CN113621528B

CN113621528B - Saccharomyces cerevisiae strain with low yield of fusel and high yield of ester and application of saccharomyces cerevisiae strain in fermented food

Info

Publication number: CN113621528B
Application number: CN202110891798.3A
Authority: CN
Inventors: 毛健; 刘双平; 赵禹宗; 韩笑
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: 2023-06-16
Anticipated expiration: 2041-08-04
Also published as: CN113621528A

Abstract

The invention discloses a saccharomyces cerevisiae strain with low yield of fusel and high yield of ester and application of the saccharomyces cerevisiae strain in fermented foods, belonging to the field of fermentation engineering and biotechnology. The saccharomyces cerevisiae jiangnan No. 1 has excellent stress resistance and good fermentation performance, can meet the industrial production requirements of low-yield fusel and high-yield ester of different alcoholic beverages and fermented foods, has the fermentation characteristics of low-yield urea and low-yield ethyl carbamate, has the urea content of 18.60+/-0.53 mg/L and the ethyl carbamate content of 67.91 +/-3.06 mug/L in the fermented yellow wine, is remarkably lower than the current commonly used saccharomyces cerevisiae strain in factories, can remarkably improve the flavor of the yellow wine, meets the industrial production requirements of high-quality yellow wine with high comfort and high safety, has good application prospect, and can realize the improvement of safety and quality when applied to other fermented foods.

Description

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

Technical Field

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

Background

Saccharomyces cerevisiae is a main flavor generating microorganism of alcoholic beverages, the fermentation power of Saccharomyces cerevisiae is an important power source for brewing production, the yeast utilizes sugar in raw materials to generate alcohol, and proteins and fat are converted into organic acid, amino acid, esters and the like after the action of microorganisms such as yeast, lactobacillus and the like. The quality of yeast directly affects the production efficiency and flavor quality of the final product. Meanwhile, yeast is closely related to the generation of some non-preference substances, namely higher alcohols (fusel oil), urea and ethyl carbamate in the brewing process, the content of the substances exceeding the limit value can cause negative influence to influence the drinking comfort and safety, and the quality, the value and the price of the product are not considered preferentially at present along with the improvement of the consumption level and the consumption cognition of consumers, and the safety and the health factors of the product are paid more importance to more and more, so that the contradiction problem of mismatching of the characteristics of the product and the demands of consumers occurs.

Fusel and some acetate species are metabolites of yeast during fermentation. Alcohol ester imbalance (high content of fusel and low content of esters), and non-favored metabolic substances such as biogenic amine, urea and urethane are fermented wine such as yellow wine, cooking wine, beer, wine and fruit wine, and distilled wine such as white wine, vodka and brandy have common problems. Saccharomyces cerevisiae is involved directly or indirectly in the metabolic production of the above substances throughout the fermentation process, and how to metabolize by controlling fermenting microorganisms is extremely important for improving the quality of fermented foods.

At present, the Saccharomyces cerevisiae mutant strain obtained by means of mutagenesis can obviously reduce the content of substances such as fusel, urea and urethane, but the strain bred by means of mutagenesis is preferentially selected to meet the requirement of a single purpose, the mutagenesis is uncertain, the craving character is not easy to obtain, the mutagenesis means can change the fermentation characteristics of relatively balanced composition of different original metabolic substances of the strain, and the mutant strain cannot have excellent fermentation characteristics at the same time. The inventor constructs a high-yield ester low-yield higher alcohol saccharomyces cerevisiae engineering strain, improves the ester content in white spirit and reduces the higher alcohol content, and constructs a low-yield higher alcohol high-yield ethyl lactate saccharomyces cerevisiae strain, remarkably improves the ethyl lactate production and reduces the higher alcohol production simultaneously in the patent application with the publication number of CN 201511017931.3. In addition, the strain transformation is carried out on the wine saccharomyces cerevisiae by adopting genetic engineering abroad, only 2 transgenic wine saccharomyces cerevisiae strains ML01 and ECMO01 respectively realize the purpose transformation without generating biogenic amine and urea and are allowed to be used in a certain range, but whether the transgenic technology is absolutely safe is not clear, so that the transgenic technology is not widely accepted, and most of metabolites of the saccharomyces cerevisiae are always remained in the final product in fermented foods such as yellow wine and vinegar. The low-yield and high-yield ester saccharomyces cerevisiae obtained through natural breeding is still the most effective breeding means.

Regarding the research related to the breeding and application of low-yield and high-yield ester saccharomyces cerevisiae in fermented foods such as white spirit, yellow wine, beer, wine and the like, a strain of low-yield urea yellow wine saccharomycete and application thereof are reported in the CN201510971067.4 patent, a strain of saccharomycete for producing Shaoxing yellow wine by rapid fermentation is reported in the CN201210136289.0 patent, no effective method can fundamentally solve the problems in the research and patent of the retrievable saccharomyces cerevisiae applied to the food related field, and the safety industrial production saccharomyces cerevisiae strain with high alcohol content, low impurity alcohol content and high ester content and obviously lower than the specified limit and application thereof have not been found on the premise of meeting normal fermentation. Therefore, the low-impurity-alcohol high-ester-yield saccharomyces cerevisiae obtained through saccharomyces cerevisiae strain separation screening and systematic evaluation breeding and application thereof in fermented foods are of great significance in improving the quality, health and safety of alcoholic beverages such as yellow wine and fermented foods such as table vinegar.

Disclosure of Invention

In order to solve the problems, the saccharomyces cerevisiae with excellent fermentation performance and high safety is aimed at the fact that the prior production of fermented food has no low-yield fusel and high-yield ester. The invention provides a saccharomyces cerevisiae (Saccharomyces cerevisiae) strain with excellent stress resistance and good fermentation performance, which is low in impurity alcohol yield and high in ester yield, and application of the saccharomyces cerevisiae (Saccharomyces cerevisiae) strain in fermented foods.

The invention provides a strain jiangnan1#, which is a strain of Saccharomyces cerevisiae with low yield of fusel and high yield of ester, and is preserved in China Center for Type Culture Collection (CCTCC) in the year 2021, the month 13, and the preservation number is CCTCC NO: m2021523, the preservation address is China, the university of Wuhan, and Wuhan.

In one embodiment of the invention, the morphology of the monoclonal colony of Saccharomyces cerevisiae jiangnan No. 1 is milky white, oval or elliptic, convex, smooth, moist and glossy on the surface and neat on the edge of the strain.

The invention also provides an application mode of the low-impurity-alcohol-yield and high-ester-yield saccharomyces cerevisiae in fermented foods.

The invention also provides a composition prepared by compounding the microbial agent of the saccharomyces cerevisiae jiangnan1# with other microorganisms.

