A method for increasing volatile flavour compounds in kiwifruit wine

Abstract

The application discloses a method for improving volatile flavor compounds of kiwi fruit wine and belongs to the field of fruit wine brewing. 127 strains are obtained through TTC culture medium screening, and one Pichia kudriavzevii FPW7-6 is obtained through screening of the 127 strains. The fruit wine prepared by fermentation of kiwi fruit by using the Pichia kudriavzevii FPW7-6 exhibits higher fruitiness, sweetness, sourness and floral aroma than AQ. In the kiwi fruit wine, the content of beta-damascenone reaches 3.4 mu / g / L, the content of linalool is 338.3 mu / g / L, the content of 2-methyl butyric acid ethyl ester is 117.7 mu / g / L, the content of gamma-butyrolactone is 254.0 ug / L, and the content of phenylethanol is 63265.6 mu / g / L. The Pichia kudriavzevii FPW7-6 is suitable for kiwi fruit wine fermentation and can make the kiwi fruit wine produce more excellent flavor.

Description

A method for improving the volatile flavor compounds in kiwifruit wine Technical Field

[0001] This invention relates to a method for improving the volatile flavor compounds in kiwifruit wine, belonging to the field of fruit wine brewing. Background Technology

[0002] Kiwifruit, sweet and delicious, is an excellent raw material for making fruit wine. Traditionally, kiwifruit wine is mainly fermented using Saccharomyces cerevisiae. For example, patent application CN103160405A discloses a wild kiwifruit wine and its brewing method, which involves preparing kiwifruit pulp, using Saccharomyces cerevisiae, and undergoing primary fermentation, secondary fermentation of the juice, and aging to obtain kiwifruit wine. Another example is patent CN102776098A, which discloses a kiwifruit wine and its brewing method, made from kiwifruit, white sugar, potassium bicarbonate, sulfur dioxide, pectinase, and Saccharomyces cerevisiae through processes such as kiwifruit pulp preparation, primary fermentation, pressing and separation, secondary fermentation, and aging. However, kiwifruit wine prepared using Saccharomyces cerevisiae has a bland flavor and lacks distinctive characteristics, and there is a lack of yeast strains specifically designed to enhance the flavor of kiwifruit wine during fermentation. Therefore, screening for non-brewing yeasts that can accelerate fermentation and enhance the volatile flavor compounds in kiwifruit is of great significance for improving the quality and value of kiwifruit wine. Summary of the Invention

[0003] To address the shortcomings of the existing technology, this invention provides a method for improving the volatile flavor compounds in kiwifruit wine, aiming to solve the technical problem of the lack of yeast strains specifically designed to enhance the flavor of kiwifruit wine during fermentation.

[0004] The first technical solution provided by this invention is a strain of Pichia kudriavzevii FPW7-6, which was deposited at the China Center for Type Culture Collection on July 1, 2024, with accession number CCTCC NO:M 20241440.

[0005] The second technical solution provided by the present invention is a microbial preparation containing Pichia pastoris FPW7-6 as described in the first technical solution.

[0006] In some embodiments, the amount of Pichia pastoris FPW7-6 added to the microbial preparation is at least 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

[0007] Furthermore, the amount of Pichia pastoris FPW7-6 added to the microbial preparation is at least 1×10⁻⁶. 8 CFU / mL or 1×10 8 CFU / g.

[0008] The third technical solution provided by the present invention is a fermentation agent containing Pichia pastoris FPW7-6 as described in the first technical solution or the microbial preparation described in the second technical solution.

[0009] In some embodiments, the amount of Pichia pastoris FPW7-6 added to the fermentation agent is at least 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

[0010] Furthermore, the amount of Pichia pastoris FPW7-6 added to the fermentation agent is at least 1×10⁻⁶. 8 CFU / mL or 1×10 8 CFU / g.

[0011] The fourth technical solution provided by this invention is the application of Pichia pastoris FPW7-6 as described in the first technical solution, or the microbial preparation described in the second technical solution, or the third fermentation agent in fruit wine brewing.

[0012] In some embodiments, the fruit wine is kiwi fruit wine.

[0013] The fifth technical solution provided by the present invention is a method for brewing kiwifruit wine, wherein the method uses kiwifruit juice to ferment Pichia pastoris FPW7-6 as described in the first technical solution, the microbial preparation as described in the second technical solution, or the fermentation agent as described in the third technical solution to prepare kiwifruit wine.

