Wickerhamomyces anomalus JG-WA1 and application of Wickerhamomyces anomalus JG-WA1 in aroma-enhancing fruit wine
By using the abnormal Wickham yeast JG-WA1 strain selected from the yeast starter for stepwise fermentation, the problem of insufficient fruit wine flavor was solved, and the aroma and flavor of the fruit wine were enriched and enhanced, especially in cider.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- JINAN INST OF FRUIT PRODS CHINA GENERAL SUPPLY & MARKETING COOP
- Filing Date
- 2026-01-12
- Publication Date
- 2026-05-12
AI Technical Summary
In existing fruit wine fermentation processes, the unique aromatic substances of the fruit itself are not fully released and transformed, resulting in insufficient complexity of fruit wine flavor and distinct fruit aroma. Furthermore, existing compound yeast fermentation processes exhibit inter-strain antagonism, which affects the fermentation effect.
A stepwise fermentation was carried out using the abnormal Wickham yeast JG-WA1 strain selected from the yeast starter. The abnormal Wickham yeast JG-WA1 was inoculated first, followed by brewer's yeast. The stepwise fermentation method was used to improve the aroma content of the fruit wine.
It significantly improves the flavor and quality of fruit wine, especially the aroma layers of apple cider, enhances fruit aroma and nutrient retention, and improves the overall taste of fruit wine.
Smart Images
Figure CN122012262A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of fermentation engineering technology, and specifically relates to an abnormal Wickham yeast JG-WA1 and its application in flavoring fruit wines. Background Technology
[0002] In recent years, fruit wine, as an important representative of low-alcohol beverages, has become increasingly popular among consumers and in the market due to its natural ingredients, mild taste, and nutritional benefits. Fruit wines generally have a low alcohol content, combining fruit flavors with a mellow aroma, meeting modern consumers' dual needs for healthy and functional foods. However, the fruit wine industry still faces some common challenges: many products use relatively traditional fermentation processes with single fermentation strains, resulting in the insufficient release and transformation of the fruit's unique aromatic substances. The finished product still has room for improvement in flavor complexity, fruit aroma intensity, and nutrient retention. To enhance the flavor complexity of fruit wines, current technologies often employ compound yeasts for fermentation, but this can negatively impact fermentation results due to antagonistic interactions between strains.
[0003] Abnormal Wickham yeast ( Wickerhamomycesanomalus This yeast is an ascomycete yeast belonging to the order Yeastales and family Yeastaceae. It is widely found in environments such as fruits, nectar, yeast starters, and natural soil. This yeast has a variety of functional properties, and its fermentation ability has been utilized in the fermentation of products such as wine, beer, and bread in the past. It can contribute unique flavors and improve the quality of the products. For example, patent application number 202411504047.1 describes the preparation of a compound yeast agent by combining abnormal Wickham yeast with brewing yeast and applying it to the fermentation of passion fruit wine. However, the simultaneous fermentation of the compound yeast agent may lead to antagonistic effects between the strains, which is not conducive to maintaining the vitality of the strains. Patent application number 202510650407.7 discloses the application of an abnormal Wickham yeast strain with high esterase production in solid-state fermentation koji to improve the esterase activity of koji. Its main application scenario is the esterification and aroma enhancement of high-proof grain distilled spirits. Patent application number 202510958944.8 discloses an abnormal Wickham yeast strain with high floral and fruity aroma and its cultivation method, and describes its application in the brewing of soy sauce and other soy sauce-flavored foods such as baijiu, huangjiu, and soy sauce.
[0004] Therefore, screening an abnormal Wickham yeast strain with high aroma-enhancing activity and adjusting the fermentation process to enrich the aroma layers of fermented fruit wine and improve its quality is of great practical significance. Summary of the Invention
[0005] To address the problems of bland taste and insufficient aroma in current fruit wines on the market, this invention provides an abnormal Wickham yeast strain JG-WA1 screened from yeast starter, its application in flavor-enhancing fruit wines, and its application method. Using the abnormal Wickham yeast JG-WA1 provided by this invention for stepwise fermentation of apple juice can significantly improve the aroma and enrich the flavor profile of apple wine, showing broad application prospects.
