CN116286411A

CN116286411A - Brewing method for co-fermentation of non-saccharomyces cerevisiae and saccharomyces cerevisiae

Info

Publication number: CN116286411A
Application number: CN202211475786.3A
Authority: CN
Inventors: 毛健; 李信; 刘双平; 朱胜虎; 赵禹宗; 崔鹏景
Original assignee: Industrial Technology Research Institute Of Jiangnan University Shaoxing; Jiangsu Hengshun Vinegar Industry Co ltd; Jiangnan University
Current assignee: Industrial Technology Research Institute Of Jiangnan University Shaoxing; Jiangsu Hengshun Vinegar Industry Co ltd; Jiangnan University
Priority date: 2022-11-23
Filing date: 2022-11-23
Publication date: 2023-06-23

Abstract

The invention discloses a brewing method for co-fermentation of non-saccharomyces cerevisiae and saccharomyces cerevisiae, belonging to the field of fermentation engineering and biotechnology. The invention provides candida tropicalis CS8 and the strain is used for co-fermentation with saccharomyces cerevisiae. The alcohol content, the impurity alcohol content and the amino acid content of the yellow wine prepared by co-fermenting the candida tropicalis CS8 and the saccharomyces cerevisiae are not obviously different from those of the yellow wine prepared by the common fermentation method, the organic acid content is reduced, the content of ethyl ester substances is improved, the produced yellow wine alcohol ester is coordinated, the bitter taste is reduced, the fragrance and the taste are more balanced, the content of guaiacol in vinyl is obviously increased (P is less than 0.001), the fragrance of the herbal medicine is prominent, and the flavor and the sensory diversity of the yellow wine are increased.

Description

Brewing method for co-fermentation of non-saccharomyces cerevisiae and saccharomyces cerevisiae

Technical Field

The invention relates to a brewing method for co-fermenting non-saccharomyces cerevisiae and saccharomyces cerevisiae, belonging to the field of fermentation engineering and biotechnology.

Background

The traditional yellow wine brewing process is complex, and the wheat starter and the yeast are used as the ferment, so that a unique double-side fermentation process of saccharification and fermentation is provided, and the whole fermentation process is multi-strain and open fermentation. The process is characterized in that the process adopts single-side fermentation for producing wine and beer of three ancient wines in the world, and the core of the process difference is that the yellow wine fermentation synchronously utilizes different microorganisms to synchronously decompose raw materials and form flavors, so that the efficient labor division cooperation of complex community metabolism is realized. The starter (wheat starter and wine medicine) used in traditional yellow wine production provides rich brewing microorganisms including bacteria, fungi and yeast, and the modern yellow wine production uses pure culture yeast instead of wine medicine. Compared with the two modes, the traditional yellow wine has relatively good quality, which shows that other microorganisms besides the saccharomyces cerevisiae have great contribution to the flavor quality formation of the yellow wine. Many studies analyzed the microbial diversity of yellow wine fermenters and their effect on the fermentation process using High Throughput Sequencing (HTS) technology (bacterial 16S rRNA gene, fungal ITS2 and metagenome), and a large number of saccharomyces cerevisiae microorganisms in addition to saccharomyces cerevisiae were also screened based on culture methods. How to utilize and exert the value of the microorganisms in the mechanized yellow wine production process is important for improving the quality of the yellow wine.

The alcohol metabolism speed is high in the main fermentation stage of mechanized yellow wine production, and most strains which have positive contribution to fermentation cannot play a role due to the problem of alcohol tolerance. non-Saccharomyces cerevisiae is an important microorganism in brewing yellow wine, and can affect the quality of yellow wine (ethanol production, volatile flavor compounds, organic acids, amino acids, etc.). The non-saccharomyces cerevisiae can secrete extracellular enzymes such as protease, pectase, glucosidase, lipase hydrolase, cellulase and the like, and acts on related substrates in the raw materials to generate substances such as alcohols, esters, acids, terpenes and the like, thereby influencing the quality and flavor of alcoholic beverages. A large number of documents prove that the non-saccharomyces cerevisiae is rich in the yellow wine fermentation process, but the non-saccharomyces cerevisiae cannot play more positive roles due to tolerance problems such as ethanol and the like.