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

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

In one embodiment, the microbial agent contains living cells of the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# bacterial cells, freeze-dried low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# dry bacterial cells obtained by freeze drying, the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# bacterial cells obtained by a solidification technology, liquid bacterial agents of the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# or solid bacterial agents of the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# bacterial strains existing in any other forms.

In one embodiment, the microbial preparation has a number of Saccharomyces cerevisiae jiangnan1 ∈1×10 ⁶ CFU/g。

In one embodiment, the other microorganisms include, but are not limited to, bacteria or fungi.

In one embodiment, the method of application of the compound with other microorganisms refers to the steps of compounding the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# with other microorganisms, wherein 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 formulation is prepared as follows: inoculating the strain into a rice saccharification liquid culture medium, and carrying out shaking culture for 20-24 hours at the culture temperature of 28+/-2 ℃ to obtain primary seed liquid; inoculating the first seed solution into new rice saccharification liquid culture medium at a ratio of 5% -10%, and shake culturing at 28deg.C+ -2deg.C for 36-48 hr to obtain second seed solution with yeast number not less than 1×10 ⁷ CFU/mL, and the budding rate is more than or equal to 30 percent.

In one embodiment, the strengthening wine is that the saccharomyces cerevisiae jiangnan1# or the microbial agent is added into the wine in a compounding way or any other way in the manufacturing process of the wine, so that the purpose that the fermentation characteristic of the wine can be strengthened when the wine is used is achieved.

In one embodiment, the reinforced distiller's yeast means that the Saccharomyces cerevisiae jiangnan1# or the microbial agent is added into the distiller's yeast in any form in the preparation process of the distiller's yeast, so that the purpose that the distiller's yeast can strengthen the fermentation characteristic of the distiller's yeast when in use is achieved.

The invention also provides application of the low-impurity-alcohol-yield and high-ester-yield saccharomyces cerevisiae in fermented food.

In one embodiment, the use of the fermented food comprises the use of the fermented food in 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 rice wine, fruit vinegar, table vinegar, white wine, beer, tobacco leaf, fermented ice cream, etc.

In one embodiment, the application is for yellow wine brewing, and the low-impurity-alcohol-yield high-ester-yield saccharomyces cerevisiae is quick brewing yeast.

In one embodiment, the yellow wine brewing is to use the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# as a quick brewing parent, add the quick brewing parent into raw materials (rice, millet, corn, millet and the like) which are steamed or gelatinized according to the adding amount of 5% -15%, and ferment, squeeze, decoct, age, filter, sterilize and fill the yellow wine.

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

In one embodiment, the cooking wine brewing is to prepare the cooking wine by first fermenting the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# serving as a quick brewing parent to obtain the yellow wine and then using the yellow wine.

In one embodiment, the application is for rice wine brewing, and the low-yield and high-yield ester saccharomyces cerevisiae jiannnan 1# is used as a starter.

In one embodiment, the rice wine brewing is to brew the low-impurity-alcohol high-ester-yield saccharomyces cerevisiae jiangnan1# and distiller's yeast as fermenting agents, add the low-impurity-alcohol high-ester-yield saccharomyces cerevisiae jiangnan1# and distiller's yeast into the steamed rice raw material according to the adding amount of 0.5% -1.5%, and obtain the rice wine through the processes of steaming, adding yeast (adding the saccharomyces cerevisiae), saccharification, fermentation, squeezing and the like.

In one embodiment, the application is for sweet rice brewing, and the low-impurity-alcohol high-yield ester saccharomyces cerevisiae jiangnan1# is used as a starter.

In one embodiment, the rice wine brewing is obtained by brewing the low-impurity-alcohol high-ester-yield saccharomyces cerevisiae jiangnan1# and distiller's yeast as fermenting agents, adding the low-impurity-alcohol high-ester-yield saccharomyces cerevisiae jiangnan1# and distiller's yeast into steamed raw materials (rice, millet, corn, millet and the like) according to the adding amount of 0.5% -1.5%, and performing the processes of steaming, adding yeast (adding the saccharomyces cerevisiae), saccharification, fermentation and the like.

In one embodiment, the application is for vinegar brewing.

In one embodiment, the application is for brewing vinegar by using the low-impurity-alcohol high-ester-yield saccharomyces cerevisiae as a yeast to ferment first to obtain yellow wine and then using the yellow wine as an acetic acid fermentation raw material.

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

In one embodiment, the brewing white spirit is that the low-yield miscellaneous alcohol high-yield ester saccharomyces cerevisiae jiangnan1# is additionally added when the white spirit is fermented into a pool for fermentation.

In one embodiment, the added amount of the low-impurity-alcohol-yield high-ester-yield saccharomyces cerevisiae jiangnan1# is 1% (v/v) of the raw material system when the white wine is brewed.

In one embodiment, the use is for brewing wine.

In one embodiment, the brewed wine is additionally added with the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# in the alcohol fermentation stage.

In one embodiment, the low-impurity-alcohol high-ester-yield Saccharomyces cerevisiae jiangnan1# is added in an amount of 0.5% (v/v) to 1.5% (v/v) of the total volume of the fermentation system when brewing the wine.

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

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

In one embodiment, the brewed fruit wine comprises the following processes: and (3) treatment before fermentation, and inoculating the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# into clear juice after juice clarification.

In one embodiment, the low-impurity-alcohol high-ester-producing Saccharomyces cerevisiae jiangnan1# is inoculated in an amount of 1% (v/v) to 5% (v/v) of the total volume of the juice when the fruit wine is brewed.

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

In one embodiment, the application is that the low-yield and high-yield ester saccharomyces cerevisiae jiannnan 1# is firstly utilized to ferment to obtain fruit wine, then the fruit wine is utilized as a fermentation raw material to be inoculated with acetic acid strain, and acetic acid fermentation is carried out to brew the fruit vinegar.

In one embodiment, the use is for brewing beer.

In one embodiment, the application is for brewing beer, wherein the low-impurity-alcohol high-ester-yield saccharomyces cerevisiae jiangnan1# is used as pure yeast, and added into wort according to the addition amount of 0.2% -1% after the cultivation is expanded, and the beer is obtained through malt treatment, brewing, filling and other processes.

In one embodiment, the use is for making cigarettes.

In one embodiment, the application is used for making cigarettes, and the low-yield and high-yield ester saccharomyces cerevisiae jiannnan 1# culture solution is directly used for preparing tobacco flavor; or taking pear, grape, sweet osmanthus, tobacco extract and the like as culture medium raw materials, inoculating the low-yield fusel high-yield ester saccharomyces cerevisiae jiannnan 1# for fermentation to prepare fermented tobacco flavor, and then directly adding the obtained tobacco flavor into a tobacco rolling group according to the adding amount of 1% -5% of the mass of the tobacco rolling group.

