CN116376729A - Wick yeast, microbial preparation and medlar western style wine and brewing method thereof - Google Patents
Wick yeast, microbial preparation and medlar western style wine and brewing method thereof Download PDFInfo
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/165—Yeast isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12G3/023—Preparation of other alcoholic beverages by fermentation of botanical family Solanaceae, e.g. potato
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12R2001/00—Microorganisms ; Processes using microorganisms
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- C12R2001/85—Saccharomyces
- C12R2001/865—Saccharomyces cerevisiae
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- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
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Abstract
The invention discloses a Wilker's yeast, a microbial preparation and medlar western beating wine and a brewing method thereof, wherein the preservation numbers of the Wilker's yeast BYBC2.22146 and the Saccharomyces cerevisiae BYBC2.21113 are CGMCC No.26360 and CGMCC No.23129 respectively; the brewing method comprises the following steps: mixing fructus Lycii juice with water and/or crushing dried fructus Lycii with water, homogenizing, and sterilizing to obtain fermentation liquid; inoculating the activated and cultured abnormal Wick yeast into the liquid to be fermented for liquid fermentation, and then inoculating the activated and cultured Saccharomyces cerevisiae for liquid fermentation; aging to obtain fructus Lycii western medicated liquor. The invention utilizes the mixed fermentation of the abnormal Wick ham yeast and the saccharomyces cerevisiae to brew the medlar western style wine, greatly improves the total ester content of the medlar western style wine, obviously improves the taste and the flavor of the medlar western style wine, and effectively improves the fermentation degree and the quality of the medlar western style wine.
Description
Technical Field
The invention relates to the technical field of microorganism application and food brewing, in particular to Wick ham yeast, a microorganism preparation, medlar Xiya wine and a brewing method thereof.
Background
The medlar serving as a traditional Chinese medicinal material for both medicine and food has rich nutrition ingredients and high medicinal value, is rich in various active nutrition ingredients such as medlar polysaccharide, medlar acid, flavone, polyphenol, betaine and the like, and has the health care effects of resisting oxidation, resisting fatigue, reducing blood fat, reducing hyperlipidemia, reducing hypertension, hyperglycemia, hyperlipidemia, hyperglycemia, hyperlipidemia, diabetes, hypertension, hyperlipidemia, diabetes, hypertension, diabetes mellitus and the like. However, fresh fruits of Lycium barbarum are small and juicy, and are easy to age and spoil after picking, and have short storage time, so the Lycium barbarum is sold in a dry form in many cases.
Along with the enhancement of health care consciousness of people, medlar porridge, tea and soup are cooked, so that the health care effects of clearing liver and improving vision, nourishing yin and moisturizing lung, nourishing kidney and replenishing vital essence are achieved. The wolfberry industry is also focused on the exploration and development of wolfberry, is focused on the fields of basic research, application research, cultural propagation and the like of the wolfberry, is continuously innovated in the field of deep processing of the wolfberry in order to improve the social and economic benefits of the wolfberry, and has important significance and value in widening the variety of deep processed products of the wolfberry.
The western style wine is a low-alcohol juice beverage which is brewed by adopting special-grade medlar as a raw material and through a low-temperature fermentation-natural aging process, combines the advantages of beer and medlar puree, and has clear and mellow taste and rich nutrition. Yeast is an important factor in the fermentation process of the Xiya wine, directly influences the flavor of the Xiya wine, and determines the quality of the Xiya wine. At present, most of China uses single Saccharomyces cerevisiae to ferment Xiya wine, so that the Xiya wine obtained by brewing often has the problems of low wine body, insufficient fragrance and the like.
In view of this, the present invention has been made.
Disclosure of Invention
In order to solve the defects in the technology, the invention provides abnormal Wick ham yeast, a microbial preparation, medlar Xiya wine and a brewing method thereof.
In order to achieve the above purpose, the technical scheme of the invention is as follows:
in one aspect, the invention provides an abnormal Wick ham yeastWickerhamomyces anomalus) BYBC2.22146 is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.26360 in the year 12 and 30 of 2022.
Specifically, the abnormal Wick ham yeastWickerhamomyces anomalus) BYBC2.22146 can be used for fermenting the Chinese wolfberry western style wine to solve the problems of low wine body, insufficient fragrance and the like.
