CN112457941B - Aronia melanocarpa fruit wine and brewing method thereof - Google Patents
Aronia melanocarpa fruit wine and brewing method thereof Download PDFInfo
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- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
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- C12G3/024—Preparation of other alcoholic beverages by fermentation of fruits other than botanical genus Vitis
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- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
- C12G3/026—Preparation of other alcoholic beverages by fermentation with health-improving ingredients, e.g. flavonoids, flavones, polyphenols or polysaccharides, added before or during the fermentation stage; with flavouring ingredients added before or during the fermentation stage
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- C12H—PASTEURISATION, STERILISATION, PRESERVATION, PURIFICATION, CLARIFICATION OR AGEING OF ALCOHOLIC BEVERAGES; METHODS FOR ALTERING THE ALCOHOL CONTENT OF FERMENTED SOLUTIONS OR ALCOHOLIC BEVERAGES
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Abstract
The invention discloses aronia melanocarpa fruit wine and a brewing method thereof, and belongs to the technical field of food engineering. The invention discloses a process for fermenting aronia melanocarpa fruit wine by using mixed bacteria, which comprises the steps of adopting aronia melanocarpa fruits or fruit residues or fruit powder as raw materials, carrying out enzyme treatment on the raw materials, sequentially adding Candida divaversasa Z3 and saccharomyces cerevisiae to carry out primary fermentation, feeding, separation of skin and residue after fermentation, secondary fermentation, separation of skin and residue after secondary fermentation, and aging to obtain the aronia melanocarpa fruit wine. The method adopts a specific fruit wine preparation process, so that the characteristic of single-thin aroma in the traditional brewing process of the black fruit and burley flower fruit wine is improved, the prepared fruit wine is mellow in taste and rich in aroma, the acidity and astringency of the fruit wine are well reduced, and the development and utilization of the black fruit and burley flower fruit wine in the field of foods are promoted.
Description
Technical Field
The invention relates to aronia melanocarpa fruit wine and a brewing method thereof, belonging to the technical field of food engineering.
Background
Aronia melanocarpa (Michx.) Elliott, Rosaceae, Photinia foliacea bush. The fruits have the purposes of medicine, food, dye and the like; native to the united states. In the 90 s of the last century, the plants began to be planted in Liaoning areas of China. By 2015, the total planting area reaches 2000-2500 mu. At present, the aronia melanocarpa fruit is mainly used for medicine and production of health care products.
The Aronia melanocarpa fruit contains abundant polyphenols (about 2-5 g/kg), anthocyanin content of about 20% of total phenols, and large amount of bioactive substances such as gallic acid, catechin, epicatechin gallate, etc. in the fruit juice. The phenolic substances not only can remove free radicals and resist aging, but also have good treatment and prevention effects on arteriosclerosis, prostate, arthritis and the like; in addition, the aronia melanocarpa fruit contains rich polyphenol and anthocyanin, and the anthocyanin in the aronia melanocarpa has good antioxidation and also has beneficial effects on obesity, diabetes, cardiovascular diseases (such as hyperlipidemia, hypertension and the like) and the like.
Organic acid in the fruit wine is an important flavor substance in the fruit wine, and the existence of the organic acid can accelerate the transformation of polysaccharide and the decomposition of pectin substances and promote the aging and clarification of the fruit wine; can be esterified with alcohol to generate lipid, the wine has lipid fragrance, the fruit wine has pleasant taste due to proper amount of acid, and the excessive acid can reduce the flavor of the wine. The aronia melanocarpa has too high total acid content in the fruit, so that the fruit wine has too much acid, rough taste and reduced comfort level.
The main current deacidification methods are: physical deacidification, chemical deacidification and biological deacidification are not widely applied because the physical deacidification can only be carried out singly aiming at the special physical property of a certain acid; other chemical reagents are generally required to be introduced into the chemical deacidification, so that the final separation and purification steps are involved besides potential safety hazards; therefore, biological methods are the mainstream acid reduction measures at present due to their safety and green color. For example, it has been reported that biological deacidification using lactic acid bacteria or yeast not only reduces the acidity of wine, but also increases the stability and quality of wine.
However, in the chinese patent application with publication No. CN106939273A, the inventor of the present invention only improved the bitterness and astringency of fruit wine by using the mixed fermentation method of lactic acid bacteria and yeast, but did not improve the acidity of fruit wine.
In addition, the brewed fruit wine has thinner fragrance, so that the flavor of the brewed fruit wine is not accepted by most of people, and the development and utilization rate of the brewed fruit wine in the field of food is not high.
The taste of the brewed fermented wine is poor due to the high total acid content in the aronia melanocarpa fruit, and meanwhile, the fermented wine has thinner fragrance and serious bitterness; therefore, the method also becomes a primary problem which troubles the development of the black fruit fermented wine.
