester production screening plate;

[0031] Figure 3 This is a graph showing the ester production ability of different yeast strains;

[0032] Figure 4 is a growth curve of Meyerozyma guilliermondii YT-60;

[0033] Figure 5 Schematic diagram of the pH tolerance of Meyerozyma guilliermondii YT-60;

[0034] Figure 6Schematic diagram of the tolerance of Meyerozyma guilliermondii YT-60 to sugar content;

[0035] Figure 7 Schematic diagram of the tolerance of Meyerozyma guilliermondii YT-60 to ethanol;

[0036] Figure 8 Schematic diagram of the tolerance of Meyerozyma guilliermondii YT-60 to SO2;

[0037] Figure 9 This is a morphological diagram of Meyer yeast;

[0038] (a) shows the colony morphology of Meyerozyma guilliermondii YT-60; (b) shows the microscopic morphology of Meyerozyma guilliermondii YT-60;

[0039] Figure 10 This is the phylogenetic tree of Meyerozyma guilliermondii YT-60. DETAILED DESCRIPTION

[0040] In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with examples. It should be understood that the specific examples described herein are merely used to explain the present invention and are not intended to limit the present invention. In addition, it should be understood that after reading the content of the present invention, those skilled in the art can make various changes and modifications to the present invention, but these equivalent forms also fall within the scope defined by the appended claims of the application.

[0041] In this application, % not otherwise specified represents weight percentage, i.e. wt%; unqualified reagents are all commercially available; methods or steps not otherwise specified are conventional steps.

[0042] Example 1:

[0043] Yeast enrichment

[0044] Under sterile conditions, 25 g of mature lemon peel was minced and placed in 225 mL of sterile saline, shaken evenly, and 25 mL of the homogenate was pipetted into 225 mL of YPD liquid medium, and placed in an incubator at 28°C for enrichment culture for 36 h.

[0045] Yeast screening

[0046] YPD liquid medium: 20 g peptone, 10 g yeast extract, 20 g glucose, 1 L distilled water (solid medium should be supplemented with 20 g agar). Sterilize at 121°C for 20 min. To eliminate bacterial interference during screening, add 0.06 g chloramphenicol.

[0047] The enriched culture fluid was diluted with sterile physiological saline and 10 -4 , 10 -5 , 10 -6 Place 100 μL of each of the three gradient dilutions (or other suitable gradients) onto a plate containing YPD solid medium containing chloramphenicol and spread evenly with a spreader until the surface of the medium is dry. Place the coated plate in a 28°C incubator for 36 hours. Select strains of varying morphology from the YPD plate for purification. Initially screen for strains with aroma using the olfactory method. These strains are then inoculated into YPD liquid medium for repeated fermentation to observe whether they still produce aroma.

[0048] Identification of ester-producing ability of yeast

[0049] Ester production screening medium: 0.04 g bromocresol purple, 10 mL tributyrin, 10 g yeast extract powder, 20 g peptone, 20 g glucose, 20 g agar, 1 L distilled water. Sterilize at 121°C for 20 min. This experiment must be performed in the dark.

[0050] Eight strains with aroma were selected from the YPD plate. The eight strains were activated and streaked on the ester production screening plate and cultured at 28°C for 36 hours. The results of their ester production ability were as follows: Figure 2 shown.

[0051] The darker the yellow color of the strain on the ester production screening plate, the stronger its ester production ability. Figure 2 It can be seen that all eight strains have a certain ability to produce esters, and strains YT-23 and YT-60 are slightly darker in color than other strains.

[0052] Screening of high ester-producing yeast

[0053] The activated 8 strains were inoculated into YPD liquid culture medium at an inoculum size of 2%, cultured at 28°C for 5 days, and then the total esters were determined.

[0054] The method for determining total esters is the saponification reflux method, and the specific method is as follows:

[0055] (1) Sample pretreatment: Measure 100 ml of fermented YPD medium into a 500 ml round-bottom flask and add 100 ml of distilled water. Connect a condenser and use a 100 ml conical flask to collect the liquid. Turn on the cooling water and slowly heat and distill. When the distillate is collected close to the scale line, remove the conical flask and seal it with sealing film. Cool to room temperature to obtain the sample to be tested, and let it stand for use.

[0056] (2) Determination of total esters: Accurately measure 25 mL of the sample to be tested in a 250 mL round-bottom flask, add 25 mL of deionized water, add 2 drops of 5% phenolphthalein indicator solution, and titrate with 0.1 mol / L sodium hydroxide standard solution until the color turns slightly red and does not fade for 30 seconds. Record the number of milliliters of sodium hydroxide standard solution consumed. Accurately add 20 mL of 0.1 mol / L sodium hydroxide standard solution and record it as V0. After shaking, install a condenser and saponify and reflux in a boiling water bath for 0.5 h. After the liquid cools to room temperature, remove the round-bottom flask. Titrate with 0.1 mol / L hydrochloric acid standard solution until the red color just disappears completely as the endpoint. Record the volume of hydrochloric acid standard solution consumed as V1.

