CN113604372A

CN113604372A - Saccharomyces cerevisiae with low yield of fusel and high yield of ester and application of saccharomyces cerevisiae in production of fermented food

Info

Publication number: CN113604372A
Application number: CN202110892850.7A
Authority: CN
Inventors: 毛健; 刘双平; 赵禹宗; 姬中伟
Original assignee: Industrial Technology Research Institute Of Jiangnan University Shaoxing; Jiangnan University
Current assignee: Industrial Technology Research Institute Of Jiangnan University Shaoxing; Jiangnan University
Priority date: 2021-08-04
Filing date: 2021-08-04
Publication date: 2021-11-05

Abstract

The invention discloses a saccharomyces cerevisiae with low fusel yield and high ester yield and application thereof in fermented food production, belonging to the field of fermentation engineering and biotechnology. The invention provides a saccharomyces cerevisiae jiangnan2# which is a safe strain for industrial production, has excellent stress resistance, good fermentation performance and different fermentation performance, can meet the requirements of different alcoholic beverages and fermented foods on low yield of fusel and high yield of ester, and the yellow wine produced by fermenting the strain has urea and ethyl carbamate contents lower than the limit regulation in the yellow wine, and has organic acid, amino acid and other physical and chemical indexes meeting the national standard requirements of the yellow wine. The saccharomyces cerevisiae provided by the invention can obviously improve the flavor of yellow wine, meet the industrial production requirements of high-quality yellow wine with high comfort and safety, obviously improve the flavor of yellow wine, meet the industrial production requirements of high-comfort high-quality yellow wine, has good application prospect, and can realize the improvement of safety and quality when being applied to other fermented foods.

Description

Saccharomyces cerevisiae with low yield of fusel and high yield of ester and application of saccharomyces cerevisiae in production of fermented food

Technical Field

The invention relates to a saccharomyces cerevisiae with low fusel yield and high ester yield and application thereof in fermented food production, belonging to the field of fermentation engineering and biotechnology.

Background

The yeast converts sugar in the raw materials into alcohol, and the protein and fat are converted into organic acid, amino acid, ester, phenol, polysaccharide, peptide and the like through the action of microorganisms such as yeast, lactobacillus and the like. The quality of yeast performance directly influences the production efficiency and flavor quality of the final product. Meanwhile, the yeast is closely related to the generation of some non-favorite substances such as higher alcohol (fusel oil), urea, ethyl carbamate and the like in the brewing process, and the content of the substances exceeding the limited value can cause negative influence and influence the drinking comfort and safety.

The low fusel high ester, comfortable taste, unique flavor, rich nutrition, health care function, safe product drinking and the like are the development requirements of future high-quality fermented food. Fusel alcohols and some acetates are metabolites of yeast during fermentation. The substances with safety hazards, such as alcohol ester imbalance (high fusel content and low ester content), biogenic amine, urea, ethyl carbamate and the like are the common problems of fermented wines, such as yellow wine, cooking wine, beer, wine, fruit wine and the like, and distilled wines, such as white spirit, vodka, brandy and the like. The saccharomyces cerevisiae directly or indirectly participates in the metabolism generation of the substances through the whole fermentation process, and how to improve the quality of fermented food is very important by controlling the metabolism of fermentation microorganisms.

The genetic engineering means is utilized to modify the saccharomyces cerevisiae to definitely meet different target requirements, for example, in patent applications with publication numbers of CN201511017931.3 and CN201610012975.5, an inventor constructs a high-ester-yield and low-yield higher alcohol saccharomyces cerevisiae engineering bacterium, improves the ester content and reduces the higher alcohol content in white spirit, constructs a high-ethyl lactate saccharomyces cerevisiae strain with low-yield and high-yield higher alcohol, and remarkably improves the production of ethyl lactate and simultaneously reduces the production of higher alcohol. In addition, foreign wine brewing yeast is also transformed by adopting genetic engineering, 2 transgenic wine yeast strains ML01 and ECMO01 realize the purpose of transformation without producing biogenic amine and urea and the allowable use in a certain range, but the absolute safety of the transgenic technology is not clear, so that the transgenic technology is not widely accepted, and especially in fermented foods such as yellow wine, table vinegar and the like, most metabolites of the wine brewing yeast are usually retained in final products.

In order to improve the comfort and safety of fermented foods, the saccharomyces cerevisiae with low fusel yield, high ester content or characteristic flavor must be screened and obtained, and good saccharomyces cerevisiae strains with low urea and ethyl carbamate production are used as leaven to prepare the fermented foods. In the research on the breeding of low-yield fusel and high-yield ester saccharomyces cerevisiae in fermented foods such as white wine, yellow wine, beer, wine and the like, the related application of the saccharomyces cerevisiae, and the research and patent of the searchable saccharomyces cerevisiae applied to the related food field, no effective method for fundamentally and systematically solving the problems exists at present, and the safe industrial production saccharomyces cerevisiae strain which has high alcohol content, low fusel and high ester content, remarkably low urea and ethyl carbamate content and a specified limit and the application of the safe industrial production saccharomyces cerevisiae strain are not found on the premise of meeting the normal fermentation. Therefore, the low-yield fusel high-ester-yield saccharomyces cerevisiae obtained by separating, screening and systematically evaluating and breeding saccharomyces cerevisiae strains and the application of the saccharomyces cerevisiae in fermented food have important significance for improving the quality, health and safety of alcoholic beverages such as yellow wine and the like and fermented food such as table vinegar and the like.

Disclosure of Invention

Aiming at solving the problems, the saccharomyces cerevisiae with low yield of fusel and high yield of ester, excellent fermentation performance and high safety is not produced in the current fermented food production. The invention provides saccharomyces cerevisiae (S.cerevisiae) with excellent stress resistance, good fermentation performance, low yield of fusel and high ester yield and application thereof in fermented food.

The invention provides Saccharomyces cerevisiae (Saccharomyces cerevisiae) jiangnan2# with low fusel yield and high ester yield, which is preserved in China center for type culture Collection with the preservation number of CCTCC M2021524 and the preservation address of university of China, Wuhan and Wuhan.

In one embodiment, the saccharomyces cerevisiae monoclonal colony is characterized by milky white, oval or elliptical, convex, smooth surface, wetness and luster, and regular strain edges.

The invention also provides an application mode of the saccharomyces cerevisiae with low yield of fusel and high ester yield in fermented food.

The invention also provides a composition prepared by compounding the microbial agent of the saccharomyces cerevisiae jiangnan2# with other microorganisms and the like.

In one embodiment, the composition includes, but is not limited to, a fortified wine or fortified wheat starter.

In one embodiment, the microbial agent is a solid microbial agent or a liquid microbial agent.

In one embodiment, said microbial agent comprises live cells of said saccharomyces cerevisiae jiangnan2# biomass, dried saccharomyces cerevisiae jiangnan2# biomass obtained by freeze-drying, said saccharomyces cerevisiae jiangnan2# cells obtained by a solidification technique, a liquid microbial agent of said saccharomyces cerevisiae jiangnan2#, a solid microbial agent of said saccharomyces cerevisiae jiangnan2#, or said saccharomyces cerevisiae jiangnan2# strain in any other form.

