Composite microbial inoculant prepared by adding functional strains into vinegar koji microorganisms special for fruit vinegar and preparation method of composite microbial inoculant

Abstract

The invention provides a fruit vinegar special-purpose vinegar koji microorganism composite microbial inoculant added with a functional strain and a preparation method of the fruit vinegar special-purpose vinegar koji microorganism composite microbial inoculant. The composite microbial agent comprises vinegar koji powder, a protective agent and a composite functional strain microbial agent, and aims to improve the production efficiency and product quality of fruit vinegar. The preparation process comprises the following steps: mixing and crushing the cereals and the traditional Chinese medicines, inoculating the compound leavening agent for fermentation, and performing final freeze-drying treatment. By using a specific microorganism combination and accurately controlling fermentation conditions, the production efficiency of fruit vinegar can be improved, the stability of product quality is ensured, and meanwhile, the stability of the complex microbial inoculant is improved.

Description

Technical Field

[0001] The invention relates to the field of microorganisms, and in particular to a composite bacterial agent of vinegar koji microorganisms added with functional strains for fruit vinegar and a preparation method thereof. Background Art

[0002] The vinegar koji is a key factor in the production of fruit vinegar, which affects the flavor, quality and production efficiency of fruit vinegar. The vinegar koji is a composite strain composed of a variety of microorganisms, including yeast, acetic acid bacteria, etc., which jointly participate in the fermentation process of fruit vinegar. In the production of fruit vinegar, the role of vinegar koji is crucial because it not only affects the fermentation speed and efficiency, but also determines the taste and aroma of the final product.

[0003] Vinegar koji plays a key role in the vinegar brewing process, but due to its characteristics and production process, there are also some inevitable shortcomings. The types of microorganisms in traditional vinegar koji are complex and limited in number, and the fermentation process is often long, which will lead to a longer production cycle and affect production efficiency. This shortcoming is particularly obvious in industrial large-scale production. The fermentation process of vinegar koji is affected by a variety of environmental factors such as temperature and humidity. These conditions are difficult to control in actual production and can easily cause unstable product quality. At the same time, because fermentation involves the interaction of multiple microorganisms, the slightest carelessness may lead to fermentation failure. Traditional vinegar koji has fewer metabolites during the fermentation process, resulting in a relatively single flavor of the final product. Modern consumers have increasingly higher requirements for the flavor of condiments, and this single flavor is difficult to meet diverse needs. The production of vinegar koji requires specific technology and experience, especially in maintaining the activity and diversity of microorganisms. This is a big challenge for inexperienced producers. Vinegar koji is easily affected by environmental changes during storage. If the temperature and humidity are not properly controlled, it is easy to cause the vinegar koji to mold or fail, thus affecting the use effect. Summary of the invention

[0004] In view of the above reasons, the present invention provides a composite bacterial agent for fruit vinegar with added functional bacteria species of vinegar koji microorganisms and a preparation method thereof.

[0005] To achieve the above object, the present invention is implemented by the following technical scheme: a composite bacterial agent for adding functional bacteria to vinegar koji microorganisms for fruit vinegar and a preparation method thereof, comprising the following steps:

[0006] S1: Crush corn, wheat bran, rice husk and other cereals, and then sieve through a 100-mesh sieve to ensure fineness, then mix them evenly to obtain cereal powder, add water to the cereal powder, and then steam for 30 to 50 minutes to soften the cereals, and cool to room temperature to obtain cereal porridge; mix almonds, siler, costusroot, lotus leaves, lily and amomum, and then crush them, and sieve through a 100-mesh sieve to obtain Chinese medicine powder; mix the cereal porridge and the Chinese medicine powder evenly to obtain koji material.

[0007] S2: Inoculate the composite fermentation agent into the koji material prepared in S1, with the inoculation amount accounting for 2% to 5% of the weight of the koji material. Mix the composite fermentation agent and the koji material evenly, and then press them into rectangular koji embryos. Stack the koji embryos in the koji room, control the temperature of the koji room to be 28℃ to 32℃, and the relative humidity to be 60℃ to 70℃, and ferment for 24 to 30 days to obtain vinegar koji.

