CN115820371B - Fruit wine and fermentation method thereof - Google Patents
Fruit wine and fermentation method thereof Download PDFInfo
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- CN115820371B CN115820371B CN202211616248.1A CN202211616248A CN115820371B CN 115820371 B CN115820371 B CN 115820371B CN 202211616248 A CN202211616248 A CN 202211616248A CN 115820371 B CN115820371 B CN 115820371B
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Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the technical field of wine brewing, and particularly relates to fruit wine and a fermentation method thereof. The fruit wine is prepared by mixing and inoculating first non-saccharomyces cerevisiae Hanseniaspora guilliermondii NF and second non-saccharomyces cerevisiae Hanseniaspora thailandica YLL16 into fruit juice for fermentation; the preservation numbers of the first non-saccharomyces cerevisiae and the second non-saccharomyces cerevisiae are respectively CCTCC NO: m20221615 and CCTCC NO: m2018140. The mixed fermentation method provided by the invention has the advantages that on one hand, the fermentation capacity and the yield of alcohol are obviously improved, on the other hand, the problem of insufficient aroma and sour taste of the wine body is effectively avoided, the problem of lower content of vitamin C, lactic acid, hesperetin and nobiletin is also solved, the mouthfeel and health care effect of the fruit wine are further improved, and the additional utilization value of citrus fruit juice is improved.
Description
Technical Field
The invention belongs to the technical field of wine brewing, and particularly relates to fruit wine and a fermentation method thereof.
Background
Citrus fruits such as ponkan and the like, nanfeng orange and the like are common fruits with rich nutrition and fragrant and sweet taste, have long planting history in China and have high yield. The citrus fruits are processed into fruit juice and fruit wine, so that the problems of excessive stock and fruit spoilage caused by high yield can be solved, and the purposes of prolonging the shelf life and increasing the added value of the citrus fruits are achieved. The citrus fruit juice is raw juice extracted from citrus fruit. The citrus fruit wine is a fermented low-alcoholicity fruit juice beverage, has a clear and mellow taste, is rich in nutrients such as multiple vitamins, amino acids, calcium, magnesium and the like and organic acids, can help human metabolism, controls internal balance and is deeply favored by consumers.
At present, saccharomyces cerevisiae (Saccharomyces cerevisiae) is the most commonly used fermentation strain for commercial fruit wine fermentation, and has stable fermentation performance, but the fermentation product has single flavor, so that the development of fruit wine is greatly limited.
non-Saccharomyces cerevisiae is an important microorganism in the process of brewing fruit wine, plays a decisive role in the formation of fermentation flavor substances of fruit wine, and researches in recent years find that the addition of non-Saccharomyces cerevisiae can produce a plurality of extracellular enzymes such as collagenase, proteinase, glucanase, xylanase and the like, and the enzymes act on related substrates of fruit juice so as to increase aroma components of the fruit wine, so that the flavor of the fruit wine is richer, more mellow and more convenient. However, when the Saccharomyces cerevisiae is not fermented alone, the fermentation performance is unstable, the fermentation capacity is low, the fermentation process is difficult to complete, and the actual production requirements are difficult to meet.
Disclosure of Invention
In recent years, a part of technicians found that a fermentation mode of a commercial Saccharomyces cerevisiae (Saccharomyces cerevisiae) and a non-Saccharomyces cerevisiae (Hanseniaspora guilliermondii) through sequential fermentation can solve the problems of using Saccharomyces cerevisiae alone and using non-Saccharomyces cerevisiae alone; however, the content of vitamin C, lactic acid, hesperetin and nobiletin is low, and the mouthfeel is not as good as that of the fruit wine obtained by fermenting the fruit wine by solely adopting non-saccharomyces cerevisiae; moreover, there are studies reporting that Saccharomyces cerevisiae has an inhibitory effect on non-Saccharomyces cerevisiae during fermentation.