In one embodiment, the microbial preparation comprises Saccharomyces cerevisiae jiangnan2# in an amount of 1 × 10⁶CFU/g。

In one embodiment, the other microorganism includes, but is not limited to, Saccharomyces cerevisiae jiangnan2 #.

The saccharomyces cerevisiae jiangnan2# is preserved in China Center for Type Culture Collection (CCTCC) No. M2021524 in 13.05.2021, and the preservation address is university of China, Wuhan and Wuhan.

In one embodiment, the said application mode of combination with other microorganisms means that the low-fusogenic high-ester-yielding saccharomyces cerevisiae jiangnan2# is combined with other microorganisms, and the other microorganisms comprise saccharomyces cerevisiae and non-saccharomyces cerevisiae; the non-saccharomyces cerevisiae includes, but is not limited to, lactic acid bacteria.

In one embodiment, the microbial preparation is prepared as follows: inoculating the strain into a rice saccharification liquid culture medium, and performing shake culture for 20-24 hours at the culture temperature of 28 +/-2 ℃ to obtain a first-level seed liquid; inoculating the first-stage seed liquid to a new rice saccharified liquid culture medium at a ratio of 5% -10%, and performing shake culture at 28 + -2 deg.C for 36-48 h to obtain a second-stage seed liquid with the number of yeast not less than 1 × 10⁷CFU/mL, germination rate is not less than 30%.

In one embodiment, the wine fortification is that the saccharomyces cerevisiae jiangnan2# or the microbial agent is compounded with other microorganisms or added into the wine in any other form during the manufacturing process of the wine, so that the purpose of strengthening the fermentation characteristic of the wine during the use of the wine is achieved.

In one embodiment, the enhanced koji means that the saccharomyces cerevisiae jiangnan2# or the microbial agent is added into the koji in any other form during the koji making process so as to achieve the purpose of enhancing the fermentation characteristics of the koji when the koji is used.

The invention also provides application of the saccharomyces cerevisiae with low yield of fusel and high ester yield in fermented food.

In one embodiment, the fermented food product comprises all the application modes of brewing wine with low yield of fusel and high yield of ester.

In one embodiment, the fermented food includes, but is not limited to, yellow wine, cooking wine, rice wine, sweet fermented rice, wine, fruit wine, vinegar, white spirit, beer, tobacco leaf, or fermented nutritional ice cream, etc.

In one embodiment, the application is for yellow wine brewing, and the low-fusogenic high-ester-yielding saccharomyces cerevisiae jiangnan2# is a quick brewing yeast.

In one embodiment, the yellow wine brewing is yellow wine obtained by adding the low-yield fusel high-yield ester-producing saccharomyces cerevisiae jiangnan2# as a quick brewing yeast to a steamed or gelatinized raw material (rice, husked millet, corn, millet and the like) according to the addition amount of 5-15%, fermenting, squeezing, wine decocting, aging, filtering, sterilizing and filling.

In one embodiment, the use is for cooking wine brewing.

In one embodiment, the cooking wine brewing is to firstly use the low-yield fusel high-yield ester-producing saccharomyces cerevisiae jiangnan2# as a quick brewing yeast to ferment to obtain yellow wine, and then use the yellow wine to prepare the cooking wine.

In one embodiment, said use is for sake brewing, using said low-fusogenic, high-ester-yielding saccharomyces cerevisiae jiangnan2# as a starter.

In one embodiment, the rice wine brewing is implemented by brewing the low-yield fusel high-yield ester-producing saccharomyces cerevisiae jiangnan2# and distiller's yeast as leaven, adding the leaven into the cooked rice raw material according to the addition amount of 0.5-1.5%, and performing the processes of cooking, yeast adding (adding the saccharomyces cerevisiae), saccharification, fermentation, squeezing and the like.

In one embodiment, the application is for brewing sweet rice, and the low-fusogenic high-ester-producing saccharomyces cerevisiae jiangnan2# is used as a leavening agent.

In one embodiment, the rice wine brewing is implemented by brewing the low-yield fusel high-yield ester-producing saccharomyces cerevisiae jiangnan2# and distiller's yeast as leaven, adding the leaven into a cooked raw material (rice, millet, corn, millet and the like) according to the addition amount of 0.5-1.5%, and performing the processes of cooking, yeast adding (adding the saccharomyces cerevisiae), saccharification, fermentation and the like.

In one embodiment, the use is for vinegar brewing.

In one embodiment, the application is used for brewing vinegar, wherein the low-yield fusel high-yield ester-producing saccharomyces cerevisiae is used as yeast to be fermented to obtain yellow wine, and then the yellow wine is used as an acetic acid fermentation raw material to brew the vinegar.

In one embodiment, the use is for brewing white wine.

In one embodiment, the brewing white spirit is that the low-yield fusel high-ester-yield saccharomyces cerevisiae jiangnan2# is additionally added when white spirit is fermented in a pool for fermentation.

In one embodiment, the addition amount of the saccharomyces cerevisiae jiangnan2# with low yield of fusel and high yield of ester is 1% (v/v) of the raw material system when brewing white spirit.

In one embodiment, the use is for brewing wine.

In one embodiment, said brewed wine is prepared by adding said low-fusogenic high-ester-producing Saccharomyces cerevisiae jiangnan2# additionally during the alcoholic fermentation stage.

In one embodiment, the low-fusel high-ester-yielding Saccharomyces cerevisiae is added in an amount of 0.5% (v/v) to 1.5% (v/v) of the total volume of the fermentation system.

In one embodiment, the use is for brewing wine.

In one embodiment, the brewed fruit wine includes, but is not limited to, any one of plum wine, waxberry wine, kiwi wine, green plum wine, hawthorn wine, pomegranate wine, lemon wine, loquat wine.

In one embodiment, the brewing of the wine comprises the following processes: and (3) treating before fermentation, namely inoculating the low-yield fusel high-yield ester-producing saccharomyces cerevisiae jiangnan2# into clarified clear juice of the fruit juice.

In one embodiment, the inoculation amount of the saccharomyces cerevisiae jiangnan2# with low yield of fusel and high yield of ester is 1% (v/v) to 5% (v/v) of the total volume of the clear juice when brewing the fruit wine.

In one embodiment, the use is for brewing fruit vinegar.

In one embodiment, the application is used for brewing fruit vinegar by firstly fermenting the low-yield fusel high-yield ester-producing saccharomyces cerevisiae jiangnan2# to obtain fruit wine, and then inoculating acetic acid strain to the fruit wine as a fermentation raw material to perform acetic acid fermentation.

In one embodiment, the use is for brewing beer.

In one embodiment, the application is used for brewing beer, the low-fusel high-ester-yield saccharomyces cerevisiae jiangnan2# is used as a pure yeast, the pure yeast is added into wort according to the addition amount of 0.2% -1% after propagation, and the beer is obtained through the processes of malting, brewing, filling and the like.

In one embodiment, the use is for making cigarettes.

In one embodiment, the application is used for manufacturing cigarettes, and the tobacco flavor is directly prepared from the culture solution of saccharomyces cerevisiae jiangnan2# with low yield of fusel and high ester yield; or pear, grape, osmanthus fragrans, tobacco extract and the like are taken as culture medium raw materials, the saccharomyces cerevisiae jiangnan2# culture is inoculated for fermentation to prepare the fermentation type tobacco flavor, and then the obtained tobacco flavor is directly added into the cigarette leaf group according to the addition amount of 1-5% of the mass of the cigarette leaf group.

