CN112592841A - Non-saccharomyces cerevisiae and fermentation method for increasing flavor of Prunus salicina fruit wine by using same - Google Patents
Non-saccharomyces cerevisiae and fermentation method for increasing flavor of Prunus salicina fruit wine by using same Download PDFInfo
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Abstract
The invention discloses a non-saccharomyces cerevisiae and a fermentation method for increasing the flavor of hollow plum wine by using the yeast, wherein the non-saccharomyces cerevisiae has a deposit number of 20905. The invention also discloses a fermentation method for increasing the aroma of the hollow plum wine by using the yeast, which is characterized in that the hollow plum is taken as a raw material, the hollow plum is cleaned and crushed to obtain juice, the juice is inoculated by non-saccharomyces cerevisiae and saccharomyces cerevisiae according to a certain proportion and sequence at intervals and then fermented, the fermentation is stopped, centrifuged, bottled and the like, and the prepared hollow plum wine is pure in color and luster, outstanding in aroma, harmonious in aroma, clear and transparent in wine liquid, rich in flavor, full in wine body and balanced in acid and astringent taste, the types and the content of aroma substances of the hollow plum wine are obviously improved, the aim of obviously increasing the aroma of the hollow plum wine is fulfilled, and the aroma quality of the product is outstanding. The brewing method is simple and easy to operate, low in cost and easy to industrialize.
Description
Technical Field
The invention relates to the technical field of brewing, in particular to a fermentation method for enhancing aroma of hollow plum wine.
Background
The history of cultivation of sand hollow plums along rivers has been over one hundred years, the unique geographical environment dependence enables the plums to be produced independently and to be produced in sand towns in autonomous counties along rivers, and meat and stones are separated after the plums are matured. The meat is sweet, crisp and tasty. The Prunus salicina is rich in protein, fat, carbohydrate, calcium, phosphorus, iron, carotene, riboflavin, nicotinic acid, ascorbic acid and the like, and particularly contains rich selenium elements beneficial to the human body; and has effects of clearing away summer-heat, promoting appetite, caring skin, and prolonging life. However, the sand hollow plums are easy to mildew and rot, the preservation and storage technology falls behind, and abundant finely and deeply processed products are not available, so that the added value of the products is low. At present, sand hollow plums are sold mainly by fresh fruits in the market, the development of sand hollow plum products is concentrated on preserved fruits, fruit wine and the like at the present stage, and the sand hollow plum fruit wine in the market is mostly brewed by commercial saccharomyces cerevisiae, the fruit wine homogenization problem is more prominent, the content of flavor substances is lower, and the obtained product has single flavor.
The aroma is a key factor determining the sensory quality of the Prunus salicina wine, is a result of interaction of various compounds, and has three main sources: the aroma of the Prunus salicina fruits, the fermentation aroma generated by the metabolism of the fermentation microorganisms, and the aging aroma generated during the aging. The fermentation aroma is an extremely important factor, and the participation of various yeast strains (including saccharomyces cerevisiae and non-saccharomyces cerevisiae) can not only convert glucose in fruits into alcohol, but also generate a plurality of complex metabolites which greatly contribute to the final taste and aroma of the hollow plum wine, thereby having a determining effect on the final quality of the hollow plum wine. The mixed fermentation of non-saccharomyces cerevisiae and saccharomyces cerevisiae strains is one of effective ways for improving the quality and specificity of the fruit wine.
The natural fermentation process of fruit wine is essentially the result of the interaction of various microorganisms. The yeasts of different genera participate together to complete the natural fermentation of the fruit wine, including saccharomyces cerevisiae and non-saccharomyces cerevisiae. Researches show that unpredictable compounds can be produced by adopting saccharomyces cerevisiae and non-saccharomyces cerevisiae to mix and ferment the fruit wine, or fermentation products with different contents influence the aroma quality of the fruit wine. The non-saccharomyces cerevisiae exists in orchard soil, fruit epidermis and fruit wine brewing environment, can produce a large amount of metabolites such as glycerol, esters and the like, improves aroma components of fruit wine by metabolizing sugar and autolysis in fruit juice, and participates in the formation of complex flavor substances of the fruit wine. In the fermentation process of the fruit wine, non-saccharomyces cerevisiae can produce a plurality of extracellular enzymes, if collagenase, protease, glucanase, xylanase, amylase, lipase and the like, the non-saccharomyces cerevisiae has certain extracellular enzyme activity, and the enzymes act on related substrates in fruit juice to further influence the components and flavor substances of the fruit wine. Meanwhile, by inoculating different strains, the complex taste and aroma of the wine can be increased, the defects of process conditions can be made up, and the fermentation capacity of the strains is improved. The aroma characteristic of the Prunus salicina fruit wine can be improved by adding the non-Saccharomyces cerevisiae to perform mixed fermentation of the non-Saccharomyces cerevisiae and the Saccharomyces cerevisiae, so that the method is an effective fermentation method for enhancing the aroma of the Prunus salicina fruit wine.