Therefore, in order to solve the problems of single taste, low content of vitamin C, lactic acid, hesperetin and nobiletin and the like of the traditional fermented Nanfeng orange fruit wine, the invention provides a mixed fermentation method of specific non-saccharomyces cerevisiae strains obtained by screening two strains, which comprises the following steps: mixing and inoculating the first non-saccharomyces cerevisiae and the second non-saccharomyces cerevisiae into fruit juice for fermentation to obtain the fruit wine; the first non-saccharomyces cerevisiae is named Hanseniaspora guilliermondii NF and is preserved in China center for type culture collection (China center for type culture collection) 10-20 days in 2022, the preservation address is the university of Wuhan in Wuhan, and the preservation number is CCTCC NO: m20221615; the second non-saccharomyces cerevisiae is named Hanseniaspora thailandica YLL and is preserved in China center for type culture collection (China center for type culture collection) in 3 months and 18 days, wherein the preservation address is the university of Wuhan in Wuhan, and the preservation number is CCTCC NO: m2018140.
The inventor finds that the two specific non-saccharomyces cerevisiae strains can be mutually cooperated for fermentation, so that the fermentation capacity of the non-saccharomyces cerevisiae is greatly improved, and meanwhile, compared with the fruit wine obtained by singly using the commercial saccharomyces cerevisiae or the mixed fermentation mode of the commercial saccharomyces cerevisiae and the non-saccharomyces cerevisiae, the fruit wine prepared by the invention has better taste and higher contents of vitamin C, lactic acid, hesperetin and nobiletin. Methods related to non-saccharomyces cerevisiae mixed fermentation of citrus fruit wine in fermentation technology have not been reported yet. At present, the development and production of the preparation of the citrus fruit wine by non-saccharomyces cerevisiae aiming at the citrus fruit juice are less, and the preparation of the citrus fruit wine by using the citrus fruit juice has better prospect.
According to the invention, by selecting effective fermentation strains and fermentation modes, better fruit wine sensory quality and higher content of bioactive substances are obtained, theoretical support is provided for fermenting citrus fruit wine by using non-saccharomyces cerevisiae, and practical application of citrus fruit (wide orange such as ponkan orange, nanfeng orange, navel orange and the like) fruit wine is promoted.
In some preferred embodiments, the first non-Saccharomyces cerevisiae and the second non-Saccharomyces cerevisiae are added in a ratio of (0.5-1): (1-10). Too low or too high a ratio results in slow fermentation process and reduced sugar alcohol conversion. More preferably 1:1, too low or too high a ratio results in slow fermentation process and reduced sugar alcohol conversion. More preferably, the inoculation amount of the first non-saccharomyces cerevisiae and the second non-saccharomyces cerevisiae is 7% -9% of the total volume of the system, and too low proportion can lead to slow fermentation process, too high proportion can influence the production of fermentation products, and the sensory quality of the fruit wine is poor.
In some preferred embodiments, the juice has a sugar content of 9 DEG Brix-21 DEG Brix and a pH of 3.8-4.2 in the fruit wine fermentation method. More preferably, the juice has a Brix of 20 ° and a pH of 3.8. Too low a sugar level results in a slow fermentation process and too high a sugar level inhibits fermentation. Depending on the optimal pH of the first non-Saccharomyces cerevisiae and the second non-Saccharomyces cerevisiae, too low a pH may inhibit fermentation, too high a pH may increase the possibility of contamination with bacteria and inhibit fermentation. The sugar degree adjusting method is to add drinkable water into the Nanfeng orange juice, and the pH adjusting method is to add citric acid or edible baking soda for adjustment.
The fruit juice is subjected to bacteriostasis and clarification before inoculation fermentation, and then is kept stand for 20-24 hours. The bacteriostasis operation is specifically as follows: adding food-grade sodium metabisulfite to make effective SO in Nanfeng orange juice 2 The content range is 65-75 mg/L, and the effective SO 2 Means that a certain amount of sodium metabisulfite is added to completely react in an acid solution to generate a preservative effect which is equivalent to a certain amount of SO 2 Is a corrosion-resistant effect. The clarifying operation is to add pectase to the fruit juice, and the adding amount of pectase is 55-65 mg/L.