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

In one embodiment, the application is used for preparing fermented nutritional ice cream, which is obtained by fermenting raw materials (fruit juice, sugar, honey, whole milk powder and the like, sweet potato, purple sweet potato, corn and other starches) with 5-10% of additive amount by taking the low-impurity-alcohol high-ester saccharomyces cerevisiae jiangnan1# as a fermenting agent, adding other food additives, and then mixing, pasteurizing, homogenizing, cooling and aging, freezing and stirring, injection molding, freezing, demolding and other processes.

The invention also provides a product obtained by distilling, blending or adding the saccharomyces cerevisiae jiangnan1# fermentation product.

The invention has the beneficial effects that:

(1) The invention provides a strain of low-impurity-alcohol high-ester-yield saccharomyces cerevisiae jiangnan1#, which has excellent stress resistance, ethanol tolerance as high as 20% (v/v), and good growth condition under the osmotic pressure condition of 1.6mol/L sodium chloride.

(2) Yellow wine fermentation experiments show that the jiangnan No. 1 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 ℃.

(3) When the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# provided by the invention is adopted for brewing mechanized yellow wine, compared with 85# saccharomycetes commonly used in the yellow wine industry disclosed by patent CN 105385613B, the jiangnan1# strain is quick in fermentation starting, the alcohol content of the yellow wine obtained after the fermentation is more than 13% (v/v) for 48 hours, the alcohol content in the yellow wine obtained after the fermentation is 16.5% (v/v), the impurity alcohol content is 398.68 +/-13.25 mg/L, the 2-phenethyl alcohol content is 79.43+/-5.21 mg/L, the content of ethyl ester (ethyl acetate, isoamyl acetate and phenethyl acetate) is reduced by more than 20%, the low-yield and high-yield ester fermenting characteristics of the strain are more obvious, and the finally obtained yellow wine after squeezing, blending and filtering has softer and more coordinated fragrance and taste, good drinking comfort, and is not easy to drink up, and sobering up after drinking.

(4) When the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# provided by the invention is used for brewing mechanized yellow wine, the urea content is 18.60+/-0.53 mg/L, the ethyl carbamate content is 67.91 +/-3.06 mug/L, which is obviously lower than the current common strain in factories, the amount of ethyl carbamate in the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# fermentation age Xie Shengcheng is less, and the safety of fermenting foods by taking the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# as a fermentation strain can be improved.

Preservation of biological materials

Saccharomyces cerevisiae (Saccharomyces cerevisiae) jiangnan1#, which is classified and named as Saccharomyces cerevisiae (Saccharomyces cerevisiae) jiangnan1#, is preserved in China Center for Type Culture Collection (CCTCC) No. M2021523 in the year 2021, month 05 and 13, and has a preservation address of China, wuhan university.

Drawings

FIG. 1 shows the growth curves of Saccharomyces cerevisiae # 1 of the present invention at various temperatures.

FIG. 2 analysis of correlation of yeast isolates with flavor substances.

FIG. 3 is a diagram showing the aroma and taste profile of yellow wine brewed with Saccharomyces cerevisiae # 1 of the present invention.

Detailed Description

And (3) detecting physical and chemical indexes of the yellow rice wine: the measurement of alcohol content, amino acid nitrogen and total acid is carried out by referring 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 and the like are detected by gas chromatography-mass spectrometry (GC-MS). The determination of the reducing sugar content adopts a DNS method. The concentration of urea and bacterial liquid is measured by spectrophotometry. The higher alcohol (also called as fusel) in the yellow wine mainly comprises 4 types of n-propanol, isobutanol, isovaleryl alcohol and 2-phenethyl alcohol, a dispersion liquid microextraction technology (DLLME) is adopted, GC-MS detection is utilized, 4-methyl-2-pentanol is used as an internal standard, and an external standard curve is established for quantitatively measuring the fusel content.

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

Sensory evaluation: the difference between the samples and the control was compared after 10 persons (5 men and 5 women) of the evaluator aged between 20 and 40 years who had the sensory evaluation experience of the alcoholic beverage were selected and presented to the evaluator according to the sensory requirements (appearance, aroma, taste and style characteristics) of the alcoholic beverage in the corresponding national standards.

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

YPD medium: 10g yeast extract, 20g peptone, 20g glucose, 1000mL of water, 2% agar, and autoclaved at 121 ℃ for 20min, and cooled for later use.

Rice saccharification liquid culture medium: taking proper amount of high-quality rice raw materials, soaking the rice for 30min in a water bath at the constant temperature of 60 ℃ and steaming for 20min under normal pressure, then respectively adding 150U/g-300U/g of saccharifying enzyme and 200U/g-400U/g of liquefying enzyme based on the rice raw materials, 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 above 13Brix, sub-packaging for 121 ℃ and sterilizing for 15min-20min under high pressure, and cooling for later use.

Example 1 selection and identification of Saccharomyces cerevisiae Strain selection and selection of Saccharomyces cerevisiae with Low yield of fusogenic high yield of esters

1. Sample preparation, strain isolation and preservation

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

2. Primary screening based on yellow wine fermentation of Saccharomyces cerevisiae isolates: in order to select Saccharomyces cerevisiae with excellent fermentation performance capable of meeting the requirements of brewing yellow wine, all the separated strains are respectively prepared into quick brewing mother and then subjected to yellow wine fermentation, physicochemical indexes such as alcohol degree and the like are measured after fermentation is finished, the strains are subjected to primary screening, and the measured impurity and flavor substances with alcohol degree meeting national standards are screened according to different batches. The method comprises the following specific steps:

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

According to 100% glutinous rice (basis): 500g;125% water: 625g;12% -15% wheat starter: 60g-75g;10% -15% of yeast triangular flask culture solution: 50mL-75mL.

The manufacturing method of the quick brewing master comprises the following steps: slant preparation of Saccharomyces cerevisiae glycerol pipe preservation strain, inoculating slant cultured Saccharomyces cerevisiae (S.cerevisiae) jiangnan No. 1 strain into rice saccharification liquid culture medium, shake culturing at 28deg.C+ -2deg.C for 24 hr to obtain primary seed liquid (bacterial liquid concentration 10) ⁶ -10 ⁸ CFU/mL), inoculating the primary seed liquid to a new rice saccharification liquid culture medium at a ratio of 5% -10%, and carrying out shaking culture for 36-48 h at a culture temperature of 28+/-2 ℃ to obtain a secondary seed liquid, wherein the obtained secondary seed liquid is used for measuring the viable count and the budding rate of the yeast, and the number of the yeast is more than or equal to 1 multiplied by 10 ⁷ CFU/mL, the budding rate is more than or equal to 30%, and the rice wine brewing is carried out by taking the rice wine as a quick brewing master.