The invention also provides a microbial preparation which comprises the abnormal Wickham yeastWickerhamomyces anomalus）BYBC 2.22146。
The invention also provides the abnormal Wick ham yeast with the preservation number of CGMCC No.26360Wickerhamomyces anomalus) Use of BYBC2.22146 in brewing a lyceum berry western style wine.
The invention also provides a brewing method of the wolfberry western style wine, which comprises the following steps:
s1, adding water into the raw wolfberry pulp, mixing and/or adding water into dried wolfberry fruits, crushing, homogenizing, and sterilizing to obtain a to-be-fermented liquid;
s2, firstly, performing activation culture on the abnormal substancesWeikeham yeastWickerhamomyces anomalus) BYBC2.22146 is inoculated into the liquid fermentation to be fermented, and then the activated and cultured saccharomyces cerevisiae is subjected to liquid fermentationSaccharomyces cerevisiae) BYBC2.21113 is inoculated and subjected to liquid fermentation;
and S3, ageing to obtain the medlar western beating wine.
In the above technical solution, in step S2,
abnormal Weikeham yeastWickerhamomyces anomalus) BYBC2.22146 inoculated at 1X 10 each 6 - 8×10 6 CFU/ml and 3.5-6 wt%, the temperature and time of liquid state fermentation are 16-24 ℃ and 45-54 h respectively;
saccharomyces cerevisiaeSaccharomyces cerevisiae) BYBC2.21113 inoculated at 1X 10 each 6 - 8×10 6 CFU/ml and 3.5-6. 6 wt%, the temperature and time of liquid fermentation are 16-24 ℃ and 66-78 h, respectively.
In detail, in the technical scheme, the abnormal Wick yeast is firstly inoculated, and then the saccharomyces cerevisiae is inoculated after fermentation, the growth of the abnormal Wick yeast is inhibited by adding the saccharomyces cerevisiae, more volatile aroma components can be generated by inoculating the abnormal Wick yeast after inoculating 48 h, the fragrance of the wolfberry western beating wine is improved, and then the fermentation efficiency is improved and the advantages of the abnormal Wick yeast and the saccharomyces cerevisiae are fully exerted due to inoculating the saccharomyces cerevisiae.
Specifically, in the above technical scheme, in step S2, abnormal wilms yeast @Wickerhamomyces anomalus) The activation culture of BYBC2.22146 is specifically as follows:
activating the abnormal Wick ham microzyme preserved at-80 ℃ for 15-30min at 28 ℃, dipping the abnormal Wick ham microzyme into a solid wort culture medium flat plate for streaking, then culturing the abnormal Wick ham microzyme at 28 ℃ for 48 h, picking a single colony in a 100 mL wort liquid culture medium by using the inoculating loop, and culturing the abnormal Wick ham microzyme at a constant temperature of 28 ℃ and 200 rpm for 22 h.
Specifically, in the above technical scheme, in step S2, saccharomyces cerevisiae @ is usedSaccharomyces cerevisiae) The activation culture of BYBC2.21113 is specifically as follows:
activating Saccharomyces cerevisiae preserved at-80deg.C for 15-30min at 28deg.C, streaking with inoculating loop, culturing at 28deg.C for 48 h, picking single colony with inoculating loop in 100 mL wort liquid medium, and culturing at 28deg.C and 200 rpm under constant temperature shaking for 24 h.
Further, in the above technical solution, in step S3, the aging specifically includes:
taking fermentation liquor with residual sugar concentration of 6.0-8.0 g/L, cooling to 4 ℃ at constant speed at a rate of 10 ℃ every 24-h, and storing 7-d.
In detail, in the above technical scheme, the brewing starts when the concentration of residual sugar is reduced to 6-8 g/L, and the required alcoholicity is reached and the sensory score is optimal; the temperature is gradually reduced in stages, so that the thalli enters a dormant state in a relatively gentle environment, and the thalli dies, autolyzes and breaks to enable content substances to flow into the wine body due to severe changes of the environment.
Further, in the above technical solution, in step S1,
the initial sugar degree of the to-be-fermented liquid is 8-16 Brix%.
In addition, the invention also provides the wolfberry western style wine brewed by the brewing method of the wolfberry western style wine.
Specifically, in the technical scheme, the alcoholic strength, the actual fermentation degree, the alcohol substances and the ester substances of the medlar sirocci wine are respectively 2.26-3.73 v%, 62-68%, 32.39-35.21% and 56.00-63.43%.
