CN107022467B - Brewing method of high-free-state amino acid vinegar - Google Patents
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12J—VINEGAR; PREPARATION OR PURIFICATION THEREOF
- C12J1/00—Vinegar; Preparation or purification thereof
- C12J1/04—Vinegar; Preparation or purification thereof from alcohol
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Abstract
The invention belongs to the technical field of vinegar production and brewing, and provides a method for brewing high-free-state amino acid vinegar, aiming at solving the problems that the protein content in Shanxi mature vinegar precipitate is high, the precipitate is difficult to solve, the content of free amino acid nitrogen in the vinegar is low, the utilization rate of protein in raw materials is low and the like. The sorghum is used as main grain for puffing treatment, yeast for making hard liquor, aspergillus niger which is a bacterium producing acid protease and bacillus subtilis which is a bacterium producing neutral protease are added for alcoholic fermentation, and acetic acid bacteria are added for normal vinegar fermentation, so that amino acid nitrogen and delicate fragrance of vinegar are increased, and the content of protein in mature vinegar precipitate and vinasse is reduced. The prepared vinegar has reduced protein content in precipitate, and reduced precipitation rate of edible vinegar during production. The content of amino acid nitrogen reaches more than 0.42 g/100ml, which is improved by more than 180% compared with the common Shanxi mature vinegar sold in the market, the precipitation rate of vinegar is reduced by 55%, the utilization rate of protein in the precipitate is improved by 59%, and the utilization rate of protein in the vinegar residue is improved by 77%.
Description
Technical Field
The invention belongs to the technical field of vinegar production and brewing, and particularly relates to a brewing method of high-free-state amino acid vinegar.
Background
The problem of vinegar precipitation is a ubiquitous problem in the vinegar industry and is not solved effectively all the time. The vinegar precipitation not only affects the shelf life, but also affects the appearance of the product and reduces the value of the product.
The Shanxi mature vinegar precipitate has rich nutrition, the highest protein content is 14.96%, the types of amino acids are complete, 7 amino acids necessary for human bodies are contained, the composition is relatively reasonable, the necessary amino acids account for 22.12-28.28% of the total amino acid content, the ratio is close to the ratio (35.38%) recommended by a WHO/FAO mode, the ratio of the umami amino acids is high and exceeds 50%, in addition, the contents of total polyphenol and total flavone are high and reach 10.44 mg/g and 14.73 mg/g respectively, and the Shanxi mature vinegar precipitate has development and utilization values. (analysis and evaluation of nutrient components of Shanxi mature vinegar precipitate, Jun Zhao Rui Huan, Kanjijie, Tianjinrui, Junxian, food science 2014.6.15, 219 Bu 222).
The amino acid is one of the main sources of the fresh flavor of the vinegar product, and the higher the content of the amino acid is, the better the fresh flavor of the vinegar is. Therefore, the macromolecular protein is effectively decomposed into the amino acid and the functional small peptide in the vinegar fermentation process, so that the content of free amino acid in vinegar is further improved, the content of the macromolecular protein can be reduced, the precipitation amount in the production process is reduced, the problem of vinegar precipitates is effectively solved, the vinegar flavor can be promoted, the nutritive value of vinegar products can be improved, and the added value of products is improved.
Disclosure of Invention
The invention provides a brewing method of high-free-state amino acid vinegar, aiming at solving the problems that the existing Shanxi mature vinegar precipitate has high protein content and is difficult to effectively solve the precipitation, the vinegar has low content of amino acid nitrogen, the vinegar residue has low protein utilization rate and the like.
The invention is realized by the following technical scheme: a method for brewing high free state amino acid vinegar comprises taking puffed sorghum as main grain for fermentation, adding Daqu 60 wt% of the main grain and mixed solid bacteria preparation 20 wt% of the main grain, performing alcohol fermentation, and adding acetic acid bacteria 12-15 wt% of vinegar grains for normal vinegar fermentation; wherein the mixed solid bacteria preparation is acid protease producing bacteria Aspergillus nigerAspergillus niger) Solid bacterial preparation and neutral protease producing bacterium Bacillus subtilis (B.) (Bacillus subtilis) The solid bacteria preparation is prepared by mixing the solid bacteria preparation according to the weight ratio of 3: 2.