Experimental group: the Saccharomyces cerevisiae strain obtained by separation is used as pure yeast to prepare quick Saccharomyces cerevisiae.

Control group: saccharomyces cerevisiae strain # 85 (disclosed in patent CN 105385613B) for factory production was used as a seed yeast for making instant brewing yeast.

(2) Traditional yellow wine brewing process

a) Preparation of fermented raw material rice: the raw rice with the production dosage is added with water to be soaked until the water exceeds the liquid level by more than 10cm, the acidity of the rice slurry of the soaked rice reaches more than 4.5g/L for 3-5 days, the water is drained to obtain wet rice, the wet rice is steamed for 20-30 min at the temperature of 121 ℃ in a rice steaming cabinet until the rice is cooked but not transparent, white cores are not arranged in the rice grains, the rice has sour taste and has 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:

the method comprises the following specific steps of:

s1, respectively marking the strains stored in the glycerol pipe in a freezing way to form monoclonal colonies on a flat plate, and picking the monoclonal colonies to inoculate on an inclined plane to obtain a saccharomyces cerevisiae inclined plane culture medium capable of being stored at a low temperature;

s2, transferring the strain of S1 from the inclined plane to a rice saccharification liquid culture medium, and culturing at 28+/-2 ℃ until the order of magnitude is more than or equal to 1 multiplied by 10 ⁷ The germination rate is more than 30 percent, and the germination rate is used as an inoculation liquid;

s3, adding raw material rice and water into a sterilized fermentation container, adding the inoculation liquid obtained in the step S2 accounting for 10-15% of the mass of the raw material rice into a rice-water mixed fermentation system, adding wheat starter accounting for 12-15% of the mass of the raw material rice, completing material mixing at 25-28 ℃, and standing for 3-5 days at 20-35 ℃ for pre-fermentation;

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

s5, squeezing the fermented mash obtained in the step S4 through a plate frame (4 times of feeding, the mash inlet pressure is 0.2-0.6MPa, the filtering area is 100m < 2 >, the filter plate diameter is 1 m), filtering by diatomite (the diatomite adding proportion is 4% -6%, the pressure is 0.3-0.5 MPa), clarifying the obtained filtrate to obtain sake, blending the sake, adding 1-3 per mill of caramel according to national standard of yellow wine, and frying the sake to obtain yellow wine.

A total of 144 isolates were taken as an example of a batch containing Saccharomyces cerevisiae (strain number CYY-661) jiangnan1# (strain number at the time of breeding). And (3) after fermentation, selecting the alcohol content of more than 16% (v/v) to perform physical and chemical index, impurity alcohol and flavor measurement. PCA analysis (FIG. 2) was performed based on the flavor and fusel index of the strain, and the strain with low fusel and high ester yield was clearly distinguished.

3. Re-screening based on yellow wine fermentation of Saccharomyces cerevisiae isolates: and (3) repeatedly re-screening the strain with excellent fermentation performance obtained in the step (2) for 3 times, and measuring physicochemical indexes, flavor and fusel after fermentation. The physicochemical indexes of the Saccharomyces cerevisiae with excellent low-yield and high-yield fusel after the fermentation of the Saccharomyces cerevisiae are in accordance with national standards of yellow wine, the fusel and ester contents are shown in table 2, and the CYY-690 strain shows the characteristics of low-yield and high-yield fusel, but has relatively low alcohol content and is not suitable for the fermentation of yellow wine. Comprehensive analysis of Saccharomyces cerevisiae jiangnan1# (CYY-661) shows excellent fermentation characteristics of low-yield fusel and high-yield ester.

Table 1 re-screening to obtain physicochemical index of Saccharomyces cerevisiae with excellent low yield of fusel and high yield of ester at fermentation end

Table 2 obtaining good impurity alcohol and total ester content at the end of fermentation of high ester producing Saccharomyces cerevisiae with low impurity alcohol production by double sieves

4. And (3) verifying the stability of yellow wine fermentation based on saccharomyces cerevisiae isolates: and (3) repeating fermentation for 1 time for stability verification on the strain jiangnan1# (CYY-661) obtained by re-screening, and measuring physicochemical indexes, flavors and fusel after fermentation. Because of the quality difference between raw material batches, certain fluctuation exists in acid and ester production results of each batch, the difference of saccharomyces cerevisiae jiangnan1# and a control strain in terms of producing fusel and ester is focused, and in order to verify the low fusel production capability of the strain, the comparison is carried out by adopting the total fusel amount (the ratio of the total fusel amount to the ethanol content) of unit alcohol degree. The result shows that the total amount of the 85# hetero alcohol after the fermentation is 432.12 +/-22.44 mg/L and the total ester is 126.77 +/-7.84 mg/L; the saccharomyces cerevisiae jiangnan1# is 378.82 +/-7.35 mg/L, the total ester is 326.79 +/-66.2 mg/L, the main esters (ethyl acetate, ethyl lactate and the like) in the saccharomyces cerevisiae jiangnan1#1 strain fermented yellow wine are higher than the control strain 85#, wherein the contents of ethyl acetate, phenethyl acetate and isoamyl acetate are respectively increased by 1.6 times, 53.85% and 2.25 times, and the jiangnan1# has the brewing characteristics of low-yield and high-yield ester of fusel and is a low-yield and high-yield ester strain.

Table 3 physical and chemical indexes of Saccharomyces cerevisiae with excellent low yield of fusel and high yield of ester at the end of fermentation are obtained by re-screening

Table 4 Compound sifting to obtain the main impurity alcohol and ester content after the fermentation of the excellent low-impurity alcohol high-ester Saccharomyces cerevisiae

5. Identification of strains

Morphological characteristics of the strain, the ITS sequence of Saccharomyces cerevisiae jiangnan1# is shown as SEQ ID NO.1 through ITS sequencing. The NCBI database alignment shows that the strain is more than 99% homologous to Saccharomyces cerevisiae and can be identified as Saccharomyces cerevisiae (S.cerevisiae). The strain is preserved in the China center for type culture Collection (CCTCC M2021523) at the preservation address of Wuhan, wuhan and university in China on the 13 th year of 2021.

Example 2 preparation of low-heteroalol high-ester-yielding Saccharomyces cerevisiae jiangnan1# starter

(1) Taking quick brewing yeast as an example, quick brewing yeast containing saccharomyces cerevisiae jiangnan1# is prepared for brewing yellow wine.