Disclosure of Invention
In order to solve the problems that the utilization rate of the existing aronia melanocarpa fruit in the field of food is low, and the fruit wine brewed by a common method has poor flavor and is not accepted by consumers, the invention provides a method for producing aronia melanocarpa fruit wine by using specific yeast, which is mainly finished by taking fresh aronia melanocarpa as a raw material through multiple steps of removing stalks, cleaning, juicing, adding water for uniformly mixing, inoculating, fermenting, supplementing materials and the like. The special yeast, namely the acid-reducing yeast and the commercial saccharomyces cerevisiae are creatively utilized to carry out mixed fermentation according to the proportion, and the fermentation method can effectively decompose the citric acid in the aronia melanocarpa wine to reduce the acidity of the fruit wine; the compound with combined aroma in the fruit can be converted into a free state, so that the characteristic of single thin aroma of the aronia melanocarpa wine is greatly improved, the fruit wine is mellow in taste, rich in fruit aroma and proper in oak aroma; the method comprises the following steps:
the Aronia melanocarpa fruit or fruit residues or fruit powder is used as a raw material, and is subjected to cleaning, crushing, uniformly mixing, enzyme treatment, primary fermentation by sequentially inoculating Candida divaricata Z3 and saccharomyces cerevisiae, material supplementing, skin and residue separation after fermentation, secondary fermentation, skin and residue re-separation after secondary fermentation and aging to obtain the Aronia melanocarpa fruit wine.
The Candida divansa Z3 is preserved in China center for type culture collection with the preservation number of CCTCC NO: m2015400, and the preservation date is 2015, 06 months and 24 days.
In one embodiment of the present invention, Candida divaricata Z3 can be cultured in yeast extract peptone glucose medium (YPD) with an optimum temperature of 30 ℃ and an aerobic condition; the culture medium is composed of 2% glucose, 1% peptone, 0.5% yeast extract, 2% agar and distilled water.
The Candida divaricata Z3 has the main function of reducing the acidity of the fruit wine by decomposing citric acid in fruit juice, thereby reducing the total acid content.
The Candida divaricata Z3 not only has the function of reducing acid, but also has better glycosidase activity, can convert glucoside-combined aroma compounds in fruits into free aroma compounds, and greatly improves the defect of single aroma of the aronia melanocarpa wine.
In one embodiment of the invention, the method comprises the steps of:
(1) cleaning: cleaning with sterile water to remove soil and dust, sieving to remove mildewed and rotten fruits, and air drying;
(2) crushing and uniformly mixing: crushing the cleaned fruits, adding sterile water, and mixing uniformly to obtain a aronia melanocarpa crushing liquid; directly adding sterile water into the raw materials of the fruit residues and the fruit powder without cleaning and crushing, and uniformly mixing;
(3) enzyme treatment: filling the crushed aronia melanocarpa liquid into a sterilized fermentation tank, adding pectinase, standing for 3-4h, and performing enzyme treatment to obtain aronia melanocarpa juice;
(4) primary fermentation: sequentially inoculating activated Candida albicans Candida divansa Z3 and saccharomyces cerevisiae into the aronia melanocarpa juice obtained in the step (3) for primary fermentation, wherein the inoculation ratio of the Candida albicans Candida divansa Z3 to the saccharomyces cerevisiae is 1: 1;
(5) feeding: adding white granulated sugar when the fermentation temperature of the primary fermentation is 25-27 ℃ and the fermentation is carried out for 18-30 h, and continuing fermenting for 10-12 days to obtain a primary fermentation mixture;
(6) and (3) separating skin and residue after fermentation: filtering the primary fermentation mixture obtained in the step (5);
(7) and (3) secondary fermentation: performing secondary fermentation on the filtrate obtained in the step (6), wherein the fermentation temperature is 18-22 ℃, and the fermentation time is 12-14 days, so as to obtain a secondary fermentation mixture;
(8) and (3) separating the peel and the residue after secondary fermentation: filtering the secondary fermentation mixture obtained in the step (7);
(9) aging: and (4) standing and ageing the filtrate obtained in the step (8) at the temperature of 18-20 ℃ to obtain the aronia melanocarpa fruit wine.
In one embodiment of the present invention, the Candida divaricata Z3 and Saccharomyces cerevisiae in step (4) are inoculated in the following order: firstly, inoculating activated Candida albicans Candida divansa Z3 into fruit juice, fermenting for 22-26h, and then inoculating saccharomyces cerevisiae.
In one embodiment of the present invention, in step (4), the inoculation amount of Candida albicans Candida divarsa Z3 is: 0.1-0.3% (v/v); the inoculation amount of the saccharomyces cerevisiae is 0.1-0.3% (v/v).
In one embodiment of the present invention, in step (4), the Candida davidi Z3 is inoculated with a concentration of at least 1X 106cfu/mL; the concentration of inoculated Saccharomyces cerevisiae is at least 1 × 106cfu/mL。
In one embodiment of the present invention, in step (4), the primary fermentation conditions are: and (3) performing open tank transfer twice every day on the 1 st to 3 rd days after the fermentation is started at the fermentation temperature of 25-27 ℃, and performing open tank transfer once every day on the 4 th day after the fermentation is started for 12-14 days.
In one embodiment of the present invention, in the step (3), the addition amount of the pectinase is: adding pectinase according to 2% of the weight of the juice after being uniformly mixed, standing for 3-4h to break cells and improve the juice yield.
In one embodiment of the present invention, in the step (3), the enzyme activity of the pectinase is at least 60000U/mL.