The method specifically comprises the following steps:
(1) culturing a microbial seed solution: acid protease producing bacterium Aspergillus nigerAspergillus niger) The seeds are cultured in PDA liquid culture medium at 30 deg.C and 180rpm at natural pH for 48 hr with viable count not less than 108CFU/mL is the Aspergillus niger seed liquid; bacillus subtilis as neutral protease producing bacterium (A)Bacillus subtilis) The seed adopts MRS liquid culture medium, the culture conditions are 35 ℃, 100rpm, the constant temperature culture is carried out for 48h under natural pH, the viable count is more than or equal to 108CFU/mL is the bacillus subtilis seed solution;
(2) culturing a mixed solid bacterium preparation: the culture medium raw materials and the weight ratio of the mixed solid bacteria preparation are as follows: 55% of bran, 45% of soybean meal and 70% of water, naturally adjusting pH, sterilizing at 121 deg.C for 20-30 min; inoculating acid protease producing bacteria Aspergillus niger into the culture medium under aseptic conditionAspergillus niger) And neutral protease producing bacterium Bacillus subtilis (B.) (Bacillus subtilis) The inoculum size of the seed liquid is 5%, the Aspergillus niger is statically cultured for 48h at 30 ℃, and the viable count is more than or equal to 1.0 × 109CFU/g to obtain Aspergillus niger solid preparation, standing and culturing Bacillus subtilis at 35 deg.C for 48 hr with viable count not less than 1.0 × 109And (3) obtaining a bacillus subtilis solid preparation according to CFU/g, and then obtaining an aspergillus niger solid preparation: mixing the bacillus subtilis solid preparations in a weight ratio of 3:2 to obtain a mixed solid preparation;
(3) puffing main grains: the sorghum is crushed to the granularity of 1.5-1.7 mm and then is placed in a bulking machine for bulking under the following conditions: the pressure of the puffing gun head is 1.8-2.0 MPa, the puffing temperature is 110-120 ℃, and the feeding speed is 6 Hz; the swelling degree is controlled to be 160-200 g/L, the gelatinization degree is 75-80%, and the water content is 5-6%;
(4) alcohol fermentation: inoculating the mixed solid bacterium preparation obtained in the step (2) into the main grain puffed in the step (2) according to the amount of 20% of the weight of the main grain, and then adding yeast and water for alcohol fermentation, wherein the using amount of the yeast is 60% of that of the main grain, and the using amount of the water is 3 times of that of the main grain; raking once every morning and evening, controlling the temperature to be less than 35 ℃, fermenting for 9 days to obtain fermented mash, wherein the alcoholic strength of the fermented mash is more than or equal to 9 degrees;
(5) acetic acid fermentation: the raw materials for acetic fermentation and the parts by weight are as follows: 21.5 parts of fermented glutinous rice, 4 parts of bran, 5 parts of bran coat, 5 parts of rice hull and 15 parts of water, which are obtained in the step (3), and then the acetic fermentation raw material is uniformly stirred into vinegar mash, wherein the alcoholic strength of the vinegar mash is 3.5-4.2, and the water content is 63% -66%; inoculating acetic acid bacteria with the weight of 12-15% of the fermented vinegar, wherein the acidity of the acetic acid bacteria liquid is 1.2-1.5; controlling the temperature to be 30-32 ℃ to carry out acetic fermentation, turning over the fermented grains 1-2 times every day, wherein the fermentation time is 8 days, and the acidity of the vinegar grains is 3.8-4.0; pouring vinegar after acetic acid fermentation is finished, taking first hogwash after soaking for 12-15h during vinegar pouring, and reserving second hogwash and third hogwash for later use; boiling the first hogwash at 100 deg.C for 30min, and sterilizing;
wherein: the acid protease producing bacterium Aspergillus nigerAspergillus niger) Purchased from China center for culture collection and management of Industrial microorganisms (CICC), and the strain numbers are as follows: CICC 2377; bacillus subtilis as neutral protease producing bacterium (A)Bacillus subtilis) Purchased from China center for culture collection and management of Industrial microorganisms (CICC), and the strain numbers are as follows: CICC 20076.