Compared with the prior art, the invention has the following advantages:
(1) The invention utilizes the provided abnormal Wick ham yeastWickerhamomyces anomalus) BYBC2.22146 and Saccharomyces cerevisiaeSaccharomyces cerevisiae) BYBC2.21113 mixed fermentation is adopted to brew the Chinese wolfberry western style wine, and a specific brewing process is adopted, so that the total ester content of the Chinese wolfberry western style wine is greatly improved, the taste and the flavor of the Chinese wolfberry western style wine are obviously improved, and the fermentation degree and the quality of the Chinese wolfberry western style wine are effectively improved;
(2) The invention adopts fresh medlar magma and special medlar as raw materials, and the western style wine directly brewed by adding yeast has no additive, not only has the beer flavor of beer, but also has the characteristic of low alcohol degree, in addition, the brewed medlar western style wine contains various minerals, amino acids, vitamins, trace elements, organic acids and the like beneficial to human bodies, has the advantages of low sugar content, abundant nutrient content, low alcohol degree and the like, has unique taste, has the functions of resisting oxidation, reducing blood fat, regulating human immunity, whitening, delaying organism aging and the like, and has great development and utilization value.
Drawings
FIG. 1 is a flow chart of a brewing process of the Lycium barbarum western beaten wine in the embodiment of the invention;
FIG. 2 is a total ion flow chromatogram of aroma components of Lycium barbarum western style wine obtained in example 1 of the present invention;
FIG. 3 is a graph showing total ion flow chromatograms of aroma components of a blank (prepared from a yeast-free stock of Lycium barbarum of example 1);
FIG. 4 is a graph showing the measurement of antioxidant efficacy in the examples of the present invention;
FIG. 5 is a standard graph of sodium glycocholate and sodium taurocholate in an example of the present invention;
FIG. 6 shows the results of measuring the cholate binding ability of the Lycium barbarum puree of the present invention and the Lycium barbarum Sida wine obtained in example 1;
FIG. 7 is a graph showing the results of measurement of tyrosinase inhibitory activity of the raw wolfberry fruit pulp of the present invention and the Sichuan wolfberry fruit wine obtained in example 1.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent.
It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
In the examples, all means used are conventional in the art unless otherwise specified.
The terms "comprising," "including," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, step, method, article, or apparatus.
In addition, the technical features of the embodiments of the present invention described below may be combined with each other as long as they do not collide with each other.
In the following examples of the present invention, all raw materials used were commercial products, which were used without special treatment.
The saccharomyces cerevisiae in the embodiment of the invention is screened out in the fresh medlar fruits by combining thallus enrichment with gradient dilution and flat coating in the earlier stage; the abnormal Wikimann yeast is screened from the dyed medlar puree which is not stored properly.
Specifically, under the aseptic condition, a certain amount of fresh fruits of Chinese wolfberry are taken and added into a 500 mL triangular flask which is sterilized and filled with 100 mL of YEPD liquid culture medium, 24 h is cultivated in a shaking table with the rotation speed of 200 rpm at the temperature of 28 ℃ to enrich thalli, then the enriched bacterial liquid is diluted step by step, bacterial liquid with proper gradient is coated on a YEPD culture medium flat plate added with penicillin, 48 h is cultivated in a constant temperature incubator at the temperature of 28 ℃, single bacterial colony is selected, and repeated separation and purification microscopic examination is carried out, and strain identification is carried out to obtain the saccharomyces cerevisiae strain.
Specifically, under the aseptic condition, taking a certain amount of Chinese wolfberry magma with bacteria, adding the Chinese wolfberry magma into a 500 mL triangular flask which is sterilized and filled with 100 mL of YEPD liquid culture medium, culturing 24 h in a shaking table with the rotation speed of 200 rpm at the temperature of 28 ℃ to enrich thalli, gradually diluting the enriched bacterial liquid, taking bacterial liquid with proper gradient, coating the bacterial liquid on a YEPD culture medium flat plate added with penicillin, culturing 48 h in a constant temperature incubator at the temperature of 28 ℃, picking single bacterial colony, repeatedly separating and purifying for microscopic examination, and obtaining the abnormal Weikeham yeast strain through strain identification.
Description of biological preservation
Abnormal Weikeham yeastWickerhamomyces anomalus) BYBC2.22146, 12/30 of 2022The preservation address is stored in the China general microbiological culture Collection center of the China Committee for culture Collection of microorganisms: the collection number is CGMCC No.26360, which is the national academy of sciences of China, no. 3 of North Chen West Lu, the Korean region of Beijing.