However, at present, non-saccharomyces cerevisiae and saccharomyces cerevisiae are mixed together and inoculated to a fermented product together to be fermented in a mutually competitive manner, so that aroma substances are mainly added to the non-saccharomyces cerevisiae, and the saccharomyces cerevisiae mainly generates products such as alcohol, but the non-saccharomyces cerevisiae and saccharomyces cerevisiae are mixed together and inoculated to the fermented product together, so that the inactivation probability of the non-saccharomyces cerevisiae is high, and the generated products such as alcohol can inhibit the fermentation process of the non-saccharomyces cerevisiae after the saccharomyces cerevisiae starts fermentation, so that the aroma substances in the fermented product of the fruit wine are low in content, and the prominent fruit wine quality cannot be formed.
Disclosure of Invention
The invention mainly aims to overcome the problems that the fruit wine in the prior art is insufficient in aftertaste and flavor due to fermentation and the problem that the flavor is not prominent due to the fact that non-saccharomyces cerevisiae and saccharomyces cerevisiae are simultaneously mixed and fermented, and aims to brew the hollow plum wine through the steps of inoculating the non-saccharomyces cerevisiae and the saccharomyces cerevisiae according to a certain proportion and sequence at intervals, fermenting, terminating the fermentation, centrifuging, bottling and the like. The brewing method is simple and easy to operate, low in cost and easy to industrialize.
In order to achieve the technical purpose, the invention adopts the following technical scheme:
providing a non-saccharomyces cerevisiae F119, wherein the non-saccharomyces cerevisiae F119 has a deposit number of 20905:
further, a fermentation method for increasing the flavor of the prunus salicina wine is provided, which comprises the following steps:
(1) raw material treatment: crushing fresh mature hollow plum fruit, squeezing to obtain juice, adding potassium metabisulfite or SO into hollow plum juice2；
(2) Adjusting fermentation liquor: after measuring the pH value and the sugar degree value of the fermentation liquor, adjusting the pH value and the sugar of the fermentation liquor;
(3) inoculating and fermenting: YPD solid culture of activated Saccharomyces cerevisiae and non-Saccharomyces cerevisiae, shake culture of Saccharomyces cerevisiae and non-Saccharomyces cerevisiae, adding non-Saccharomyces cerevisiae and Saccharomyces cerevisiae at certain ratio into the fermentation liquid at certain interval, starting fermentation, and controlling fermentation temperatureMonitoring fermentation progress with alcohol hydrometer and saccharimeter, ending main fermentation when saccharinity is lower than a certain value, adding potassium metabisulfite or SO2；
(4) Fining and clarifying: naturally clarifying the wine after fermentation, separating the wine liquid on the upper layer, and adding the diatomite for fining and clarifying;
(5) filtering and bottling: blending the wine liquid after the glue discharging and clarification, filtering, sterilizing and bottling after the wine liquid is stabilized.
Further, the non-saccharomyces cerevisiae and the saccharomyces cerevisiae are added in the form of seed liquid.
Further, the fermentation temperature is 18 +/-2 ℃, and the condition of finishing the main fermentation is that the sugar degree is lower than 4 g/L.
Further, after the fermentation is finished, the wine is placed at 0-4 ℃ for natural clarification for 10 days, and after the diatomite is added, the wine is placed at 0-4 ℃ for 14 days for glue clarification.
Further, the method for adding the non-saccharomyces cerevisiae and the saccharomyces cerevisiae into the fermentation liquor at certain time intervals in sequence comprises the following steps: when inoculating and fermenting, non-saccharomyces cerevisiae is inoculated for 4-7 days, and then saccharomyces cerevisiae is inoculated.
Further, in the inoculation fermentation, non-saccharomyces cerevisiae is inoculated for 7 days, and then saccharomyces cerevisiae is inoculated.
Further, the ratio of the non-saccharomyces cerevisiae to the saccharomyces cerevisiae is 10:1-1:1 during the inoculation and fermentation.
Further, the potassium metabisulfite or SO2The amount of (B) added was 50 mg/L.
Further, the pH value of the fermentation liquor is 3.5, and the sugar degree of the fermentation liquor is 10-30 DEG Brix.
Further, the activation mode of the saccharomyces cerevisiae and the non-saccharomyces cerevisiae is as follows: dissolving glucose, peptone, yeast extract powder and agar in deionized water at 20-25 deg.C, sterilizing in vertical automatic pressure steam sterilizing pot at 120 deg.C for 20min, inoculating Saccharomyces cerevisiae and non-Saccharomyces cerevisiae in ultraclean bench by streaking, culturing at 28 deg.C for 48 hr, and storing at 4 deg.C for use.
Further, the expanding culture mode of the saccharomyces cerevisiae and the non-saccharomyces cerevisiae is as follows: dissolving glucose, peptone and yeast extract powder in deionized water at 20-25 deg.C, sterilizing at 120 deg.C for 20min in vertical automatic pressure steam sterilization pot, inoculating Saccharomyces cerevisiae and non-Saccharomyces cerevisiae in ultra-clean bench, culturing at 28 deg.C for 48 hr, and storing at 4 deg.C for use.
Furthermore, in the activation mode of the saccharomyces cerevisiae and the non-saccharomyces cerevisiae, the dosages of glucose, peptone, yeast extract powder and agar are respectively 20g/L, 10g/L and 20 g/L.
Furthermore, in the amplification culture mode of the saccharomyces cerevisiae and the non-saccharomyces cerevisiae, the dosage of glucose, peptone and yeast extract powder is 20g/L, 20g/L and 20g/L respectively.