[0057] (3) Data analysis: The total ester content in the sample was calculated according to the following formula.

[0058]

[0059] Where:

[0060] X——total ester content in the fermentation broth sample, expressed as mass concentration (calculated as ethyl acetate), g / L;

[0061] C0——actual molar concentration of sodium hydroxide standard solution, mol / L;

[0062] V0——the volume of sodium hydroxide standard solution added during saponification, mL;

[0063] C1——molar concentration of hydrochloric acid standard solution, mol / L;

[0064] V1——the volume of hydrochloric acid standard solution consumed during titration, mL;

[0065] 25——sampling volume, mL;

[0066] 0.088 – The mass of ethyl acetate in grams equivalent to 1 mL of standard sodium hydroxide solution (C = 1 mol / L).

[0067] The calculation result should be rounded to two decimal places.

[0068] The total ester production capacity of the 8 strains was as follows Figure 3As shown in the figure, the strains with ester production greater than 1.00 g / L are YT-13, YT-23, YT-36, YT-60, and YT-61. Among them, the ester production of YT-60 can reach the maximum of 2.60 g / L, indicating that the ester production ability of YT-60 is significantly higher than that of other strains.

[0069] Determination of growth curve

[0070] The seed solution of the screened yeast YT-60 was activated. 1% of the seed solution was added to 200 ml of YPD liquid medium and cultured at 28°C and 150 rpm for 24 h. The blank YPD liquid medium was used as a control. The absorbance at OD600 nm was measured every 2 h. The results were as follows: Figure 4 As shown, YT-60 began to enter the stable phase at 14 h of culture.

[0071] Determination of pH tolerance

[0072] Activate YT-60 and absorb 1% of the seed solution into 10 ml YPD liquid medium with pH values ​​of 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, and 5.0, respectively. Cultivate at 28°C and 180 rpm for 24 h and measure the absorbance at OD600 nm. The results are as follows: Figure 5 shown.

[0073] Determination of sugar tolerance

[0074] Activate YT-60 and absorb 1% of the seed liquid into 10 ml YPD liquid medium with sugar content (g / L) of 100, 150, 200, 250, 300, 350, and 400 respectively. Cultivate at 28°C and 180 rpm for 24 h and measure the absorbance at OD600 nm. The results are as follows: Figure 6 shown.

[0075] Determination of ethanol tolerance

[0076] Activate YT-60, take 1% of the seed solution and add it to 10ml YPD liquid medium with 95% ethanol added in amounts of 3, 6, 9, and 12% respectively, and culture it at 28℃ and 180rpm for 24h. Measure the absorbance at OD600nm. The results are as follows: Figure 7 shown.

[0077] Determination of SO2 tolerance

[0078] Activate YT-60, absorb 1% of the seed solution and add it to 10ml YPD liquid medium with SO2 addition amount (mg / L) of 100, 200, 300, 400, and 500 respectively, culture it at 28℃ and 180rpm for 24h, and measure the absorbance at OD600nm. The results are as follows: Figure 8 shown.

[0079] Morphological characteristics identification

[0080] The strain was streaked onto YPD agar medium and cultured at 28°C for 48 h. The colony morphology, color, and surface smoothness were observed. Figure 9 As shown in the figure, the YT-60 strain has large colonies, milky white, round, with neat edges, opaque, smooth or rough surface, moist texture, and is easy to pick up; Gram staining is positive (blue-purple), and the colonies are oval or spherical.

[0081] Molecular biology identification

[0082] The strain was streaked onto YPD agar and incubated at 28°C for 48 hours. Total DNA extraction, ITS rDNA amplification, and sequencing were performed by Shanghai Jie Li Biotechnology Co., Ltd. The resulting ITS rRNA spliced ​​sequence was collated and then analyzed by BLAST analysis against the NCBI GenBank database. The species relationship of the strain was determined based on sequence homology.

[0083] The ITS rRNA splicing sequence is shown in SEQ ID NO: 1:

[0084] .

[0085] The phylogenetic tree of strain YT-60 constructed by MEGA 11, as shown Figure 10 As shown in the figure. It can be seen from the figure that the affinity between strain YT-60 and Meyerozyma guilliermondii KY104252.1 reaches 98%. Therefore, it can be determined that the YT-60 strain is Meyerozyma guilliermondii, and the strain was deposited in the Guangdong Provincial Microbiological Culture Collection on April 3, 2025, with the deposit number GDMCC No: 66091. The deposit address is Guangdong Institute of Microbiology, 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Province, and the deposit address postal code is: 510070.