Saccharomyces cerevisiaeSaccharomyces cerevisiae) BYBC2.21113, deposited at the China general microbiological culture Collection center, accession number: the collection number is CGMCC No.23129, which is No. 3 of North West Lu No. 1 (China academy of sciences microbiological institute) in the Korean region of Beijing city.
The activation, culture and expansion processes of the abnormal Wikipedia are as follows:
taking abnormal Wikimann yeast preserved at-80 deg.CWickerhamomyces anomalus) BYBC2.22146 is placed in a constant temperature incubator at 28deg.C for 15-30min, inoculated with an inoculating loop, streaked onto a solid wort medium plate, then cultured at 28deg.C for 48 h, inoculated with an inoculating loop, picked up as a single colony in 100 mL wort liquid medium, cultured at 28deg.C under shaking at 200 rpm shaking table constant temperature for 22 h, and finally diluted 25 times (OD 600 =0.491), and the seed liquid is obtained.
The activating, culturing and expanding processes of the saccharomyces cerevisiae are as follows:
collecting Saccharomyces cerevisiae preserved at-80deg.CSaccharomyces cerevisiae) BYBC2.21113 is placed in a constant temperature incubator at 28deg.C for 15-30min, inoculated with an inoculating loop, streaked onto a solid wort medium plate, then cultured at 28deg.C for 48 h, inoculated with an inoculating loop, picked up as a single colony in 100 mL wort liquid medium, cultured at 28deg.C under shaking at 200 rpm shaking table constant temperature for 24 h, and finally diluted 25 times (OD 600 =0.529), and a seed liquid thereof is obtained.
Example 1
The embodiment of the invention provides a brewing method of medlar western beating wine, as shown in figure 1, which specifically comprises the following steps:
s1, adding water into the medlar original pulp, mixing, adjusting the sugar degree to be 10 Brix, and sterilizing to obtain a to-be-fermented liquid;
s2, preparing abnormal Wick ham yeastWickerhamomyces anomalus) Adding BYBC2.22146 seed solution into the liquid to be fermented (inoculum size is 5%), fermenting 48 h, and adding Saccharomyces cerevisiaeSaccharomyces cerevisiae) Seed liquid of BYBC2.21113 is added (inoculum size is 5%), liquid fermentation is 72 h, and inoculation concentration of both bacteria is (1-8) x 10 6 CFU/mL, the total fermentation process is 5d, in the fermentation process, sufficient oxygen is provided for the liquid to be fermented in advance for yeast proliferation, then the pressure in a pressure tank is controlled to be 0.12 MPa, the constant temperature fermentation is carried out at 18 ℃, fermentation liquid starts to be clarified after 5d fermentation, thalli starts to be condensed at the bottom of the tank, and the concentration of residual sugar is 6.0-8.0 g/L;
s3, cooling to 4 ℃ at a constant speed at a rate of 10 ℃ every 24-h, storing 7-d, filtering out residues and canning to obtain the Chinese wolfberry western style wine.
Example 2
The embodiment of the invention provides a brewing method of medlar western beating wine, which specifically comprises the following steps:
s1, crushing and homogenizing dried fruits of Chinese wolfberry with water with the volume of 5 times, extracting 24 h at 60+/-0.5 ℃, adjusting the sugar degree to be 12 Brix, and sterilizing to obtain a to-be-fermented liquid;
s2, preparing abnormal Wick ham yeastWickerhamomyces anomalus) Adding BYBC2.22146 seed solution into the liquid to be fermented (inoculum size is 5%), fermenting 48 h, and adding Saccharomyces cerevisiaeSaccharomyces cerevisiae) Seed liquid of BYBC2.21113 is added (inoculum size is 5%), liquid fermentation is 72 h, and inoculation concentration of both bacteria is (1-8) x 10 6 CFU/mL, the total fermentation process is 5d, in the fermentation process, sufficient oxygen is provided for the liquid to be fermented in advance for yeast proliferation, then the pressure in a pressure tank is controlled to be 0.12 MPa, the constant temperature fermentation is carried out at 21 ℃, fermentation liquid starts to be clarified after fermentation for 5d, thalli starts to be condensed at the bottom of the tank, and the concentration of residual sugar is 6.0-8.0 g/L;
s3, cooling to 4 ℃ at a constant speed at a rate of 10 ℃ every 24-h, storing 7-d, filtering out residues and canning to obtain the Chinese wolfberry western style wine.