Research shows that the reasonable use of non-saccharomyces cerevisiae can effectively improve the quality of alcoholic beverages, the created more diversified styles of alcoholic beverages are favored by consumers and the brewing industry, the non-saccharomyces cerevisiae is widely applied to both the wine industry and the beer industry, different types of non-saccharomyces cerevisiae can possibly contribute to the quality of alcoholic beverages, the application of non-saccharomyces cerevisiae in alcoholic beverages is still a research hotspot at present, but the research of non-saccharomyces cerevisiae in yellow wine is less due to the complexity and uniqueness of the yellow wine production process, and the non-saccharomyces cerevisiae is basically blank. Therefore, the invention establishes a non-saccharomyces cerevisiae and saccharomyces cerevisiae co-fermentation yellow wine brewing method aiming at the current yellow wine production process, evaluates different types of non-saccharomyces cerevisiae, carries out yellow wine fermentation on the excellent strain obtained by breeding, and has important significance for improving the flavor diversity and quality of modern yellow wine by establishing and applying the non-saccharomyces cerevisiae and saccharomyces cerevisiae co-fermentation yellow wine brewing method.

Disclosure of Invention

In order to solve the problems, no brewing process using non-saccharomyces cerevisiae and saccharomyces cerevisiae co-fermentation is available in the traditional brewing food production at present. The invention provides a non-saccharomyces cerevisiae and saccharomyces cerevisiae co-fermentation brewing method.

The invention provides candida tropicalis CS8 which is a wild non-saccharomyces cerevisiae separated from a water sample rich in citric acid in Yixing citric acid factory, is classified and named as candida tropicalis (Candida tropicalis) CS8, and is preserved in China Center for Type Culture Collection (CCTCC) No. M20221651 in the 10 month 25 of 2022, and the preservation address is China, wuhan and university of Wuhan.

The invention also provides a starter containing the candida tropicalis CS 8.

In one embodiment, the starter comprises candida tropicalis and saccharomyces cerevisiae.

In one embodiment, the Saccharomyces cerevisiae is Saccharomyces cerevisiae jiangnan1#, accession number CCTCC NO: M2021523, published in CN 113621528A.

In one embodiment, the method of preparing the starter is:

(1) Mixing raw rice and water at a mass ratio of 1:4, adding liquefying enzyme, saccharifying enzyme and raw wheat starter, saccharifying at 60deg.C for 4-5 hr until sugar degree is greater than 13 ° Brix, filtering, and sterilizing at 115deg.C for 20min to obtain rice saccharifying liquid culture medium;

(2) Inoculating Saccharomyces cerevisiae jiangnan1# or candida tropicalis CS8 into the rice saccharification liquid culture medium prepared in the step (1), and performing shake cultivation for 36 hours at 28 ℃ to obtain a Saccharomyces cerevisiae jiangnan1# or candida tropicalis CS8 culture solution.

The invention also provides a method for co-fermenting the candida tropicalis CS8 and the saccharomyces cerevisiae jiangnan1#.

In one embodiment, the method is to combine the saccharomyces cerevisiae jiangnan1# with the candida tropicalis CS8 at 1: the proportion of (1-1000) participates in fermentation.

In one embodiment, the method involves fermenting the Saccharomyces cerevisiae jiangnan1# with the Candida tropicalis CS8 in a ratio of 1:1, or 1:10, or 1:100.