[0006] The technical solution of the present invention is as follows: The first aspect of this invention is the provision of an abnormal Wickham yeast strain JG-WA1, isolated from brewing koji, which has the following phenological characteristics: colonies are usually round, medium-sized, reaching 1-3 mm in diameter after 2-3 days of cultivation, typically milky white to cream-colored, with a smooth, moist, viscous surface and neat edges. Based on these phenological characteristics and gene sequence determination, it was identified as an abnormal Wickham yeast (JG-WA1). Wickerhamomycesanomalus This strain was deposited on September 9, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Biology, Chinese Academy of Sciences, with accession number CGMCC NO.35866.
[0007] A second aspect of the present invention is the application of the aforementioned abnormal Wickham yeast JG-WA1 in flavoring fruit wines, wherein the fruit wines include, but are not limited to, any one of apple wine, pear wine, grape wine, blueberry wine, mulberry wine, mango wine, pineapple wine, lychee wine, passion fruit wine, lemon wine, peach wine, and plum wine; preferably, any one of apple wine and pear wine; more preferably, apple wine.
[0008] Preferably, the abnormal Wickham yeast JG-WA1 enhances the aroma of fruit wine by increasing the content of the following components: 2-pentanol, ethyl propionate, propyl acetate, 3-methyl-2-pentanone, 3-penten-2-one, hexyl hexanoate, ethyl hexanoate, methyl acetate, heptanal, 2-octanone, and 2,3-dimethylpyrazine.
[0009] Of the above ingredients, 2,3-dimethylpyrazine has nutty and coffee aromas, which can significantly enhance the aroma of cider. Apart from that, the other ingredients all have a sweet fruity aroma.
[0010] The third aspect of this invention is to provide a fermentation method for flavoring cider using *Saccharomyces cerevisiae* JG-WA1, specifically employing a stepwise fermentation method, the specific operation of which is as follows: Crush the apples and press to extract the juice. Add 0.5% to 1.5% potassium metabisulfite by weight of the apple juice. After 20 to 30 minutes, inoculate 3 to 10 mL of abnormal Wickham yeast JG-WA1 into the apple juice. Let it ferment at 20 to 30°C for 12 to 72 hours. Then, add 0.2% to 0.3% active dry yeast of Saccharomyces cerevisiae by weight of the apple juice and let it ferment at 20 to 30°C.
[0011] In this invention, the fermentation order of *Saccharomyces cerevisiae* JG-WA1 and *Saccharomyces cerevisiae* is crucial. If *Saccharomyces cerevisiae* is inoculated first and then *Saccharomyces cerevisiae* JG-WA1 is inoculated, the alcohol production capacity of the strain is weak, the alcohol content is insufficient, and the variety of aroma components in the fermented fruit wine is not rich enough. However, if *Saccharomyces cerevisiae* JG-WA1 is inoculated first and then *Saccharomyces cerevisiae* is inoculated, the fermentation process of *Saccharomyces cerevisiae* can be accelerated, the alcohol content of the fermented fruit wine can be increased, and the aroma enhancement effect is also more significant. Compared with the above fermentation process, after fermentation using the process of inoculating *Saccharomyces cerevisiae* JG-WA1 first and then *Saccharomyces cerevisiae*, a variety of volatile aroma components, including 2-pentanol, ethyl propionate, propyl acetate, 3-methyl-2-pentanone, 3-penten-2-one, hexanoate, ethyl hexanoate, methyl acetate, heptanal, 2-octanone, and 2,3-dimethylpyrazine, are detected in the fermented fruit wine, which significantly improves the flavor quality of the fruit wine.
[0012] Preferably, the abnormal Wickham yeast JG-WA1 bacterial culture is obtained by culturing abnormal Wickham yeast JG-WA1 preserved on slant culture medium for two generations at 25~30℃ and 120~180 r / min, and then resuspending the cells in 10~30 mL of physiological saline.