The invention utilizes the cooperation of acid and neutral protease producing bacteria to act on the whole mature vinegar fermentation process. The vinegar production process is a process of continuously reducing pH. In the vinegar brewing process, the pH value of the wine mash after alcohol fermentation reaches about 4.0, and is reduced to 3.6-3.8 through acetic acid fermentation, so that the acid protease producing bacteria or the neutral protease producing bacteria are only inoculated to hardly play a role. Therefore, the mixed inoculation culture of the acidic protease producing bacteria and the neutral protease producing bacteria has great significance for producing the brewed vinegar with high amino acid content.
The acid protease producing strain Aspergillus niger CICC of the invention: 2377 and neutral protease producing bacteria Bacillus subtilis CICC: 20076 the solid pure culture is mixed at a weight ratio of 3:2, and inoculated with Daqu together into staple food for alcoholic fermentation, and further acetic fermentation, so as to increase amino acid nitrogen and flavor of the brewed vinegar, reduce protein content in mature vinegar precipitate, and reduce protein content in distiller's grains. The high amino acid vinegar prepared by the method is a reddish brown clear liquid, has acid aroma and special smell of solid fermentation, and is sour, sweet and palatable. Wherein: the content of amino acid nitrogen reaches more than 0.42 g/100ml, which is improved by more than 180 percent compared with the common Shanxi mature vinegar sold in the market.
The invention can effectively reduce the precipitation rate of the mature vinegar and the protein content in the precipitate, and simultaneously improve the utilization rate of the protein in the vinegar residue. Compared with the traditional process, the method can reduce the vinegar precipitation rate by 55%, improve the protein utilization rate in the precipitate by 59% and improve the protein utilization rate in the vinegar residue by 77%.
Drawings
FIG. 1 is a flow chart of the high amino acid vinegar brewing process of the invention.
Detailed Description
A method for brewing high free state amino acid vinegar comprises taking puffed sorghum as main grain for fermentation, adding Daqu 60 wt% of the main grain and mixed solid bacteria preparation 20 wt% of the main grain, performing alcohol fermentation, and adding acetic acid bacteria 12-15 wt% of vinegar grains for normal vinegar fermentation; wherein the mixed solid bacteria preparation is acid protease producing bacteria Aspergillus nigerAspergillus niger) Solid bacterial preparation and neutral protease producing bacterium Bacillus subtilis (B.) (Bacillus subtilis) The solid bacteria preparation is prepared by mixing the solid bacteria preparation according to the weight ratio of 3: 2.
The method specifically comprises the following steps:
(1) culturing a microbial seed solution: acid protease producing bacterium Aspergillus nigerAspergillus niger) The seeds are cultured in PDA liquid culture medium at 30 deg.C and 180rpm at natural pH for 48 hr with viable count not less than 108CFU/mL is the Aspergillus niger seed liquid; bacillus subtilis as neutral protease producing bacterium (A)Bacillus subtilis) The seed adopts MRS liquid culture medium, the culture conditions are 35 ℃, 100rpm, the constant temperature culture is carried out for 48h under natural pH, the viable count is more than or equal to 108CFU/mL is the bacillus subtilis seed solution;
(2) culturing a mixed solid bacterium preparation: the culture medium raw materials and the weight ratio of the mixed solid bacteria preparation are as follows: 55% of bran, 45% of soybean meal and 70% of water, naturally adjusting pH, sterilizing at 121 deg.C for 20-30 min; inoculating acid protease producing bacteria Aspergillus niger into the culture medium under aseptic conditionAspergillus niger) And neutral protease producing bacterium Bacillus subtilis (B.) (Bacillus subtilis) The inoculum size of the seed liquid is 5%, the Aspergillus niger is statically cultured for 48h at 30 ℃, and the viable count is more than or equal to 1.0 × 109CFU/g to obtain Aspergillus niger solid preparation, standing and culturing Bacillus subtilis at 35 deg.C for 48 hr with viable count not less than 1.0 × 109CFObtaining a bacillus subtilis solid preparation according to U/g, and then obtaining an aspergillus niger solid preparation: mixing the bacillus subtilis solid preparations in a weight ratio of 3:2 to obtain a mixed solid preparation;