[0014] In some embodiments, the fermentation parameters are: fermentation temperature of 20~30℃ and fermentation time of not less than 48h.

[0015] The sixth technical solution provided by the present invention is a method for improving the volatile flavor compounds of kiwifruit wine. The method involves fermenting kiwifruit juice using Pichia pastoris FPW7-6 as described in the first technical solution, the microbial preparation as described in the second technical solution, or the fermentation agent as described in the third technical solution.

[0016] In some embodiments, the sugar content of the kiwi juice in the fermentation system is 20-22%.

[0017] In some embodiments, the fermentation parameters are: fermentation temperature of 20~30℃ and fermentation time of not less than 48h.

[0018] In some embodiments, the volatile flavor compounds include β-damascone, linalool, ethyl 2-methylbutyrate, γ-butyrolactone, and phenylethanol.

[0019] The technical effects of this invention are as follows:

[0020] This invention obtained 127 strains of Pichia pastoris through screening on TTC medium, and further screened these 127 strains to obtain Pichia pastoris FPW7-6. Kiwi wine prepared using Pichia pastoris FPW7-6 exhibits higher levels of fruit flavor, sweetness, acidity, and floral aroma compared to AQ wine. The kiwi wine contains 3.4 μg / L of β-damascone, 338.3 μg / L of linalool, 117.7 μg / L of ethyl 2-methylbutyrate, 254.0 μg / L of γ-butyrolactone, and 63265.6 μg / L of phenylethanol. Pichia pastoris FPW7-6 is suitable for kiwi wine fermentation and can produce a superior flavor.

[0021] Certificate of Preservation of Biological Materials

[0022] A strain of Pichia kudriavzevii FPW7-6, taxonomically named Pichia kudriavzevii FPW7-6, was deposited on July 1, 2024, at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, China, with accession number CCTCC NO: M 20241440. Attached Figure Description

[0023] Figure 1 shows TTC staining (a) and colony morphology (b) of some strains.

[0024] Figure 2 shows the sensory evaluation of different kiwi wines.

[0025] Figure 3 shows the morphological characteristics of yeast strain FPW7-6. a: Single colony morphology on TTC solid medium cultured at 28℃ for 48 h; b: Colony morphology under a microscope.

[0026] Figure 4 shows the phylogenetic tree of FPW7-6 yeast based on ITS rDNA sequence data neighbor-joining analysis.

[0027] Figure 5 shows the aroma pleasantness (Figure A) and the content of important aroma compounds in kiwi wine fermentation with different yeasts; B: β-damascone; C: linalool; D: ethyl 2-methylbutyrate; E: γ-butyrolactone; F: phenylethanol. Detailed Implementation

[0028] The preferred embodiments of the present invention are described below. It should be understood that the embodiments are for better explanation of the present invention and are not intended to limit the present invention.

[0029] Test method:

[0030] 1. The determination of total acidity, ascorbic acid and reducing sugar content was carried out in accordance with the national standard GB / T 15038-2006 "General Analytical Methods for Wine and Fruit Wine".

[0031] 2. The pH value was measured by a pH meter.

[0032] 3. Total phenols were determined using the Folin-Ciocalteu colorimetric method: Standard curve preparation: Gallic acid solutions of different concentrations were prepared by adding 1000, 980, 960, 940, 920, and 900 μL of distilled water to 0, 20, 40, 60, 80, and 100 μL of gallic acid, respectively. OD values ​​were measured at 765 nm using a mixture of 50 μL gallic acid, 50 μL Folin-Ciocalteu, 50 μL sodium carbonate, and 100 μL distilled water.

[0033] Experimental group: 50 μL sample + 50 μL Folin-Ciocalteu + 50 μL 7.5% sodium carbonate + 100 μL distilled water, let stand for 30 min, and measure the OD value at 765 nm.

[0034] 4. The alcohol content was measured using a handheld alcohol meter.

[0035] 5. The sugar content was measured using a handheld saccharimeter.