Further, the preparation method of the non-saccharomyces cerevisiae and saccharomyces cerevisiae seed liquid specifically comprises the following steps: 1) activating strains, inoculating ultralow-temperature-preserved non-saccharomyces cerevisiae and saccharomyces cerevisiae to YPD solid culture medium respectively, and culturing at 28 ℃ for 48h for later use; 2) Preparing seed liquid, namely respectively transferring the non-saccharomyces cerevisiae and the saccharomyces cerevisiae activated in the step 1) into a YPD liquid culture medium. Culturing at 28 ℃ and 120-180 rpm for 20-28 h to logarithmic phase. Diluting the bacteria solution with sterilized normal saline to 600nm, wherein the light absorption value is 1, and the bacteria solution concentration is 108cfu/mL。
Further, the sterilization method in the filtration and bottling steps is pasteurization.
Has the advantages that:
compared with the prior art, the invention has the following beneficial effects:
according to the invention, non-saccharomyces cerevisiae and saccharomyces cerevisiae are inoculated into the hollow plum juice for fermentation at certain intervals according to a certain proportion and sequence, so that the good characteristic of natural fermentation of the hollow plum juice is maintained while the national production standard of fruit wine is achieved, wherein the saccharomyces cerevisiae consumes sugar to produce alcohol; the non-saccharomyces cerevisiae has low alcohol yield and high fragrance substances, and the combination of the non-saccharomyces cerevisiae and the saccharomyces cerevisiae ensures the proper alcohol content of the plum wine, obviously improves the content of the fragrance compounds such as esters, aldehydes and ketones and the like, and also obviously improves the fragrance characteristic of the plum wine. Endows the hollow plum wine with more intense flower and fruit fragrance, sweet fragrance and baking fragrance, effectively improves the fragrance characteristic of the hollow plum wine, and achieves the aim of enhancing the fragrance of the hollow plum wine.
The method is simple and feasible, can obtain fruit wine with various aroma characteristics, effectively increases the variety of the hollow plum fruit wine, meets the requirements of different consumers, and simultaneously avoids the vicious competition of the homogeneity of the hollow plum fruit wine in the market. In addition, compared with the prior art, the invention adopts non-saccharomyces cerevisiae and saccharomyces cerevisiae to inoculate, mix and ferment according to a certain proportion and sequence and at intervals of a certain time, thereby not only ensuring the inhibition to mixed bacteria in the fermentation process, but also enriching the flavor of the fermented product. Through GC-MS analysis and determination, compared with the fruit wine fermented by single commercial saccharomyces cerevisiae or the fruit wine fermented by mixing the non-saccharomyces cerevisiae and the saccharomyces cerevisiae simultaneously, the hollow plum fruit wine inoculated and fermented by the non-saccharomyces cerevisiae and the saccharomyces cerevisiae at certain proportion and sequence interval has richer fragrance.
Drawings
FIG. 1 shows colony morphology (A) and cell morphology (B) of non-brewing yeast F119;
FIG. 2 is a histogram of the content of esters in each experimental group in the specific examples;
FIG. 3 is a histogram of the alcohol content of each experimental group in the specific examples;
FIG. 4 is a histogram of the contents of acids of each experimental group in the examples;
FIG. 5 is a bar graph of aromatic content for each experimental group in the examples;
FIG. 6 is a histogram of the amounts of other species in each experimental group in the example;
FIG. 7 is a bar graph of the total volatile aroma component content of each experimental group in the examples.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.
Commercial Saccharomyces cerevisiae (Saccharomyces cerevisiae) strain X16: company, name: lafport.
Hansenula polymorpha (Hanseniaspora uvarum) strain F119 with deposit number 20905;
preservation information:
storage unit and code: china general microbiological culture Collection center (CGMCC);
address: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, North;
whether survival is carried out: survival;
and (3) classification and naming: hansenula polymorpha is a grape juice;
the preservation date is as follows: 10 and 19 days in 2020.
Commercial non-saccharomyces cerevisiae 32349: (China Industrial microorganism culture Collection management center No. 32349).
The preparation process of the hollow plum juice comprises the following steps: selecting mature and fresh hollow plum fruits, washing off impurities such as surface microorganisms, silt and the like by clean running water, crushing and juicing, and adding 50mg/L potassium metabisulfite into the hollow plum juice.
Preparation of seed liquid
1. Activating strains, inoculating Saccharomyces cerevisiae X16 stored at ultralow temperature to a YPD solid culture medium, and culturing at 28 deg.C for 48 hr;
2. and (3) preparing seed liquid, namely transferring the saccharomyces cerevisiae X16 activated in the step 1 into a YPD liquid culture medium. Culturing at 28 ℃ and 120-180 rpm for 20-28 h to logarithmic phase. When the sterilized normal saline is used for diluting the bacterial liquid concentration to 600nm, the light absorption value is 1, and the bacterial liquid concentration is 108 cfu/mL. Obtaining the saccharomyces cerevisiae X16 seed liquid.
3. And (3) replacing saccharomyces cerevisiae X16 with the non-saccharomyces cerevisiae Hansenula polymorpha strain F119, and performing operation according to the steps 1 and 2 to obtain a non-saccharomyces cerevisiae F119 seed solution.