In some preferred embodiments, the conditions of the conditional fermentation of the fermentation in the fruit wine fermentation process described above include: according to the optimal fermentation temperature of the first non-saccharomyces cerevisiae and the second non-saccharomyces cerevisiae, the fermentation temperature is selected to be 27-29 ℃, the ventilation frequency is 1/d-2/d, the growth of the non-saccharomyces cerevisiae is not favored by the excessively high or excessively low temperature, the ventilation can promote the growth of the strain, but the possibility of impurity bacteria pollution is increased due to the excessively high frequency.
In some preferred embodiments, the first non-saccharomyces cerevisiae and the second non-saccharomyces cerevisiae are further activated by the strain before mixed inoculation, and the specific process includes: and respectively inoculating the strain 1 and the strain 2 into YPD liquid culture medium, culturing at 28 ℃ until the logarithmic growth later stage, inoculating the bacterial liquid after the first generation of activation into the YPD liquid culture medium, and culturing at 28 ℃ until the logarithmic growth later stage.
YPD medium was prepared as follows: mixing 10g peptone, 5g yeast extract powder, 20g glucose and 1L water uniformly, and sterilizing at 121deg.C for 30min. YPD culture medium is a composite culture medium for conventional growth of various yeasts, and has the advantages of comprehensive nutrition, high cost performance and the like compared with other yeast culture mediums such as SC, YNB and the like.
After the activation of the strain, a centrifugation operation was performed at 6000rpm for 20min to collect the strain, and the lower layer deposit (strain) obtained by the centrifugation operation was inoculated.
After the fermentation process is finished (the sugar degree is not changed or is changed very little any more, and a person skilled in the art can judge according to the conventional technology), filtering and sterilizing to obtain the final finished product of the fruit wine.
The beneficial effects of the invention are as follows: the invention provides a mixed fermentation method of specific non-saccharomyces cerevisiae strains obtained by screening, which has the advantages that on one hand, compared with the existing single non-saccharomyces cerevisiae fermentation, the fermentation capacity is obviously improved, the yield of alcohol is improved, the probability of bacterial contamination in the fermentation process is reduced, so that the loss is reduced, on the other hand, the problems of insufficient aroma and sour taste of wine body caused by single fermentation of traditional commercial saccharomyces cerevisiae are effectively avoided, in addition, the problem that the contents of vitamin C, lactic acid, hesperetin and nobiletin are lower in the sequential fermentation of the traditional commercial saccharomyces cerevisiae and the non-saccharomyces cerevisiae are also overcome, the mouthfeel and health care effect of fruit wine are further improved, the additional utilization value of the Nanfeng orange juice is improved, and the problem of diapause caused by the fact that the supply of Nanfeng orange on the market is greater than the demand is improved.
Drawings
FIG. 1 is a diagram showing the ethanol content of fermented Nanfeng orange fruit wine by different strains and fermentation modes;
FIG. 2 is a schematic diagram showing antioxidant activity of various strains and fermented Nanfeng orange fruit wine;
FIG. 3 is a graph showing the vitamin C content of the fermented Nanfeng orange fruit wine by different strains and fermentation modes;
FIG. 4 is a graph showing lactic acid content of fermented Nanfeng orange fruit wine by different strains and fermentation modes;
FIG. 5 is a graph showing the hesperetin content of the fermented Nanfeng orange fruit wine by different strains and fermentation modes;
FIG. 6 is a diagram showing the content of nobiletin in the fermented Nanfeng orange fruit wine by different strains and fermentation modes;
FIG. 7 is a schematic diagram showing the content of volatile aroma compounds in the fermented Nanfeng orange fruit wine by different strains and fermentation modes.