In one embodiment, the amount of candida tropicalis CS8 in the fermentation system is not less than 1×10 ⁶ CFU/mL。

In one embodiment, the Saccharomyces cerevisiae jiangnan1# is present in an amount of 1X 10 or more in the fermentation system ⁶ CFU/mL。

In one embodiment, the method comprises:

(1) Adding a starter of Saccharomyces cerevisiae jiangnan1# and candida tropicalis CS8 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 primary fermentation;

(2) Lowering the temperature of the fermentation tank to 10-15 ℃, and standing for 15-20 days for post fermentation;

(3) Squeezing the fermented mash obtained in the step (2) through a plate frame, filtering with diatomite (the adding proportion of diatomite is 4% -6%, and the pressure is 0.3-0.5 MPa), clarifying the obtained filtrate to obtain sake, blending the sake, and decocting the sake according to national standard of yellow wine with 1-3 per mill of caramel color to obtain yellow wine.

The invention also provides the candida tropicalis CS8, or the leaven, or the application of the method in the aspect of producing fermented seasonings.

In one embodiment, the fermented flavoring includes, but is not limited to, yellow wine, cooking wine, or vinegar.

In one embodiment, the yellow wine is prepared by using Saccharomyces cerevisiae jiangnan1# and non-Saccharomyces cerevisiae CS8 as quick brewing yeast according to the following formula 1:1 or 1:10, and adding 5% -15% of the total additive amount into the steamed or gelatinized raw materials, fermenting, squeezing, decocting, aging, filtering, sterilizing and packaging.

In one embodiment, the cooking wine is prepared by fermenting the Saccharomyces cerevisiae jiangnan1# and the non-Saccharomyces cerevisiae CS8 serving as quick brewing yeast to obtain yellow wine, and then using the yellow wine.

In one embodiment, the application is for brewing vinegar by first fermenting the Saccharomyces cerevisiae jiangnan1# and the non-Saccharomyces cerevisiae CS8 serving as quick brewing yeast to obtain yellow wine and then using the yellow wine as an acetic acid fermentation raw material.

The beneficial effects are that:

(1) The brewing method for co-fermenting non-saccharomyces cerevisiae and saccharomyces cerevisiae, which is established by the invention, can be used as an evaluation mode for rapidly breeding the non-saccharomyces cerevisiae suitable for brewing yellow wine, table vinegar and cooking wine.

(2) The excellent candida tropicalis (Candida tropicalis) CS8 obtained by breeding grows well in the pH range of 3-5 with the alcohol content being less than 12%vol, can exert the fermentation effect, has no obvious difference in alcohol content, impurity alcohol content and amino acid content when being used in yellow wine, compared with the single inoculation of saccharomyces cerevisiae jiangnan1#, the candida tropicalis CS8 and the saccharomyces cerevisiae jiangnan1#, reduces the organic acid content, and improves the ethyl ester substance content by 22.95%.

(3) Compared with yellow wine alcohol ester produced by inoculating Saccharomyces cerevisiae alone, the co-fermentation group has advantages of balanced fragrance and taste, remarkably increased content of vinylguaiacol from 9.16 to 92.64 μg/L (P < 0.001), prominent fragrance of herbal medicine, increased content of ethyl ester by 22.95%, and increased content of vinylguaiacol by more than 10 times.

Preservation of biological materials

Candida tropicalis (Candida tropicalis) CS8, which is classified and named as candida tropicalis (Candida tropicalis) CS8, is preserved in China Center for Type Culture Collection (CCTCC) No. M20221651 in the 10 th month of 2022, and has a preservation address of China, the university of Wuhan and Wuhan.

Drawings

FIG. 1 is a flow chart of a non-Saccharomyces cerevisiae application process.

FIG. 2 shows colonies of non-Saccharomyces cerevisiae CS8 on YPD plates.

FIG. 3 is a sensory evaluation of CS8 co-fermented with jiannan1# and jiannan1# fermented yellow wine alone.

FIG. 4 shows the amino acid content of CS8 co-fermented with jiannan1# and jiannan1# fermented yellow wine alone.

FIG. 5 shows the fusel content of candida tropicalis CS8 and Saccharomyces cerevisiae jiangnan1# fermented yellow wine.

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 the volatile flavor substance is detected by gas chromatography-mass spectrometry (GC-MS) with 2-octanol as an internal standard. The determination of the reducing sugar content adopts a DNS method. 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-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 to quantitatively determine the fusel content. The strain information used in the establishment of the method is shown in Table 1.