In one embodiment, the application is for making a fermented nutritional ice cream bar.

In one embodiment, the application is used for making fermented nutritional ice cream, and the fermented nutritional ice cream is obtained by fermenting raw materials (fruit juice, sugar, honey, whole milk powder and the like, sweet potato, purple sweet potato, corn and other starch) by taking the low-yield fusel high-yield ester-producing saccharomyces cerevisiae jiangnan2# as a leavening agent in an adding amount of 5% -10%, adding other food additives, and then performing processes such as mixing, pasteurization, homogenization, cooling aging, coagulating stirring, injection molding, freezing, mold stripping and the like.

The invention also provides a product obtained by distilling, blending or adding the low-yield fusel high-yield ester-production saccharomyces cerevisiae jiangnan2# fermentation product.

The invention has the beneficial effects that:

(1) the invention provides a saccharomyces cerevisiae jiangnan2# with low fusel yield and high ester yield, which has better temperature tolerance, and the strain still has better growth condition within the range of 15-45 ℃, and can meet the requirements of different types of fermented food.

(2) The strain jiangnan2# of the invention grows better under the condition of 20% (v/v) ethanol, still has better growth condition under the condition of 1.6mol/L sodium chloride osmotic pressure, and the temperature tolerance range is 15-45 ℃.

(3) Yellow wine fermentation experiments show that jiangnan2# can show good fermentation performance under the fermentation conditions that the main fermentation temperature is 20-35 ℃ and the post-fermentation temperature is 10-15 ℃, and the rapid-brewing yeast strain prepared by jiangnan2# is used for brewing the northern husked millet yellow wine, compared with Angel yeast, after the fermentation is finished, the alcoholic strength is 18% (v/v) -19.8% (v/v), the content of fusel is in the ranges of 398.68 +/-13.25 mg/L and 419.77 +/-4.09 mg/L, the alcoholic strength is reduced by 20-25.86%, the content of 2-phenethyl alcohol is about 74.65 +/-0.45 mg/L, the alcoholic strength is reduced by 48.55%, the content of total ester is 241.42 +/-12.50 mg/L, and the alcoholic strength is increased by more than 50%.

(4) When the saccharomyces cerevisiae jiangnan2# with low fusel yield and high ester yield provided by the invention is adopted for mechanical yellow wine brewing, compared with 85# yeast commonly used in the yellow wine industry disclosed in the patent CN 105385613B, the alcoholic strength of yellow wine obtained after the fermentation of the strain jiangnan2# is 16.15% (v/v), the fusel content is 416.05 +/-10.25 mg/L, the 2-phenethyl alcohol content is 96.63 +/-5.21 mg/L, the reduction is more than 15%, the content of ethyl ester (ethyl acetate, isoamyl acetate and phenethyl acetate) is increased by more than 50%, the fermentation characteristic of the low-fusel high-ester yield is more remarkable, and the yellow wine obtained after squeezing, blending and filtering is softer and more harmonious in aroma and taste, good in drinking comfort level, and is not easy to get up after being drunk in a proper amount, and is fast in sobering after being drunk.

(5) When the saccharomyces cerevisiae jiangnan2# with low yield of fusel and high ester yield is adopted for mechanical yellow wine brewing, the urea content is 19.80 +/-0.53 mg/L, the ethyl carbamate content is 73.02 +/-2.41 mu g/L, the content is obviously lower than that of the conventional strains in factories, the amount of ethyl carbamate generated by metabolism during fermentation of the saccharomyces cerevisiae jiangnan2# is less, and the safety of fermented food by taking the saccharomyces cerevisiae jiangnan2# as a fermentation strain can be improved.

Biological material preservation

Saccharomyces cerevisiae (Saccharomyces cerevisiae) jiangnan2#, which is classified and named as Saccharomyces cerevisiae (Saccharomyces cerevisiae) jiangnan2#, and has been preserved in China center for type culture collection (CCTCC NO: M2021524) at 13.05.2021, with the preservation address being China, Wuhan and Wuhan university.

Drawings

FIG. 1 is a graph showing the growth of Saccharomyces cerevisiae (S. cerevisiae) jiangnan2# of the present invention at different temperatures.

FIG. 2 correlation analysis of yeast isolates with flavour materials.

Detailed Description

Detecting physical and chemical indexes of yellow wine: the alcohol content, the amino acid nitrogen and the total acid are measured according to GB/T13662-2018 yellow wine. The content of organic acid and amino acid is detected by High Performance Liquid Chromatography (HPLC), and volatile flavor substances such as ethyl carbamate, higher alcohol, esters, etc. are detected by gas chromatography-mass spectrometry (GC-MS). The content of reducing sugar is measured by a DNS method. The concentration of urea and bacterial liquid is measured by spectrophotometry. The high-grade alcohol (also called fusel) in the yellow wine mainly comprises 4 types of n-propanol, isobutanol, isoamylol and 2-phenethyl alcohol, and the content of the fusel is quantitatively determined by adopting a dispersion liquid microextraction technology (DLLME), utilizing GC-MS (gas chromatography-mass spectrometry) to detect and using 4-methyl-2-pentanol as an internal standard curve.

Fusel per unit alcohol content: the unit alcohol content fusel is the ratio of the total amount of 4 kinds of fusel to the alcohol content in the alcohol metabolism process, namely the content of fusel corresponding to 1% (v/v), and the unit is mg/L.

Sensory evaluation: the sensory evaluation of alcoholic beverages was performed by selecting 10 persons (5 men and 5 women) who had experienced sensory evaluation of alcoholic beverages and had an age of 20 to 40 years, and comparing the difference between the samples and the control after introducing the sensory requirements (appearance, aroma, taste and style characteristics) of alcoholic beverages to the evaluators in accordance with the corresponding national standards.

The following describes embodiments of the present invention with reference to the drawings. The experimental methods used in the examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are commercially available unless otherwise specified.

YPD medium: 10g of yeast extract powder, 20g of peptone and 20g of glucose, adding water to 1000mL, adding 2% agar, sterilizing at 121 ℃ for 20min under high pressure, and cooling for later use.

Rice saccharification liquid culture medium: taking a proper amount of high-quality rice raw material, soaking the rice in water bath at the constant temperature of 60 ℃ for 30min, cooking for 20min under the normal pressure condition, then respectively adding saccharifying enzyme which takes the rice raw material as the reference and is 150U/g-300U/g1 per thousand and liquefying enzyme which takes the rice raw material as the reference and 200U/g-400U/g2 per thousand, adding raw wheat starter with the mass of 10% of rice, saccharifying for 4h-5h under the condition of 55 ℃ to 65 ℃ until the sugar degree is more than 13Brix, subpackaging, sterilizing for 15min-20min under the high pressure of 121 ℃, and cooling for later use.

Example 1 selection and characterization of Saccharomyces cerevisiae strains

(1) Sample preparation and strain isolation

Collecting yellow wine fermentation liquor from the traditional manual yellow wine brewing process in Shaoxing region, storing at 4 ℃, taking 1mL of uniformly mixed sample, and performing gradient dilution with sterile water (10)^-1-10^-5) Taking 100 mu L of diluted sample, coating the sample on a YPD plate, inverting the coated plate, standing and culturing at 28 ℃ for 48h, selecting the dilution degree with a monoclonal colony, selecting a strain with colony morphology, color and appearance conforming to the physiological morphology of yeast, carrying out multiple streaking to purify the strain, and finally numbering and preserving the obtained pure strain.