In one embodiment of the invention, in the step (5), white granulated sugar is added according to the final alcoholic strength of 10-14 degrees after the fruit wine fermentation is started, namely, 1-degree alcohol is generated according to 17g of sugar, the initial sugar degree of the mixed raw materials is subtracted from the total sugar, and the concentration of the added white granulated sugar is 90-120 g/L.
In one embodiment of the invention, the additional white granulated sugar has a concentration of 96 g/L.
In one embodiment of the invention, in the step (4), the saccharomyces cerevisiae is one or more of angel K1 saccharomyces cerevisiae, angel D254 saccharomyces cerevisiae, angel F5 saccharomyces cerevisiae, angel F15 saccharomyces cerevisiae, and angel RW saccharomyces cerevisiae.
In one embodiment of the present invention, in step (1), the aronia melanocarpa is harvested: picking after the aronia melanocarpa fruits are ripe; cleaning and removing stems: carefully removing bad fruits and fruit stalks, and removing the fruit stalks cleanly, wherein the fruit stalks can be soaked with a large amount of tannin in the fermentation process, the astringent feeling of the fruit wine is higher as the content of the tannin is higher, and then the aronia melanocarpa is dried by using sterile water after being washed for 3 times.
In one embodiment of the invention, in the step (2), the aronia melanocarpa fruit is crushed, and the crushed aronia melanocarpa fruit is 2 parts by weight and the sterile water is 1 part by weight.
In one embodiment of the present invention, in the step (2), the crushing manner is: breaking cell wall, pulverizing, squeezing juice, or manually mashing.
In one embodiment of the present invention, in step (3), the volume of the Aronia melanocarpa crushing liquid charged into the fermentation tank does not exceed 2/3 of the volume of the fermentation tank.
In one embodiment of the present invention, in the step (3), the step of sterilizing the crushed black chokeberry liquid after filling into the fermentation tank comprises: adding flaky potassium sulfate into a fermentation tank, and standing for 10-12 h after the final concentration of the flaky potassium sulfate is 60ppm, wherein in the step (2), the yeast activation method comprises the following steps: respectively weighing 0.1-0.3% (v/v) Candida divansa Z3 and 0.1-0.3% (v/v) saccharomyces cerevisiae, and respectively activating for 20-40 min at 35-37 ℃ by using sucrose solutions with volume fractions of 2-6%.
In one embodiment of the invention, in the step (4), the secondary fermentation mixture is filtered and transferred into an oak barrel, meanwhile, flaky potassium sulfate with the final concentration of 40-50 mg/L is added into the oak barrel, the oak barrel is aged for 9-12 months at 20 ℃, and the middle aging process is carried out by paying attention to supplement wine consumed in the oak barrel, so that the oak barrel is always kept in a full barrel state, and the problem that the fruit wine is seriously oxidized and the taste is influenced due to too much air is avoided.
The invention also provides the aronia melanocarpa fruit wine prepared by the brewing method.
Has the advantages that:
the total acid content of the fruit wine obtained by the brewing method is 4.98g/L, the acidity of the fruit wine is reduced by 37% compared with the fruit wine obtained by only using saccharomyces cerevisiae, and in the fruit wine obtained by the brewing method, more volatile substances are detected, and the content of the volatile compounds in the fruit wine is increased in the categories of esters, lactones, alcohols, aldehydes, ketones, aromatics, furans, nitrogen-containing compounds and terpenes compared with the category of the fruit wine obtained by only using saccharomyces cerevisiae, particularly ester compounds with fruity aroma characteristics and terpene compounds with flowery aroma characteristics.
Biological material preservation
A Candida albicans strain Candida divansa Z3, which is taxonomically named as Candida divansa Z3, is deposited in China Center for Type Culture Collection (CCTCC) 24.06.2015, and has a preservation number of CCTCC NO: m2015400, the preservation address is Wuhan, Wuhan university in China.
Drawings
FIG. 1: a process flow chart of aronia melanocarpa fruit wine.
FIG. 2: an analysis chart of an electronic tongue of aronia melanocarpa fruit wine.
FIG. 3: the concentration of volatile compounds in the aronia melanocarpa fruit wine.
Detailed Description
The yeast for reducing acid of Angel yeast D254, 71B involved in the examples of the present invention was purchased from Angel yeast GmbH.
The media involved in the following examples are as follows:
a yeast extract peptone glucose medium (YPD) culture medium (according to mass ratio): 2% glucose, 1% peptone, 0.5% yeast extract, 2% agar and distilled water.
WL medium: accurately weighing 4g of yeast powder, 5g of peptone, 50g of anhydrous glucose and 20g of agar; adding 40mL of stock solution A and B1mL of stock solution B, adjusting the pH to about 6.5, dissolving with deionized water, and fixing the volume to 1000 mL; sterilizing at 121 deg.C for 20 min; stock C1mL was added after sterilization was complete.