TABLE 1 Strain information

Specific embodiments of the present invention are described below with reference to the accompanying drawings. The experimental methods used in the implementation 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.

EXAMPLE 1 isolation and screening of candida tropicalis (Candida tropicalis) CS8

(1) Sample preparation and strain isolation

Collecting water sample rich in citric acid from Yixing citric acid plant, and performing gradient dilution with sterile water (10 ^-1 -10 ^-5 ) Taking 100 mu L of diluted sample, performing plate coating on a YPD plate, performing inversion stationary culture at 28 ℃ for 48 hours, selecting the dilution of monoclonal bacterial colony, selecting bacterial strain with colony morphology, color and appearance conforming to the physiological morphology of yeast, and performing repeated streaking for bacteriaAnd (5) purifying the strain, and numbering and preserving the obtained pure strain finally.

(2) Identification of strains

Morphological characteristics of the strain, the homology of the strain to candida tropicalis can be compared with 99% in NCBI database by ITS sequencing, and candida tropicalis can be identified. The strain is preserved in the China center for type culture Collection (CCTCC M20221651) at the 10 th and 28 th of 2022 and the preservation address is China, wuhan and university of Wuhan.

Example 2 preparation of non-Saccharomyces cerevisiae and Saccharomyces cerevisiae co-ferment System starter

(1) Liquid culture of yeast strains: a10. Mu.L inoculating loop is used for taking a loop of yeast S.cerevisiae jiangnan1# preserved by glycerol, after 3 division lines are formed on a YPD plate, a monoclonal colony is obtained, and the monoclonal colony is selected and cultured for 24 hours in 100mL YPD culture medium at 37 ℃ to obtain yeast S.cerevisiae jiangnan1# bacterial liquid.

(2) Soaking rice: weighing polished round-grained glutinous rice according to the requirement, adding tap water, mixing rice water, and soaking rice in water bath at 60deg.C for 20min, wherein the water exceeds 7cm above the liquid surface to fully absorb water.

(3) Steaming rice: boiling water in the steamer, filtering out the rice-soaking water after soaking rice, uniformly spreading wet rice on 2 layers of gauze, steaming for 20min, and spraying with hot water at 80 ℃.

(4) And (3) airing: the steamed rice is spread and cooled to a temperature below 60 ℃.

(5) Rice saccharification liquid culture medium: mixing raw rice and water (1:4 mass ratio), adding liquefying enzyme, saccharifying enzyme and raw wheat starter at a mass fraction of 2%of rice mass, 1%of saccharifying enzyme and 10% of raw wheat starter, controlling the temperature at 60 ℃ for water bath, saccharifying and liquefying for 4-5h, measuring the sugar degree by a refractometer until the sugar degree is greater than 13 DEG Brix, filtering by a filter bag after saccharification, sub-packaging, sterilizing at 115 ℃ for 20min, and cooling for later use.

(6) Preparation of Yeast culture solution: inoculating the yeast liquid-cultured in the step (1) into the rice saccharification liquid culture medium prepared in the step (5) at a ratio of 1% ^-1 -10 ^-7 100 mu L of 10 ^-5 、10 ^-6 And 10 ^-7 Coating the yeast culture solution on YPD plates, culturing at 28 ℃ for 36 hours, performing plate counting to determine the number of yeasts of the triangular flask yeast culture solution, and simultaneously taking 1.5mL of the yeast cultured by the liquid in the step (1) to store at low temperature (4 ℃) in an EP tube as seed liquid for subsequent inoculation;

(7) Preparing a fermentation agent: inoculating Saccharomyces cerevisiae jiangnan1# culture solution obtained in step (6) into rice saccharification liquid culture medium respectively at a ratio of 1%o, shake culturing at 28deg.C for 36 hr to obtain pure yeast starter containing only non Saccharomyces cerevisiae or S.cerevisiae jiangnan1% ⁸ CFU/mL, non-Saccharomyces cerevisiae concentration of 1×10 ⁷ CFU/mL～1×10 ⁸ CFU/mL。