(3) puffing main grains: the sorghum is crushed to the granularity of 1.5-1.7 mm and then is placed in a bulking machine for bulking under the following conditions: the pressure of the puffing gun head is 1.8-2.0 MPa, the puffing temperature is 110-120 ℃, and the feeding speed is 6 Hz; the swelling degree is controlled to be 160-200 g/L, the gelatinization degree is 75-80%, and the water content is 5-6%;
(4) alcohol fermentation: inoculating the mixed solid bacterium preparation obtained in the step (2) into the main grain puffed in the step (2) according to the amount of 20% of the weight of the main grain, and then adding yeast and water for alcohol fermentation, wherein the using amount of the yeast is 60% of that of the main grain, and the using amount of the water is 3 times of that of the main grain; raking once every morning and evening, controlling the temperature to be less than 35 ℃, fermenting for 9 days to obtain fermented mash, wherein the alcoholic strength of the fermented mash is more than or equal to 9 degrees;
(5) acetic acid fermentation: the raw materials for acetic fermentation and the parts by weight are as follows: 21.5 parts of fermented glutinous rice, 4 parts of bran, 5 parts of bran coat, 5 parts of rice hull and 15 parts of water, which are obtained in the step (3), and then the acetic fermentation raw material is uniformly stirred into vinegar mash, wherein the alcoholic strength of the vinegar mash is 3.5-4.2, and the water content is 63% -66%; inoculating acetic acid bacteria with the weight of 12-15% of the fermented vinegar, wherein the acidity of the acetic acid bacteria liquid is 1.2-1.5; controlling the temperature to be 30-32 ℃ to carry out acetic fermentation, turning over the fermented grains 1-2 times every day, wherein the fermentation time is 8 days, and the acidity of the vinegar grains is 3.8-4.0; pouring vinegar after acetic acid fermentation is finished, taking first hogwash after soaking for 12-15h during vinegar pouring, and reserving second hogwash and third hogwash for later use; boiling the first hogwash at 100 deg.C for 30min, and sterilizing;
wherein: the acid protease producing bacterium Aspergillus nigerAspergillus niger) Purchased from China center for culture collection and management of Industrial microorganisms (CICC), and the strain numbers are as follows: CICC 2377; bacillus subtilis as neutral protease producing bacterium (A)Bacillus subtilis) Purchased from China center for culture collection and management of Industrial microorganisms (CICC), and the strain numbers are as follows: CICC 20076.
Experimental example 1: preparation of solid pure culture of microorganism:
acid protease producing bacterium Aspergillus nigerAspergillus niger) Andbacillus subtilis as neutral protease producing bacterium (A)Bacillus subtilis) The culture medium raw materials and the weight ratio of the fungus solid-state fungus preparation are as follows: 55% of bran, 45% of soybean meal and 70% of water, naturally adjusting pH, sterilizing at 121 deg.C for 20-30 min; inoculating Aspergillus niger (A.niger) of acidic protease producing bacterium into the culture medium under aseptic conditionsAspergillus niger) And neutral protease producing bacterium Bacillus subtilis (B.) (Bacillus subtilis) The inoculum size of the seed liquid is 5 percent, aspergillus niger is kept still and cultured for 48 hours at the temperature of 30 ℃, bacillus subtilis is kept still and cultured for 48 hours at the temperature of 35 ℃, the bacterial load is measured after 48 hours, and the result is shown in table 1.