Wherein the stock solution A, B, C consists of:
stock solution a: accurately weighing KH2PO45.5g, KCl 4.25g, anhydrous CaCl21.25g, anhydrous MgSO41.25g, dissolving to a constant volume of 400mL, and storing in a refrigerator at 4 ℃ for later use; and (4) liquid storage B: accurately weighing FeCl3·6H2O 0.25g，MnSO4·H2Dissolving 0.25g of O, fixing the volume to 100mL, and storing in a refrigerator at 4 ℃ for later use; stock solution C: accurately weighing 0.44g of bromocresol green, dissolving the bromocresol green in a mixed solution of 10mL of deionized water and 10mL of absolute ethyl alcohol, and storing the mixed solution in a refrigerator at 4 ℃ for later use; in order to eliminate the interference of bacteria in the screening process, 50-100 mg of chloramphenicol can be added into the sterilized WL culture medium.
Yeast enrichment medium: weighing 50g of anhydrous glucose, 1g of urea and (NH)4)2SO4 1g，KH2PO4 2.5g，Na2HPO4 0.5g，MgSO4·7H2O 1g，FeSO4·7H20.1g of O and 0.5g of yeast powder, dissolving with deionized water, fixing the volume to 1000mL, and sterilizing at 121 ℃ for 20min for later use.
Acid reducing culture medium: the components of the preparation method are the same as those of the yeast enrichment culture medium, except that the anhydrous glucose in the yeast enrichment culture medium is replaced by citric acid; the sterilization conditions were the same as for the yeast enrichment medium.
The detection method according to the following example is as follows:
the method for detecting the total acid content in the aronia melanocarpa fruit wine comprises the following steps:
according to the national standard GB/T15038-2006, 10mL of sample is sucked into a 100mL beaker, 50mL of water is added, an electrode is inserted, a rotor is arranged on the beaker, the beaker is arranged on an electromagnetic stirrer, stirring is started, and titration is carried out by using a sodium hydroxide standard solution. The volume of sodium hydroxide standard solution consumed was recorded and a blank experiment was performed.
X＝[c×(V1-V0)×75]/V2
X represents the total acid content (calculated by tartaric acid) in the sample, g/L; c is the concentration of a sodium hydroxide standard solution, and the unit mol/L; v0The volume of the sodium hydroxide standard solution consumed in the blank experiment is expressed in unit mL; v1For consumption of the sampleThe concentration of the sodium hydroxide standard solution is in unit mL; v2Unit mL for the volume of sample aspirated; and 75 is the molar mass value of tartaric acid, unit g/moL.
The method for detecting the content of volatile substances in the aronia melanocarpa fruit wine comprises the following steps:
5mL of the wine sample and 1.5g of NaCl were placed in a 20mL empty bottle, 10. mu.L of an internal standard (2-methoxy-D3-phenol, final mass concentration 609.76. mu.g/L) was added, and after sealing with a hollow magnetic metal cap with a PTFE/blue silica gel spacer, HS-SPME operation was performed by a multifunctional autosampler system MPS 2. The sample is balanced for 5min at 50 ℃, extracted for 45min, and the rotating speed is set to be 250 r/min. After extraction, the extraction head with the volatile compounds and semi-volatile compounds adsorbed thereon is desorbed at 250 deg.C for 5min without flow splitting at the GC sample inlet, and subjected to GC × GC-TOFMS analysis. Each sample was injected 3 times under the same conditions.
GC × GC-TOFMS analysis method and mass spectrum conditions:
the GC x GC-TOFMS analysis method and the mass spectrum condition refer to the method established by optimization in the laboratory. GC-MS column oven temperature program: the initial temperature of 45 ℃ was maintained for 2min, and the temperature was raised to 230 ℃ at a rate of 4 ℃/min and maintained for 15 min. The modulator modulation time is 4s, the thermal modulation time is 1s, and the modulation compensation temperature is 20 ℃.
Two-dimensional column oven temperature rise program: the initial temperature was maintained at 40 ℃ for 2min, and the temperature was raised to 250 ℃ at 5 ℃/min and maintained for 5 min. High purity helium (> 99.995%) as carrier gas, constant flow mode, flow rate: 1 mL/min.
Time-of-flight mass spectrometry conditions: the EI ion source is adopted, the temperature of the ion source is 230 ℃, the temperature of a transmission line is 280 ℃, and the voltage is controlled to be 70 eV. The mass number range of the detector is 35-400 amu, the collection frequency is 100spec-trum/s, and the voltage of the detector is 430V. Data were collected from the LECO company Pega-sus4D workstation.
The method for detecting the physical and chemical indexes of the aronia melanocarpa fruit wine comprises the following steps:
the alcohol content is measured according to national standard GB/T15038-2006, namely 100mL of wine sample is taken, nonvolatile substances in the sample are removed by a distillation method, distilled water is used for supplementing to 100mL, the volume fraction of alcohol is measured by an alcohol meter method, and the temperature of the alcohol meter and an alcohol content conversion table are searched.
Total sugar determination method:
firstly, decomposing total sugar into reducing sugar, measuring a certain amount of sample V1In a 100mL volumetric flask, the sugar content is adjusted to 0.2-0.4g, 5mL of hydrochloric acid solution is added, water is added to 20mL, and the mixture is shaken up. Hydrolyzing in water bath at 68 + -1 deg.C for 15min, cooling, neutralizing with sodium hydroxide to neutrality, adjusting temperature to 20 deg.C, adding water, and metering to volume2And (5) standby. V2/V1Is the dilution multiple of the wine sample. Taking 0.4mL of diluted sample, adding 0.6mL of LDNS reagent into a test tube, boiling for 5min, cooling, adding 3mL of water, and measuring the light absorption value by using a spectrophotometer at the wavelength of 540 nm. The total sugar concentration in g/L was calculated using a standard curve plotted on glucose.