Example 3 sequential inoculation Co-fermentation Process with non-Saccharomyces cerevisiae

non-Saccharomyces cerevisiae and S.cerevisiae jiangnan1# were inoculated sequentially for co-fermentation: determining Saccharomyces cerevisiae jiangnana1# starter (bacterial concentration 1×10) according to dilution plate count result ⁸ CFU/mL), candida tropicalis starter (bacterial concentration 1X 10) ⁸ CFU/mL) at 1:1 (25 mL+25 mL), respectively; 1:10 (5mL+45 mL); 1:100 (0.5mL+50 mL) was inoculated into a 1.5L fermentation system, with jiangnan1# (50 mL) single inoculation, and non-Saccharomyces cerevisiae (50 mL) single inoculation as a control.

In order to exert the potential value of yellow wine brewing applications of non-Saccharomyces cerevisiae, a yeast saccharification liquid culture medium (secondary seed liquid) prepared by different yeast strains was used as a starter for evaluating non-Saccharomyces cerevisiae according to the method of example 2. Firstly, a sequential inoculation mode is discussed, and candida tropicalis is inoculated firstly for fermentation according to the proportion of saccharomyces cerevisiae jiangnan1# and candida tropicalis CS8 (the volume ratio is 1:1;1:10 and 1:100), and then S.cerevisiae jiangnan1# is inoculated after 24 hours of inoculation.

The yellow wine starter was prepared in the same manner as in example 2, and the yellow wine fermentation was performed in the following manner:

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) The method comprises the following specific steps of:

s1, adding raw materials of rice and water into a sterilized fermentation container, adding 50mL of prepared leaven into a rice-water mixed fermentation system (1.5L), respectively inoculating candida tropicalis according to the proportion of saccharomyces cerevisiae jiangnan1# and candida tropicalis CS8 (the volume ratio is 1:1;1:10 and 1:100), fermenting, inoculating s.cerevisiae jiangnan1# after 24 hours of inoculation, adding wheat starter accounting for 12% -15% of the mass of raw materials of rice, completing material mixing at 25-28 ℃, and standing for 3-5 days at 28 ℃;

s2, reducing the temperature of the fermentation tank to 15 ℃, and standing for 15-20 days for post fermentation;

s3, squeezing the fermented mash obtained in the step S2 through a plate frame (4 times of feeding, the mash feeding 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, and adding 1-3 per mill of caramel color to obtain yellow wine according to the national standard of yellow wine.

The results show that the alcohol content after fermentation of candida tropicalis CS8 and then saccharomyces cerevisiae jiangnan1# is less than 14%vol, the total acid content is more than 8.0mg/L, and the saccharification and liquefaction of starch (residual sugar is more than 50.0 g/L) are normal. The reason for this phenomenon may be that the ethanol content produced by the yeast in the early stage of fermentation (24 hours before) is low, and the growth of other flora cannot be inhibited, while the acid concentration produced by the acid-producing bacteria is too high, which may cause rancidity in a sequential inoculation mode of inoculating the non-saccharomyces cerevisiae and then inoculating the s.cerevisiae jiangnan1#.

EXAMPLE 4 determination of the manner and ratio of inoculation of the co-fermentation System of candida tropicalis and Saccharomyces cerevisiae

The preparation method of the yellow wine starter is the same as that in the example 2, and yellow wine is producedThe fermentation was performed as in example 3, except that the starter was inoculated in a simultaneous inoculation: fermenting S.cerevisiae jiangnan1# starter (bacterial concentration 1×10) ⁸ CFU/mL), candida tropicalis starter (bacterial concentration 1X 10) ⁸ CFU/mL) are inoculated together according to the adding proportion of 1:1,1:10 and 1:100 respectively, and small-system yellow wine fermentation is carried out, and the specific steps are as follows:

b) The method comprises the following specific steps of:

s1, adding raw materials of rice and water into a sterilized fermentation container, adding 50mL of a fermentation agent (the volume ratio of bacterial liquid is 1:1,1:10 or 1:10) of Saccharomyces cerevisiae jiangnan1# and candida tropicalis CS8 into a 1.5L rice water mixed fermentation system, adding wheat starter accounting for 12% -15% of the mass of the raw materials of rice, completing material mixing at 25-28 ℃, standing at 20-35 ℃ and performing pre-fermentation for 3-5 days;

s2, reducing the temperature of the fermentation tank to 10-15 ℃, and standing for 15-20 days for post fermentation;