(2) Identification of strains

Morphological characteristics of the strain are determined by ITS sequencing, and the ITS sequence of Saccharomyces cerevisiae jiangnan2# is shown as SEQ ID NO. 1. The strain is compared and analyzed in NCBI database to have homology of more than 99% with Saccharomyces cerevisiae, and can be identified as Saccharomyces cerevisiae (S. The strain is preserved in China center for type culture Collection (CCTCC M2021524) at 13.05.2021, jiangnan2# with preservation number of China, Wuhan university.

Example 2 selection of Low-Productivity Heterohol high-ester-production Saccharomyces cerevisiae (S. cerevisiae) Strain jiangnan2#

1. Primary screening based on yellow wine fermentation of saccharomyces cerevisiae isolates: in order to select saccharomyces cerevisiae with excellent fermentation performance and capable of meeting the brewing requirement of yellow wine, all separated strains are respectively made with quick brewing yeast and then are subjected to yellow wine fermentation, physicochemical indexes such as alcoholic strength and the like are measured after fermentation is finished, strains are preliminarily screened, and substances with the alcoholic strength meeting the national standard, namely, fusel and flavor substances are screened according to different batches. The method comprises the following specific steps:

(1) the raw material ratio of the traditional yellow wine fermentation selected in the embodiment

100 percent of glutinous rice as a reference, and the raw materials comprise 125 percent of water, 12 to 15 percent of wheat starter and 10 to 15 percent of quick brewing yeast. The method comprises the following steps of:

500g of glutinous rice, 625g of water, 60-75 g of wheat koji, and quick brewing yeast: 50mL-75 mL.

The preparation method of the quick brewing yeast comprises the following steps: the saccharomyces cerevisiae glycerol tube preservation strain is subjected to slant preparation, the saccharomyces cerevisiae jiangnan2# strain cultured by slant is inoculated in a rice saccharification liquid culture medium, and is subjected to shaking culture for 24 hours under the condition that the culture temperature is 28 +/-2 ℃ to obtain a first-class seed liquid (the concentration of the bacterial liquid is 10)⁶-10⁸CFU/mL) is added, the first-stage seed liquid is inoculated to a new rice saccharification liquid culture medium according to the proportion of 5 percent to 10 percent, the oscillation culture is carried out for 36h to 48h under the condition that the culture temperature is 28 +/-2 ℃ to obtain the second-stage seed liquid, the viable count and the budding rate of the yeast are measured by the obtained second-stage seed liquid, and the yeast number is more than or equal to 1 multiplied by 10⁷And the budding rate is more than or equal to 30 percent, and the rice wine is brewed as the quick brewing yeast.

Experimental groups: the separated saccharomyces cerevisiae strain is used as pure yeast to make the quick brewing yeast for brewing yellow wine.

Control group: the saccharomyces cerevisiae strain 85# for factory production (disclosed in patent CN 105385613B) is used for preparing the quick brewing yeast by preparing the pure yeast for brewing yellow wine.

(2) Traditional yellow wine brewing process

a) Preparing fermented raw material rice: adding water into raw rice until the amount of the raw rice exceeds the liquid level by more than 10cm, soaking the raw rice for 3-5 days until the acidity of rice slurry of the soaked raw rice reaches more than 4.5g/L, draining the water to obtain wet rice, steaming the wet rice in a rice steaming cabinet at 121 ℃ for 20-30 min until the rice is cooked but not penetrated, the rice has no white heart, the rice has sour taste and rice fragrance, and the rice yield is 140-160%.

b) Blanking and fermenting according to the raw material proportion of the traditional yellow wine fermentation:

blanking and fermenting according to the raw material proportion of the traditional yellow wine fermentation, and specifically comprising the following steps:

s1, respectively streaking strains frozen and preserved in a glycerol tube to generate monoclonal colonies, and selecting the monoclonal colonies to inoculate on a slope to obtain a saccharomyces cerevisiae slope culture medium capable of being preserved at a low temperature;

s2, transferring the strain S1 to the rice saccharification liquid culture medium from the inclined plane for 2 times, culturing at 28 +/-2 ℃ to the order of magnitude of more than or equal to 1 multiplied by 10⁷The germination rate is more than 30 percent and is used as inoculation liquid;

s3, adding raw materials of rice and water into a sterilized fermentation container, inoculating an inoculation liquid obtained in S2 with the proportion of 10% -15% of the raw material rice as a reference into a rice-water mixed fermentation system, adding wheat starter with the proportion of 12% -15% of the raw material rice, mixing the materials at 25-28 ℃, standing at 20-35 ℃ for fermentation for 3-5 days;

s4, reducing the temperature of the fermentation tank to 10-15 ℃, standing for 15-20 days for after-fermentation;

s5, the fermented mash obtained from S4 is pressed by a plate frame (4 times of feeding, the pressure of the mash is 0.2-0.6MPa, the filtering area is 100m2, the diameter of a filter plate is 1m), diatomite is filtered (the adding proportion of the diatomite is 4-6%, and the pressure is 0.3-0.5MPa), the obtained filtrate is clarified to obtain the sake, and the sake is blended to be decocted into yellow wine according to the national standard of yellow wine after 1-3 thousandths of caramel is added.

144 isolates, exemplified are batches comprising Saccharomyces cerevisiae (S.cerevisiae) jiangnan2# (strain number TP-554 at the time of selection). After the fermentation is finished, the physical and chemical indexes, the fusel and the flavor of the wine with the alcoholic strength of more than 16% (v/v) are selected for measurement. PCA analysis (FIG. 2) was performed based on the strain's flavor and fusel indices, and strains with low fusel and high ester yields were clearly distinguished.

2. Re-screening based on yellow wine fermentation of the saccharomyces cerevisiae isolate: and (4) carrying out 3 repeated screenings on the preliminarily screened strains with excellent fermentation performance, and determining physicochemical indexes, flavors and fusel after the fermentation is finished. The physical and chemical indexes of the saccharomyces cerevisiae with excellent low-yield fusel and high ester yield after fermentation meet the national standard of yellow wine, the contents of fusel and ester are shown in table 2, and saccharomyces cerevisiae (S. cerevisiae) jiangnan2# shows excellent fermentation characteristics of low-yield fusel and high ester yield.

TABLE 1 physical and chemical indexes of double-screening to obtain excellent low-yield fusel high-yield ester Saccharomyces cerevisiae at the end of fermentation

TABLE 2 rescreening to obtain good Low-yield Heterohol high-yield esters content Heterohol and Total esters content at the end of fermentation of Saccharomyces cerevisiae

3. Stability verification based on yellow wine fermentation of saccharomyces cerevisiae isolates: and (4) carrying out stability verification on the bacterial strain obtained by re-screening by repeating fermentation for 1 time, and determining physicochemical indexes, flavor and fusel after the fermentation is finished. After fermentation, the total amount of the fusel of No. 85 is 381.85 +/-12.45 mg/L, and the total ester is 126.77 +/-7.84 mg/L; the saccharomyces cerevisiae (S.cerevisiae) jiangnan2# is 349.29 +/-7.35 mg/L, the total ester is 142.16 +/-16.2 mg/L, and the main esters (ethyl acetate, ethyl lactate and the like) in the jiangnan2# strain fermented yellow wine are higher than the reference strain 85#, which shows that jiangnan2# has the brewing characteristics of low-yield fusel and high-yield ester, and is a low-yield fusel and high-yield ester strain.