[0086] Example 2

[0087] A fermented lemon wine processing technology comprises the following steps:

[0088] 1. Preparation of Bacterial Suspension

[0089] According to the inoculation ratio of 2%, the seed liquid of Meyerozyma guilliermondii YT-60 preserved in Example 1 was inoculated into YPD liquid medium, and then the inoculated culture liquid was cultured at 28°C and 150 rpm for 14 h to obtain the fermentation stock solution with a strain concentration of 3×10 8 CFU / ml.

[0090] The fermentation stock solution was centrifuged at 8000 rpm and 4°C for 8 min, the supernatant was removed, and the solution was washed three times with sterile saline. Finally, the solution was resuspended with an equal volume of saline to obtain a bacterial suspension, i.e., the bacterial suspension for YT-60 fermentation.

[0091] 2. Fermentation process

[0092] Wash the fresh lemons, remove the seeds and peel, and squeeze the juice. Then add 2 times the amount of purified water and an appropriate proportion of lemon peel without white pulp (the amount of lemon peel without white pulp added is 1 / 9 of the volume of fresh lemon juice). Then add 0.1g / L potassium metabisulfite for sterilization, and then add 0.2g / L pectinase for enzymatic hydrolysis at a constant temperature of 45℃ for 2h. Adjust the sugar content to 23Brix with white sugar.

[0093] 3% YT-60 bacterial suspension and 0.5 g / L Angel Wine Yeast SY were added to the treated lemon juice and fermented at 20°C for 10 days.

[0094] The fermented lemon wine was filtered through a filter and put into sterile glass bottles, sterilized at 80°C for 20 minutes, and aged in a refrigerator at 4°C for 1 month.

[0095] The supernatant of the aged lemon wine is sucked out with a siphon, placed in a sterile glass bottle and sealed to obtain the final lemon wine.

[0096] Example 3

[0097] Flavor determination of lemon wine

[0098] Headspace solid phase microextraction (HS-SPME)

[0099] 4 mL of lemon wine was added to a 15 mL vial, along with 60 μL of 2-octanol (32 g / L) as the internal standard, 1 g of NaCl, and a magnetic stir bar. Headspace extraction was performed at 50°C for 20 minutes. After extraction, the extraction head was inserted into the GC inlet and desorbed for 5 minutes.

[0100] Gas chromatography-mass spectrometry (GC-MS)

[0101] GC conditions: chromatographic column DB-5MS (30m×0.25mm, 0.25mm); heating program: 40℃ for 3min, increase to 160℃ at a rate of 3℃ / min, maintain for 2min, then increase to 230℃ at 8℃ / min, maintain for 3min; He flow rate 1.0mL / min; splitless injection; injection port temperature: 250℃.

[0102] MS conditions: electron ionization source; ion source temperature 250°C; interface temperature 250°C; scan rate 35-450 m / z throughout.

[0103] Qualitative and quantitative analysis: GC-MS experimental data were qualitatively analyzed by similarity search against the NIST 2011 database. 2-Octanol was used as the internal standard for semi-quantification, and the concentrations of each compound were calculated according to the following formula:

[0104]

[0105] The types and contents of volatile flavor compounds in lemon wine fermented by Angel wine yeast SY alone (CK group) and lemon wine fermented by YT-60 strain and Angel wine yeast SY mixed culture (EG group) are shown in Table 1.

[0106] Table 1 Comparison of volatile flavor substances in single-bacteria fermentation and mixed-bacteria fermentation Unit: ug / L

[0107] Flavor substances CK EG Alcohols Phenylethyl alcohol 3248.91±1.23 4529.47±2.30 Isoamyl alcohol 2371.28±1.75 2814.59±0.92 Nerolidol 12.18±0.88 22.45±1.67 Linalool 6.15±0.97 25.38±2.35 4-terpene alcohol 366.79±1.77 632.54±1.39 α-Terpineol 524.84±1.75 859.32±0.55 Geraniol - 22.78±0.25 Citronellol 428.95±1.33 513.76±1.36 glycerin - 497.23± Acids bitter 398.11±2.91 253.75±1.14 Esters Ethyl acetate 587.27±0.78 1834.33±1.86 Ethyl nonanoate 1.88±0.05 16.51±0.75 Ethyl isovalerate 42.65±0.41 127.24±1.62 Ethyl decanoate 7.53±1.09 36.82±2.02 Ethyl laurate 0.22±0.01 1.70±0.30 Ethyl palmitate - 0.35±0.05 Aldehydes Nonanal 4.28±1.09 8.13±2.01 Terpenes Ocimene 2.43±0.95 13.62±2.17 D-limonene 62.83±1.30 128.75±1.81

[0108] Note: “-” means not detected.