Example 3
The embodiment of the invention provides a brewing method of medlar western beating wine, which specifically comprises the following steps:
s1, adding water into the medlar original pulp, mixing, adjusting the sugar degree to 15 Brix, and sterilizing to obtain a to-be-fermented liquid;
s2, preparing abnormal Wick ham yeastWickerhamomyces anomalus) Adding BYBC2.22146 seed solution into the liquid to be fermented (inoculum size is 5%), fermenting 48 h, and adding Saccharomyces cerevisiaeSaccharomyces cerevisiae) Seed liquid of BYBC2.21113 is added (inoculum size is 5%), liquid fermentation is 96 h, and inoculation concentration of both bacteria is (1-8) x 10 6 CFU/mL, the total fermentation process is 6d, in the fermentation process, sufficient oxygen is provided for the liquid to be fermented in advance for yeast proliferation, then the pressure in a pressure tank is controlled to be 0.12 MPa, the constant temperature fermentation is carried out at 21 ℃, the fermentation liquid starts to be clarified after fermentation for 6d, thalli starts to be condensed at the bottom of the tank, and the concentration of residual sugar is 6.0-8.0 g/L;
s3, cooling to 4 ℃ at a constant speed at a rate of 10 ℃ every 24-h, storing 7-d, filtering out residues and canning to obtain the Chinese wolfberry western style wine.
Comparative example 1
The embodiment of the invention provides a brewing method of medlar western beating wine, which specifically comprises the following steps:
s1, adding water into the medlar original pulp, mixing, adjusting the sugar degree to be 10 Brix, and sterilizing to obtain a to-be-fermented liquid;
s2, mixing Saccharomyces cerevisiaeSaccharomyces cerevisiae) BYBC2.21113 seed solution (10% inoculum size) was added and the inoculum concentration of the bacteria was (1-8). Times.10 6 CFU/mL, liquid fermentation 72 h, in the fermentation process, providing sufficient oxygen for the liquid to be fermented in advance for yeast proliferation, then controlling the pressure in a pressure tank to be 0.12 MPa, fermenting at a constant temperature of 18 ℃, clarifying the fermented liquid after fermentation, and condensing thalli at the bottom of the tank, wherein the concentration of residual sugar is 6.0-8.0 g/L;
s3, cooling to 4 ℃ at a constant speed at a rate of 10 ℃ every 24-h, storing 7-d, filtering out residues and canning to obtain the Chinese wolfberry western style wine.
Comparative example 2
The embodiment of the invention provides a brewing method of medlar western beating wine, which specifically comprises the following steps:
s1, adding water into the medlar original pulp, mixing, adjusting the sugar degree to be 10 Brix, and sterilizing to obtain a to-be-fermented liquid;
s2, preparing abnormal Wick ham yeastWickerhamomyces anomalus) Adding BYBC2.22146 seed solution into the liquid to be fermented (inoculation amount is 10%), and inoculating bacteria with concentration of (1-8) x 10 6 CFU/mL, liquid fermentation 48 h, in the fermentation process, providing sufficient oxygen for the liquid to be fermented in advance for yeast proliferation, then controlling the pressure in a pressure tank to be 0.12 MPa, fermenting at a constant temperature of 18 ℃, clarifying the fermented liquid after fermentation, and condensing thalli at the bottom of the tank, wherein the concentration of residual sugar is 6.0-8.0 g/L;
s3, cooling to 4 ℃ at a constant speed at a rate of 10 ℃ every 24-h, storing 7-d, filtering out residues and canning to obtain the Chinese wolfberry western style wine.