The total phenol determination method comprises the following steps:
diluting the wine sample with distilled water 50 times by Folin-Ciocaltcu method, taking 0.5mL sample, adding 1mL Folin phenol reagent, and adding 3mL 10% Na2CO3The solution is mixed evenly and reacted for 40min at 37 ℃ in a dark place, and the light absorption value is measured under the wavelength of 760 nm. Drawing a standard curve with gallic acid as standard, calculating the concentration of total phenols in the sample, and expressing with gallic acid equivalent (mg GAE. L)-1)。
The determination method of anthocyanin comprises the following steps:
and (3) measuring the content of the total anthocyanin in the fruit wine by adopting a pH differential method. 0.1mL of the wine sample was placed in a test tube, and pH 1.0(KCl-HCl) buffer and pH 4.5 (CH) were added to the test tube, respectively3COONa) to 5mL, shaking up, keeping out of the sun, stabilizing for 15-20min, measuring absorbance at 520nm and 700nm respectively, and completing measurement within 30 min. The total anthocyanin calculation formula is as follows:
ΔA＝(A520-A700)pH1-(A520-A700)pH4.5
W(mg/L)＝ΔA×MW×DF×1000/(ε×L)
in the formula: w represents the concentration of anthocyanin in the sample; DF represents dilution factor; MW represents the relative molecular mass of cyanidin-3-glucoside (Cy-3-Glu), 449.2; ε represents the extinction coefficient of cyanidin-3-glucoside, 26900; l represents the thickness of the liquid layer, cm.
Example 1: isolation, purification, screening and molecular biological characterization of Candida divaricata DiversaZ3
1. Separation, purification and screening of Candida divansaZ 3
(1) Accurately sampling and weighing samples (5.00 g of each soil in an orchard of water chestnut red bayberry and Dongkui red bayberry, 2.00g of each soil in branch and leaf of red bayberry and 10.00g of each fruit of red bayberry) at multiple points, respectively placing the obtained samples into 250mL conical flasks, adding 100mL of physiological saline (0.9% NaCl solution) and a little glass beads, respectively placing the marked samples into a 30-DEG C shaking table at 200r/min, shaking and shaking uniformly for about 2h, and standing. And (3) accurately sucking 5mL of the supernatant respectively, inoculating the supernatant into a yeast enrichment culture medium (50 mL of culture solution is filled in each 250mL conical flask) sterilized in advance, and culturing for 1 day in a shaking table at 30 ℃ and 200r/min to obtain a pretreatment culture solution.
(2) The pretreated culture solution obtained in the step (1) is stood and then is subjected to gradient dilution (10)-1～10-6) 3 appropriate dilution gradients (10) were selected-3、10-4、10-5) 0.1mL of each dilution was pipetted and applied to WL medium plates, and each dilution was repeated 3 times, and incubated at a constant temperature in an incubator at 30 ℃ for 3 days.
(3) And (3) respectively inoculating single colonies with complete colony morphology growth in the WL culture medium obtained in the step (2) into a deacidification test tube containing the deacidification culture medium, culturing for 4 days at the temperature of 30 ℃ and at the speed of 200r/min, observing the growth condition of thalli in the test tube every 12 hours, and recording the growth condition after the culture is finished.
(4) And selecting test tubes which become turbid after culture, namely test tubes with bacteria growing, and numbering the test tubes. And (4) streaking and separating on the sterilized deacidified solid culture medium, and purifying the strain.
2. Screening of Candida divansa Z3
Inoculating the pure strain obtained by separation into sterilized YPD liquid culture medium with the inoculation amount of 100 μ L, and performing shake bed activation culture at 30 deg.C and 200r/min for 24 h. Inoculating the activated bacterial liquid into an acid reduction culture medium containing 10% (v/v) of absolute ethyl alcohol, wherein the inoculation amount is 6% (v/v). Under the conditions of 30 ℃ and 200r/min,performing shake culture; inoculating the obtained bacterial liquid of the turbid test tube to a culture tube containing 100ppm SO2The amount of the inoculum in the acid-reducing medium of (4) is 6% (v/v). Performing shake culture at 30 ℃ and 200 r/min; the resulting turbid bacterial solution of the test tube was inoculated into a 250mL Erlenmeyer flask in an amount of 6% (v/v) in a deacidification medium (liquid). Shaking culture at 30 deg.C and 200 r/min. Sampling 1mL every 12 hours, centrifuging for 1min under 10000r/min, taking supernatant, diluting to 10mL, and determining the residual acid amount in the culture solution.