[0008] S3: Grind the vinegar koji prepared in S2 to a particle size of less than 2.0 mm to obtain vinegar koji powder, mix the vinegar koji powder with a protective agent and a composite functional bacterial strain inoculant in a certain proportion, and freeze-dry to obtain a composite bacterial strain of vinegar koji microorganisms added with functional bacterial strains for fruit vinegar.

[0009] The koji material is composed of the following raw materials in parts by mass: 60-70 parts of corn, 20-30 parts of wheat bran, 12-20 parts of rice husk, 100-100 parts of water, 1-3 parts of almonds, 1-2 parts of siler, 1-2 parts of costusroot, 0.4-0.9 parts of lotus leaves, 0.5-0.7 parts of lilies, and 0.3-0.5 parts of amomum.

[0010] The composite fermentation agent consists of acetobacter pasteurianus, saccharomyces cerevisiae and lactobacillus acidophilus, and the weight ratio of acetobacter pasteurianus, saccharomyces cerevisiae and lactobacillus acidophilus is 3:3:4.

[0011] The Acetobacter pasteurianus, Saccharomyces cerevisiae, and Lactobacillus acidophilus are expanded and diluted with sterile water to make the concentration of the bacteria or spores reach 1 to 3×10 8 Pieces / mL.

[0012] The mass ratio of the vinegar koji powder, the protective agent and the composite functional bacterial strain inoculant is 7:2:1.

[0013] The protective agent consists of beta-cyclodextrin, trehalose and glycerol, and the mass ratio of beta-cyclodextrin, trehalose and glycerol is 4:3:1.

[0014] The composite functional bacterial strain agent consists of Bacillus Velezii and Lactobacillus delbrueckii subspecies bulgaricus, and the mass ratio of Bacillus Velezii to Lactobacillus delbrueckii subspecies bulgaricus is 3:7.

[0015] The Velez subsp. velezii and Lactobacillus delbrueckii subsp. bulgaricus are expanded and then diluted with sterile water to make the concentration of the bacteria or spores reach 1 to 3×10 8 Pieces / mL.

[0016] The present invention provides a composite bacterial agent for adding functional bacteria to vinegar koji microorganisms for fruit vinegar and a preparation method thereof. It has the following beneficial effects:

[0017] 1. The composite bacterial agent prepared by the invention with the special vinegar koji microorganisms and functional bacteria added can improve the production efficiency of industrial large-scale production of fruit vinegar and reduce the production cost.

[0018] 2. The present invention accurately controls the inoculation amount of microorganisms and fermentation conditions, and the composite bacterial agent prepared by the present invention with added functional bacteria for the vinegar koji microorganisms for fruit vinegar is helpful to achieve the stability and consistency of the quality of the fruit vinegar product.

[0019] 3. By using microbial protective agents and specific preparation methods, the activity of microorganisms is effectively protected and the stability of the composite bacterial agent is improved. DETAILED DESCRIPTION

[0020] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0021] In the following embodiments, Saccharomyces cerevisiae may be Saccharomyces cerevisiae CICC 1001. Saccharomyces cerevisiae CICC 1001 is a strain with a collection number of CICC 1001 in China Industrial Microbiological Culture Collection Center (CICC) (address: Building 6, No. 24, Jiuxianqiao Middle Road, Chaoyang District, Beijing, zip code 100015).

[0022] Acetobacter pasteurianus in the following embodiments may be Acetobacter pasteurianus CI CC 20761. Acetobacter pasteurianus CI CC 20761 is a strain with a collection number of CI CC 20761 in China Industrial Microbiological Culture Collection Center (CICC) (address: Building 6, No. 24, Jiuxianqiao Middle Road, Chaoyang District, Beijing, zip code 100015).

[0023] The Lactobacillus acidophilus in the following embodiments may be Lactobacillus acidophilus CI CC 6074. Lactobacillus acidophilus CI CC 6074 is a strain with a collection number of CI CC 6074 from China Industrial Microbiological Culture Collection (CICC) (address: Building 6, No. 24, Jiuxianqiao Middle Road, Chaoyang District, Beijing, zip code 100015).