Comparative example 3
The embodiment of the invention provides a brewing method of medlar western beating wine, which specifically comprises the following steps:
s1, adding water into the medlar original pulp, mixing, adjusting the sugar degree to be 10 Brix, and sterilizing to obtain a to-be-fermented liquid;
s2, mixing Saccharomyces cerevisiaeSaccharomyces cerevisiae) Adding BYBC2.21113 seed solution (with an inoculum size of 5%), fermenting in liquid state for 48 h, and adding abnormal Wick ham yeastWickerhamomyces anomalus) Adding BYBC2.22146 seed solution into the liquid to be fermented (inoculum size is 5%), liquid fermenting 72 h, and inoculating two bacteria at concentration of (1-8) x 10 6 CFU/mL, the total fermentation process is 5d, in the fermentation process, sufficient oxygen is provided for the liquid to be fermented in advance for yeast proliferation, then the pressure in a pressure tank is controlled to be 0.12 MPa, the constant temperature fermentation is carried out at 18 ℃, fermentation liquid starts to be clarified after 5d fermentation, thalli starts to be condensed at the bottom of the tank, and the concentration of residual sugar is 6.0-8.0 g/L;
s3, cooling to 4 ℃ at a constant speed at a rate of 10 ℃ every 24-h, storing 7-d, filtering out residues and canning to obtain the Chinese wolfberry western style wine.
The evaluation criteria for the sensory scores are shown in table 1 below.
Table 1 evaluation criteria for sensory scores
The alcohol content is detected by using a GB/T4928-2008 medium density bottle method.
The actual fermentation degree is detected and calculated by adopting the following formula:
true fermentation = (W-W) 1 ）/W×100%；
Wherein:
w: concentration of fermentation broth before fermentation (%)
W 1 : concentration (%) of fermentation broth after removal of alcohol after fermentation.
The content of aroma components (alcohols and esters) is detected by gas chromatography-mass spectrometer.
FIGS. 2 and 3 are graphs showing total ion flow patterns of aroma components of the Lycium barbarum western style wine obtained in example 1 and the blank (the Lycium barbarum primary pulp without yeast in example 1) and the Lycium barbarum western style wine obtained in examples 1 to 3 and the comparison examples 1 to 3 and the blank (the Lycium barbarum primary pulp without yeast in example 1) respectively, and the results of detecting the contents of aroma components (alcohols and esters) are shown in Table 2 below.
TABLE 2 detection results of the content of aroma components (alcohols and esters)
In the wolfberry western style wine obtained in the examples 1-3, the content of ester substances is obviously improved, so that the wine body is more plump and mellow, and in addition, the quality of the wolfberry western style wine is improved due to the improvement of the fermentation degree; the Lycium barbarum western beaten wine (single Saccharomyces cerevisiae fermentation) obtained in comparative example 1 has high alcoholicity, low ester content, and insufficient fragrance; the Chinese wolfberry western style wine (single abnormal Wick ham yeast fermentation) obtained in the comparative example 2 has low fermentation degree, reduces the quality of the Chinese wolfberry western style wine, but obviously improves the content of esters; after the saccharomyces cerevisiae 2d is added in the comparative example 3, the saccharomyces cerevisiae is dominant in fermentation broth, and the growth of the saccharomyces cerevisiae can be obviously inhibited by the abnormal connection of the saccharomyces cerevisiae, so that the content of ester substances is not improved.
The alcohol substances are important volatile aroma substances in the yeast fermentation process, and isoamyl alcohol, 2-methyl butanol, furfuryl alcohol, phenethyl alcohol and alpha-terpineol are main higher alcohols detected in the medlar siren wine, so that compared with pure fermentation of saccharomyces cerevisiae, the content of the higher alcohols in mixed fermentation is reduced, and the introduction of abnormal Wick ham yeast in the fermentation process is indicated, so that the synthesis of the higher alcohols is affected to a certain extent. The ester substance has the characteristics of flower fragrance and fruit fragrance, the ethyl acetate has fruit fragrance, and the phenethyl acetate has sweet fragrance, so that the ester substance is remarkably improved in the medlar western beating wine fermented by mixed bacteria; isoamyl acetate imparts primarily sweetness, fruity and banana-like odors; the addition of the abnormal Wick ham yeast ensures that the wine body has stronger fermentation flavor.
The examples 1 to 3 of the Lycium barbarum western style wine, the comparative examples 1 to 3 of the Lycium barbarum western style wine and the blank control group (the Lycium barbarum primary pulp without yeast in example 1) were taken and tested, and the test results are shown in Table 3 below.