The manufacturing method of the quick brewing master comprises the following steps: slant preparation of Saccharomyces cerevisiae glycerol tube deposited strain, inoculating Saccharomyces cerevisiae jiangnan1# strain selected in slant culture example 1 into rice saccharification liquid culture medium, shake culturing at 28deg.C+ -2deg.C for 24 hr to obtain primary seed liquid (bacterial liquid concentration 10) ⁶ -10 ⁸ CFU/mL), inoculating the primary seed liquid to a new rice saccharification liquid culture medium at a ratio of 5% -10%, and carrying out shaking culture for 36-48 h at a culture temperature of 28+/-2 ℃ to obtain a secondary seed liquid, wherein the obtained secondary seed liquid is used for measuring the viable count and the budding rate of the yeast, and the number of the yeast is more than or equal to 1 multiplied by 10 ⁷ CFU/mL, the germination rate is more than or equal to 30%, and the CFU/mL is used as a quick brewing master (starter) for brewing yellow wine.

(2) Preparation of starter for Saccharomyces cerevisiae jiangnan1# for other types of fermented foods

And (3) using the instant brewing yeast containing the pure strain prepared in the step (1) for fermentation production of food according to different fermentation food requirements to prepare a fermented alcoholic beverage containing the saccharomyces cerevisiae, wherein the culture medium can select various culture mediums meeting the food fermentation production C and N nutrition requirements such as rice saccharification liquid, wort, sugar solution and the like to perform activation culture on the saccharomyces cerevisiae jiangnan1# to reach corresponding colony numbers meeting the requirements.

Example 3 application of low-impurity-alcohol-yield high-ester-yield Saccharomyces cerevisiae jiangnan1# in yellow wine industry

(1) Application in traditional yellow wine production

A fast brewing yeast is prepared by taking low-yield hybrid alcohol high-yield ester saccharomyces cerevisiae jiangnan1# and a factory production strain 85# as experimental strains and control strains respectively according to the method of the embodiment 2 (1), and the fast brewing yeast is used for mechanical fermentation according to the method of the embodiment 1 step (2), and physical and chemical properties such as alcohol degree, hybrid alcohol and flavor, urea content, carbamate, amino acid, organic acid and the like are measured after fermentation is finished.

The alcohol content in the yellow wine obtained after the fermentation of the saccharomyces cerevisiae jiangnan1# is 16.5% (v/v), the impurity alcohol content is 398.68 +/-13.25 mg/L, the 2-phenethyl alcohol content is 79.43+/-5.21 mg/L, compared with the strain 85#, the 2-phenethyl alcohol content in the yellow wine obtained after the fermentation of the saccharomyces cerevisiae jiangnan1# is reduced by more than 20%, the ethyl ester (ethyl acetate, isoamyl acetate and phenethyl acetate) content is 196.5mg/L, the fermentation characteristics of the strain with low impurity alcohol and high yield are more remarkable, the fragrance and the taste of the yellow wine body obtained finally after the squeezing, blending and filtering are softer and more coordinated (figure 3), the drinking comfort is good, the drinking is not easy to top, the wine sobering is fast after the excessive drinking, the urea content is 18.60+/-0.53 mg/L, the urethane content is 67.91 +/-3.06 mug/L, the content is remarkably lower than that of the strain 85%, and other indexes such as amino acid, organic acid and the like are all in line with national standard GB/51-2011#.62-8.

(2) Application of Saccharomyces cerevisiae jiangnan1# in 20 ton mechanized millet yellow rice wine

Preparing quick brewing yeast according to the method of the step (1) of the embodiment 1, and brewing traditional yellow wine according to the method of the step (2) of the embodiment 1, wherein the inoculating liquid of the step S2 is replaced by saccharomyces cerevisiae jiangnan1# bacterial liquid or Angel brewing high activity dry yeast liquid respectively; and S5, carrying out plate and frame squeezing and diatomite filtration on the fermented mash, and clarifying the obtained filtrate to obtain the sake, and decocting the sake to obtain the yellow wine (without adding caramel color).

The mechanized millet yellow wine is prepared from the following raw materials:

experimental group: millet: 5500kg; water: 6600kg (L); raw wheat starter: 550kg-82.5kg; cooked wheat starter; 99kg; quick brewing mother: 62.7L;

control group: 5500kg of millet; 6600kg (L) of water; 11kg of Angel brewing high-activity dry yeast, 27.5kg of Angel brewing yeast, and 150L of sugar water containing 20g/L glucose with the water temperature of 30-35 ℃ are used for activating Angel brewing high-activity dry yeast and Angel brewing yeast for 20-30 min in advance, and are used as an activating fermentation agent, wherein the number of yeasts in the activating fermentation agent is more than 10 ⁸ CFU/mL。

The final yellow wine alcohol degree obtained by fermenting the strain jiannnan 1# after the fermentation is finished is 19.6% (v/v), the content of the unit alcohol content fusel and the 2-phenethyl alcohol is 20.5mg/L and 4.34mg/L respectively, the total amount of esters is 229.15mg/L, and the content of the ethyl carbamate is 82.25+/-1.26 mug/L; the control group fermented yellow wine is 19.4% (v/v), the content of unit alcohol and 2-phenethyl alcohol are 29.7mg/L and 12.0mg/L respectively, the total amount of esters is 173.33mg/L, the unit alcohol content is reduced by 30.98%, the unit alcohol content is reduced by 63.8% by 2-phenethyl alcohol, the effect of reducing alcohol is remarkable, the total amount of esters is increased by 32.2%, and other indexes all meet national standard GB/T13662-2018 of yellow wine.

(3) Application of saccharomyces cerevisiae jiangnan1# in millet yellow rice wine

The millet wine fermentation raw materials selected in the embodiment are as follows:

millet (benchmark): 500g; water: 500g (mL) -625g (mL); raw wheat starter: 50g-75g; cooked wheat starter: 9g; saccharomyces cerevisiae jiangnan1# fast brewing yeast prepared in example 2: 57mL; when the control group uses Angel brewing high activity dry yeast and Angel brewing yeast, the active starter is used to replace raw wheat yeast, cooked wheat yeast and quick brewing yeast.

Experimental group: the method of example 2, step (1), was followed using a low-fusogenic high-producing ester Saccharomyces cerevisiae jiangnan1# as a master yeast for rapid brewing.

Control group 1: a quick-acting Saccharomyces cerevisiae was produced by using a plant-produced Saccharomyces cerevisiae strain 85# as a pure yeast in the same manner as in step (1) of example 2.