TABLE 3 physical and chemical indexes of good low-yield fusel high-ester-yield Saccharomyces cerevisiae at the end of fermentation

EXAMPLE 3 preparation of Low-yield Heterohol high-yield ester-producing Saccharomyces cerevisiae jiangnan2# leaven

(1) Taking the quick brewing yeast as an example, the quick brewing yeast containing the saccharomyces cerevisiae jiangnan2# for brewing yellow wine is prepared.

The preparation method of the quick brewing yeast comprises the following steps: the saccharomyces cerevisiae glycerol tube preservation strain is subjected to slant preparation, the saccharomyces cerevisiae jiangnan2# strain screened in the slant culture example 1 is inoculated in a rice saccharification liquid culture medium, and is subjected to shaking culture for 24 hours under the condition that the culture temperature is 28 +/-2 ℃ to obtain a first-grade seed liquid (the concentration of the bacterial liquid is 10)⁶-10⁸CFU/mL) is added, the first-stage seed liquid is inoculated to a new rice saccharification liquid culture medium according to the proportion of 5 percent to 10 percent, the oscillation culture is carried out for 36h to 48h under the condition that the culture temperature is 28 +/-2 ℃ to obtain the second-stage seed liquid, the viable count and the budding rate of the yeast are measured by the obtained second-stage seed liquid, and the yeast number is more than or equal to 1 multiplied by 10⁷CFU/mL, the budding rate is more than or equal to 30 percent, and the obtained product is used as a quick brewing yeast (leaven) to brew yellow wine.

(2) Preparation of leaven from Saccharomyces cerevisiae jiangnan2# for other types of fermented food

And (2) according to different fermented food requirements, the quick brewing yeast containing the pure strain prepared in the step (1) is used for fermentation production of food, fermented alcoholic beverages containing saccharomyces cerevisiae are prepared, and various culture mediums meeting the nutritional requirements of food fermentation production C and N, such as rice saccharification liquid, wort, sugar solution and the like, can be selected as culture mediums to perform activation culture on the saccharomyces cerevisiae jiangnan2# to meet the corresponding colony number of the requirements.

Example 4 yellow wine Industrial application of Saccharomyces cerevisiae jiangnan2# with Low yield of fusel and high yield of esters

(1) Application of saccharomyces cerevisiae jiangnan2# with low yield of fusel and high yield of ester in glutinous rice yellow wine

The industrial production and application are carried out in an enlarged way by the yellow wine fermentation in the embodiment 2.

According to the method of the embodiment 3, the low-fusel high-ester-yield saccharomyces cerevisiae jiangnan2# and the factory production strain 85# are respectively used as an experimental strain and a control strain to prepare the quick brewing yeast, the method of the embodiment 2 is used for mechanized traditional yellow wine brewing, and the physicochemical properties such as alcohol degree, fusel and flavor, urea content, ethyl carbamate, amino acid, organic acid and the like are measured after the fermentation is finished.

The alcohol content of yellow wine obtained after the fermentation of saccharomyces cerevisiae jiangnan2# is 16.15% (v/v), the fusel content is 416.05 +/-10.25 mg/L, the 2-phenethyl alcohol content is 96.63 +/-5.21 mg/L, compared with 85#, 2-phenethyl alcohol is reduced by more than 15%, the content of ethyl ester (ethyl acetate, isoamyl acetate and phenylethyl acetate) is increased by more than 50%, the fermentation characteristic of the low-yield fusel and high-yield ester of the strain is more obvious, the aroma and the taste of the yellow wine finally obtained after squeezing, blending and filtering are softer and more coordinated, the drinking comfort is good, the yellow wine is not easy to get on the head after being drunk in a proper amount, the sobering is fast after being drunk in an excessive amount, the urea content is 19.80 +/-0.53 mg/L, the content of ethyl carbamate is 73.02 +/-2.41 mu g/L and is lower than 85#, and other indexes such as amino acid, organic acid and the like of the strain commonly used in the factory at present all meet the national standard GB/T13662 and 2018 of yellow wine.

(2) Application of low-yield fusel high-yield ester-producing saccharomyces cerevisiae jiangnan2# in 20-ton mechanized husked millet yellow wine

Yellow wine brewing is carried out according to the traditional yellow wine brewing process in the embodiment 2, except that the inoculation liquid in the step S2 is replaced by saccharomyces cerevisiae jiangnan2# bacterial liquid or Angel yeast activation leaven respectively; step S5, performing plate-and-frame squeezing and diatomite filtration on the fermented mash to obtain a filtrate, and clarifying the filtrate to obtain sake, wherein the sake is decocted into yellow wine (no caramel color is added).

The mechanized husked millet yellow wine fermentation comprises the following raw materials in percentage by weight:

experimental groups: husked millet: 5500 kg; water: 6600kg (L); raw wheat koji: 550kg-82.5 kg; cooked wheat koji; 99 kg; saccharomyces cerevisiae jiangnan2# quick brewing yeast: 62.7L;

control group: 5500kg of husked millet; 6600kg (L) of water; 11kg of Angel brewing high-activity dry yeast and 27.5kg of Angel brewing yeast, wherein the Angel brewing high-activity dry yeast and the Angel brewing yeast are activated by sugar water containing 20g/L glucose with water temperature of 150L and water temperature of 30-35 ℃ for 20-30 min in advance to be used as an activation leaven, and the yeast number in the activation leaven is more than 10⁸CFU/mL。

After fermentation, the final alcoholic strength of yellow wine obtained by fermenting saccharomyces cerevisiae jiangnan2# with low yield of fusel and high yield of ester is 19.8% (v/v), the content of fusel and 2-phenethyl alcohol in unit alcoholic strength is 21.9mg/L and 4.95mg/L respectively, the total amount of ester is 254.14mg/L, and the content of ethyl carbamate is obviously lower than that of a control group within a limited range; the contrast group is 19.4 percent (v/v), the content of fusel with unit alcoholic strength and the content of 2-phenethyl alcohol are respectively 29.7mg/L and 12.0mg/L, and the total amount of esters is 173.33mg/L, compared with the contrast group, the experimental group saccharomyces cerevisiae jiangnan2# unit alcoholic strength fusel is reduced by 26.26 percent, the unit alcoholic strength fusel 2-phenethyl alcohol is reduced by 58.75 percent, the fusel reduction effect is obvious, the total amount of esters is increased by 46.62 percent, and other indexes all accord with the yellow wine national standard GB/T13662-2018.

(3) Application of saccharomyces cerevisiae (S.cerevisiae) strain jiangnan2# in millet yellow wine

The millet yellow wine selected in the embodiment is fermented by the following raw materials:

millet (base): 500 g; water: 500g (mL) -625g (mL); raw wheat koji: 50g-75 g; cooked wheat koji: 9g of a mixture; saccharomyces cerevisiae jiangnan2# quick brewing yeast: 57 mL; when Angel brewing high-activity dry yeast and Angel brewing yeast are used, the activating leaven is used to replace raw wheat yeast, cooked wheat yeast and quick brewing yeast.