[0036] 6. Headspace Solid-Phase Microextraction (HS-SPME): HS-SPME combined with gas chromatography-mass spectrometry (GC-MS) (Agilent 8890B GC, 5977B MS; Agilent Technologies, Santa Clara, CA, USA) was used to analyze volatile compounds produced at the end of kiwi wine fermentation. Accurately aspirate 5 mL of sample and add 1 mL of saturated NaCl solution to a 15 mL SPME flask. Then add 25 μL of toluene-d8 (2.024 mg / L) and 50 μL of ethyl acetate-d3 (19.7 mg / L) as internal standards. After sealing, equilibrate at 40 °C for 15 min, insert the aged extraction head, and extract at 300 rpm / min for 30 min. After extraction, insert the syringe into the headspace vial (22 mm depth), incubate at 40 °C for 10 min, and collect the volatile substances for GC-MS analysis. GC-MS conditions: A DB-5MS column (60 m × 250 μm, film thickness 0.25 μm; Agilent, Santa Clara, CA, USA) was used with helium as the carrier gas at a flow rate of 1.0 mL / min. The column oven temperature was initially set at 40 °C for 1 min, then increased to 230 °C at a rate of 4 °C / min and held for 15 min. Mass spectrometry analysis was performed at an ion source temperature of 230 °C, a mass range of m / z 29–500, and an electron ionization energy of 70 eV. Identification of volatile compounds involved comparison of mass spectrometry and retention index (Rl). The relative retention times of volatile compounds were calculated using a modified Kovats method based on the linear retention times of the n-alkanes (C, ConDB-WAX) column.

[0037] 7. Qualitative and quantitative analysis of aroma compounds: Detected unknown compounds were matched with the National Institute of Standards and Technology (NIST) mass spectrometry library and the Wiley Library. Substances with high matching scores were selected as effective aroma components for qualitative comparison. Semi-quantitative analysis was performed using the internal standard method. Toluene-d8 and ethyl acetate-d3 were used as internal standards. The mass concentration of the analytes was calculated after correction by comparing the peak area ratio of each component with the internal standards.

[0038] Raw materials used in the examples:

[0039] 1. Angel Yeast (Brewing Yeast, abbreviated as AQ): Angel Yeast Co., Ltd.

[0040] 2. Kiwifruit (MHT), loquat (PP), mulberry (SS), blueberry (LM), Shine Muscat (PTX), and Jasmine grapes (TZ) were purchased from Guida Fresh Fruit Shop in Laochaoyang Village, Huaxi District (Guizhou Province, Guiyang). Commercial pickled peppers (PJ) were purchased from Huimin Fresh Supermarket in Laochaoyang Village, Huaxi District (Guizhou Province, Guiyang).

[0041] 3. YPD liquid culture medium (100 mL): Weigh 1 g of yeast extract, 2 g of tryptone, and 2 g of glucose, add distilled water to make up to 100 mL, autoclave at 121℃ for 15 min and set aside.

[0042] 4. Kiwi juice YPD liquid culture medium (100 mL): Weigh 1 g of yeast extract and 2 g of tryptone, add 20% kiwi juice with sugar content to make up to 100 mL, autoclave at 121℃ for 15 min and set aside.

[0043] 5. Yeast Extract Peptone Glucose Agar (YPD) Solid Medium (100 mL): Weigh 4.9 g of sample, add 100 mL of distilled water, heat to dissolve, and autoclave at 115℃ for 30 min before use.

[0044] 6. TTC medium (100 mL): Weigh 3.3 g of TTC nutrient agar, add 100 mL of distilled water, stir and heat until completely dissolved, and autoclave at 121℃ for 15 min.

[0045] Example 1

[0046] I. Preliminary Screening Using the TTC Method

[0047] Rinse the purchased fruits and pickled peppers with YPD liquid culture medium in a clean bench. Add the obtained bacterial culture to a shaker tube (3-5 mL), centrifuge at 5000 rpm for 5 min, discard the supernatant, and dilute with sterile water to 10. -3 and 10 -4 Gradient method. Spread 20 μL of each culture onto TTC solid medium for yeast screening and incubate at 28 ℃ for 48-72 h. Observe the color development of the medium, pick single colonies with good growth and purify them by streaking TTC plates, and observe the colony morphology.

[0048] II. Duchenne tubule method for secondary screening

[0049] After the strains obtained from the initial screening by the TTC method were activated by inoculation into YPD medium, the inoculation amount was 2% (1×10⁻⁶). 8(CFU / mL) was inoculated into 5 mL of kiwi juice YPD liquid medium containing DuPont tubes and cultured at 28℃ and 50 rpm / min for 72 h. The bubble production was observed and recorded every 24 h, and the number of the strain with excellent gas production performance was recorded.