Control group 2: 1g of Angel brewing high activity dry yeast and 2.5g of Angel brewing yeast are used for preparing activated fermentation according to the method of example 3 (2)The agent is prepared by activating Angel brewing high activity dry yeast and Angel brewing yeast with sugar water containing 20mg/mL glucose at 30-35deg.C for 20-30 min, and is used as activating ferment with yeast number > 10 ⁸ CFU/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 millet in water 2-3 times of the weight of millet raw rice at normal temperature for 1-3 days to obtain wet rice with acidity greater than 3mg/L, draining water to obtain wet rice, steaming the wet rice in a rice steaming cabinet at 121deg.C for 30-50 min until the rice is cooked without penetration, without clamping, and with rice yield of 140-160%.

b) Cooling the rice of the experimental group and the control group treated in the step a) to 25-28 ℃ according to the raw material proportion of millet yellow wine fermentation, performing primary fermentation, wherein the primary fermentation temperature is 20-35 ℃, fermenting for 3-7 days, entering secondary fermentation, performing secondary fermentation for 10-15 ℃, and performing secondary fermentation for 15-20 days.

c) The fermented mash is subjected to plate and frame squeezing and diatomite filtration in factory production, the obtained filtrate is clarified to obtain sake, the sake is prepared into yellow wine after the sake is decocted, and medlar, red date, honey, astragalus mongholicus, mountain ginseng, preserved plums and the like can be optionally added for blending after ageing. The experimental example is a laboratory scale small system simulated fermentation, which is carried out by adopting a gauze filtration and centrifugation mode.

After the fermentation is finished, the alcohol content of the final millet fermented mash obtained by fermenting the control group and the experimental group strain jiannnan 1# is 10% (v/v) -15% (v/v), the alcohol content of the strain jiannnan 1# is lower than 400mg/L, the alcohol content of the unit alcohol and the 2-phenethyl alcohol are reduced by more than 15% compared with the control group 1 and the control group 2, the total ester content is 80mg/L-160mg/L, the total ester content is increased by more than 20% compared with the control group 1 and the control group 2, the urethane content is less than 100 mu g/L, other indexes meet the national standard of yellow wine, the millet yellow wine obtained by fermenting the strain jiannnan 1# is transparent, bright in color and luster and fresh in taste, and the final millet yellow wine obtained by fermenting the strain jiannnan 1# has fruit fragrance and ester fragrance, and can meet the requirements of consumers on low alcohol content, high ester content and nutrition and health.

(4) Application of saccharomyces cerevisiae jiangnan1# in reduction of red yeast rice wine

The red yeast rice wine production and application are carried out according to the method of the step (2) of the example 1, wherein the red yeast rice is used for replacing wheat yeast in the raw materials of the step S3, the fermentation of the step S5 is carried out for 15-20 days, the fermented mash is subjected to plate and frame squeezing and diatomite filtration, and the obtained filtrate is clarified to obtain the clear wine, and the clear wine is the red yeast rice wine (without adding caramel color).

The method comprises the steps of preparing quick brewing yeast by taking low-impurity-alcohol-yield high-ester brewing yeast jiannnan 1# as an experimental strain in the manner of example 2, preparing an activated starter by taking Angel brewing high-activity dry yeast and Angel brewing yeast as a control group to replace red yeast and quick brewing yeast, respectively fermenting the red yeast yellow wine according to the raw material ratio, and measuring physicochemical properties such as alcohol degree, impurity alcohol and flavor, urea content, carbamic acid, amino acid, organic acid and the like after fermentation is finished.

The red rice yellow wine obtained by fermenting the control group and the experimental group after the fermentation is finished has the alcohol content of 15% (v/v) -18% (v/v), the red rice yellow wine body obtained by fermenting is brown or dark brown, the taste is fresh and clear, no peculiar smell exists, the red rice yellow wine obtained by fermenting the strain jiannnan 1# has stronger fruit fragrance and ester fragrance, and compared with the control group, the red rice yellow wine obtained by fermenting the control group and the experimental group has the alcohol content of 30.6mg/L and the 2-phenethyl alcohol content of 13.5mg/L respectively, the total amount of esters is 165.28mg/L, the alcohol content of the unit is reduced by 32.12%, the alcohol content of the unit is reduced by 55.20%, the alcohol reducing effect is obvious, the total amount of esters is increased by 35.12%, and other indexes meet national standards of yellow wine.

Example 4 application of low-impurity-alcohol-yield high-ester-yield Saccharomyces cerevisiae jiangnan1# in cooking wine

The yellow wine is obtained by first fermenting in the manner of the embodiment 3 (1), wherein the difference is that the rice water ratio is 1:1, the adding proportion of other raw materials is unchanged, 10% of edible salt is added into a part of yellow wine sample, edible water, spice and caramel color can be added according to the product requirement, the low-yield and high-yield ester saccharomyces cerevisiae (S.cerevisiae) strain jiangnan1# is obtained after sterilization treatment, the cooking wine prepared by taking yellow wine as the main raw material has the alcoholic strength of 10% (v/v) -15% (v/v), the amino nitrogen content is higher than 0.5g/L, the ethyl carbamate content is 80.5 mu g/L, the brewed cooking wine has high ester content, rich amino acid, better flavor and improved safety, and the product accords with SB/T10416-2007 seasoning cooking wine.

Example 5 application of low-impurity-alcohol-yield high-ester-yield Saccharomyces cerevisiae jiangnan1# in rice wine

Taking rice as a main raw material, washing with clear water, soaking for about 4 hours at normal temperature until water is fully absorbed, and kneading the rice with hands after soaking until the rice is fragile and has no hard rice core; draining the soaked rice grains, and cooking for about 30min under normal pressure until no white core exists in the rice; steaming rice, cooling to 25-28deg.C with cold boiled water, adding sweet wine yeast nest with mass of 0.4-0.8% of rice, saccharifying at 28deg.C+ -2deg.C for 36-42 h, inoculating 5-10% of low-impurity alcohol high-ester Saccharomyces cerevisiae jiangnan1# prepared according to example 2, stirring, adding drinking water with mass of 1-1.5 times of raw material rice, fermenting at 26-34 deg.C for 3-5 days, reducing temperature to 10-15deg.C, fermenting for 10-15 days, squeezing, and filtering to obtain rice wine. The rice wine obtained by fermentation is rich in rice aroma, has low higher alcohol content, has the impurity alcohol content of less than 20mg/L per unit alcohol degree, and is not easy to drunk after drinking.