[0013] Preferably, the culture conditions for the abnormal Wickham yeast JG-WA1 are 28°C and 180 r / min for 12 h.
[0014] As a preferred method, the bacterial cells are resuspended in 15 mL of physiological saline.
[0015] Preferably, 3-6 mL of abnormal Wickham yeast JG-WA1 bacterial culture is added to apple juice and fermented statically at 25-28°C for 20-30 h; more preferably, the inoculation volume of abnormal Wickham yeast JG-WA1 bacterial culture is 5 mL and the fermentation conditions are fermentation at 25°C for 24 h.
[0016] As a preferred method, active dry yeast of brewing yeast is added at 0.25‰ of the apple juice mass, and fermentation is carried out at 25℃.
[0017] In addition, the present invention provides a fermented apple cider, which is produced by fermenting with abnormal Wickham yeast JG-WA1 according to the fermentation method described above.
[0018] The present invention has the following advantages and effects compared with the prior art: This invention screened a flavor-enhancing abnormal Wickham yeast strain JG-WA1 from brewing yeast. By inoculating this strain into apple juice for initial fermentation, followed by secondary fermentation with brewer's yeast, a flavor-enhancing apple cider was prepared. The apple cider contained significantly increased levels of fruit flavor components such as 2-pentanol, ethyl propionate, propyl acetate, 3-methyl-2-pentanone, 3-penten-2-one, hexanoate, ethyl hexanoate, methyl acetate, heptanal, 2-octanone, and 2,3-dimethylpyrazine. The resulting apple cider had a distinct ester aroma, a bright color, and significantly improved flavor and quality. Attached Figure Description
[0019] Figure 1 This is a graph showing the changes in CO2 weight loss during fermentation using different fermentation processes according to the present invention; Figure 2 Bar graph showing the alcohol content of cider obtained by different fermentation processes according to the present invention; Figure 3 This is a visualized dot matrix fingerprint of cider obtained by different fermentation processes in this invention. Detailed Implementation
[0020] To enable those skilled in the art to better understand the present invention, the present invention will now be further described in conjunction with specific embodiments.
[0021] Example 1 The fermentation method for flavoring cider using the unusual Wickham yeast JG-WA1 is as follows: (1) Activation of abnormal Wickham yeast JG-WA1 Atypical Wickham yeast preserved on slant culture was inoculated into 5 mL of PCA medium and cultured at 28℃ and 180 r / min for 12 h to prepare primary seed culture. The primary seed culture was then inoculated into 45 mL of PCA medium and cultured at 28℃ and 180 r / min for 12 h to prepare secondary seed culture. The secondary seed culture was centrifuged at 4℃ and 12000 r / min, the supernatant was removed, and the bacterial sludge was obtained. The bacterial cells were resuspended in 15 mL of physiological saline to obtain abnormal Wickham yeast JG-WA1 bacterial culture. (2) Cider brewing Apples were crushed and pressed to extract juice. One part per ten thousand of potassium metabisulfite was added to the apple juice. After 30 minutes, 5 mL of abnormal Wickham yeast JG-WA1 was added to the apple juice. The mixture was allowed to ferment at 25°C for 24 hours. Then, active dry yeast of Saccharomyces cerevisiae was added at 0.25‰ of the apple juice weight. The mixture was allowed to ferment at 25°C for another 6 days.
[0022] Comparative Example 1 Unlike Example 1, in (2), the fermentation step of abnormal Wickham yeast JG-WA1 is omitted. The specific operation is as follows: crush and press apples to extract juice, add 1 / 10,000 potassium metabisulfite according to the weight of apple juice, add 0.25‰ active dry yeast of brewing yeast to apple juice after 30 min, and let it ferment at 25℃ for 6 days.