4. And (3) replacing Saccharomyces cerevisiae X16 with non-Saccharomyces cerevisiae 32349, and operating according to the steps 1 and 2 to obtain non-Saccharomyces cerevisiae 32349 seed liquid.
Secondly, fermentation
The operation was divided into the following 9 groups:
group 1(X16 fermentation alone, X16): inoculating the Saccharomyces cerevisiae X16 seed solution prepared in the first step into hollow plum juice (10 can be specifically inoculated in each 1mL of hollow plum juice)7Individual cfu s.cerevisiae X16), the initial sugar degree of the fermentation broth was 28 ° Brix, and 50mg/L of potassium metabisulfite was added. And (3) fermentation process: controlling the fermentation temperature to be 18 +/-2 ℃, monitoring the fermentation progress by using an alcohol hydrometer and a brix meter during the fermentation, finishing the main fermentation when the brix is lower than 4g/L, and adding 50mg/L potassium metabisulfite to terminate the fermentation. After fermentation, the wine is naturally clarified at 0-4 ℃ for 10 days, and after the upper layer wine liquid is separated, the diatomite is added for fining clarification at 0-4 ℃ for 14 days. Blending the wine liquid after the glue discharging and clarification, filtering, sterilizing and bottling after the wine liquid is stabilized.
Group 2(X16 inoculated fermentation 7 days apart from F119, F119-F-1): inoculating the Saccharomyces cerevisiae X16 seed solution prepared in the first step into hollow plum juice (10 can be specifically inoculated in each 1mL of hollow plum juice)7C, inoculating the cfu saccharomyces cerevisiae X16), and inoculating the non-saccharomyces cerevisiae F119 seed liquid prepared in the step one into a fermentation system after 7 days (10 can be specifically inoculated in each 1mL of hollow plum juice)8Cfu non-s.cerevisiae F119), the initial sugar degree of the fermentation broth was 28 ° Brix, and 50mg/L of potassium metabisulfite was added. And (3) fermentation process: controlling the fermentation temperature to be 18 +/-2 ℃, monitoring the fermentation progress by using an alcohol hydrometer and a brix meter during the fermentation, finishing the main fermentation when the brix is lower than 4g/L, and adding 50mg/L potassium metabisulfite to terminate the fermentation. After fermentation, the wine is naturally clarified at 0-4 ℃ for 10 days, and after the upper layer wine liquid is separated, the diatomite is added for fining clarification at 0-4 ℃ for 14 days. Blending the wine liquid after the glue discharging and clarification, filtering, sterilizing and bottling after the wine liquid is stabilized.
Group 3(X16 and 32349 with 7 days apart for inoculum fermentation, 32349-F-2): inoculating the Saccharomyces cerevisiae X16 seed solution prepared in the first step into hollow plum juice (10 can be specifically inoculated in each 1mL of hollow plum juice)7C, Saccharomyces cerevisiae X16), 7 days laterInoculating the non-Saccharomyces cerevisiae 32349 seed solution prepared in the first step into a fermentation system (10 can be specifically inoculated into each 1mL of hollow plum juice)8Cfu non-saccharomyces cerevisiae 32349), the initial sugar degree of the fermentation broth was 28 ° Brix, and 50mg/L potassium metabisulfite was added. And (3) fermentation process: controlling the fermentation temperature to be 18 +/-2 ℃, monitoring the fermentation progress by using an alcohol hydrometer and a brix meter during the fermentation, finishing the main fermentation when the brix is lower than 4g/L, and adding 50mg/L potassium metabisulfite to terminate the fermentation. After fermentation, the wine is naturally clarified at 0-4 ℃ for 10 days, and after the upper layer wine liquid is separated, the diatomite is added for fining clarification at 0-4 ℃ for 14 days. Blending the wine liquid after the glue discharging and clarification, filtering, sterilizing and bottling after the wine liquid is stabilized.
Group 4(F119 inoculated fermentation with X16, F119-G): inoculating the non-Saccharomyces cerevisiae F119 seed solution prepared in the first step and Saccharomyces cerevisiae X16 seed solution into the hollow plum juice (10 can be specifically inoculated in each 1mL of hollow plum juice)8Cfu non-Saccharomyces cerevisiae F119 and 107Individual cfu s.cerevisiae X16), the initial sugar degree of the fermentation broth was 28 ° Brix, and 50mg/L of potassium metabisulfite was added. And (3) fermentation process: controlling the fermentation temperature to be 18 +/-2 ℃, monitoring the fermentation progress by using an alcohol hydrometer and a brix meter during the fermentation, finishing the main fermentation when the brix is lower than 4g/L, and adding 50mg/L potassium metabisulfite to terminate the fermentation. After fermentation, the wine is naturally clarified at 0-4 ℃ for 10 days, and after the upper layer wine liquid is separated, the diatomite is added for fining clarification at 0-4 ℃ for 14 days. Blending the wine liquid after the glue discharging and clarification, filtering, sterilizing and bottling after the wine liquid is stabilized.