Experimental groups: the method of example 3(1) was followed, and Saccharomyces cerevisiae jiangnan2# with low production of fusel and high production of ester was used as a fast brewing yeast strain.

Control group 1: according to the method of example 3(1), a quick brewing yeast strain prepared by using Saccharomyces cerevisiae strain No. 85 as a pure yeast was used in the factory.

Control group 2: 1g of Angel brewing high-activity dry yeast and 2.5g of Angel brewing yeast, and activating Angel with 30mL of sugar water containing 20g/L glucose at 30-35 deg.CHigh-activity dry yeast for brewing wine and Angel yeast for brewing wine for 20-30 min, and the active yeast is more than 10⁸CFU/mL。

The millet yellow wine brewing process comprises the following steps:

a) the preparation method of the fermented raw material rice comprises the following steps: soaking the millet raw rice in water 2-3 times the weight of the millet raw rice at normal temperature for 1-3 days, wherein the acidity is more than 3mg/L, draining the water to obtain wet rice, and steaming the wet rice in a rice steaming cabinet at 121 ℃ for 30-50 min until the rice is cooked but not completely cooked, and the rice yield is 140-160%.

b) Respectively cooling the experimental group and the control group to 25-28 ℃ according to the raw material ratio of millet yellow wine fermentation, performing primary fermentation at the temperature of 20-35 ℃, performing secondary fermentation for 3-7 days, performing secondary fermentation at the temperature of 10-15 ℃, and performing secondary fermentation for 15-20 days.

c) The factory production comprises plate-frame squeezing fermented liquor, filtering with diatomite, clarifying the obtained filtrate to obtain sake, blending sake with sake to obtain yellow wine, aging, and optionally adding fructus Lycii, fructus Jujubae, Mel, radix astragali, radix Ginseng Indici, and preserved plum. This example is a laboratory scale small system simulated fermentation, which is performed by gauze filtration and centrifugation.

After fermentation, the final millet yellow wine obtained by fermentation of a control group and an experimental group has the alcoholic strength of 10% (v/v) -15% (v/v), the content of fusel and 2-phenylethyl alcohol in unit alcoholic strength of a strain jiangnan2# is lower than that of the control group, the total ester content is higher than that of the control group, the content of fusel in the strain jiangnan2# is lower than 420mg/L, the content of fusel and 2-phenylethyl alcohol in unit alcoholic strength is reduced by more than 15% compared with that of the control group 1 and the control group 2, the total ester content is 100mg/L-160mg/L, the content is increased by more than 20% compared with that of the control group 1 and the control group 2, the content of ethyl carbamate is less than 100 mu g/L, other indexes all accord with the national standard of yellow wine, the millet yellow wine obtained by fermentation has transparent, bright color and mouthfeel, and the final millet yellow wine obtained by fermentation of the strain jiangnan2# has fruity flavor, the ester fragrance can meet the requirements of consumers on low fusel, high ester, nutrition and health of alcoholic beverages.

(4) Application of saccharomyces cerevisiae (S.cerevisiae) strain jiangnan2# in reduction of red yeast yellow wine

The application of the red yeast rice yellow wine production is carried out according to the method in the step (2) in the embodiment 2, except that red yeast rice is used to replace wheat starter in the raw material in the step S3, after the fermentation is carried out for 15-20 days in the step S5, the fermented mash is pressed by a plate frame and filtered by diatomite, and the obtained filtrate is clarified to obtain the sake and fried wine which is the red yeast rice yellow wine (without adding caramel).

Taking saccharomyces cerevisiae jiangnan2# with low yield of fusel and high yield of ester as an experimental strain, preparing quick brewing yeast in the mode of the embodiment 3, using Angel brewing high-activity dry yeast and Angel brewing yeast to replace red yeast and quick brewing yeast in the mode of the

embodiment

4 and 2 to prepare an activated leavening agent as a comparison group, respectively fermenting red yeast yellow wine according to the raw material proportion, and measuring the physicochemical properties such as alcoholic strength, fusel, flavor, urea content, ethyl carbamate, amino acid, organic acid and the like after the fermentation.

After fermentation, the alcohol content of the red yeast rice yellow wine obtained by fermentation of a control group and an experimental group is 15% (v/v) -18% (v/v), the red yeast rice yellow wine obtained by fermentation is brown or dark brown, the mouth feel is fresh and cool, no peculiar smell is generated, the wine body is harmonious, the red yeast rice yellow wine obtained by fermentation of the strain jiangnan2# has stronger fruit aroma and ester aroma, compared with the control group, the fusel alcohol content and the 2-phenethyl alcohol content per unit alcohol content are respectively 34.2mg/L and 14.5mg/L, the total ester content is 176.52mg/L, the fusel alcohol content per unit alcohol content is reduced by 27.12%, the 2-phenethyl alcohol content per unit alcohol content is reduced by 25.50%, the fusel alcohol reduction effect is obvious, the total ester content is increased by 38.17%, and other indexes all meet the national standards of yellow wine.

Example 5 application of Saccharomyces cerevisiae jiangnan2# with low yield of fusel and high ester yield in cooking wine

The yellow wine is obtained by fermentation according to the mode in the embodiment 2(2), the difference is that the rice water ratio is 1:1, the adding proportion of other raw materials is unchanged, part of the yellow wine sample is taken and added with 10% of edible salt, the edible water, spice and caramel color can be added according to the product requirement, the cooking wine prepared by taking the yellow wine brewed by the saccharomyces cerevisiae (S.cerevisiae) strain jiangnan2# with low yield and high ester yield as the main raw material is obtained after sterilization treatment, the alcoholic strength is 10% (v/v) -15% (v/v), the amino nitrogen content is higher than 0.5g/L, the ethyl carbamate content is 84 mu g/L, the brewed cooking wine has high ester content, is rich in amino acid, has better flavor and improved safety, and the product meets the requirements of SB/T10416-type 2007 seasoning cooking wine.

Example 6 application of Saccharomyces cerevisiae jiangnan2# with low yield of fusel and high yield of ester in rice wine

Washing rice as main raw material with clear water, soaking at room temperature for about 4 hr until water is absorbed sufficiently, and kneading with hand until the soaked rice is fragile and has no hard rice core; draining off water from the soaked rice grains, and steaming at normal pressure for about 30min until no white core exists in the rice; steaming rice, cooling with cold boiled water to about 25-28 deg.C, adding sweet distiller's yeast with 0.4-0.8% of rice mass, saccharifying at 28 + -2 deg.C for 36-42 h, inoculating 5-10% of the low-yield fusel high-yield ester-producing Saccharomyces cerevisiae jiangnan2# quick brewing yeast prepared according to example 3 after saccharification, stirring uniformly, adding drinking water with 1-1.5 times of the raw material rice mass, fermenting at 26-34 deg.C for 3-5 days, reducing the temperature to 10-15 deg.C, fermenting for 10-15 days, squeezing, and filtering to obtain rice wine. The rice wine obtained by fermentation is rich in rice aroma and low in higher alcohol content, the content of fusel in unit alcohol degree is less than 20mg/L, and the rice wine is not easy to get drunk after being drunk.