Detailed Description
The conception and the technical effects produced by the present invention will be clearly and completely described below with reference to the embodiments and the drawings to fully understand the objects, aspects and effects of the present invention. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
Example 1:
two strains of non-saccharomyces cerevisiae are adopted for mixed fermentation of Nanfeng orange juice, and the fermentation modes respectively comprise: hanseniaspora guillie rmondii-Hanseniaspora thailandica sequential fermentation, hanseniaspora guilliermondii: candida ethanolic a (from Minghu Biotechnology Co.) 1:1 mixed fermentation, hanseniaspora guilliermondii: hanseniaspor a thailandica1:1 mixed fermentation and Hanseniaspora guilliermondii: hanseniaspora thailandica1:100 mixed fermentation, the process steps Hanseniaspora guilliermondii and Hanseniaspora thailandica comprising:
(1) Preparing Nanfeng orange juice: performing bacteriostasis and clarification operation on the Nanfeng orange juice, wherein the bacteriostasis operation is to add food-grade sodium metabisulfite into the Nanfeng orange juice to ensure that the Nanfeng orange juice has effective SO 2 The content is about 70mg/L, and the clarifying operation is to add pectase with the addition amount of 60mg/L into Nanfeng orange juice; adding drinkable water into Nanfeng orange juice to make the sugar degree of Nanfeng orange juice be about 20 DEG Brix, adding citric acid or edible sodium bicarbonate into Nanfeng orange juice to adjust the pH to be about 3.8, and then standing for 24 hours;
(2) Inoculating and fermenting: centrifuging the activated two strains of non-saccharomyces cerevisiae, inoculating the two strains of non-saccharomyces cerevisiae into the Nanfeng orange juice obtained in the step (1) according to different mixed fermentation modes, and carrying out mixed fermentation, wherein the fermentation conditions are as follows: the temperature is about 28 ℃, and the ventilation frequency is 2 times/d; during fermentation, sugar degree and alcohol degree are monitored every day; the method for activating the strain comprises the following steps: respectively inoculating non-Saccharomyces cerevisiae strains preserved at ultralow temperature into YPD liquid culture medium, culturing at 28deg.C to logarithmic growth late stage, inoculating the bacterial liquid after the first generation of activation into YPD liquid culture medium, culturing at 28deg.C to logarithmic growth late stage, and making into seed liquid for use; wherein, the YPD medium is prepared as follows: mixing 10g peptone, 5g yeast extract powder, 20g glucose and 1L water uniformly, and sterilizing at 121deg.C for 30min; during centrifugation, the rotating speeds are 6000rpm, and the time is 20min; after centrifugation, inoculating the obtained lower sediment;
(3) Preparing a finished product: and when the sugar degree is not changed, ending fermentation, filtering and sterilizing to obtain the Nanfeng orange fruit wine, wherein the filtering and sterilizing method is sterile membrane filtration, and physical and chemical indexes such as total acid, reducing sugar, chromaticity, organic acid and the like in the Nanfeng orange fruit wine are measured.
Comparative example 1:
the difference from example 1 is that: the single strain fermentation was performed using Saccharomyces cerevisiae strain CICC 1750 alone, non-Saccharomyces cerevisiae strain (Hanseniaspora guilliermondii) alone, non-Saccharomyces cerevisiae strain (Hanseniaspora thailandica) alone, H.gullier mondii and commercial Saccharomyces cerevisiae (CICC 1750, available from Angel Yeast Co., ltd.) in sequence, as fermentation broth, respectively, comprising the steps of:
(1) Preparing Nanfeng orange juice: performing bacteriostasis and clarification operation on the Nanfeng orange juice, wherein the bacteriostasis operation is to add food-grade sodium metabisulfite into the Nanfeng orange juice to ensure that the Nanfeng orange juice has effective SO 2 The content is about 70mg/L, and the clarifying operation is to add pectase with the addition amount of 60mg/L into Nanfeng orange juice; adding drinkable water into Nanfeng orange juice to make the sugar degree of Nanfeng orange juice be about 20 DEG Brix, adding citric acid or edible sodium bicarbonate into Nanfeng orange juice to adjust the pH to be about 3.8, and then standing for 24 hours;