Table 1: effective viable count of culture
Experimental example 2: and (3) detecting the activity of the protease: the protease activity is measured by adopting a Folin phenol method, and the specific implementation scheme is as follows:
(1) preparing an enzyme solution: 10g of the Aspergillus niger and Bacillus subtilis solid microbial inoculum prepared in experimental example 1 are respectively dried at 40 ℃, crushed, sampled, added with deionized water and leached in water bath at 40 ℃ for 30 min. Filtering the leaching solution with filter paper, and taking the filtrate to fix the volume to 10ml for the activity determination of the protease.
(2) Protease activity determination: the reaction substrate was a 2% casein solution as determined by the Folin phenol method (Zhangzhen. enzyme preparations industries [ M ]. scientific Press, 1984.). The results are shown in Table 2.
Table 2: culture protease activity detection result
Note: and (3) protease activity calculation: the amount of enzyme (g) producing 1mg of tyrosine in 1min is one activity unit.
Experimental example 3: an optimization experiment of the proportion of aspergillus niger and bacillus subtilis in the mixed solid bacterial preparation comprises the following steps:
the raw materials and the mixture ratio used for alcohol fermentation are as follows: 10kg of sorghum, 6kg of Daqu and 30kg of water, and 5 experimental groups are set, wherein the formula of the mixed bacteria preparation in each experimental group is as follows: the mixing ratio of the acid protease producing aspergillus niger to the neutral protease producing bacillus subtilis is respectively 8: 2; 7: 3; 6: 4; 5: 5; 4:6, inoculating, wherein the inoculation amount of the mixed bacteria preparation is 2kg (20% of the sorghum weight), the control is that the inoculation amount of Aspergillus niger and Bacillus subtilis is 0, and 2kg of bran is added, and the specific experimental formula and the dosage are shown in Table 3.
Before fermentation, firstly carrying out puffing treatment on sorghum, then carrying out size mixing according to an alcohol fermentation formula shown in table 3, raking once every morning and evening, controlling the temperature to be less than 35 ℃, and fermenting for 18 days to obtain fermented glutinous rice, wherein the alcoholic strength of the fermented glutinous rice is more than or equal to 9 degrees;
table 3: alcohol fermentation formula and proportion
After alcohol fermentation is carried out to obtain fermented mash, carrying out acetic fermentation, and mixing the fermented mash according to the acetic fermentation formula and the proportion shown in table 4, wherein the alcoholic strength of the fermented mash is 3.5-4.2, and the water content is 63% -66%; controlling the temperature to be 30-32 ℃ to carry out acetic fermentation for 8 days, and turning over the fermented grains 1-2 times every day; pouring vinegar after acetic acid fermentation is finished, taking first hogwash after soaking for 12-15h during vinegar pouring, and reserving second hogwash and third hogwash for later use; boiling the first hogwash at 100 deg.C for 30min, and sterilizing.
Table 4: formula proportion of acetic fermentation
After alcohol fermentation, the alcohol content, the amino acid nitrogen content and the total acid content of each experimental group and the control group are detected, and the results are shown in table 5; the total acid and amino acid nitrogen contents were measured after acetic acid fermentation, and the results are shown in table 6; the total acid and amino acid nitrogen content was also tested after sterilization and the results are shown in table 7.
After the alcoholic fermentation is finished, the contents of amino acid nitrogen in the batches 3, 4 and 5 are higher than those in the batches 1 and 2 and the comparison batch, which shows that in the alcoholic fermentation stage, the pH value is gradually changed from neutral to acidic, the fermentation time in the stage is longer, and the neutral protease-producing bacillus subtilis plays a main role.
After the acetic acid fermentation is finished, the content of amino acid nitrogen in the batches 1 and 2 is obviously improved, which shows that the acid-producing lipase Aspergillus niger plays a main role in the acetic acid fermentation stage (pH3.6-3.8).
Compared with the control group, the content of amino acid nitrogen in the vinegar produced in the batch 3 is obviously improved by 210 percent. Therefore, the mixing ratio of the acid-producing protease Aspergillus niger to the neutral protease Bacillus subtilis is 6:4, namely 3:2, the solid mixed bacteria preparation can better play the roles of neutral lipase and acid lipase in different stages of acetic acid fermentation.