[0109] As shown in Table 1, GC-MS analysis detected a total of 20 volatile compounds in both the single-bacteria and mixed-bacteria fermentation groups, with 17 in the single-bacteria fermentation group (CK) and 20 in the mixed-bacteria fermentation group (EG). The EG group contained 9 alcohols, 1 acid, 6 esters, 1 aldehyde, and 2 terpenes, an increase of 1 ester and 2 alcohols compared to the CK group. The overall content of these compounds increased slightly, with the ethyl acetate content increasing significantly.

[0110] The geraniol added to the EG group has a tropical fruit and grassy flavor, while glycerol can make the wine more rounded, and ethyl palmitate gives the lemon wine a unique creamy aroma.

[0111] The above-described embodiments merely represent specific implementation methods of the present application. While the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of protection of the present application. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the technical concept of the present application, and all such variations and improvements fall within the scope of protection of the present application.

[0112] This background section is provided to generally present the context of the invention, and the work of the presently named inventors, the work to the extent described in this background section, and aspects of the description in this section that did not constitute prior art at the time of filing are neither explicitly nor implicitly admitted to be prior art to the present invention.

Claims

1. A strain of Meyerozyma guilliermondii YT-60 was deposited in the Guangdong Provincial Microbial Culture Collection on April 3, 2025, with the deposit number GDMCC NO: 66091. The depository address is Guangdong Institute of Microbiology, 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

2. Use of the Meyerozyma guilliermondii YT-60 according to claim 1 in preparing fermented lemon wine.

3. A product containing the Meyerozyma guilliermondii YT-60 according to claim 1, characterized in that: Therefore, the products include but are not limited to a bacterial suspension containing Meyerozyma guilliermondii YT-60, a bacterial slurry containing Meyerozyma guilliermondii YT-60, and a bacterial agent containing Meyerozyma guilliermondii YT-60.

4. A fermented lemon wine, characterized in that The method comprises using the Meyerozyma guilliermondii YT-60 according to claim 1 or the product containing the Meyerozyma guilliermondii YT-60 according to claim 3.

5. The method for preparing fermented lemon wine according to claim 4, wherein The following steps are involved: 1) Fresh lemons are washed, seeded, peeled, and juiced, and then purified water and lemon peel without white pulp are added. Potassium metabisulfite is then added for sterilization, followed by pectinase enzymolysis, and the sugar concentration is adjusted; 2) preparing a bacterial suspension containing Meyerozyma guilliermondii YT-60; then adding the bacterial suspension and Angel Wine Yeast SY to the lemon juice treated in step 1) for static fermentation to obtain lemon wine; 3) filtering the lemon wine in step 2) through a filter screen and then filling it into a sterile glass bottle, sterilizing it, aging it, and collecting the supernatant to obtain a finished lemon wine.

6. The method for preparing fermented lemon wine according to claim 5, wherein: In step 1), the amount of purified water added is 1-3 times the volume of fresh lemon juice; the amount of lemon peel without white pulp added is 1 / 9-1 / 10 the volume of fresh lemon juice; the amount of potassium metabisulfite added is 0.08-0.1 g / L; the amount of pectinase added is 0.2-0.4 g / L; enzymolysis is carried out under constant temperature conditions, the enzymolysis temperature is 45-55° C., the time is 2-3 hours, and the sugar concentration is adjusted to 23 Brix with white sugar.

7. The method for preparing fermented lemon wine according to claim 5, wherein: The preparation steps of the bacterial suspension containing Meyerozyma guilliermondii YT-60 in step 2) are as follows: The seed liquid was inoculated into YPD liquid medium to obtain culture solution; the culture solution was then fermented to obtain fermentation stock solution. At this time, the strain concentration was 2×10 8 CFU / mL-6×10 8 CFU / mL; The fermentation stock solution was centrifuged at 8000 rpm and 4° C. for 8 min, the supernatant was removed, and the solution was washed three times with sterile saline and finally resuspended with an equal volume of saline to obtain a bacterial suspension.

8. The method for preparing fermented lemon wine according to claim 5, wherein: In step 2), the amount of the bacterial suspension containing Meyerozyma guilliermondii YT-60 added is 3% of the volume of the treated lemon juice; the amount of Angel Wine Yeast SY added is 0.2-0.8 g / L; the static fermentation temperature is 20-25° C., and the fermentation time is 10-12 days.

9. The method for preparing fermented lemon wine according to claim 5, wherein: In step 3), the sterilization temperature is 80° C. and the time is 20 minutes; the aging temperature is 4° C. and the time is 1 month.

10. A fermented lemon wine prepared by the method according to any one of claims 5 to 9, characterized in that: Fermented lemon wine contains 20 volatile compounds, with a high content of ethyl acetate. The geraniol it contains gives the fermented lemon wine a tropical fruit and grassy flavor; the glycerol it contains can make the wine body more rounded, and the ethyl palmitate it contains gives the lemon wine a unique creamy aroma.

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