3. Identification of Candida divaricata Z3
By subjecting the 1 strain obtained above to resistance to both alcohol and SO2Meanwhile, the strain with higher deacidification rate extracts the whole genome, the processed gene fragment is sent to Shanghai gene data research institute for sequencing after PCR amplification, and DNA man and NCBI database data comparison are carried out on the returned sequencing result. The results of the comparison of NCBI data show that the screened yeasts are Candida CEC Y111 type, the homology of the genes is 99%, the nucleotide sequence is shown in SEQ ID NO.1, and the yeasts can be determined to be Candida. And finally determining the Candida as the Candida by integrating morphological feature observation, physiological and biochemical tests and molecular biological sequencing results, wherein the Candida is named Candida divarsansa Z3.
Example 2: brewage of aronia melanocarpa fruit wine
The specific steps are as follows (as shown in figure 1):
(1) preparing the aronia melanocarpa fruit: picking after the aronia melanocarpa fruits are ripe; cleaning and removing stems: bad fruits and fruit stalks are carefully removed, because a large amount of tannin can be soaked in the fruit stalks in the fermentation process, silt and dust are cleaned by sterile water, water is dried in the air, and the cleaning time is not suitable to be too long.
Crushing and uniformly mixing: because the aronia melanocarpa has low water content, the crushed aronia melanocarpa fruits and sterile water are mixed according to the mass ratio of 2:1 to obtain a crushed black chokeberry liquid, and filling the crushed black chokeberry liquid into a fermentation tank, wherein the volume of the crushed black chokeberry liquid is not more than 2/3.
Sterilizing the aronia melanocarpa crushing liquid: adding SO in the form of potassium sulfate flake into the fermentation tank2Standing at final concentration of 60ppm for 12 hr to obtain crude juice; SO (SO)2Has the main function of sterilization.
Enzyme treatment: adding pectase into the crude fruit juice according to 2% of the crude fruit juice, wherein the enzyme activity of the pectase is 60000U/mL, and standing for 3-4h to obtain an enzymatic hydrolysate; the pectinase is added to mainly act on breaking cells by an enzymolysis method so as to improve the juice yield; filtering the obtained enzymolysis solution to obtain Aronia melanocarpa juice.
(2) Activating the yeast: respectively inoculating the Candida albicans Z3 and the Angel yeast D254 obtained in the example 1 into a yeast extract peptone glucose medium (YPD) solid medium, culturing for 8-12 h at 30 ℃ to respectively obtain a Candida albicans Z3 bacterial colony and an Angel yeast D254 bacterial colony, respectively selecting Candida albicans Z3 and an Angel yeast single bacterial colony, inoculating the single bacterial colony into a sucrose solution with the volume fraction of 2-6%, and activating for 20min at 37 ℃; respectively obtaining the concentration of the bacteria of 1 multiplied by 106The seed liquid and the bacterial concentration of cfu/mL Candida divarsia Z3 are 1 multiplied by 107cfu/mL Angel yeast D254 seed liquid.
Inoculation: sequentially inoculating activated Candida albicans Candida divansa Z3 and Angel yeast D254 to the Aronia melanocarpa juice obtained in the step (1), wherein the inoculation amount of the Angel yeast is 0.2% (v/v), and the inoculation amount of the Candida albicans Candida divasa Z3 is 0.2% (v/v); the method comprises the steps of inoculating activated Candida albicans Candida divansa Z3 into fruit juice, fermenting for 22-26h at 20 ℃, and inoculating Angel yeast D254 to obtain an inoculated raw material.
Primary fermentation (namely main fermentation): fermenting the inoculated raw materials in a fermentation tank; wherein the fermentation liquor in the fermentation tank accounts for 2/3, the air accounts for 1/3, and the tank opening of the fermentation tank ensures the air to enter and exit;
on the 2 nd day after the fermentation is started, after the fermentation is full, adding white granulated sugar (supplementary material) according to the final alcoholic strength of 13 degrees, namely subtracting the initial sugar degree of the mixed raw materials from the total sugar which is calculated according to the alcohol with 1 degree generated by 17g of sugar, wherein the initial sugar degree is 125g/L, and therefore 96g/L of white granulated sugar is respectively supplemented into the fermentation tank; in the fermentation process, the temperature is 27 ℃, the open tank transfer is carried out twice every day on the 1 st to 3 rd days after the start of the fermentation, the open tank transfer is carried out once every day on the 4 th day after the start of the fermentation, the fermentation time is 12 days, the first fermentation mixture is obtained, the specific gravity is reduced to 0.998, and the calculation method of the specific gravity is as follows: the specific gravity and the fermentation degree of the wine are used for measuring, so that whether the fermentation is normal or not and whether the fermentation is finished or not are judged.
(3) Separating peel and slag: filtering the primary fermentation mixture, and separating skin and residue;
placing the filtrate in a stainless steel storage tank, storing in full volume, and performing secondary fermentation (i.e. after-fermentation) on the filtrate; fermenting at 20 deg.C for 14 days to obtain secondary fermentation mixture.
(4) Aging: filtering the secondary fermentation mixture, transferring the filtrate into an oak barrel for aging, simultaneously adding sheet potassium sulfate with final concentration of 50mg/L into the oak barrel, and aging at 20 deg.C for 10 months to obtain Aronia melanocarpa fruit wine 1. In the middle aging process, the liquor consumed in the oak barrel is supplemented, so that the oak barrel is always kept in a full state. Avoid the serious oxidation of the fruit wine caused by too much air. The oak barrel can provide a micro-aerobic environment for the aronia melanocarpa fruit wine, promote the aggregation and agglutination of tannin, and reduce the astringent feeling of the fruit wine.