[0024] The Bacillus velezensis in the following embodiments may be Bacillus velezensis CI CC 22774. Bacillus velezensis CI CC 22774 is a strain with a collection number of CI CC 22774 in China Industrial Microbiological Culture Collection Center (CICC) (address: Building 6, No. 24, Jiuxianqiao Middle Road, Chaoyang District, Beijing, zip code 100015).

[0025] Lactobacillus delbrueckii subsp. bulgaricus in the following embodiment can be Lactobacillus delbrueckii subsp. bulgaricus (Lactobacillus delbrueckii subsp. bulgaricus) CI CC 6097. Lactobacillus delbrueckii subsp. bulgaricus (Lactobacillus delbrueckii subsp. bulgaricus) CI CC 6097 is the bacterial strain of China Industrial Microorganism Culture Collection (CICC) (address: Building 6, No. 24, Jiuxianqiao Middle Road, Chaoyang District, Beijing, zip code 100015) collection number CICC 6097. The specific selection of bacterial strain and the adjustment of consumption belong to the general knowledge category of those skilled in the art, and therefore, the present invention does not set special limiting conditions to these parameters. In addition, unless otherwise specified, in all embodiments of the present invention, described ratio is weight ratio.

[0026] An embodiment of the present invention provides a method for processing a fiber polymer spraying material, comprising the following steps:

[0027] Example 1

[0028] S1: 60 parts of corn, 20 parts of wheat bran, and 12 parts of rice husks are crushed and passed through a 100-mesh sieve to ensure fineness, and then mixed evenly to obtain grain powder, an equal amount of water is added to the grain powder, and then steamed for 30 minutes to soften the grains, and cooled to room temperature to obtain grain porridge, 1 part of almond, 1 part of siler, 1 part of costus root, 0.4 part of lotus leaf, 0.5 part of lily, and 0.3 part of amomum villosum are mixed and crushed, and passed through a 100-mesh sieve to obtain traditional Chinese medicine powder, and the grain porridge and the traditional Chinese medicine powder are evenly mixed to obtain koji material;

[0029] S2: inoculating the composite starter into the koji material prepared in S1, the inoculation amount accounting for 2% of the weight of the koji material, mixing the composite starter and the koji material evenly, and then pressing them into rectangular koji embryos, stacking the koji embryos in a koji room, controlling the temperature of the koji room to be 28°C, the relative humidity to be 60°C, and fermenting for 24 days to obtain vinegar koji;

[0030] S3: The vinegar koji prepared in S2 is crushed to a particle size of less than 2.0 mm to obtain vinegar koji powder, and the vinegar koji powder is evenly mixed with a protective agent and a composite functional bacterial agent in a mass ratio of 7:2:1, and freeze-dried to obtain a composite bacterial agent with added functional bacterial species of vinegar koji microorganisms for fruit vinegar.

[0031] Example 2

[0032] S1: 65 parts of corn, 25 parts of wheat bran, and 16 parts of rice husks are crushed and passed through a 100-mesh sieve to ensure fineness, and then mixed evenly to obtain grain powder, an equal amount of water is added to the grain powder, and then steamed for 40 minutes to soften the grains, and cooled to room temperature to obtain grain porridge, 2 parts of almonds, 1.5 parts of siler, 1.5 parts of costus root, 0.7 parts of lotus leaves, 0.6 parts of lily, and 0.4 parts of amomum villosum are mixed and crushed, and passed through a 100-mesh sieve to obtain traditional Chinese medicine powder, and the grain porridge and the traditional Chinese medicine powder are evenly mixed to obtain koji material;

[0033] S2: inoculating the composite starter into the koji prepared in S1, the inoculation amount accounting for 3.5% of the weight of the koji, mixing the composite starter and the koji evenly, and then pressing them into rectangular koji embryos, stacking the koji embryos in a koji room, controlling the temperature of the koji room at 30°C and the relative humidity at 65°C, fermenting for 27 days, and obtaining vinegar koji;

[0034] S3: The vinegar koji prepared in S2 is crushed to a particle size of less than 2.0 mm to obtain vinegar koji powder, and the vinegar koji powder is evenly mixed with a protective agent and a composite functional bacterial agent in a mass ratio of 7:2:1, and freeze-dried to obtain a composite bacterial agent with added functional bacterial species of vinegar koji microorganisms for fruit vinegar.