(2) Inoculating and fermenting: centrifuging the activated two strains of non-saccharomyces cerevisiae, inoculating the two strains of non-saccharomyces cerevisiae into the Nanfeng orange juice obtained in the step (1) according to different mixed fermentation modes, and carrying out mixed fermentation, wherein the fermentation conditions are as follows: the temperature is about 28 ℃, and the ventilation frequency is 2 times/d; during fermentation, sugar degree and alcohol degree are monitored every day; the method for activating the strain comprises the following steps: respectively inoculating non-Saccharomyces cerevisiae strains preserved at ultralow temperature into YPD liquid culture medium, culturing at 28deg.C to logarithmic growth late stage, inoculating the bacterial liquid after the first generation of activation into YPD liquid culture medium, culturing at 28deg.C to logarithmic growth late stage, and making into seed liquid for use; wherein, the YPD medium is prepared as follows: mixing 10g peptone, 5g yeast extract powder, 20g glucose and 1L water uniformly, and sterilizing at 121deg.C for 30min; during centrifugation, the rotating speeds are 6000rpm, and the time is 20min; after centrifugation, inoculating the obtained lower sediment;
(3) Preparing a finished product: and when the sugar degree is not changed, ending fermentation, filtering and sterilizing to obtain the Nanfeng orange fruit wine, wherein the filtering and sterilizing method is sterile membrane filtration, and physical and chemical indexes such as total acid, reducing sugar, chromaticity, organic acid and the like in the Nanfeng orange fruit wine are measured.
Example 2:
to further illustrate that the present invention can achieve the above-described technical effects, the physicochemical index test was performed on the Nanfeng orange fruit wine prepared in example 1 and comparative example 1, all of the wine samples inoculated with Saccharomyces cerevisiae or non-Saccharomyces cerevisiae in the tables and figures were respectively referred to as Sc, hg and Ht, the wine samples of H.gullimonondii and C.ethane 1:1 mixed fermentation were referred to as Hg: ce (1:1), the wine samples of H.gullimonondii and H.thailandica 1:1 mixed fermentation were referred to as Hg: ht (1:1), the wine samples of H.gullimonondii and H.thailandica 1:100 mixed fermentation were referred to as Hg: ht (1:100), the wine samples of H.gullimonondii and H.thailandica mixed fermentation were referred to as Hg-Ht, and the wine samples of H.gullimonondii and commercial yeast mixed fermentation were referred to as Hg-Ht.
1. Ethanol content in the fermented Nanfeng orange fruit wine by different strains and fermentation modes:
measurement of alcohol content was performed using a hand-held refractometer. As shown in FIG. 1, the final alcoholic strength of the wine sample prepared in example 1 was significantly higher than that of the non-Saccharomyces cerevisiae single strain fermentation and non-Saccharomyces cerevisiae other mixed fermentation fruit wine samples, similar to that of the conventional commercial Saccharomyces cerevisiae and the commercial Saccharomyces cerevisiae non-Saccharomyces cerevisiae mixed fermentation.
Compared with the prior art, the method provided by the invention effectively promotes the fermentation efficiency of the ethanol in the wine sample, obviously improves the final conversion rate of the ethanol, has high final yield of the ethanol concentration, is beneficial to reducing the probability of bacterial contamination in the fermentation process, reduces the loss caused by bacterial contamination, and has certain economic value.
2. Antioxidant activity of Nanfeng orange fruit wine fermented by different strains and fermentation modes:
antioxidant activity was measured using ABTS radical scavenging method. Specifically, an ABTS (7.0 mM,10 mL) solution was mixed with potassium persulfate (140 mM, 176. Mu.L) and incubated in the dark at room temperature for 14h as follows. The mixed ABT S solution was diluted with an appropriate amount of ethanol to reach an absorbance of 0.70±0.02 at 734 nm. After diluting the sample to a suitable concentration using absolute ethanol, 200. Mu.L of the sample dilution was added to 3.8mL of the mixed ABTS solution, and after incubation at room temperature for 6min, absorbance was measured at 734 nm. Each sample was assayed in 3 replicates. The formula for calculating the clearance rate of the ABTS free radicals is as follows: ABTS radical clearance (%) = (1-AS/Ab) ×100%; wherein AS is the absorbance of the sample and Ab is the absorbance of the blank.