Example 6 application of low-impurity-alcohol-yield high-ester-yield Saccharomyces cerevisiae jiangnan1# in sweet rice brewing

Taking rice as a main raw material, washing with clear water, soaking for about 4 hours at normal temperature until water is fully absorbed, and kneading the rice with hands after soaking until the rice is fragile and has no hard rice core; draining the soaked rice grains, and cooking for about 30min under normal pressure until no white core exists in the rice; pouring cold boiled water into rice to 25-28 ℃ after the rice is steamed, transferring the rice to a fermentation tank, adding Angel sweet distiller's yeast with the mass of 0.4-0.8% of that of raw rice, inoculating low-yield hybrid alcohol high-yield ester Saccharomyces cerevisiae jiangnan No. 1 quick Saccharomyces cerevisiae prepared in example 2 with the mass of 0.5-1.5%, stirring uniformly, and digging a groove in the center of the rice to ensure a certain dissolved oxygen amount; adding 1-1.5 times of drinking water, fermenting at 26-34 deg.C for 36-72 hr, and providing wine fragrance. The brewing accuracy of the obtained sweet rice is 2% -4% (v/v), the aroma is coordinated, the sweet rice is rich, the texture is uniform, and the taste is sour and sweet and palatable.

Example 7 application of low-impurity-alcohol-yield high-ester-yield Saccharomyces cerevisiae jiangnan1# in vinegar

Acetic fermentation was carried out using yellow wine obtained in the method described in (1) of example 3 as a raw material.

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

Example 8 application of low-impurity-alcohol-yield high-ester-yield Saccharomyces cerevisiae jiangnan1# in white spirit

Taking sorghum as a raw material, carrying out 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 husk, 10% -15% of yeast and 6% -10% of bran, inoculating 1% of low-yield and high-yield ester saccharomyces cerevisiae strain jiangnan1# pure strain prepared in the embodiment 2, carrying out first round closed fermentation for 30 days, and then distilling. Adding medium-temperature Daqu 10% -15% by secondary fermentation, continuously adding 1% of jiangnan1# quick brewing yeast or jiangnan1# -containing bacterial liquid prepared in the embodiment 2, continuously fermenting for 15 days, distilling, mixing 2 rounds of distilled white spirit to obtain white spirit with the alcoholic strength of 60% (v/v), wherein the content of acetate in the white spirit is increased, mainly ethyl acetate, isoamyl acetate and phenethyl acetate, the total amount is increased by more than 65%, the content of fusel is reduced by more than 25%, the content of carbamate is reduced by more than 60%, and the safety is improved.

Example 9 application of low-impurity-alcohol-yield high-ester-yield Saccharomyces cerevisiae jiangnan1# in wine

Taking 100kg Cabernet Sauvignon grape with complete fruit grain and good maturity as raw material, removing peduncles, crushing, packaging in 150L fermentation tank, adding 20mg/L pectase (enzyme activity 20000U/g) and 50mg/L SO ₂ Mixing, and soaking at 4deg.C for about 24 hr. Then respectively and uniformly mixing the components and inoculating the components to the final concentration of 1 multiplied by 10 when the temperature is increased to 20 DEG C ⁶ CFU/mL of the manufacturer of LAFFORT, franceSaccharomyces cerevisiae F15, and a final concentration of 1X 10 ⁶ CFU/mL of the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# pure brewing yeast prepared in the example 2 is pumped and circulated and mixed uniformly to perform mixed fermentation; controlling the fermentation temperature to 25-27 ℃, sampling and monitoring the specific gravity and the temperature of the fermentation liquid after 2 times of beating circulation every day in the morning and evening. When the specific gravity no longer decreases, it is regarded as the end of alcoholic fermentation, and the storage is sealed at 4℃during which several blowdown is carried out to separate the precipitate. There are 2 parallel fermentors per group. The alcohol content of the wine obtained by fermenting the saccharomyces cerevisiae added with low-yield fusel and high-yield ester is 12.8% (v/v) to 14% (v/v), and the total sugar (calculated by glucose)<5g/L, total acid (calculated by tartaric acid) of about 4.6g/L, volatile acid (calculated by acetic acid) of 0.15-0.4g/L, higher alcohol content of 195.5mg/L-210mg/L, total esters of 40mg/L-48.5mg/L, reduction of hetero alcohol by more than 25%, and increase of esters by more than 30%. Sensory evaluation results show that the color, the aroma and the taste of the wine fermented by the strain jiannan1 # added with the low-impurity-alcohol-yield high-ester saccharomyces cerevisiae (S.cerevisiae) are all better than those of the wine fermented by the commercial saccharomyces cerevisiae, and the fruit aroma and the flower aroma are more intense and prominent. The strain jiangnan1# of Saccharomyces cerevisiae (S.cerevisiae) can meet the normal fermentation requirement by being compounded with other microorganisms, and the indexes meet the national standard GB/T15037-2006 of wine.

Example 10 application of low-impurity-alcohol high-ester-yield Saccharomyces cerevisiae jiangnan1# in fruit wine

Sequentially cleaning fructus Pruni Salicinae, removing core, draining, crushing, adding 2-3 times of fructose-glucose syrup solution to obtain 100L mixture, subpackaging in 150L fermenter, adding 20-40 mg/L pectase (enzyme activity 20000U/g) and 40-80 mg/L potassium metabisulfite, and inoculating 1x10 at 1% (v/v) -5% (v/v) ⁷ Performing primary fermentation for 10-15 days at 20-25 ℃ and pH 2.5-3.5 under the condition of CFU/mL concentration of low-impurity-alcohol high-ester-yield saccharomyces cerevisiae (S.cerevisiae) strain jiangnan1# culture expanding liquid, and performing post fermentation for 5-10 days at 5-8 ℃ to obtain the fermented plum wine. There are 2 parallel fermentors per group. The alcoholic strength of the fermented plum is 10.5% (v/v) to 12% (v/v), and the total sugar (calculated by glucose)<40g/L. Sensory evaluation results show that the strain jiangnan1# brew added with the low-yield fusel high-yield ester saccharomyces cerevisiae (S.cerevisiae)The manufactured plum wine has harmonious wine body, soft taste and better quality. Other indexes of the fermented plum wine accord with the specification of the general technical requirements of GB 2758-2012 and QB/T5476-2020 fruit wine.

Example 11 application of low-impurity-alcohol-yield high-ester-yield Saccharomyces cerevisiae jiangnan1# in fruit vinegar

Acetic fermentation was performed using acetic acid from the fruit wine obtained in example 10. The fruit vinegar brewing adopts a liquid fermentation process: inoculating 1% -3% acetic acid bacteria, fermenting at 34 deg.C for 16 days, spraying vinegar, clarifying, degassing, and sterilizing to obtain final product. The acidity of the vinegar in the obtained fruit vinegar is 3% -8% (g/100 mL), other physical and chemical indexes are normal, the national standard of apple vinegar beverage GB/T30884-2014 is satisfied, and the fruit vinegar has rich fruit vinegar taste, soft and clean taste and unique flavor.