[0035] Example 3

[0036] S1: 70 parts of corn, 30 parts of wheat bran, and 20 parts of rice husks are crushed and passed through a 100-mesh sieve to ensure fineness, and then mixed evenly to obtain grain powder, an equal amount of water is added to the grain powder, and then steamed for 50 minutes to soften the grains, and cooled to room temperature to obtain grain porridge, 3 parts of almonds, 2 parts of siler, 2 parts of costus root, 0.9 parts of lotus leaves, 0.7 parts of lily, and 0.5 parts of amomum villosum are mixed and crushed, and passed through a 100-mesh sieve to obtain traditional Chinese medicine powder, and the grain porridge and the traditional Chinese medicine powder are evenly mixed to obtain koji material;

[0037] S2: inoculating the composite starter into the koji material prepared in S1, the inoculation amount accounting for 5% of the weight of the koji material, mixing the composite starter and the koji material evenly, and then pressing them into rectangular koji embryos, stacking the koji embryos in a koji room, controlling the temperature of the koji room to be 32°C and the relative humidity to be 70°C, and fermenting for 30 days to obtain vinegar koji;

[0038] S3: The vinegar koji prepared in S2 is crushed to a particle size of less than 2.0 mm to obtain vinegar koji powder, and the vinegar koji powder is evenly mixed with a protective agent and a composite functional bacterial agent in a mass ratio of 7:2:1, and freeze-dried to obtain a composite bacterial agent with added functional bacterial species of vinegar koji microorganisms for fruit vinegar.

[0039] Example 4

[0040] S1: 60 parts of corn, 20 parts of wheat bran, and 12 parts of rice husks are crushed and passed through a 100-mesh sieve to ensure fineness, and then mixed evenly to obtain grain powder, an equal amount of water is added to the grain powder, and then steamed for 30 minutes to soften the grains, and cooled to room temperature to obtain grain porridge, 1 part of almond, 1 part of siler, 1 part of costus root, 0.4 part of lotus leaf, 0.5 part of lily, and 0.3 part of amomum villosum are mixed and crushed, and passed through a 100-mesh sieve to obtain traditional Chinese medicine powder, and the grain porridge and the traditional Chinese medicine powder are evenly mixed to obtain koji material;

[0041] S2: inoculating the composite starter into the koji material prepared in S1, the inoculation amount accounting for 2% of the weight of the koji material, mixing the composite starter and the koji material evenly, and then pressing them into rectangular koji embryos, stacking the koji embryos in a koji room, controlling the temperature of the koji room to be 28°C, the relative humidity to be 60°C, and fermenting for 24 days to obtain vinegar koji;

[0042] S3: The vinegar koji prepared in S2 is crushed to a particle size of less than 2.0 mm to obtain vinegar koji powder, and the vinegar koji powder is evenly mixed with a protective agent and a composite functional bacterial agent in a mass ratio of 7:2:1, and freeze-dried to obtain a composite bacterial agent with added functional bacterial species of vinegar koji microorganisms for fruit vinegar.