[0050] 127 strains were obtained by screening on TTC medium. The gas production of these strains and the AQ strain after 36 h of Durham tube fermentation was summarized and compared (as shown in Table 1). Among them, strains MHT-6-11, MHT-8-49, MHT-8-58, MHT-8-66, PP-2-16, PP-2-25, PP-5-3, FPW7-6, PP-7-12, PJ-I-5, PJ-I-6, TZ-1-9, TZ-1-11, TZ-2-1, LM-1-4, LM-1-5 and AQ strains were observed to have 1 / 3 of their Durham tubes filled with gas after 24 h of culture, indicating that these strains have rapid gas production capacity. After 48 hours of cultivation, strains MHT-1-8, MHT-6-16, MHT-8-42, PP-2-5, and PP-3-1 began to produce gas, but in small volumes. At 72 hours, strains MHT-8-49, MHT-8-66, PP-2-16, PP-2-25, PP-5-3, FPW7-6, PJ-I-5, PJ-I-6, LM-1-4, and AQ Durbin tubes were filled with gas and floated completely on the surface. Therefore, through preliminary screening, 10 yeast strains with strong gas-producing capabilities suitable for kiwifruit wine fermentation were obtained.

[0051] Table 1. Gas production in the Durham tubules of some bacterial strains after 72 hours.

[0052] Strain ID 24h48h72h Strain ID 24h48h72h MHT-1-3--+PP-4-7--+MHT-1-8-++PP-4-8-++MHT-6-4--+PP-5-3++++++MHT-6-11-+++PP-6-4-++MHT-6-14--+PP-6-8-++MHT-6-16-+++PP-6-10--+MHT-6-17-+++PP-6-11-+ ++++MHT-6-25--+PP-7-6+++++++MHT-6-67--++PP-7-9--+MHT-6-68-++++PP-7-12-++MHT-6-69-++PJ -I-1-++++MHT-8-37-++++PJ-I-4-++MHT-8-42-++PJ-I-5++++++MHT-8-44-++PJ-I-6++++++MHT-8-49+ +++++PJ-I-7--+MHT-8-52-++PJ-4-28+++MHT-8-53--+PJ-4-29-++MHT-8-55--+SS-2-1--+MHT-8-58+ ++SS-3-1--+MHT-8-66++++++TZ-1-6-++MHT-8-68-+++TZ-1-9++++MHT-8-70-++TZ-1-10-+++MHT-8-7 3--+TZ-1-11++++MHT-8-80--+TZ-2-1++++PP-2-5-+++LM-1-4++++++PP-2-16++++LM-1-5++++PP-2-25++++++PTX-3-16-++++PP-2-34-++PTX-3-17-++++PP-3-1-+++PTX-4-2-++++PP-3-2-+++Angel Yeast+++++++ surface

[0053] Note: "-" indicates that no gas was produced in the Durbin tubules; "+" indicates that the gas volume is <1 cm; "++" indicates that the gas volume is 1.5~2 cm; and "+++" indicates that the gas volume is 2~3 cm.

[0054] After fermenting kiwifruit wine using 10 single-strain yeasts, the aroma characteristics of the kiwifruit wine were evaluated by 10 trained winemaking professionals aged 24-35 years. In the sensory experiment, samples were randomly coded and assigned. Six aroma attributes for evaluating the aroma of kiwifruit wine were defined as: fruity, alcoholic, sweet, acidic, medicinal, floral, and herbal. Judges scored the intensity of aroma characteristics: 0 = no aroma characteristic, 1 = very weak, 2 - weak, 3 = medium, 4 - strong, 5 = very strong. Each tested wine sample was analyzed twice. Sensory results showed that AQ and 5 non-Sacchariform yeasts scored higher. The results are shown in Figure 2. It can be seen that FPW7-6 and PP-5-3 groups exhibited higher levels of fruity, sweet, acidic, and floral flavors compared to AQ. LM-1-4 showed a significant acidic palate characteristic, but its fruity and sweet aroma intensity was not significant; excessive acidity negatively impacted the flavor of the wine sample. M-8-49 exhibits different chemical properties, but its overall aroma intensity is relatively low. In conclusion, FPW7-6 fermented kiwifruit wine improves the sensory quality of kiwifruit wine to a certain extent and has promising market prospects.