Group 5(32349 simultaneous inoculation fermentation with X16, 32349-G): inoculating the non-saccharomyces cerevisiae 32349 seed liquid and saccharomyces cerevisiae X16 seed liquid prepared in the step one into the hollow plum juice (10 can be specifically inoculated in each 1mL of the hollow plum juice)8Cfu non-Saccharomyces cerevisiae 32349 and 107Individual cfu s.cerevisiae X16), the initial sugar degree of the fermentation broth was 28 ° Brix, and 50mg/L of potassium metabisulfite was added. And (3) fermentation process: controlling the fermentation temperature to be 18 +/-2 ℃, monitoring the fermentation progress by using an alcohol hydrometer and a brix meter during the fermentation, finishing the main fermentation when the brix is lower than 4g/L, and adding 50mg/L potassium metabisulfite to terminate the fermentation. After fermentation, naturally clarifying at 0-4 deg.C for 10 days, and separating to obtain upper layer wineAfter the solution is obtained, adding the diatomite at 0-4 ℃ for 14 days for fining and clarifying. Blending the wine liquid after the glue discharging and clarification, filtering, sterilizing and bottling after the wine liquid is stabilized.
Group 6(F119 and X16 were inoculated for fermentation 7 days apart, F119-S-1): inoculating the non-Saccharomyces cerevisiae F119 seed solution prepared in the first step into hollow plum juice (10 can be specifically inoculated in each 1mL of hollow plum juice)7Cfu non-Saccharomyces cerevisiae F119), inoculating the Saccharomyces cerevisiae X16 seed solution prepared in the step one in a fermentation system after 7 days (10 can be specifically inoculated in each 1mL hollow plum juice)7Individual cfu saccharomyces cerevisiae X16), the initial sugar degree of the fermentation broth was 22 ° Brix, and 60mg/L potassium metabisulfite was added. And (3) fermentation process: controlling the fermentation temperature to be 18 +/-2 ℃, monitoring the fermentation progress by using an alcohol hydrometer and a brix meter during the fermentation, finishing the main fermentation when the brix is lower than 4g/L, and adding 50mg/L potassium metabisulfite to terminate the fermentation. After fermentation, the wine is naturally clarified at 0-4 ℃ for 10 days, and after the upper layer wine liquid is separated, the diatomite is added for fining clarification at 0-4 ℃ for 14 days. Blending the wine liquid after the glue discharging and clarification, filtering, sterilizing and bottling after the wine liquid is stabilized.
Group 7(32349 inoculated fermentation 7 days apart from X16, 32349-S-1): inoculating the non-Saccharomyces cerevisiae 32349 seed solution prepared in the first step into hollow plum juice (10 can be specifically inoculated in each 1mL of hollow plum juice)7C, cfu non-saccharomyces cerevisiae 32349), inoculating the saccharomyces cerevisiae X16 seed liquid prepared in the step one into a fermentation system after 7 days (10 can be specifically inoculated in each 1mL hollow plum juice)7Individual cfu s.cerevisiae X16), the initial sugar degree of the fermentation broth was 22 ° Brix, and 50mg/L of potassium metabisulfite was added. And (3) fermentation process: controlling the fermentation temperature to be 18 +/-2 ℃, monitoring the fermentation progress by using an alcohol hydrometer and a brix meter during the fermentation, finishing the main fermentation when the brix is lower than 4g/L, and adding 50mg/L potassium metabisulfite to terminate the fermentation. After fermentation, the wine is naturally clarified at 0-4 ℃ for 10 days, and after the upper layer wine liquid is separated, the diatomite is added for fining clarification at 0-4 ℃ for 14 days. Blending the wine liquid after the glue discharging and clarification, filtering, sterilizing and bottling after the wine liquid is stabilized.
Group 8(F119 and X16 were inoculated for fermentation 7 days apart, F119-S-2): inoculating the non-saccharomyces cerevisiae F119 seed liquid prepared in the step one into the hollow plum juice(specifically, 10 can be inoculated per 1mL hollow plum juice8Cfu non-Saccharomyces cerevisiae F119), inoculating the Saccharomyces cerevisiae X16 seed solution prepared in the step one in a fermentation system after 7 days (10 can be specifically inoculated in each 1mL hollow plum juice)7Individual cfu s.cerevisiae X16), the initial sugar degree of the fermentation broth was 22 ° Brix, and 50mg/L of potassium metabisulfite was added. And (3) fermentation process: controlling the fermentation temperature to be 18 +/-2 ℃, monitoring the fermentation progress by using an alcohol hydrometer and a brix meter during the fermentation, finishing the main fermentation when the brix is lower than 4g/L, and adding 50mg/L potassium metabisulfite to terminate the fermentation. After fermentation, the wine is naturally clarified at 0-4 ℃ for 10 days, and after the upper layer wine liquid is separated, the diatomite is added for fining clarification at 0-4 ℃ for 14 days. Blending the wine liquid after the glue discharging and clarification, filtering, sterilizing and bottling after the wine liquid is stabilized.