[0043] Example 5

[0044] S1: 70 parts of corn, 30 parts of wheat bran, and 20 parts of rice husks are crushed and passed through a 100-mesh sieve to ensure fineness, and then mixed evenly to obtain grain powder, an equal amount of water is added to the grain powder, and then steamed for 50 minutes to soften the grains, and cooled to room temperature to obtain grain porridge, 3 parts of almonds, 2 parts of siler, 2 parts of costus root, 0.9 parts of lotus leaves, 0.7 parts of lily, and 0.5 parts of amomum villosum are mixed and crushed, and passed through a 100-mesh sieve to obtain traditional Chinese medicine powder, and the grain porridge and the traditional Chinese medicine powder are evenly mixed to obtain koji material;

[0045] S2: inoculating the composite starter into the koji material prepared in S1, the inoculation amount accounting for 5% of the weight of the koji material, mixing the composite starter and the koji material evenly, and then pressing them into rectangular koji embryos, stacking the koji embryos in a koji room, controlling the temperature of the koji room to be 32°C and the relative humidity to be 70°C, and fermenting for 30 days to obtain vinegar koji;

[0046] S3: The vinegar koji prepared in S2 is crushed to a particle size of less than 2.0 mm to obtain vinegar koji powder, and the vinegar koji powder is evenly mixed with a protective agent and a composite functional bacterial agent in a mass ratio of 7:2:1, and freeze-dried to obtain a composite bacterial agent with added functional bacterial species of vinegar koji microorganisms for fruit vinegar.

[0047] Comparative Example 1

[0048] The main difference between Comparative Example 1 and Example 1 is that no composite microbial agent containing multiple microorganisms is added in Comparative Example 1. In addition, Comparative Example 1 is consistent with Example 1 in other aspects, including but not limited to raw material ratio, preparation method, operating conditions, etc., and all follow the specific steps and parameters described in Example 1.

[0049] Performance testing:

[0050] The effective viable count of the composite bacterial agent of the special vinegar koji microorganism added functional bacteria obtained in Examples 1 to 5 and Comparative Example 1 was detected, and the storage conditions were 20 to 30°C, humidity 40% to 50% and shading. The detection method is to take different groups of newly prepared composite bacterial agents of special vinegar koji microorganisms added functional bacteria for effective viable count 5g in a test tube, add 20ml of sterile water respectively, shake at 35°C, 120rpm oscillation mixing 2h, cultivate at 28°C for 48h, use the plate counting method to count the effective viable count A1 in the composite bacterial agent of special vinegar koji microorganisms added functional bacteria in different groups of fruit vinegar, and then use the same method to detect the effective viable count A2 of the composite bacterial agent of special vinegar koji microorganisms added functional bacteria in different groups of fruit vinegar after 3 months, and calculate the viable bacteria retention rate (viable bacteria retention rate = A2 / A1×100%), and the results are shown in Table 1.

[0051] Table 1

[0052] <![CDATA[A1(×10 8 CFU / mL)]]> <![CDATA[A2(×10 8 CFU / mL)]]> Live bacteria retention rate (%) Example 1 5.93 4.65 0.78 Example 2 5.71 4.38 0.77 Example 3 5.27 4.17 0.79 Example 4 5.19 4.15 0.80 Example 5 5.67 4.27 0.75 Comparative Example 1 5.09 3.01 0.60

[0053] The method of preparing apple cider vinegar by using the composite bacterial agent of the vinegar koji microorganism added with functional bacteria obtained in Examples 1 to 5 comprises the following steps: washing and crushing apples, controlling the sugar content at 12% to 15%, and obtaining apple pulp; adding 0.5% to 2% of the mass of the apple pulp of the composite bacterial agent of the vinegar koji microorganism added with functional bacteria and 60% to 80% of the mass of the apple pulp of water to the apple pulp, controlling the room temperature at 20° C. to 30° C., and performing closed fermentation for 5 to 7 days to obtain apple wine; and then performing aerobic fermentation, controlling the room temperature at 20° C. to 30° C., and performing aerobic fermentation for 6 to 9 days to obtain apple cider vinegar.

[0054] Control example: apples are washed and crushed, and the sugar content is controlled at 12% to 15% to obtain apple pulp; 0.02% to 0.05% of brewer's yeast by weight of the apple pulp and 60% to 80% of water by weight of the apple pulp are added to the apple pulp, the room temperature is controlled at 20° C. to 30° C., and apple cider is obtained after closed fermentation for 5 to 7 days; 0.02% of Acetobacter pasteurianus by weight of apple juice is added for aerobic fermentation, the room temperature is controlled at 20° C. to 30° C., and apple cider vinegar is obtained after aerobic fermentation for 12 to 15 days.