It can be found that co-fermentation groups with high S.cerevisiae jiangnan1# and non-Saccharomyces cerevisiae inoculation ratios (1:1 and 1:10) can mostly complete fermentation, different mixed fermentation ratios can have different effects on yellow wine fermentation, and non-Saccharomyces cerevisiae single inoculation fermentation can produce more flavor substances, but rancidity usually occurs, and yellow wine fermentation cannot be completed; under the condition that the inoculation ratio of the cerevisiae to the non-saccharomyces cerevisiae is small (1:100), the small quantity of the cerevisiae can cause too slow rise of the alcoholic strength in the earlier stage of main fermentation, can not inhibit the growth of mixed bacteria, has low alcoholic strength and high total acid after the fermentation is finished, and has the risk of fermentation stagnation or fermentation termination; when the adding ratio of the cerevisiae to the non-saccharomyces cerevisiae is 1:1 and 1:10, the candida tropicalis CS8 can be fermented normally with the saccharomyces cerevisiae, the physicochemical indexes meet the national standard of yellow wine (the alcoholicity is more than 14% vol, the total acid is 4-8g/L, the amino acid nitrogen content is more than 0.5 g/L), as shown in the table 2, compared with the contrast, the alcohol content of the yellow wine prepared by co-fermenting the candida tropicalis CS8 and the saccharomyces cerevisiae is not obviously different or higher than the contrast (P < 0.05), and other physicochemical indexes (total acid, ammonia nitrogen and reducing sugar) meet the national standard requirements. The candida tropicalis does not influence the fermentation completion under the condition of proper inoculation concentration (1:1 and 1:10), which shows that the strain has the possibility of co-fermentation with yellow wine. Wherein CS8 (J-Non-Sc 21) has no significant differences in alcoholicity, total acids and ammonia nitrogen compared to control inoculated with jiangnan1# alone at inoculation ratios of 1:1 and 1:10.

TABLE 2 physical and chemical indexes of co-fermentation of candida tropicalis CS8 and S.cerevisiae jiangnan1#

Note that: values are mean ± standard deviation of at least three independent assays, representing P<0.05， ^ns Representing no significant difference.

Example 5 application of candida tropicalis (Candida tropicalis) CS8 in yellow wine production

The mixed bacteria inoculation fermentation process of candida tropicalis CS8 and S.cerevisiae jiangnan1#: s.cerevisiae jiangnana1# was determined based on the results of the dilution plate count (bacterial concentration 1X 10) ⁸ CFU/mL), candida tropicalis CS8 (bacterial concentration 1X 10) ⁸ CFU/mL) is inoculated according to the ratio of the number to the ratio of 1:1; candida tropicalis CS8 and Saccharomyces cerevisiae jiangnan1# were inoculated alone as controls.

A medium was prepared as in example 2, and the conditions for yellow wine fermentation were the same as in example 4.

The results show that after 2 days of fermentation with candida tropicalis CS8 alone, rancidity (total acid > 10 g/L) occurs, so that no subsequent fermentation experiments were performed. Inoculation of S.cerevisiae jiangnan1# and non-Saccharomyces cerevisiae CS8 (1:1) can meet the fermentation requirement of yellow wine, no obvious difference in physical and chemical indexes exists, the total amount of 4 main fusel is no obvious difference, the volatile substances are 22.95% of ethyl ester content, the content of vinylguaiac is obviously increased, and the content of vinylguaiac is improved by more than 10 times. Inoculation s.cerevisiae jiangnan1# has no significant difference in amino acid content compared to non-saccharomyces cerevisiae CS8, and succinic, lactic and acetic acid content is significantly reduced compared to the control (P < 0.05), tartaric and citric acid content is higher than the control (P < 0.05). Sensory results show that the overall taste and overall aroma are lower than those of a control (P < 0.05), the bitter taste and mellow aroma are obviously reduced (P < 0.05), the smoke aroma, the burnt aroma, the fruit aroma and the honey aroma are obviously reduced (P < 0.05), the herbal aroma is obviously increased (P < 0.05), the produced yellow wine alcohol ester is coordinated, and the aroma and the taste are more balanced.