Group 9(32349 inoculated fermentation 7 days apart from X16, 32349-S-2): inoculating the non-Saccharomyces cerevisiae 32349 seed solution prepared in the first step into hollow plum juice (10 can be specifically inoculated in each 1mL of hollow plum juice)8C, cfu non-saccharomyces cerevisiae 32349), inoculating the saccharomyces cerevisiae X16 seed liquid prepared in the step one into a fermentation system after 7 days (10 can be specifically inoculated in each 1mL hollow plum juice)7Individual cfu s.cerevisiae X16), the initial sugar degree of the fermentation broth was 22 ° Brix, and 50mg/L of potassium metabisulfite was added. And (3) fermentation process: controlling the fermentation temperature to be 18 +/-2 ℃, monitoring the fermentation progress by using an alcohol hydrometer and a brix meter during the fermentation, finishing the main fermentation when the brix is lower than 4g/L, and adding 50mg/L potassium metabisulfite to terminate the fermentation. After fermentation, the wine is naturally clarified at 0-4 ℃ for 10 days, and after the upper layer wine liquid is separated, the diatomite is added for fining clarification at 0-4 ℃ for 14 days. Blending the wine liquid after the glue discharging and clarification, filtering, sterilizing and bottling after the wine liquid is stabilized. Third, brewing characteristic detection
1. And (3) taking the hollow plum wine obtained in each group in the step two to carry out the following detection:
(1) ethanol content: alcohol meter method (reference: GB/T15038 + 2006 general analysis method for wine and fruit wine);
(2) types and contents of volatile aroma compounds: detection was carried out using a TQ8040NX gas chromatograph-mass spectrometer (Shimadzu, Japan). The headspace solid phase microextraction method comprises the following extraction conditions: adding 8mL of hollow plum wine sample, 1.0g of NaCL and 50 μ L of cyclohexanone (internal standard final concentration: 237.5mg/L) into a 20mL headspace bottle for semi-quantification, balancing in a water bath at 40 ℃ for 15min, extracting with a 1cm-50/30 μm DVB/CAR/PDMS fiber extraction head at 40 ℃ for 30min, inserting the extraction head into an injection port after extraction is finished, desorbing at 240 ℃ for 2min, and performing GC-MS analysis. Aging the fiber head at 250 deg.C for 30 min; the GC-MS analysis conditions were: InertCap Wax capillary column, 60m × 0.25mm × 0.25 μm, high purity (99.999%) helium as carrier gas, constant flow of 1.88mL/min, temperature program: starting from 40 ℃, keeping for 3min, heating to 230 ℃/min at 3 ℃/min, keeping for 2min, and injecting the sample: 240 ℃, 250 ℃ of MS interface temperature, no-flow-splitting sample injection, 2min of solvent delay, EI ion source, 200 ℃ of ion source, 70eV of electron energy, scanning mass-to-charge ratio range: 29-500 m/z.
2. Calculation results of physicochemical index and OAV value of main volatile aroma component of Prunus Cerasifera wine
The results are shown in Table 1, FIGS. 2-7 and Table 2.
TABLE 1 ethanol content of the hollow plum wine sample after completion of the alcohol fermentation in each experimental group
TABLE 2 fragrance activity values of fragrant substances of Prunus salicina wine samples after completion of alcoholic fermentation in each experimental group
As can be seen from the data in Table 1, the basic physicochemical indexes of the fermentation experimental groups are within the national standard control range. The alcohol content of the hollow plum wine is reduced by the co-inoculation mixed fermentation of the non-saccharomyces cerevisiae and the saccharomyces cerevisiae, and the alcohol content of the hollow plum wine is improved by increasing the inoculation proportion when the non-saccharomyces cerevisiae and the saccharomyces cerevisiae are sequentially inoculated, mixed and fermented.
As can be seen from FIG. 2, from the perspective of volatile aroma components, the total content of volatile aroma components of non-Saccharomyces cerevisiae F119, 32349 and Saccharomyces cerevisiae X16 inoculated simultaneously and sequentially mixed fermented Prunus salicina wine (experimental groups 4-9) is higher than that of Saccharomyces cerevisiae X16 inoculated alone fermented Prunus salicina wine, and is much higher than that of non-Saccharomyces cerevisiae F119 and Saccharomyces cerevisiae X16 inoculated reversely fermented groups (group 2, group 3: firstly Saccharomyces cerevisiae X16 is inoculated, and after fermentation is started for 7 days, non-Saccharomyces cerevisiae F119 is inoculated), wherein the content of esters, acids, aromatics and other compounds is higher than that of Saccharomyces cerevisiae X16 inoculated alone fermented Prunus salicina wine (group 1) and that of non-Saccharomyces cerevisiae F119 and Saccharomyces cerevisiae X16 inoculated reversely (group 2, group 3), which shows that the way of non-Saccharomyces cerevisiae and Saccharomyces cerevisiae fermented Prunus salicina wine has certain advantages, the simultaneous inoculation fermentation has certain advantages compared with the independent inoculation fermentation of the saccharomyces cerevisiae and the reverse inoculation fermentation of the non-saccharomyces cerevisiae and the saccharomyces cerevisiae, and compared with the simultaneous inoculation fermentation mode, the sequential inoculation fermentation mode of the non-saccharomyces cerevisiae and the saccharomyces cerevisiae has more remarkable advantages.