[0055] The total acid content, ethyl acetate content, malic acid content, citric acid content and lactic acid content of the apple vinegar prepared in Examples 1-5 and the comparative example were measured. The total acid was tested according to the method specified in GB / T 12456, the malic acid, citric acid, tartaric acid and succinic acid were tested according to the method specified in GB / T 5009.157, the lactic acid was tested according to the method specified in SN / T 2007-2007, and the taste was tested. The test results are shown in Table 2:

[0056] Table 2

[0057]

[0058] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a composite microbial agent of vinegar koji microorganisms for fruit vinegar with added functional strains, characterized in that: The following steps are involved: S1: Grind corn, wheat bran and rice husk, pass through a 100-mesh sieve and mix them evenly to obtain mixed grain powder, add water to the mixed grain powder and steam for 30 to 50 minutes, cool to room temperature, and obtain grain porridge; mix almonds, fangfeng, costusroot, lotus leaves, lily and amomum, grind them, pass through a 100-mesh sieve to obtain traditional Chinese medicine powder; mix the grain porridge and the traditional Chinese medicine powder to obtain koji material; S2: inoculating the composite starter into the koji material prepared in S1, the inoculation amount being 2% to 5% of the weight of the koji material, mixing the composite starter and the koji material evenly, and then pressing them into rectangular koji embryos, stacking the koji embryos in a koji room, controlling the room temperature of the koji room to be 28°C to 32°C, the relative humidity to be 60°C to 70°C, and fermenting for 24 to 30 days to obtain vinegar koji; S3: Grind the vinegar koji prepared in S2 to a particle size of less than 2.0 mm to obtain vinegar koji powder, and mix the vinegar koji powder, protective agent, and composite functional bacterial strains and inoculants in proportion.

2. The method for preparing a composite microbial agent of a special vinegar koji microorganism with added functional strains for fruit vinegar according to claim 1, characterized in that: The koji material is composed of the following raw materials in parts by mass: 60-70 parts of corn, 20-30 parts of wheat bran, 12-20 parts of rice husk, 1-3 parts of almond, 1-2 parts of siler, 1-2 parts of costusroot, 0.4-0.9 parts of lotus leaf, 0.5-0.7 parts of lily and 0.3-0.5 parts of amomum.

3. The method for preparing a composite microbial agent of a special vinegar koji microorganism with added functional strains for fruit vinegar according to claim 1, characterized in that: The composite fermentation agent consists of acetobacter pasteurianus, saccharomyces cerevisiae and lactobacillus acidophilus, and the weight ratio of acetobacter pasteurianus, saccharomyces cerevisiae and lactobacillus acidophilus is 3:3:

4.

4. The method for preparing a composite microbial agent of a special vinegar koji microorganism with added functional strains for fruit vinegar according to claim 1, characterized in that: The mass ratio of the vinegar koji powder, the protective agent and the composite functional bacterial strain inoculant is 2:

1.

5. The method for preparing a composite microbial agent of a special vinegar koji microorganism with added functional strains for fruit vinegar according to claim 1, characterized in that: The protective agent consists of beta-cyclodextrin, trehalose and glycerol, and the mass ratio of the beta-cyclodextrin, trehalose and glycerol is 4:3:

1.

6. The method for preparing a composite microbial agent of a special vinegar koji microorganism with added functional strains for fruit vinegar according to claim 1, characterized in that: The composite functional bacterial strain agent consists of Bacillus Velezii and Lactobacillus delbrueckii subspecies bulgaricus, and the mass ratio of Bacillus Velezii to Lactobacillus delbrueckii subspecies bulgaricus is 3:

7.

7. A composite bacterial agent for fruit vinegar with added functional bacteria, characterized in that: The invention is prepared by the preparation method of the composite bacterial agent of the vinegar koji microorganisms for fruit vinegar added with functional bacteria as described in claims 1 to 6.

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