[0055] III. Morphological Identification of High-Gasein-Producing Yeast Strains

[0056] Observe the colony morphology and color on the culture medium, and observe the cell morphology of single colonies with typical yeast characteristics under a microscope.

[0057] 1. Results of strain morphology identification

[0058] Based on the color and morphology of the colonies, single colonies were picked and placed on a glass slide. The characteristics of the picked cells were observed under a 100x microscope. The microscopic examination results of strain FPW7-6 are shown in Figure 3.

[0059] 2. Results of molecular identification of the strain

[0060] The ITS1 and ITS4 gene regions of four non-Saccharomyces cerevisiae strains were amplified. Sequences of selected strains representing potential species were obtained from GenBank and multiple alignments were performed. Phylogenetic analysis was conducted using MEGA7 software. As shown in Figure 4, FPW7-6 was identified as *Pichia kudriavzevii*, showing the highest homology (100%) with *Pichia kudriavzevii*, and belonging to the same evolutionary branch as *Pichia kudravzevii* SA-NS2 (OR672915). Many studies have shown that non-Saccharomyces cerevisiae can improve the quality of kiwifruit wine. Therefore, using *Pichia kudriavzevii* FPW7-6 as the fermentation strain for kiwifruit wine is of great significance.

[0061] Example 2

[0062] After activating the PP-5-3, FPW7-6, M-8-49, and LM-1-4 strains from Example 1, as well as AQ, inoculated them into YPD medium, the kiwifruit wine was prepared using the following method:

[0063] 1. Select kiwifruit that are 8-9 ripe, wash, peel, juice, and sterilize to obtain the raw material.

[0064] 2. Add potassium metabisulfite (65mg) and pectinase (500mg) to the raw materials, and enzymatically hydrolyze them in a constant temperature water bath at 35℃ for 2 hours to obtain kiwi juice. Adjust the sugar content by adding glucose to make the sugar content reach 20 Brix, and then sterilize.

[0065] 3. Inoculate strain FPW7-6 into YPD liquid medium at a 4% (v / v) inoculation rate and incubate at 28℃ and 280 rpm / min for 8 h. Then, inoculate it into sterilized kiwifruit pulp at a 2% inoculation rate for pre-fermentation (fermentation in an interfermentation chamber, 20-22℃, 7 days). Centrifuge and collect the supernatant for post-fermentation (fermentation, 20-22℃, 10 days). Sterilize, centrifuge, collect the supernatant (i.e., kiwifruit wine), and seal and bottle it.

[0066] First, the physicochemical indicators of the kiwifruit wine were tested, and the results are shown in Table 2. The testing of these physicochemical indicators is crucial in kiwifruit wine production. It ensures the quality and flavor of the wine, assesses nutritional components (such as vitamin C and total phenols), and improves the stability and shelf life of the wine (e.g., pH and total acidity control). Furthermore, physicochemical properties directly affect consumer acceptance, providing a scientific basis for product optimization and market positioning, and helping to enhance the product's market competitiveness.

[0067] Table 2. Determination of physicochemical indicators of fermented kiwi wine by strains. Note: Data in the same column with different lowercase letters above them indicate significant differences (P<0.05), while data with any identical lowercase letters or no letters indicate no significant differences (P>0.05).

[0068] Strain number PP-5-3FPW7-6MHT-8-49LM-1-4 Angel yeast pH 3.8±0.01 3.85±0.00 3.95±0.05 3.8±0.04 3.85±0.00 Sugar content (°Brix) 7.30±0.00 d 13.00±0.12 b 11.00±0.01 c 18.00±0.02 a 7.30±1.3 d Alcohol content (%Vol): 4.70 ± 0.08 b 6.10±0.00 a 1.00±0.01 d 2.20±0.03c 5.20±0.20 ab Total acid (g / L) 8.89±0.11 bc 9.56±0.20 bc 8.74±0.40 c 13.03±0.97 a 9.78±0.12 b Total phenols (mg / mL): 393.65 ± 6.17 b 445.52±21.02 a 353.52±5.0 c 447.52±12.36 a 352.13±8.32 c Reducing sugar (g / 100g) 7.84±0.13 b 7.39±0.10 c 4.68±0.06 d 14.46±0.44 a 1.79±0.02 e Ascorbic acid (g / L) 1.13±0.02 b 1.16±0.01 a 0.96±0.03 c 1.18±0.02 a 0.18±0.00 d surface