The mixed fermentation mode with non-saccharomyces cerevisiae F119 and non-saccharomyces cerevisiae 32349 can effectively improve the aroma of the Prunus salicina wine and increase the intensity and complexity of the aroma of the Prunus salicina wine. As shown in fig. 2, 4, 5, and 6, the mixed fermentation significantly increased the content of esters, acids, aromatics and other compounds, with the highest content of esters, acids, aromatics and other compounds in the sequential inoculation fermentation groups at an inoculation ratio of 10:1 (groups 8 and 9). Esters and aromatics generally have fruity, floral, sweet, and the like.
The non-saccharomyces cerevisiae mixed fermentation hollow plum juice also improves the content of various aroma components which have larger contribution to the aroma of the fruit wine, and the contribution of the aroma compounds to the whole aroma of the fruit wine is measured by using an aroma activity value (OAV). The calculation formula is as follows:
in the formula: ciIs the content of the compound (mu g/L); OTiIs the olfactive threshold (μ g/L) of the compound in 9-16% alcohol solution.
As can be seen from the data in table 2, in the aroma substances of OAV >1 in group 4 to group 9 blueberry wines, the aroma activity values of ethyl acetate, isobutyl acetate, ethyl caproate, methyl caprylate, ethyl caprate, isoamyl caprylate, ethyl laurate, caprylic acid, capric acid, ethyl benzoate, ethyl phenylacetate, eugenol and benzothiazole are all greater than those of group 1, group 2 and group 3. Wherein the OAV values of ethyl caproate, ethyl caprylate, ethyl caprate, ethyl laurate, caprylic acid, eugenol and other compounds are far greater than those of groups 1, 2 and 3, and the OAV values of the above aroma compounds are the highest in groups 8 and 9. The content of the compounds with larger aroma contribution degree can be effectively improved by the mixed fermentation of the non-saccharomyces cerevisiae and the saccharomyces cerevisiae, the effect of sequentially inoculating the fermentation groups (group 8 and group 9) is particularly obvious, and the aroma of the blueberry fruit wine is stronger. The fermentation modes (group 2 and group 3) of inoculating saccharomyces cerevisiae to start fermentation for a period of time and then inoculating non-saccharomyces cerevisiae can not achieve the aim of flavoring the fruit wine.
In conclusion, the non-saccharomyces cerevisiae and saccharomyces cerevisiae are mixed and fermented for the hollow plum wine fermentation production, so that the alcohol fermentation task is successfully completed, the aroma quality of the hollow plum wine is improved, the phenomenon of single fermentation aroma of pure species with a brewing price is improved, the aroma of the hollow plum wine is more complex and layered, particularly, the mixed fermentation mode of sequentially inoculating and fermenting the hollow plum wine by the non-saccharomyces cerevisiae F119 and the non-saccharomyces cerevisiae 32349 and the saccharomyces cerevisiae X16 according to the inoculation proportion of 10:1 has the most obvious effect, and the content of ester compounds in the hollow plum wine is remarkably improved.
According to the invention, non-saccharomyces cerevisiae and saccharomyces cerevisiae are inoculated into the hollow plum juice for fermentation at certain intervals according to a certain proportion and sequence, so that the good characteristic of natural fermentation of the hollow plum juice is maintained while the national production standard of fruit wine is achieved, wherein the saccharomyces cerevisiae consumes sugar to produce alcohol; the non-saccharomyces cerevisiae has low alcohol yield and high fragrance substances, and the combination of the non-saccharomyces cerevisiae and the saccharomyces cerevisiae ensures the proper alcohol content of the plum wine, obviously improves the content of the fragrance compounds such as esters, aldehydes and ketones and the like, and also obviously improves the fragrance characteristic of the plum wine. Endows the hollow plum wine with more intense flower and fruit fragrance, sweet fragrance and baking fragrance, effectively improves the fragrance characteristic of the hollow plum wine, and achieves the aim of enhancing the fragrance of the hollow plum wine.
The method is simple and feasible, can obtain fruit wine with various aroma characteristics, effectively increases the variety of the hollow plum fruit wine, meets the requirements of different consumers, and simultaneously avoids the vicious competition of the homogeneity of the hollow plum fruit wine in the market. In addition, compared with the prior art, the invention adopts non-saccharomyces cerevisiae and saccharomyces cerevisiae to inoculate, mix and ferment according to a certain proportion and sequence and at intervals of a certain time, thereby not only ensuring the inhibition to mixed bacteria in the fermentation process, but also enriching the flavor of the fermented product. Through GC-MS analysis and determination in each embodiment, compared with the fruit wine fermented by single commercial saccharomyces cerevisiae or the fruit wine fermented by mixing non-saccharomyces cerevisiae and saccharomyces cerevisiae simultaneously, the hollow plum fruit wine inoculated and fermented by non-saccharomyces cerevisiae and saccharomyces cerevisiae at certain proportion and sequence interval has richer fragrance.
Although the present invention has been described with reference to the above embodiments, it should be understood that the present invention is not limited to the above embodiments, and those skilled in the art can make various changes and modifications without departing from the scope of the present invention.