[0023] Comparative Example 2 The difference from Example 1 is that the aroma-enhancing yeast *Saccharomyces cerevisiae*, also selected from the yeast starter, is used instead of the abnormal Wickham yeast JG-WA1 for fermentation and aroma enhancement. The specific operation is as follows: (1) Activation of Saccharomyces cerevisiae The slant culture of *Saccharomyces cerevisiae* was inoculated into 5 mL of PCA medium and cultured at 28℃ and 180 r / min for 12 h to prepare a primary seed culture. The primary seed culture was then inoculated into 45 mL of PCA medium and cultured at 28℃ and 180 r / min for 12 h to prepare a secondary seed culture. The secondary seed culture was centrifuged at 4℃ and 12000 r / min, and the supernatant was removed to obtain bacterial sludge. The bacterial cells were resuspended in 15 mL of physiological saline to obtain the *Saccharomyces cerevisiae* bacterial culture. (2) Cider brewing Apples were crushed and juiced. One part per ten thousand of potassium metabisulfite was added according to the weight of the apple juice. After 30 minutes, 0.25‰ of active dry yeast of Saccharomyces cerevisiae was added to the apple juice. The mixture was allowed to ferment at 25℃ for 24 hours. Then, 5 mL of yeast culture was added and the mixture was allowed to ferment at 25℃ for 6 days.
[0024] Comparative Example 3 The difference from Example 1 is that, in (2), the fermentation sequence of the abnormal Wickham yeast JG-WA1 was adjusted, specifically as follows: Apples were crushed and juiced. Potassium metabisulfite was added at 0.01% by weight of the apple juice. After 30 minutes, active dry yeast of Saccharomyces cerevisiae was added to the apple juice at 0.25‰. The mixture was allowed to ferment at 25℃ for 24 hours. Then, 5 mL of abnormal Wickham yeast JG-WA1 was added and fermented at 25℃ for 6 days.
[0025] Comparative Example 4 The difference from Example 1 is that the aroma-enhancing yeast Febian Seberlindnerella vulgaris, which was also selected from the yeast starter, was used instead of the abnormal Wickham yeast JG-WA1 for fermentation and aroma enhancement. In (1), the activation method, conditions, parameters and other aspects of Febian Seberlindnerella vulgaris were the same as those in Example 1.
[0026] (2) Apple wine making: crush apples and press them to extract juice. Add 1 / 10,000 potassium metabisulfite according to the weight of apple juice. After 30 min, add 0.25‰ active dry yeast to the apple juice. Let it ferment at 25℃ for 24 h. Then add 5 mL of Ferberis thuringiensis yeast liquid and let it ferment at 25℃ for 6 days.
[0027] Comparative Example 5 The difference from Example 1 is that *Saccharomyces cerevisiae* is used instead of *Saccharomyces cerevisiae* JG-WA1 for fermentation and flavoring. In (1), the activation method, conditions, and parameters of *Saccharomyces cerevisiae* are the same as in Example 1.
[0028] (2) Apple wine brewing: Apples are crushed and pressed to extract juice. One part of potassium metabisulfite is added according to the weight of the apple juice. After 30 min, 5 mL of saccharin-coated yeast solution is added to the apple juice. The mixture is allowed to ferment at 25℃ for 24 h. Then, active dry yeast is added at 0.25‰ of the weight of the apple juice. The mixture is allowed to ferment at 25℃ for 6 days.
[0029] Comparative Example 6 The difference from Example 1 is that Febien Seberlindnerella vaginalis is used instead of Wickham yeast JG-WA1 for fermentation and flavoring. In (1), the activation method, conditions, and parameters of Febien Seberlindnerella vaginalis are the same as in Example 1.
[0030] (2) Apple wine making: crush apples and press them to extract juice. Add 1 / 10,000 potassium metabisulfite according to the weight of apple juice. After 30 min, add 5 mL of Ferberis thuringiensis yeast liquid to the apple juice. Let it ferment at 25℃ for 24 h. Add 0.25‰ of the weight of apple juice to the active dry yeast of brewing yeast. Let it ferment at 25℃ for 6 days.
[0031] The CO2 weight loss, alcohol content, and volatile aroma components of the cider during the fermentation process of Example 1 and each comparative example were measured, and the results are as follows: Figures 1-3 As shown.