TABLE 5 detection index tracking at end of alcoholic fermentation (18 d) in mash
TABLE 6 tracking of detection indexes in vinegar grains after completion of acetic acid fermentation (26 d)
Table 7: detection results of edible vinegar obtained in different batches
Experimental example 4: determination of precipitation rate in vinegar and protein content in precipitate
In experimental example 3, batch 3 and the control vinegar were put into a jar and aged under the aging conditions: sealing the cover at the outdoor natural temperature, and ageing for 1 month. Collecting aged vinegar precipitate mixture, centrifuging at 3000 r/min, removing supernatant vinegar liquid, and storing vinegar precipitate for use. The precipitation rates of batch 3 and the control group are respectively 4.01% and 8.97%, and the protein content in the precipitates is respectively 6.06% and 14.96%, so that the mixed solid-state fungus preparation prepared according to the formula of batch 3 (acid-producing protease Aspergillus niger: neutral protease Bacillus subtilis is 6: 4) is determined, and vinegar brewing is carried out according to the process flow disclosed by the embodiment of the invention, the vinegar precipitation rate can be reduced by 55%, and the protein utilization rate in the precipitates can be improved by 59%.
The precipitation rate calculation formula is as follows: w1=m1/m2. Wherein W1: the precipitation rate; m is1The quality of the precipitate; m is2: sample mass; the protein content determination method adopts a Kjeldahl method, and refers to GB/T5009.5-2010 determination of protein in food.
Experimental example 5: determination of protein content in vinegar residue
In experimental example 3, after vinegar residue was poured into batch 3 and a control, the vinegar residue was dried at 40 ℃ and weighed, and the protein content in the vinegar residue was measured, with the protein content in batch 3 being 2.65% and the protein content in the control being 11.57%. Therefore, the mixed solid bacterial preparation is prepared according to the formula of batch 3 (acid-producing protease Aspergillus niger: neutral protease Bacillus subtilis: 6: 4), and vinegar brewing is carried out according to the process flow disclosed by the invention, so that the utilization rate of protein in vinegar residue can be improved by 77%.
The content of protein in vinegar residue is determined and calculated according to the method of GB/T5009.5-2010 'determination of protein in food'.
Experimental example 6: compared with the sensory and physicochemical data of the commercial edible vinegar
Commercial Shanxi mature vinegar (2 years mature vinegar in Shanxi of Zilin: 1.4L in package; Ninghuafu brand vinegar: 1.45L in package; and mature vinegar in sewer well: 1.45L in package) was purchased and subjected to sensory evaluation and physicochemical examination, and the results were shown in Table 8.
Table 8: compared with the sensory evaluation and quality control items of the commercial mature vinegar products
According to acid-producing protease aspergillus niger: the bacillus subtilis capable of producing neutral protease is a mixed bacterium preparation prepared at a ratio of 6:4, and all indexes of vinegar products produced by the process flow according to the embodiment of the invention are superior to those of the market-sold mature vinegar.
Claims (2)
1. Brewing method of high-free-state amino acid vinegarThe method is characterized in that: taking the puffed sorghum as main grain for fermentation, adding mixed solid bacteria preparation with 60 percent of yeast and 20 percent of main grain weight, performing alcohol fermentation, and then adding acetic acid bacteria with the weight of 12-15 percent of vinegar grains for normal vinegar fermentation; wherein the mixed solid bacteria preparation is acid protease producing bacteria Aspergillus nigerAspergillus niger) Solid bacterial preparation and neutral protease producing bacterium Bacillus subtilis (B.) (Bacillus subtilis) The solid bacterium preparation is prepared by mixing the solid bacterium preparation according to the weight ratio of 3: 2;
wherein: the acid protease producing bacterium Aspergillus nigerAspergillus niger) Purchased from China center for culture collection and management of Industrial microorganisms (CICC), and the strain numbers are as follows: CICC 2377; bacillus subtilis as neutral protease producing bacterium (A)Bacillus subtilis) Purchased from China center for culture collection and management of Industrial microorganisms (CICC), and the strain numbers are as follows: CICC 20076.