[0069] Aroma pleasantness scores were assessed for the above-mentioned fruit wines. The aroma pleasantness scores of strains PP-5-3, FPW7-6, M-8-49, and LM-1-4 were significantly higher than that of AQ (Figure 5A). Among them, FPW7-6 had an aroma pleasantness score of 3.4, which was 30.8% higher than AQ. Then, quantitative analysis of volatile compounds in these five yeast strains was performed, and the contents of five common compounds in fruit wines were compared. β-Damastone is an important terpene ketone compound in fruit wine, possessing apple and sweet aroma properties. Its aroma threshold in fruit wine is 0.05 μg / L. As shown in Figure 5B, the content of β-damastone in fruit wine prepared by strain FPW7-6 was 3.4 μg / L, which is 2.8 times that of AQ. Figure 5C shows the quantitative results of linalool. Aromatic alcohols possess floral aroma properties. The concentration in fruit wine is 15 μg / L, while the concentration in fruit wine prepared by strain FPW7-6 is 338.3 μg / L, significantly higher than the linalool content in AQ, being 1.4 times that of AQ. Ethyl 2-methylbutyrate has a typical fruity aroma. The concentration in fruit wine prepared by strain FPW7-6 is 117.7 μg / L, 1.3 times that of AQ (Figure 5D). γ-Butyrolactone has creamy and caramel aroma properties. The concentration in fruit wine prepared by strain FPW7-6 is 254.0 μg / L, 1.6 times that of AQ (Figure 5E). Phenylethanol has a rose aroma. The concentration in AQ is 24255.6 μg / L, while the concentration in FPW7-6 is 63265.6 μg / L, significantly higher than AQ, being 2.6 times that of AQ (Figure 5F).

[0070] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A strain of Pichia kudriavzevii FPW7-6, characterized in that, The Pichia pastoris FPW7-6 was deposited at the China Center for Type Culture Collection on July 1, 2024, with accession number CCTCC NO:M20241440.

2. A microbial preparation containing the Pichia pastoris FPW7-6 as described in claim 1.

3. The microbial preparation according to claim 2, characterized in that, The amount of Pichia pastoris FPW7-6 added to the microbial preparation is at least 1×10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

4. A fermenting agent, characterized in that, The fermentation agent contains Pichia pastoris FPW7-6 as described in claim 1 or the microbial preparation as described in claim 2 or 3.

5. The application of Pichia pastoris FPW7-6 as described in claim 1, or the microbial preparation as described in claim 2 or 3, or the fermenting agent as described in claim 4, in the brewing of kiwifruit wine containing volatile flavor compounds, characterized in that... The volatile flavor compounds include β-damascone, linalool, ethyl 2-methylbutyrate, γ-butyrolactone, and phenylethanol.

6. A method for brewing kiwifruit wine containing volatile flavor compounds, characterized in that, The method involves fermenting kiwifruit juice with Pichia pastoris FPW7-6 as described in claim 1, or the microbial preparation as described in claim 2 or 3, or the fermenting agent as described in claim 4 to prepare kiwifruit wine. The volatile flavor compounds include β-damascone, linalool, ethyl 2-methylbutyrate, γ-butyrolactone, and phenylethanol.

7. The method according to claim 6, characterized in that, The fermentation temperature is 20~30℃, and the fermentation time is no less than 48 hours.

8. A method for improving the volatile flavor compounds in kiwifruit wine, characterized in that, The method involves fermenting kiwifruit juice using Pichia pastoris FPW7-6 as described in claim 1, or the microbial preparation as described in claim 2 or 3, or the fermenting agent as described in claim 4; compared with Angel wine yeast, it increases the volatile flavor compounds in kiwifruit wine, wherein the volatile flavor compounds include β-damascone, linalool, ethyl 2-methylbutyrate, γ-butyrolactone, and phenylethanol.

9. The method according to claim 8, characterized in that, In the fermentation system, the sugar content of the kiwi juice is 20-22%; the fermentation parameters are: fermentation temperature of 20-30℃ and fermentation time of not less than 48h.

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