[0032] Since yeast fermentation converts fermentable sugars in the substrate into alcohol and CO2, the progress of alcoholic fermentation can be determined by the amount of CO2 lost. Figure 1It can be seen that the CO2 weight loss trend of the cider samples obtained by inoculating with Saccharomyces cerevisiae first and then with flavoring bacteria in Comparative Examples 2-4 is similar to that of the cider samples fermented with a single Saccharomyces cerevisiae strain in Comparative Example 1. However, the final total CO2 weight loss of the cider samples in Comparative Examples 2-4 is lower than that of the cider sample in Comparative Example 1. In addition, it can also be seen from the figure that in Example 1 and Comparative Examples 5-6, the CO2 weight loss was lower one day after inoculation with flavoring bacteria, indicating that the alcohol fermentation ability of the flavoring bacteria strain was weak. However, the CO2 weight loss process accelerated after inoculation with Saccharomyces cerevisiae, and the final total weight loss was similar to that of fermentation with a single Saccharomyces cerevisiae strain.
[0033] Figure 2 The alcohol content of cider produced using different fermentation processes. Figure 2 As shown, the alcohol content of the seven samples differed to some extent. The alcohol content of Comparative Example 1, which was fermented with a single Saccharomyces cerevisiae, and Examples 1, 5, and 6, which were inoculated with flavoring bacteria before Saccharomyces cerevisiae, was similar, all at 5.6% vol. However, the alcohol content of Comparative Examples 2, 3, and 4, which were inoculated with Saccharomyces cerevisiae before flavoring bacteria, was slightly lower, at 5.2% vol, 5.2% vol, and 5.4% vol, respectively. This result is consistent with the CO2 weight loss results.
[0034] Figure 3 Visualized dot matrix fingerprints of cider obtained from gas chromatography-ion mobility spectrometry (GC-IMS) for different fermentation processes. Figure 3 It can be seen that there are significant differences in the composition and content of volatile aroma components in the cider prepared by the seven fermentation processes. A total of 56 aroma components were detected in the seven samples, of which 37 substances were identified.
[0035] The fingerprint spectrum was divided into three regions: A, B, and C. Region A contains components common to apple cider fermented using seven different methods, totaling 19 substances. Of these, 17 substances were identified, including 10 esters, 2 aldehydes, 2 alcohols, 1 ketone, 1 ether compound, and 1 terpene, exhibiting aromas of fruit, apple, and pine. Area B contains substances with higher content than Comparative Example 1 (fermentation with a single Saccharomyces cerevisiae), Comparative Examples 2-4 (fermentation with Saccharomyces cerevisiae followed by aroma-enhancing bacteria), and Comparative Example 5 (fermentation with Saccharomyces cerevisiae followed by Saccharomyces cerevisiae). These substances include 15 volatile aroma components, of which 8 were identified, including 2 alcohols, 2 esters, 3 ketones, and 1 ether. Except for propyl propionate and ethyl valerate, which exhibit pleasant fruity aromas, most of the other substances, such as acetone, 4-methyl-1-pentanol, 2-butanol, dimethyl disulfide, and 3-hexanone, have unpleasant odors such as pungent alcoholic odors or musty smells. Region C contains substances with higher content in the samples of Example 1 and Comparative Example 6, which were first inoculated with aroma-enhancing bacteria and then fermented with Saccharomyces cerevisiae. A total of 22 volatile aroma components were identified, of which 12 were identified as 6 esters, 1 aldehyde, 3 ketones, 1 alcohol, and 1 pyrazine. The fingerprint spectrum shows that Example 1 had a higher and richer content of aroma components. Among these substances, 11 have pleasant fruity or herbaceous aromas, such as 2-pentanol, ethyl propionate, propyl acetate, 3-methyl-2-pentanone, 3-penten-2-one, hexanoate, ethyl hexanoate, methyl acetate, heptanal, and 2-octanone, all possessing a sweet fruity aroma. 2,3-Dimethylpyrazine has nutty and coffee aromas and can significantly enhance the aroma of cider.