Table 3 comparison table of test results
| Alcohol content/v% | True degree of fermentation/% | Sensory score/score | Alcohol content/% | Content of esters/% | |
| Example 1 | 2.28±0.07 | 62.97±0.28 | 83.93 | 32.39 | 62.21 |
| Example 2 | 2.52±0.05 | 65.07±0.36 | 86.65 | 30.08 | 64.80 |
| Example 3 | 2.68±0.01 | 66.95±0.27 | 82.91 | 31.56 | 63.73 |
| Comparative example 1 | 3.24±0.01 | 60.28±0.81 | 76.21 | 81.12 | 2.51 |
| Comparative example 2 | 0.58±0.05 | 20.27±0.73 | 50.82 | 7.36 | 78.90 |
| Comparative example 3 | 3.24±0.01 | 60.28±0.81 | 75.39 | 80.93 | 3.35 |
| Blank control group | nd | nd | nd | 5.37 | 5.83 |
Functional detection of wolfberry western style wine
The efficacy of the matrimony vine sirocco wine obtained in example 1 was examined by comparing the matrimony vine sirocco wine obtained in example 1 with a blank control group (the matrimony vine primary pulp without yeast in example 1) and a positive control (vitamin Vc).
1. Antioxidant efficacy assay
Antioxidant efficacy assay DPPH and ABTS methods of GB/T39100-2020 were used to determine antioxidant properties.
The measurement results of the antioxidant efficacy are shown in fig. 4. The lower the half-inhibition concentration of free radicals is, the stronger the antioxidation capability is; as can be seen from the graph in FIG. 4, the DPPH and ABTS free radical half-inhibition amounts of the Lycium barbarum western style wine are smaller than those of the Lycium barbarum primary pulp, which indicates that the antioxidant activity of the Lycium barbarum western style wine prepared by fermenting and brewing the Lycium barbarum primary pulp is enhanced; compared with the original juice of the medlar, the DPPH and ABTS inhibition effect of the medlar western beating wine is respectively improved by 23.23 percent and 19.30 percent; the positive control (vitamin Vc) had half inhibitory concentrations of DPPH and ABTS of 0.0225 g/L and 0.03239 g/L.
2. In vitro hypolipidemic cholate binding capacity efficacy determination
The cholate content is determined by a sulfuric acid colorimetric method:
(1) Drawing a standard curve:
preparing sodium glycocholate and sodium taurocholate standard solutions with different concentrations, respectively taking 2 mL cholate standard solutions into 25 mL stoppered test tubes, adding 60% H 2 SO 4 Taking out ice bath at 70deg.C for 40 min, measuring absorbance at 387. 387nm, and respectively drawing standard graphs of sodium glycocholate and sodium taurocholate with the solution concentration as abscissa and absorbance as ordinate, as shown in FIG. 5;
(2) Determination of the samples:
putting samples with different concentrations of 3 mL into a triangular flask, adding 3 mL and 10g/L pepsin, 1 mL and 0.01 mol/L hydrochloric acid, oscillating 1h in a constant-temperature oscillating box at 37 ℃, adjusting the pH to 6.3 with NaOH, adding 4 mL and 10g/L trypsin, and oscillating 1h in the constant-temperature oscillating box at 37 ℃; adding 4 mL and 0.4 mmol/L sodium glycocholate into one part of a triangular flask, adding 4 mL and 0.5 mmol/L sodium taurocholate into the other part of the triangular flask, oscillating for 1h at a constant temperature of 37 ℃, centrifuging for 20 min at 4000 r/min, taking supernatant 2 mL, and measuring absorbance at 387 nm;
the cholate content was calculated using the formula:
ability to bind cholate= (N 1 -N 2 ）/V×A；
Wherein:
N 1 the addition amount of the cholate and the mmol are shown;
N 2 is the residual amount of cholate, mmol;
v is the volume of sample added;
a is the dilution of the sample.
The results of the in vitro hypolipidemic cholate binding capacity assay are shown in FIG. 6; as can be seen from fig. 6, the sodium glycinate-binding capacity of the fructus Lycii western style wine prepared by fermenting and brewing fructus Lycii raw pulp is higher than that of fructus Lycii raw pulp, which indicates that the fermentation has a gain effect on the cholate-binding capacity; the binding capacity of the Lycium barbarum western wine on sodium glycocholate and sodium taurocholate is respectively improved by 51.78% and 23.36%.
3. Tyrosinase inhibitory activity
Tyrosinase inhibition experiments were determined as described in T/SHRH 015-2018.
Tyrosinase is widely distributed in organisms, is a key speed limiting enzyme for catalyzing melanin synthesis, and has increased tyrosinase activity and more melanin; tyrosinase activity is inhibited and the ability of melanocytes to produce melanin is correspondingly reduced.