TABLE 3 physical and chemical indexes of non-Saccharomyces cerevisiae CS8 and S.cerevisiae jiangnan1# mixed fermentation

Note that: ^a the content of the representative total acids was calculated as lactic acid and the values were the mean ± standard deviation of at least three independent tests.

TABLE 4 content of volatile substances by fermentation of Candida tropicalis CS8 and S.cerevisiae jiangnan1#

Table 5 organic acids of CS8 and jiangnan1# co-fermented and jiangnan1# separately fermented yellow wine

Note that: values are mean ± standard deviation of at least three independent assays, representing significant differences (P < 0.05).

Example 7 use of non-Saccharomyces cerevisiae CS8 and Saccharomyces cerevisiae jiangnan1# co-fermentation in cooking wine

According to the method of example 5, yellow wine is obtained by fermentation, 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 needs of the product, the cooking wine prepared by taking yellow wine brewed by co-fermentation of non-Saccharomyces cerevisiae CS8 and Saccharomyces cerevisiae jiangnan1# as main raw materials 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 brewed cooking wine has high ester content, is rich in amino acid, has good flavor, has more coordinated aroma and taste components, and the product accords with SB/T10416-2007 seasoning wine.

Example 8 application of non-Saccharomyces cerevisiae CS8 and Saccharomyces cerevisiae jiangnan1# co-fermentation in Vinegar

Acetic fermentation was carried out using yellow wine obtained by the method of reference example 5 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 taste is soft and clean, and the flavor is unique.

Comparative example 1:

the specific embodiment is the same as in example 4, except that candida tropicalis CS8 is replaced by another candida tropicalis NL2 which is owned by the laboratory, and the result shows that the yellow wine prepared by co-fermentation has rancidity and cannot be normally fermented.

TABLE 6 comparison of physicochemical indicators of different Candida tropicalis and Jiangan # 1 co-fermentation

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.

Claims

1. A strain of candida tropicalis (Candida tropicalis) CS8 is preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of M20221651 at 10 and 25 of 2022.

2. A microbial preparation comprising the candida tropicalis CS8 of claim 1.

3. The microbial preparation according to claim 2, characterized by comprising said candida tropicalis CS8 and saccharomyces cerevisiae.

4. A microbial preparation according to claim 3, wherein the saccharomyces cerevisiae is saccharomyces cerevisiae jiangnan1#, with a preservation number of cctccc No. M2021523.

5. The microbial preparation according to any one of claims 2 to 4, wherein the concentration of cells in the microbial preparation is 1X 10 or more ⁷ CFU/mL。

6. Use of candida tropicalis CS8 according to claim 1 for co-fermentation with saccharomyces cerevisiae.

7. The use according to claim 6, wherein said s.cerevisiae and said candida tropicalis CS8 are present in a ratio of 1: the proportion of (1-1000) participates in fermentation.

8. A method for improving the flavor of yellow wine, which is characterized in that saccharomyces cerevisiae and candida tropicalis CS8 according to claim 1 are mixed according to the following ratio of 1: adding the mixture of (1-100) into a yellow wine fermentation system for fermentation.

9. Use of candida tropicalis CS8 according to claim 1, or a microbial preparation according to any one of claims 2 to 5, or a method according to claim 8 for the production of a fermented condiment.

10. Use according to claim 9, wherein the fermented flavouring comprises, but is not limited to, yellow wine, cooking wine or vinegar.