Claims (10)
1. A non-Saccharomyces cerevisiae F119, wherein the non-Saccharomyces cerevisiae F119 has a deposit number of: 20905.
2. a fermentation method for increasing the fragrance of prunus salicina wine is characterized by comprising the following steps:
(1) raw material treatment: crushing fresh mature hollow plum fruit, squeezing to obtain juice, adding potassium metabisulfite or SO into hollow plum juice2；
(2) Adjusting fermentation liquor: after measuring the pH value and the sugar degree value of the fermentation liquor, adjusting the pH value and the sugar of the fermentation liquor;
(3) inoculating and fermenting: YPD solid culture of activated Saccharomyces cerevisiae and non-Saccharomyces cerevisiae, shake culture for enlarging culture of Saccharomyces cerevisiae and non-Saccharomyces cerevisiae, adding non-Saccharomyces cerevisiae and Saccharomyces cerevisiae at a certain ratio into the fermentation liquid at certain time interval, starting fermentation, controlling fermentation temperature, monitoring fermentation progress with alcohol hydrometer and saccharimeter, ending main fermentation when saccharinity is lower than a certain value, adding potassium metabisulfite or SO2；
(4) Fining and clarifying: naturally clarifying the wine after fermentation, separating the wine liquid on the upper layer, and adding the diatomite for fining and clarifying;
(5) filtering and bottling: blending the wine liquid after the glue discharging and clarification, filtering, sterilizing and bottling after the wine liquid is stabilized.
3. The fermentation method for increasing the flavor of prunus salicina wine as claimed in claim 2, wherein the fermentation temperature is 18 ± 2 ℃, and the sugar degree is lower than 4g/L at the end of the main fermentation.
4. The fermentation method for increasing the flavor of prunus salicina wine as claimed in claim 2, wherein the wine is naturally clarified at 0-4 ℃ for 10 days after the fermentation is finished, and is subjected to degumming clarification at 0-4 ℃ for 14 days after the addition of the diatomite.
5. The method for increasing the flavor of the hollow plum wine as claimed in claim 2, wherein the non-saccharomyces cerevisiae and saccharomyces cerevisiae are added to the fermentation broth sequentially at certain time intervals by: when inoculating and fermenting, non-saccharomyces cerevisiae is inoculated for 4-7 days, and then saccharomyces cerevisiae is inoculated.
6. The fermentation method for increasing the flavor of the prunus salicina wine as claimed in claim 2, wherein the proportion of the non-saccharomyces cerevisiae and the saccharomyces cerevisiae is 10:1-1:1 during the inoculation fermentation.
7. The fermentation method for increasing the flavor of prunus salicina according to claim 2, wherein the potassium metabisulfite or SO is used as the source of the fermentation liquid2The amount of (B) added was 50 mg/L.
8. The fermentation process for increasing the flavor of a prunus salicina wine of claim 2, wherein the pH of the fermentation liquid is 3.5, and the sugar degree of the fermentation liquid is 10-30 ° Brix.
9. The fermentation method for increasing the flavor of the prunus salicina wine as claimed in claim 2, wherein the activation mode of the saccharomyces cerevisiae and the non-saccharomyces cerevisiae is as follows: dissolving glucose, peptone, yeast extract powder and agar in deionized water at 20-25 ℃, sterilizing for 20min at 120 ℃ in a vertical automatic pressure steam sterilization pot, inoculating saccharomyces cerevisiae and non-saccharomyces cerevisiae in a streak mode in an ultra-clean workbench, culturing for 48h at 28 ℃, and storing for later use at 4 ℃, wherein the expanding culture mode of the saccharomyces cerevisiae and the non-saccharomyces cerevisiae is as follows: dissolving glucose, peptone and yeast extract powder in deionized water at 20-25 deg.C, sterilizing at 120 deg.C for 20min in vertical automatic pressure steam sterilization pot, inoculating Saccharomyces cerevisiae and non-Saccharomyces cerevisiae in ultra-clean bench, culturing at 28 deg.C for 48 hr, and storing at 4 deg.C for use.
10. The fermentation method for improving the flavor of the Prunus salicina fruit wine as claimed in claim 9, wherein the activation modes of the Saccharomyces cerevisiae and non-Saccharomyces cerevisiae comprise 20g/L, 10g/L and 20g/L of glucose, peptone and yeast extract powder, and the amplification culture modes of the Saccharomyces cerevisiae and non-Saccharomyces cerevisiae comprise 20g/L, 20g/L and 20g/L of glucose, peptone and yeast extract powder.
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| CN109181985A (en) * | 2018-11-05 | 2019-01-11 | 贵州理工学院 | A kind of brewing method of hollow plum fruit wine |
| CN110923080A (en) * | 2019-10-30 | 2020-03-27 | 镇江瑞德酒业有限公司 | Flavor enhancing brewing process for fresh grape brandy |
| CN111154593A (en) * | 2020-01-15 | 2020-05-15 | 贵州理工学院 | Brewing method for improving aroma characteristic of rosa roxburghii tratt fruit wine by using non-saccharomyces cerevisiae |
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| CN109181985A (en) * | 2018-11-05 | 2019-01-11 | 贵州理工学院 | A kind of brewing method of hollow plum fruit wine |
| CN110923080A (en) * | 2019-10-30 | 2020-03-27 | 镇江瑞德酒业有限公司 | Flavor enhancing brewing process for fresh grape brandy |
| CN111154593A (en) * | 2020-01-15 | 2020-05-15 | 贵州理工学院 | Brewing method for improving aroma characteristic of rosa roxburghii tratt fruit wine by using non-saccharomyces cerevisiae |
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