As shown in figure 2, the method provided by the invention effectively improves the antioxidant activity of the fruit wine, is beneficial to improving the health care effect of the fruit wine and has certain economic value. The clearance rate of the ABTS free radical of the Nanfeng orange fruit wine (60.39% +/-0.01) prepared by Ht-1:1 mixed fermentation is highest and is obviously higher than that of the traditional fruit wine samples prepared by single fermentation of the commercial Saccharomyces cerevisiae Sc and non-Saccharomyces cerevisiae and other mixed modes of non-Saccharomyces cerevisiae.
3. Different strains and fermentation modes are used for fermenting the vitamin C content of the Nanfeng orange fruit wine:
HPLC analysis was performed on vitamin C in Nanfeng orange fruit wine using Ultimate AQ-C18 chromatographic column (4.6 mm. Times.250 mm,5 μm). 0.025% trifluoroacetic acid in methanol (95:5, V/V) as mobile phase. The flow rate is 0.8mL/min, the detection wavelength is 210nm, and the sample injection amount is 10 mu L. And (3) establishing a standard curve by using vitamin C standard substances with different concentration gradients, and quantifying the vitamin C in the Nanfeng orange fruit wine by using an external standard method.
As shown in figure 3, the method provided by the invention effectively improves the vitamin C content of the fruit wine, is beneficial to improving the health care effect of the fruit wine and has certain economic value. The vitamin C content of the Nanfeng orange fruit wine prepared by Ht-1:1 mixed fermentation (149.36 +/-6.50 mg/L) is highest and is obviously higher than that of a traditional fruit wine sample prepared by single fermentation of commercial Saccharomyces cerevisiae Sc and non-Saccharomyces cerevisiae and other mixed fermentation modes of non-Saccharomyces cerevisiae.
4. Lactic acid content of the Nanfeng orange fruit wine fermented by different strains and fermentation modes:
HPLC analysis was performed on lactic acid in Nanfeng orange fruit wine using Ultimate AQ-C18 chromatographic column (4.6 mm. Times.250 mm,5 μm). 0.025% trifluoroacetic acid in methanol (95:5, V/V) as mobile phase. The flow rate is 0.8mL/min, the detection wavelength is 210nm, and the sample injection amount is 10 mu L. And (3) establishing a standard curve by using lactic acid standard substances with different concentration gradients, and quantifying lactic acid in the Nanfeng orange fruit wine by using an external standard method.
As shown in figure 4, compared with the prior art, the method provided by the invention effectively improves the lactic acid content of the fruit wine, is beneficial to improving the taste of the fruit wine, and has certain economic value. The lactic acid content of the Nanfeng orange fruit wine prepared by Ht-1:1 mixed fermentation (8111.94 +/-108.67 mg/L) is highest and is obviously higher than that of the fruit wine samples prepared by single fermentation of traditional commercial saccharomyces cerevisiae Sc and non-saccharomyces cerevisiae and fermentation of other mixed modes of non-saccharomyces cerevisiae.
5. Different strains and fermentation modes are used for fermenting the orange peel content of the Nanfeng orange fruit wine:
the hesperetin content of the Nanfeng orange fruit wine was detected by using Aglient Eclipse XDB-C18 chromatographic column (4.6mm.times.250mm, 5 μm). HPLC was set up for a gradient elution procedure at a flow rate of 1.0 mL/min. The column temperature is 30 ℃, the quantitative wavelength is 283nm and 330nm, the scanning wavelength range is 200-400nm, and the sample injection amount is 10 mu L. The retention time is qualitative and the external standard method is quantitative. Hesperetin was analyzed at 283 nm.