Example 3: brewage of aronia melanocarpa fruit wine
1. The specific implementation manner is different from that of example 2 in that: and (2) inoculating activated saccharomyces cerevisiae into the aronia melanocarpa juice obtained in the step (1) according to 0.2 percent of the total volume of the aronia melanocarpa juice to prepare the aronia melanocarpa fruit wine 2.
2. The specific implementation manner is different from that of example 2 in that: adjusting Candida divansa Z3 to 71B acid-reducing yeast; the other conditions are completely the same as those in example 2, and the aronia melanocarpa fruit wine 3 is prepared.
3. The specific implementation manner is different from that of example 2 in that: the yeast ratio was adjusted so that the inoculum size of Angel yeast D254 was 0.2% (v/v) and the inoculum size of Candida divansa Z3 was: 0.1% (v/v), and preparing the aronia melanocarpa fruit wine 4.
Example 4: component analysis of aronia melanocarpa fruit wine
The aronia melanocarpa fruit wine prepared by the specific method of the invention not only well retains the nutrient components, but also reduces the astringent feeling and the acidity, and is better accepted by consumers.
(1) Basic physicochemical indexes are as follows: as shown in table 1:
table 1: basic physicochemical indexes of aronia melanocarpa fruit wine
As can be seen from Table 1: the black chokeberry fruit wine 1 is prepared by a specific method, wherein the alcohol content in the black chokeberry fruit wine is 12.6% (v/v), the total sugar content is 1.8g/L, the total phenol content is 2.50g/L, and the anthocyanin content is 723.25 mg/L.
The aronia melanocarpa fruit wine 2 has 13.0% (v/v) of alcohol, 1.5g/L of total sugar, 2.56g/L of total phenol and 713.28mg/L of anthocyanin.
(2) The total acid content in aronia melanocarpa is measured by an acid-base titration method, and is calculated by tartaric acid.
After detection, the method comprises the following steps: the aronia melanocarpa fruit wine 1 has the following total acid content: 4.98 g/L;
the aronia melanocarpa fruit wine 2 has the following total acid content: 7.96 g/L;
the aronia melanocarpa fruit wine 3 has the following total acid content: 6.02 g/L;
the aronia melanocarpa fruit wine 4 has the following total acid content: 5.37g/L
Therefore, the aronia melanocarpa fruit wine prepared by the preparation method has the best effect of reducing acid, and the total acid content is only 55.58% of that of single saccharomyces cerevisiae fermentation.
(3) Evaluating the electronic tongue taste of the fruit wine: the electronic tongue is used for simulating the human tongue to analyze, recognize and judge a sample to be detected, and the obtained data is processed by a multivariate statistical method, so that the sour, bitter, salty, fresh and astringent samples are quickly reflected.
Evaluated by an electronic tongue, wherein: in the aronia melanocarpa fruit wine 1, the electronic tongue scores sour taste as follows: -2.47;
in the aronia melanocarpa fruit wine 2, the electronic tongue scores sour taste as follows: -0.68;
in the aronia melanocarpa fruit wine 3, the electronic tongue scores sour taste as follows: -1.52;
the aronia melanocarpa fruit wine 4 has the following sour taste scores of the electronic tongue: -2.02
It can be known that the sour taste of the aronia melanocarpa fruit wine prepared by the specific method is-2.47, while the sour taste of the aronia melanocarpa fruit wine fermented by the common method is-0.68, and the acidity of the aronia melanocarpa fruit wine brewed by the specific method is far smaller than that of the aronia melanocarpa fruit wine fermented by the common method; the specific information is shown in figure 2.
(4) The specific information of the comparison of the volatile compounds in the examples by using HS-SPME-GC × GC-TOFMS is shown in fig. 3, wherein, compared with the aronia melanocarpa wine 2, 3, 4 brewed by other methods, the aronia melanocarpa wine 1 obtained by using the technical scheme of the present invention detects more volatile substances, and the types and contents thereof are shown in table 2:
table 2: comparison table of species of volatile compounds in Aronia Melanocarpa fruit wine
As can be seen from Table 2: 219 and 222 volatile compounds are detected respectively from the common yeast brewed Aronia melanocarpa fruit wine 2 and the common yeast brewed fruit wine 3, 267 volatile compounds are detected from the Aronia melanocarpa fruit wine 1 brewed by the method provided by the invention, and 239 volatile compounds are detected from the common yeast brewed fruit wine 4; the fruit wine 1 has more varieties of esters, lactones, alcohols, aldehydes, ketones, aromatics, furans, nitrogens and terpenes than other three groups of fruit wine, especially esters with fruity aroma characteristic and terpenes with floral aroma characteristic. Screening is carried out by utilizing two aroma banks of Flavornet Home and Flavor DB, and the compound with aroma characteristics in the locked wine is semi-quantitatively carried out by utilizing methyl caproate as an internal standard (the specific method is shown in Van der, and the research on the influence of yeast on the aroma of the Weidai ice wine), and the result is shown in Table 3.