The measurement results of tyrosinase inhibitory activity are shown in FIG. 7; as can be seen from FIG. 7, tyrosinase inhibitory activity of the Lycium barbarum western wine prepared by fermenting and brewing the Lycium barbarum primary pulp reaches 56.17%, and the inhibitory effect is improved by 69.65% compared with the Lycium barbarum primary pulp.
The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the invention.
It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (10)
1. Abnormal Wick ham yeastWickerhamomyces anomalus) BYBC2.22146, characterized in that,
the microbial strain is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.26360 in the 12 th month 30 of 2022.
2. A microbial preparation, which is characterized in that,
comprising the abnormal Weikhan yeast of claim 1Wickerhamomyces anomalus）BYBC 2.22146。
3. Abnormal Wikimann yeast with preservation number of CGMCC No.26360Wickerhamomyces anomalus) Use of BYBC2.22146 in brewing a lyceum berry western style wine.
4. A brewing method of a medlar western style wine is characterized in that,
the method comprises the following steps:
s1, adding water into the raw wolfberry pulp, mixing and/or adding water into dried wolfberry fruits, crushing, homogenizing, and sterilizing to obtain a to-be-fermented liquid;
s2, firstly activating the cultured abnormal Wikimann yeastWickerhamomyces anomalus) BYBC2.22146 is inoculated into the liquid to be fermented for liquid fermentation, and then the activated and cultured saccharomyces cerevisiae is subjected to the liquid fermentationSaccharomyces cerevisiae) BYBC2.21113 is inoculated and subjected to liquid fermentation;
and S3, ageing to obtain the medlar western beating wine.
5. The brewing method of the Chinese wolfberry western style wine according to claim 4, wherein,
in the step S2 of the process,
abnormal Weikeham yeastWickerhamomyces anomalus) BYBC2.22146 inoculated at 1X 10 each 6 - 8×10 6 CFU/ml and 3.5-6 wt%, the temperature and time of liquid state fermentation are 16-24 ℃ and 45-54 h respectively;
saccharomyces cerevisiaeSaccharomyces cerevisiae) BYBC2.21113 inoculated at 1X 10 each 6 - 8×10 6 CFU/ml and 3.5-6. 6 wt%, the temperature and time of liquid fermentation are 16-24 ℃ and 66-78 h, respectively.
6. The brewing method of the Chinese wolfberry western style wine according to claim 4, wherein,
in the step S2 of the process,
abnormal Weikeham yeastWickerhamomyces anomalus) The activation culture of BYBC2.22146 is specifically as follows:
activating the abnormal Wick ham microzyme preserved at-80 ℃ for 15-30min at 28 ℃, dipping the abnormal Wick ham microzyme into a solid wort culture medium flat plate for streaking, then culturing the abnormal Wick ham microzyme at 28 ℃ for 48 h, picking a single colony in a 100 mL wort liquid culture medium by using a inoculating loop, and culturing the abnormal Wick ham microzyme at a constant temperature of 28 ℃ and 200 rpm for 22 h;
saccharomyces cerevisiaeSaccharomyces cerevisiae) The activation culture of BYBC2.21113 is specifically as follows:
activating Saccharomyces cerevisiae preserved at-80deg.C for 15-30min at 28deg.C, streaking with inoculating loop, culturing at 28deg.C for 48 h, picking single colony with inoculating loop in 100 mL wort liquid medium, and culturing at 28deg.C and 200 rpm under constant temperature shaking for 24 h.
7. A brewing method of a Chinese wolfberry western style wine according to any one of claims 4-6, wherein,
in the step S3 of the process,
the ageing process specifically comprises the following steps:
taking fermentation liquor with residual sugar concentration of 6.0-8.0 g/L, cooling to 4 ℃ at constant speed at a rate of 10 ℃ every 24-h, and storing 7-d.
8. A brewing method of a Chinese wolfberry western style wine according to any one of claims 4-6, wherein,
in the step S1 of the process,
the initial sugar degree of the to-be-fermented liquid is 8-16 Brix%.
9. The western-style wine of wolfberry brewed by the brewing method of the western-style wine of wolfberry of any one of claims 4-8.
10. The western-style wine of claim 9 wherein,
the alcohol content, the actual fermentation degree, the alcohol content and the ester content of the medlar sirocco wine are respectively 2.26-3.73 v%, 62-68%, 32.39-35.21% and 56.00-63.43%.
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