As shown in the figure 5, compared with the prior art, the method provided by the invention effectively improves the hesperetin content of the fruit wine, is beneficial to improving the health care effect of the fruit wine, and has certain economic value. The content of hesperetin in Nanfeng orange fruit wine of Ht-1:1 mixed fermentation (3.52+/-0.13 mg/L) is highest and is obviously higher than that of single fermentation of traditional commercial saccharomyces cerevisiae Sc and non-saccharomyces cerevisiae and fermentation of fruit wine samples in other mixed modes of non-saccharomyces cerevisiae.
6. Different strains and fermentation modes are used for fermenting the content of the nobiletin in the Nanfeng orange fruit wine:
detecting the content of nobiletin in Nanfeng orange fruit wine by using Aglient Eclipse XDB-C18 chromatographic column (4.6 mm×250mm,5 μm). HPLC is provided with a gradient elution program with a flow rate of 1.0mL/min, a column temperature of 30 ℃, quantitative wavelengths of 283nm and 330nm, a scanning wavelength range of 200-400nm and a sample injection amount of 10 mu L. The retention time is qualitative and the external standard method is quantitative. The nobiletin was analyzed at 283 nm.
As shown in figure 6, compared with the prior art, the method provided by the invention effectively improves the content of the fruit wine Chuanqinensu, is beneficial to improving the health care effect of the fruit wine, and has certain economic value. The content of the nobiletin in the Nanfeng orange fruit wine is highest by Ht-1:1 mixed fermentation (0.98+/-0.02 mg/L), which is obviously higher than that of the traditional single fermentation of the commercial saccharomyces cerevisiae Sc and non-saccharomyces cerevisiae and the fermentation of fruit wine samples by other mixed modes of non-saccharomyces cerevisiae.
7. Analysis of the content of volatile aroma compounds in Nanfeng orange fruit wine:
and (3) analyzing the volatile aroma components of the Nanfeng orange fruit wine by adopting HS-SPME-GC-MS. Accurately sucking 5mL of Nanfeng orange fruit wine sample into a 20mL headspace extraction bottle, adding 1.5g of NaCl, and adding 5 mu L of 10mg/mL cyclohexanone solution as an internal standard. After 20min of equilibration at 40 ℃, the aged DVB/CAR/PDMS extraction head (Supelco, bellefonte PA, SA) was inserted into the extraction flask, the fiber head was pushed out 2cm above the sample solution and placed at 40 ℃ for adsorption for 52min, the sample was gently vortexed with a magnetic stirrer at 750rpm during equilibration and adsorption, then the extraction head was rapidly removed and inserted into the gas chromatograph sample inlet, and the gas chromatograph was started for data collection at 250 ℃ for 3 min. GC conditions: the capillary column was HP-5MS (30 m. Times.250 μm. Times.0.25 μm), the carrier gas was high purity helium gas at a flow rate of 1.2mL/min and a split ratio of 5:1. Programming temperature: keeping at 40deg.C for 2min, heating to 180deg.C (4deg.C/min), keeping for 2min, and heating to 250deg.C (10deg.C/min); the temperature of the detector and the ion source are respectively adjusted to 250 ℃ and 200 ℃; electron energy 70eV; the mass scan range is 30-500amu.
As shown in figure 7, the method provided by the invention effectively improves the content of volatile aroma compounds in the fruit wine, is beneficial to improving the sensory quality of the fruit wine and has certain economic value. The content of volatile aroma compounds of Nanfeng orange fruit wine prepared by Ht-1:1 mixed fermentation (588.42 +/-4.3 mg/L) is highest, which is obviously higher than that of fruit wine samples prepared by single fermentation of traditional commercial Saccharomyces cerevisiae Sc and non-Saccharomyces cerevisiae and other mixed modes of non-Saccharomyces cerevisiae.