Example 12 application of low-impurity-alcohol-yield high-ester-yield Saccharomyces cerevisiae jiangnan1# in beer

Barley malt (15 kg-20 kg), hops (50 g-70 g) and water (100L-120L) are used as main raw materials, specifically, a multi-step soaking sugar-out method is adopted, wort is used as a culture medium, the low-yield and high-yield ester saccharomyces cerevisiae jiangnan1# pure strain quick saccharomyces cerevisiae prepared according to the method of the embodiment 2 is added into the wort according to the addition amount of 0.2% -1%, the malt is crushed, saccharified, filtered, wort boiled, fermented and the like, the above fermentation yeast is adopted, the fermentation time is 2-3 days, the fermentation pressure is 0.15Mpa, the sugar degree per day is reduced when the sugar degree is reduced to below 5Brix, and the temperature is reduced to 5 ℃ for storage. The beer has 3% -5% (v/v) of alcohol content, total sugar (calculated by glucose) of <50g/L, prominent fragrance, no strong fragrance, malt fragrance and fruit fragrance, impurity alcohol content lower than 120mg/L, and normal other physical and chemical indexes.

Example 13 application of low-impurity-alcohol-yield high-ester-yield Saccharomyces cerevisiae jiangnan1# in cigarettes

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

Shake-culturing the low-impurity-alcohol high-ester-yield Saccharomyces cerevisiae jiangnan1# in YPD culture medium for 24h at the culture temperature of 28+/-2 DEG CThe concentration is 10 ⁶ CFU/mL-10 ⁸ Inoculating CFU/mL primary seed solution into fermentation medium containing pear juice, grape juice, flos Osmanthi Fragrantis or tobacco extract at a ratio of 5% -10%, and culturing for 48 hr to obtain Saccharomyces cerevisiae culture solution with concentration of 10 ⁸ CFU/mL-10 ⁹ CFU/mL is added spice for cigarettes, the obtained added spice for cigarettes is directly added into a tobacco leaf group according to the adding amount of 1% -5% of the mass of the tobacco leaf group, water is added to 10% -20%, then the tobacco leaf group is cultured for 4-8 hours at 25-37 ℃, finally moisture is balanced after baking, and then cigarettes are manufactured, or the added spice for cigarettes is dissolved in 75% alcohol to prepare spice liquid (0.1 g/mL-0.5 g/mL), and uniformly sprayed into the processed tobacco shred samples according to a proper proportion to manufacture cigarettes. The obtained cigarette has soft smoke, low irritation, light fruit fragrance, light flower fragrance, pleasant sweet fragrance and extract special fragrance, can improve the smoking quality of cigarettes, and is suitable for improving the special fragrance and quality of cigarettes and novel tobacco products.

Example 14 application of Saccharomyces cerevisiae jiangnan1# with low yield of fusel and high yield of ester in fermented nutritional ice cream

Mixing drinking water, squeezed juice, sugar, honey, etc. as main material in the ratio of 2-4 to 0.05-0.2 to 0.01-0.05, regulating sugar concentration to 10Brix-20Brix, regulating pH to 4-5 with food grade lactic acid, inoculating 5-10% concentration of malt juice to culture bacterial liquid at 10 ⁶ -10 ⁸ CFU/mL of low-yield and high-yield ester saccharomyces cerevisiae jiangnan1#, shaking and culturing for 24-48 hours in a constant temperature incubator at 30+/-2 ℃, centrifuging after fermentation, adding 30% -50% of whole milk powder, 1% -5% of thickening agent, 1% -5% of emulsifying agent, 1% -5% of puffing agent, 5% -10% of vegetable fat powder and other additives in proportion to replace drinking water by supernatant or yellow wine obtained in the embodiment 3 (1), mixing until the total dry matter is 20% -30%, pasteurizing, homogenizing, cooling and aging at 4 ℃ for about 1 hour, freezing and stirring the mixture, subpackaging to a mould, freezing the mould, demolding and the like to obtain the nutritional fermented ice cream. The obtained ice cream has light sweet fragrance and flower and fruit fragrance, and has soft taste and reduced effect while relieving summer heatLow ice cream irritation and sweet greasy feeling.

While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and 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 Jiangnan

Jiang Nada (Shaoxing) institute of industry technology

<120> a strain of Saccharomyces cerevisiae with low yield of fusel and high yield of ester and application thereof in fermented food

<130> BAA210949A

<160> 1

<170> PatentIn version 3.3

<210> 1

<211> 812

<212> DNA

<213> Saccharomyces cerevisiae

<400> 1

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agagcgtcta ggcgaacaat gttcttaaag tttgacctca aatcaggtag gagtacccgc 780

tgaacttaag catatcaata agcggaggaa tt 812

Claims

1. Saccharomyces cerevisiaeSaccharomyces cerevisiae) jiangnan1# was deposited in China Center for Type Culture Collection (CCTCC) at 13 th year 2021, with a deposit number of CCTCC NO: M2021523.

2. A composition comprising saccharomyces cerevisiae jiannan1# according to claim 1, wherein said composition is a microbial agent.

3. A composition comprising saccharomyces cerevisiae jiangnan1# according to claim 1, wherein said composition is a wine drug.

4. A composition comprising saccharomyces cerevisiae jiangnan1# according to claim 1, wherein said composition is wheat starter.

5. A microbial preparation comprising Saccharomyces cerevisiae jiangnan1# according to claim 1, or comprising living cells of Saccharomyces cerevisiae jiangnan1# and other microorganisms, or comprising dried cells obtained by freeze-drying Saccharomyces cerevisiae jiangnan1# and other microorganisms.

6. The microbial preparation according to claim 5, wherein the number of cells of Saccharomyces cerevisiae jiangnan1# in the microbial preparation is 1×10 or more ⁷ CFU/mL, the budding rate is more than or equal to 30%.

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

8. The use according to claim 7, wherein the fermented product is a fermented tobacco leaf or a fermented ice cream.

9. The use according to claim 7, wherein the fermented product is a fermented food product.

10. The use according to claim 7, wherein the fermented product is a fermented beverage.

11. The use according to claim 10, wherein the fermented beverage is a fermented alcoholic beverage or a fermented vinegar.

12. The use according to claim 11, wherein the fermented alcoholic beverage is: yellow wine, cooking wine, rice wine, fruit wine, beer or white wine.

13. The use according to claim 11, wherein the fermented alcoholic beverage is wine.

14. The use according to claim 11, wherein the fermented vinegar is fruit vinegar.

15. The use according to claim 11, wherein the fermented vinegar is table vinegar.