Example 7 application of Saccharomyces cerevisiae jiangnan2# for producing low-yield fusel and high-yield ester in sweet rice-brewing

Washing rice as main raw material with clear water, soaking at room temperature for about 4 hr until water is absorbed sufficiently, and kneading with hand until the soaked rice is fragile and has no hard rice core; draining off water from the soaked rice grains, and steaming at normal pressure for about 30min until no white core exists in the rice; steaming rice, spraying cold boiled water on the rice to about 25-28 ℃, transferring the rice to a fermentation tank, adding 0.4-0.8% of Angel sweet distiller's yeast based on the mass of the raw rice, inoculating 0.5-1.5% of brewing yeast Jiangnan2# quick brewing yeast with low yield of fusel and high ester yield prepared by the method in the embodiment 3, stirring uniformly, and digging a groove in the center of the rice to ensure a certain dissolved oxygen amount; adding drinking water 1-1.5 times of the original rice, and fermenting at 26-34 deg.C for 36-72 h to obtain wine flavor. The brewing precision of the sweet rice is 2-4% (v/v), the fragrance is harmonious and strong, the texture is uniform, and the taste is sweet and sour.

Example 8 application of Saccharomyces cerevisiae (S. cerevisiae) strain jiangnan2# with low yield of fusel and high ester yield in vinegar

Acetic acid fermentation was carried out using the yellow wine obtained by the method of reference example 4(1) as a raw material.

The vinegar brewing adopts a solid fermentation process: uniformly mixing yellow wine, bran and chaff according to the mass ratio of 10:4:1, inoculating vinegar grains accounting for 3-8% of the total system mass, turning the materials, keeping the fermentation temperature at 35-40 ℃, and turning the surfaces of the materials in the first 2 days. Turning over the fermented grains from top to bottom to the bottom of the material for 2-8 days, and turning over the fermented grains from bottom to top for cooling for 8-12 days. Spraying vinegar after fermentation to obtain raw vinegar, sterilizing, aging in open air, sterilizing at 85 deg.C for 30min before filling vinegar of different years, and hot packaging. After fermentation, the physical and chemical indexes of the vinegar are normal, the acetic acid content is 50-80g/L, the content of the ethyl carbamate is low, and the safety is improved.

Example 9 application of Saccharomyces cerevisiae jiangnan2# with low yield of fusel and high ester yield in white spirit

Using sorghum as a raw material, performing solid brewing of white spirit by adopting a 2-round fermentation method, cooling to about 30 ℃ after steaming the sorghum, adding 10-25% of bran koji, 8-12% of rice hulls, 10-15% of koji and 6-10% of bran, inoculating 1% of low-yield fusel high-yield ester-producing saccharomyces cerevisiae jiangnan2# pure seed quick brewing yeast prepared by the method in example 3, performing first round of closed fermentation for 30 days, and then distilling. 10-15% of medium-temperature Daqu is added in the secondary fermentation, 1% of Saccharomyces cerevisiae jiangnan2# pure seed quick brewing yeast is continuously added, the liquor is distilled after continuous fermentation for 15 days, the liquor with the alcoholic strength of 60% (v/v) is obtained after 2 rounds of distilled liquor are mixed, the acetate content of the liquor is increased, the liquor mainly comprises ethyl acetate, isoamyl acetate and phenylethyl acetate, the total amount is increased by more than 80%, meanwhile, the fusel content is reduced by more than 20%, the ethyl carbamate content is reduced by more than 50%, and the safety is improved.

Example 10 application of a combination of Saccharomyces cerevisiae jiangnan2# for low yield of fusel and high yield of esters in wine

100kg of Cabernet Sauvignon grape with complete fruit grains and good maturity is taken as a raw material, the raw material is subjected to stem removal, crushing, split charging into 150L fermentation tanks, and adding 20mg/L of pectinase (with enzyme activity of 20000U/g) and 50mg/L of SO₂Mixing, and soaking at 4 deg.C for about 24 hr. Then mixing while raising the temperature to 20 deg.CAfter homogenization, the final concentration of inoculation is 1X10⁶CFU/mL of commercial Saccharomyces cerevisiae (S.cerevisiae) F15, manufactured by LAFFORT, France, at a final concentration of 1X10⁶CFU/mL of the brewer's yeast jiangnan2# pure brewing yeast, which is prepared in the example 3, and the mixed fermentation is carried out by pumping, circulating and mixing evenly; controlling the fermentation temperature to be 25-27 ℃, and sampling and monitoring the specific gravity and the temperature of the fermentation liquid after 2 times of circulation in the morning and at night every day. When the specific gravity is not reduced any more, the alcohol fermentation is considered to be finished, and the alcohol fermentation is hermetically stored at 4 ℃, and during the storage, the precipitate is separated by discharging a plurality of times. Each set had 2 parallel fermentors. The wine obtained by adding the saccharomyces cerevisiae with low yield of fusel and high yield of ester has the precision of 12.8 percent (v/v) -14 percent (v/v) and total sugar (calculated by glucose)<5g/L, total acid (calculated by tartaric acid) is about 4.6g/L, volatile acid (calculated by acetic acid) is 0.15-0.4g/L, higher alcohol content is 205.5mg/L-230mg/L, total ester content is 45.5mg/L-55.8mg/L, fusel is reduced by more than 20%, and ester is increased by more than 30%. Sensory evaluation results show that the color, the fragrance and the taste of the wine fermented by adding the saccharomyces cerevisiae (S.cerevisiae) strain jiangnan2# with low yield, fusel and high ester yield are superior to those of the wine fermented by the commercial saccharomyces cerevisiae alone, and the fruity fragrance and the flower fragrance are more intense and prominent. The saccharomyces cerevisiae (S.cerevisiae) strain jiangnan2# can meet the normal fermentation requirement by being compounded with other microorganisms, and the index meets the national standard GB/T15037-2006 grape wine.

Example 11 application of Saccharomyces cerevisiae jiangnan2# with low yield of fusel and high ester yield in fruit wine

Washing fructus Pruni Salicinae as raw material, removing core, draining, crushing, adding high fructose syrup solution 2-3 times of the raw material weight, packaging into 150L fermentation tank, adding 20-40 mg/L pectase (enzyme activity 20000U/g) and 40-80 mg/L potassium metabisulfite, inoculating 1x10 (v/v) -5% (v/v) respectively⁷CFU/mL of low-yield fusel high-ester-yield Saccharomyces cerevisiae (S. cerevisiae) strain jiangnan2# expanding culture solution is subjected to main fermentation for 10-15 days at 20-25 ℃ and pH of 2.5-3.5, and after fermentation for 5-10 days at 5-8 ℃ to obtain the fermented plum wine. Each set had 2 parallel fermentors. The plum alcoholic strength obtained by fermentation is 10.5% (v/v) -12% (v/v), and total sugar (calculated as glucose)<40 g/L. Sensory evaluation results show that the low-yield fusel is addedThe plum wine brewed by the saccharomyces cerevisiae (S.cerevisiae) strain jiangnan2# has harmonious wine body, soft taste and better fruit wine quality. Other indexes of the fermented plum wine meet the regulations of the general technical requirements of GB 2758-.