[0036] As shown in the figure, regardless of whether the *Saccharomyces cerevisiae* strain was inoculated first or last, its aroma composition was similar to that of pure *Saccharomyces cerevisiae* fermentation. In the cider samples obtained by inoculating *Saccharomyces cerevisiae* JG-WA1 first and then *Saccharomyces cerevisiae*, the content of unpleasant odors such as acetone, 4-methyl-1-pentanol, 2-butanol, dimethyl disulfide, and 3-hexanone, which have pungent alcoholic or musty odors, was significantly reduced. Conversely, the content of flavor components with sweet fruity aromas, such as 2-pentanol, ethyl propionate, propyl acetate, 3-methyl-2-pentanone, 3-penten-2-one, hexanoate, ethyl hexanoate, methyl acetate, heptanal, and 2-octanone, was significantly increased, effectively enriching the aroma composition of the cider and improving its flavor quality.
[0037] The aroma-enhancing fruit wine prepared by the present invention using the abnormal Wickham yeast JG-WA1 has a similar alcohol content to fruit wine fermented with single brewing yeast, and has a pleasant fruit aroma and harmonious taste, effectively improving the flavor and quality of the fruit wine, and has broad application prospects.
[0038] The above are merely preferred embodiments of the present invention and do not limit the scope of the patent. All equivalent changes and modifications made within the scope of the present invention should still fall within the scope of the present invention.
Claims
1. An abnormal Wickham yeast strain JG-WA1, characterized in that, The aforementioned *Saccharomyces cerevisiae* JG-WA1 was deposited on September 9, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 35866 and classified as *Saccharomyces cerevisiae*. Wickerhamomyces anomalus ).
2. The application of the abnormal Wickham yeast JG-WA1 as described in claim 1 in flavoring fruit wines, characterized in that, The fruit wines mentioned include, but are not limited to, any one of apple wine, pear wine, grape wine, blueberry wine, mulberry wine, mango wine, pineapple wine, lychee wine, passion fruit wine, lemon wine, peach wine, and plum wine.
3. The application as described in claim 2, characterized in that, The fruit wine mentioned is either apple wine or pear wine.
4. The application as described in claim 2, characterized in that, The aforementioned abnormal Wickham yeast JG-WA1 enhances the aroma of fruit wine by increasing the content of the following components: 2-pentanol, ethyl propionate, propyl acetate, 3-methyl-2-pentanone, 3-penten-2-one, hexyl hexanoate, ethyl hexanoate, methyl acetate, heptanal, 2-octanone, and 2,3-dimethylpyrazine.
5. A fermentation method for flavoring cider using *Saccharomyces cerevisiae* JG-WA1, characterized in that... Fermentation is carried out using a step-by-step fermentation method, and the specific operation is as follows: Crush the apples and press to extract the juice. Add 0.5% to 1.5% potassium metabisulfite by weight of the apple juice. After 20 to 30 minutes, inoculate 3 to 10 mL of abnormal Wickham yeast JG-WA1 into the apple juice. Let it ferment at 20 to 30°C for 12 to 72 hours. Then, add 0.2% to 0.3% active dry yeast of Saccharomyces cerevisiae by weight of the apple juice and let it ferment at 20 to 30°C.
6. The fermentation method as described in claim 5, characterized in that, The aforementioned abnormal Wickham yeast JG-WA1 bacterial culture was obtained by continuously culturing abnormal Wickham yeast JG-WA1 preserved on slant culture medium for two generations at 25~30℃ and 120~180 r / min, and then resuspending the cells in 10~30 mL of physiological saline.
7. The fermentation method as described in claim 5, characterized in that, Add 3-6 mL of abnormal Wickham yeast JG-WA1 culture to apple juice and let it ferment at 25-28℃ for 20-30 h.
8. The fermentation method as described in claim 5, characterized in that, Inoculate with active dry brewer's yeast at 0.25‰ of the apple juice mass and let it ferment at 25℃.
9. A fermented apple cider, characterized in that, It is prepared by fermentation using any one of the fermentation methods described in claims 5 to 8.