TABLE 3 semi-quantitative results of compounds with aroma characteristics in Aronia melanocarpa fruit wine
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
SEQUENCE LISTING
<110> university of south of the Yangtze river
<120> aronia melanocarpa fruit wine and brewing method thereof
<130> BAA201003A
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 541
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Claims (11)
1. A brewing method of aronia melanocarpa fruit wine is characterized by comprising the following steps: taking Aronia melanocarpa fruit or fruit residues or fruit powder as raw materials, treating the raw materials with enzyme, and sequentially inoculating candidaCandida diversaCarrying out primary fermentation, material supplementing, skin and residue separation after fermentation, secondary fermentation, skin and residue separation after secondary fermentation and aging on the Z3 and saccharomyces cerevisiae to obtain the aronia melanocarpa fruit wine;
the CandidaCandida diversaZ3 is preserved in China center for type culture Collection with the preservation number of CCTCC NO: m2015400, and the preservation date is 2015, 06 months and 24 days.
2. The method for brewing aronia melanocarpa wine according to claim 1, which comprises the following steps:
(1) cleaning, screening out mildewed fruits and rotten fruits, and airing water;
(2) crushing and uniformly mixing: crushing the cleaned fruits, and adding water to obtain a crushed liquid of the aronia melanocarpa; directly adding sterile water into the raw materials of the fruit residues and the fruit powder without cleaning and crushing, and uniformly mixing;
(3) enzyme treatment: sterilizing the aronia melanocarpa crushing liquid, adding pectinase, standing for 3-4h, and performing enzyme treatment to obtain aronia melanocarpa juice;
(4) primary fermentation: inoculating activated candida sequentially into the aronia melanocarpa juice obtained in the step (3)Candida diversaZ3 and Saccharomyces cerevisiae, and performing primary fermentation, wherein the Candida is used as the yeastCandida diversaThe inoculation ratio of Z3 to Saccharomyces cerevisiae is 1: 1;
(5) feeding: adding white granulated sugar when the fermentation temperature of the primary fermentation is 25-27 ℃ and the fermentation is carried out for 18-30 h, and continuing fermenting for 10-12 days to obtain a primary fermentation mixture;
(6) and (3) separating skin and residue after fermentation: filtering the primary fermentation mixture obtained in the step (5);
(7) and (3) secondary fermentation: performing secondary fermentation on the filtrate obtained in the step (6), wherein the fermentation temperature is 18-22 ℃, and the fermentation time is 12-14 days, so as to obtain a secondary fermentation mixture;
(8) and (3) separating the peel and the residue after secondary fermentation: filtering the secondary fermentation mixture obtained in the step (7);
(9) aging: and (4) standing and ageing the filtrate obtained in the step (8) at the temperature of 18-20 ℃ to obtain the aronia melanocarpa fruit wine.
3. The method for brewing Aronia melanocarpa wine according to claim 2, wherein the Candida albicans in the step (4)Candida diversaThe inoculation sequence of Z3 and Saccharomyces cerevisiae was: firstly, activated candidaCandida diversaAnd Z3 is inoculated into the fruit juice, fermented for 22-26h, and then inoculated with the saccharomyces cerevisiae.
4. The method for brewing Aronia melanocarpa wine according to claim 3, wherein in the step (4), Candida sppCandida diversaThe concentration of the bacteria inoculated by Z3 is at least 1 x 106cfu/mL; the concentration of inoculated Saccharomyces cerevisiae is at least 1 × 106cfu/mL。
5. The method for brewing aronia melanocarpa wine according to claim 4, wherein in the step (3), the enzyme activity of the pectinase is at least 60000U/mL.
6. The brewing method of aronia melanocarpa fruit wine according to claim 5, characterized in that in the step (5), the concentration of the added white granulated sugar is 90-120 g/L.
7. The method for brewing aronia melanocarpa wine according to claim 6, wherein in the step (4), the saccharomyces cerevisiae is one or more of Angel K1 saccharomyces cerevisiae, Angel D254 saccharomyces cerevisiae, Angel F5 saccharomyces cerevisiae, Angel F15 saccharomyces cerevisiae and Angel RW saccharomyces cerevisiae.
8. The method for brewing aronia melanocarpa fruit wine according to claim 7, wherein in the step (2), the crushed aronia melanocarpa fruit is mixed with sterile water in a weight ratio of 2: 1.
9. The method for brewing Aronia melanocarpa wine as claimed in claim 7, wherein in the step (3), the volume of the crushed Aronia melanocarpa liquid filled into the fermentation tank does not exceed 2/3 of the volume of the fermentation tank.
10. The method for brewing aronia melanocarpa fruit wine according to claim 9, wherein in the step (3), the step of sterilizing the aronia melanocarpa crushing liquid after filling the aronia melanocarpa crushing liquid into the fermentation tank comprises: adding potassium sulfate into the fermentation tank, adding flaky potassium sulfate with a final concentration of 60ppm, adding potassium sulfate, and standing for 10-12 h.
11. Aronia melanocarpa wine obtained by the brewing method according to any one of claims 1 to 10.
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