Example 12 application of Saccharomyces cerevisiae jiangnan2# with low yield of fusel and high yield of ester in fruit vinegar

The fruit wine obtained in example 11 was used as a raw material, and acetic acid fermentation was performed using acetic acid. The fruit vinegar brewing adopts a liquid fermentation process: inoculating 1-3% acetic acid bacteria, fermenting at 34 deg.C for 16 days, pouring vinegar, clarifying, degassing, and sterilizing to obtain fruit vinegar. The acidity of the vinegar in the obtained fruit vinegar is 3-8% (g/100mL), other physical and chemical indexes are normal, the fruit vinegar meets the national standard GB/T30884-.

Example 13 application of Saccharomyces cerevisiae jiangnan2# for Low-yield fusel and high-yield ester in beer

The method is characterized in that barley malt (15kg-20kg), hops (50g-70g) and water (100L-120L) are used as main raw materials, specifically, a multi-step sugar soaking method is adopted, low-yield fusel high-yield ester-production saccharomyces cerevisiae jiangnan2# pure seed quick brewing yeast is prepared by taking wort as a culture medium according to the method in example 3, the low-yield fusel high-yield ester-production saccharomyces cerevisiae is added into the wort according to the addition amount of 0.2% -1%, and the high-yield ester-production saccharomyces cerevisiae jiangnan2# pure seed quick brewing yeast is obtained by processes of crushing, saccharifying, filtering, boiling wort, fermenting and the like, the fermentation yeast is adopted, pre-fermented at the temperature of 18% -23 ℃ for 2-3 days at the fermentation pressure of 0.15MPa, and the sugar content is reduced to be below 5Brix every day during the pre-fermentation period and then is cooled to be stored at the temperature of 5 ℃. The obtained beer has alcoholic strength of 3-5% (v/v), total sugar (calculated as glucose) of less than 50g/L, outstanding and not rich flavor, malt flavor and fruit flavor, fusel content of less than 150mg/L, and normal other physicochemical indexes.

Example 14 application of Saccharomyces cerevisiae jiangnan2# with low yield of fusel and high yield of ester in cigarette

The tobacco leaves referred to in this example are processed tobacco shred samples, and the tobacco extract is an aqueous extract or an alcohol extract, and is obtained by commercial purchase.

Culturing the saccharomyces cerevisiae jiangnan2# with low yield of fusel and high yield of ester at 28 DEG CShaking and culturing in YPD medium at + -2 deg.C for 24 hr to obtain a concentration of 10⁶CFU/mL-10⁸CFU/mL first-stage seed solution, inoculating 5% -10% of the first-stage seed solution into a fermentation culture medium containing pear juice, grape juice, sweet osmanthus or tobacco extract, and culturing for 48h to obtain Saccharomyces cerevisiae culture solution with concentration of 10⁸CFU/mL-10⁹CFU/mL is the cigarette added spice, the obtained cigarette added spice is directly added into the cigarette leaf group according to the adding amount of 1-5% of the mass of the cigarette leaf group, water is supplemented to 10-20%, then the cigarette added spice is cultured for 4-8 h at 25-37 ℃, finally the moisture is balanced after baking, and then the cigarette is manufactured, or the cigarette added spice is dissolved in 75% of alcohol to prepare a fragrant liquid (0.1-0.5 g/mL), and the fragrant liquid is uniformly sprayed in the processed tobacco shred sample according to a proper proportion to manufacture the cigarette. The obtained cigarette has soft smoke, low irritation, light fruit fragrance, light flower fragrance, pleasant sweet and mellow fragrance and special fragrance of the extract, can improve the smoking quality of the cigarette, and is suitable for improving the special fragrance and quality of the cigarette and novel tobacco products.

Example 15 application of Saccharomyces cerevisiae jiangnan2# with low yield of fusel and high yield of ester in fermented nutritional ice cream

Mixing drinking water, squeezed fruit juice, sugar, honey, etc. as main material in the ratio of (2-4): 0.05-0.2): 0.01-0.05, regulating sugar concentration to 10Brix-20Brix, regulating pH to 4-5 with food grade lactic acid, inoculating 5-10% malt juice to culture bacteria liquid concentration of 10⁶-10⁸CFU/mL Saccharomyces cerevisiae jiangnan2# with low yield of fusel and high ester yield is subjected to shaking culture in a constant-temperature incubator at 30 +/-2 ℃ for 24-48 h, after fermentation is finished, centrifugation is carried out, supernatant or traditional brewed yellow wine obtained in example 2 is used for replacing drinking water, 30-50% of whole milk powder, 1-5% of thickening agent, 1-5% of emulsifying agent, 1-5% of swelling agent, 5-10% of non-dairy creamer and other additives are added until the total dry matter is 20-30%, pasteurization and homogenization are carried out after mixing, then low-temperature cooling and aging are carried out at 4 ℃ for about 1h, the mixture is frozen and stirred, and then split charging is carried out on a die, and the nutritional fermented ice cream is obtained after freezing, demolding and the like of the die. The obtained ice cream has light sweet and mellow fragranceAnd the flower and fruit flavor is mixed, the taste is soft while the heat is removed and the summer heat is relieved, and the stimulation to the mouth and the sweet and greasy feeling of the ice cream can be reduced.

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

South of the Yangtze university (Shaoxing) industry and technology research institute

<120> saccharomyces cerevisiae with low yield of fusel and high yield of ester and application thereof in production of fermented food

<130> BAA210950A

<160> 1

<170> PatentIn version 3.3

<210> 1

<211> 815

<212> DNA

<213> Saccharomyces cerevisiae

<400> 1

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gttttaccaa ctgcggctaa tcttttttat actgagcgta ttggaacgtt atcgataaga 720

agagagcgtc taggcgaaca atgttcttaa agtttgacct caaatcaggt aggagtaccc 780

gctgaactta agcatatcaa aaaaccggga ggaaa 815

Claims

1. Saccharomyces cerevisiae Jiangnan2# which has been deposited at China center for type culture Collection at 13.05.2021 with the deposition number of CCTCC NO: M2021524.

2. A composition comprising Saccharomyces cerevisiae jiangnan2# according to claim 1.

3. The composition of claim 2, wherein the composition includes, but is not limited to, microbial agents, wine drugs, or wheat starter.

4. A microbial preparation comprising live cells of the Saccharomyces cerevisiae jiangnan2# or Saccharomyces cerevisiae jiangnan2# of claim 1 and other microorganisms, dried cells obtained by freeze-drying, or cells obtained by a solidification technique.

5. The microbial preparation of claim 4, wherein the number of yeast cells in the microbial preparation is not less than 1x10⁷CFU/mL, the germination rate is more than or equal to 30 percent.

6. Use of saccharomyces cerevisiae jiangnan2# according to claim 1, or the composition according to any one of claims 2 to 3, or the microbial preparation according to any one of claims 4 to 5 for the production of a fermented product.

7. Use according to claim 6, characterized in that the fermented product is a fermented food or a fermented beverage; the fermented beverage includes, but is not limited to, fermented alcoholic beverages or fermented vinegar.

8. The use according to claim 7, wherein said fermented alcoholic beverage comprises, but is not limited to: yellow wine, cooking wine, rice wine, sweet fermented rice, wine, fruit wine, beer or Chinese liquor; the fermented vinegar includes fruit vinegar or edible vinegar.

9. The use of claim 6, wherein the fermented product is fermented tobacco leaf or fermented ice cream.

10. The saccharomyces cerevisiae jiangnan2# of claim 1 or a product obtained by distilling or blending a fermentation product prepared by fermenting the composition of any one of claims 2 to 3.