In summary, in the embodiment 1 provided by the invention, the Nanfeng orange juice is taken as a raw material, and the Nanfeng orange fruit wine is brewed by adopting a non-saccharomyces cerevisiae mixed fermentation method, so that the high-quality Nanfeng orange fruit wine with high contents of vitamin C, lactic acid, hesperetin, nobiletin and volatile aroma compounds is obtained, the method has the characteristics of high content of bioactive components, rich and prominent aroma and coordinated and mellow taste, and the fermentation method is simple and easy to operate, fully plays the capabilities of reducing sugar and converting organic acid of non-saccharomyces cerevisiae and the capability of synergistic fermentation of non-saccharomyces cerevisiae, has high final conversion rate of alcohol, is favorable for reducing the probability of bacterial contamination in the fermentation process, and reduces the loss caused by bacterial contamination; on the other hand, the problem that the single strain of non-saccharomyces cerevisiae has weak fermentation capacity and is difficult to use for production is effectively avoided; meanwhile, the problems of insufficient aroma and sour taste of the wine body caused by the fermentation of the traditional commercial saccharomyces cerevisiae are overcome; the defects that the traditional mixed fermentation of the saccharomyces cerevisiae and the non-saccharomyces cerevisiae has lower content of vitamin C, lactic acid, hesperetin and nobiletin, and influences the health care and the taste of fruit wine are overcome, the additional utilization value of the Nanfeng orange juice is improved, the problem of diapause caused by the fact that the supply and demand of Nanfeng orange are greater than that of the market is improved, and the method has certain economic value.
The present invention is not limited to the above embodiments, but is merely preferred embodiments of the present invention, and the present invention should be construed as being limited to the above embodiments as long as the technical effects of the present invention are achieved by the same means. Various modifications and variations are possible in the technical solution and/or in the embodiments within the scope of the invention.
Claims (9)
1. A fruit wine fermentation method is characterized in thatComprising the following steps: mixing and inoculating the first non-saccharomyces cerevisiae and the second non-saccharomyces cerevisiae into fruit juice for fermentation to obtain the fruit wine; the first non-Saccharomyces cerevisiae is named asHanseniaspora guilliermondii NF6, which was preserved in China center for type culture Collection (China, university of Wuhan with a preservation address of China) and CCTCC NO: m20221615; the second non-Saccharomyces cerevisiae is namedHanseniaspora thailandica YLL16 was preserved in China center for type culture Collection (China, university of Wuhan with preservation address of Wuhan, china) at 3 months and 18 days, with preservation number of CCTCC NO: m2018140;
the adding amount ratio of the first non-saccharomyces cerevisiae to the second non-saccharomyces cerevisiae is (0.5-1): (1-10).
2. The fruit wine fermentation method according to claim 1, wherein the first non-saccharomyces cerevisiae and the second non-saccharomyces cerevisiae are added in a ratio of 1:1.
3. The fruit wine fermentation method of claim 2, wherein the inoculation amount of the first non-saccharomyces cerevisiae and the second non-saccharomyces cerevisiae is 7% -9% of the total volume of the system.
4. The method for fermenting fruit wine according to claim 1, wherein the sugar degree of the fruit juice is 19 DEG Brix-21 DEG Brix and the pH is 3.8-4.2.
5. The method for fermenting fruit wine according to claim 4, wherein the sugar degree of the fruit juice is 20 DEG Brix and the pH is 3.8.
6. The fruit wine fermentation method according to claim 1, wherein the fermentation conditions include: the temperature is 27-29 ℃, and the ventilation frequency is 1-2 times/d.
7. The fruit wine fermentation method according to claim 1, wherein the first non-saccharomyces cerevisiae and the second non-saccharomyces cerevisiae are further subjected to strain activation before mixed inoculation, and the specific process comprises: and respectively inoculating the first non-saccharomyces cerevisiae and the second non-saccharomyces cerevisiae into a YPD liquid culture medium, culturing at 28 ℃ until the logarithmic growth later stage, inoculating the bacterial liquid after the first generation of activation into the YPD liquid culture medium, and culturing at 28 ℃ until the logarithmic growth later stage.
8. A fruit wine fermentation process according to any one of claims 1 to 7 wherein the fruit juice is a citrus fruit juice.
9. Fruit wine produced by the fruit wine fermentation process according to any one of claims 1 to 8.
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CN115161142A (en) * | 2022-06-26 | 2022-10-11 | 江西科技师范大学 | Brewing process of fruit wine |
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