2. The method for brewing vinegar containing highly free amino acids according to claim 1, wherein: the method specifically comprises the following steps:
(1) culturing a microbial seed solution: acid protease producing bacterium Aspergillus nigerAspergillus niger) The seeds are cultured in PDA liquid culture medium at 30 deg.C and 180rpm at natural pH for 48 hr with viable count not less than 108CFU/mL is the Aspergillus niger seed liquid; bacillus subtilis as neutral protease producing bacterium (A)Bacillus subtilis) The seed adopts MRS liquid culture medium, the culture conditions are 35 ℃, 100rpm, the constant temperature culture is carried out for 48h under natural pH, the viable count is more than or equal to 108CFU/mL is the bacillus subtilis seed solution;
(2) culturing a mixed solid bacterium preparation: the culture medium raw materials and the weight ratio of the mixed solid bacteria preparation are as follows: 55% of bran, 45% of soybean meal and 70% of water, naturally adjusting pH, sterilizing at 121 deg.C for 20-30 min; inoculating acid protease producing bacteria Aspergillus niger into the culture medium under aseptic conditionAspergillus niger) And neutral protease producing bacterium Bacillus subtilis (B.) (Bacillus subtilis) The inoculum size of the seed liquid is 5 percent, and the aspergillus niger is at 30 DEG CStanding for 48h, the number of viable bacteria is more than or equal to 1.0 × 109CFU/g to obtain Aspergillus niger solid preparation, standing and culturing Bacillus subtilis at 35 deg.C for 48 hr with viable count not less than 1.0 × 109And (3) obtaining a bacillus subtilis solid preparation according to CFU/g, and then obtaining an aspergillus niger solid preparation: mixing the bacillus subtilis solid preparations in a weight ratio of 3:2 to obtain a mixed solid preparation;
(3) puffing main grains: the sorghum is crushed to the granularity of 1.5-1.7 mm and then is placed in a bulking machine for bulking under the following conditions: the pressure of the puffing gun head is 1.8-2.0 MPa, the puffing temperature is 110-120 ℃, and the feeding speed is 6 Hz; the swelling degree is controlled to be 160-200 g/L, the gelatinization degree is 75-80%, and the water content is 5-6%;
(4) alcohol fermentation: inoculating the mixed solid bacterium preparation obtained in the step (2) into the main grain puffed in the step (2) according to the amount of 20% of the weight of the main grain, and then adding yeast and water for alcohol fermentation, wherein the using amount of the yeast is 60% of that of the main grain, and the using amount of the water is 3 times of that of the main grain; raking once every morning and evening, controlling the temperature to be less than 35 ℃, fermenting for 9 days to obtain fermented mash, wherein the alcoholic strength of the fermented mash is more than or equal to 9 degrees;
(5) acetic acid fermentation: the raw materials for acetic fermentation and the parts by weight are as follows: 21.5 parts of fermented glutinous rice, 4 parts of bran, 5 parts of bran coat, 5 parts of rice hull and 15 parts of water, which are obtained in the step (4), and then the acetic fermentation raw material is uniformly stirred into vinegar mash, wherein the alcoholic strength of the vinegar mash is 3.5-4.2, and the water content is 63% -66%; inoculating acetic acid bacteria with the weight of 12-15% of the fermented vinegar, wherein the acidity of the acetic acid bacteria liquid is 1.2-1.5; controlling the temperature to be 30-32 ℃ to carry out acetic fermentation, turning over the fermented grains 1-2 times every day, wherein the fermentation time is 8 days, and the acidity of the vinegar grains is 3.8-4.0; pouring vinegar after acetic acid fermentation is finished, taking first hogwash after soaking for 12-15h during vinegar pouring, and reserving second hogwash and third hogwash for later use; boiling the first hogwash at 100 deg.C for 30min, and sterilizing.
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| CN109182206B (en) * | 2018-10-10 | 2021-08-27 | 内蒙古工业大学 | Bacillus subtilis capable of highly producing complex enzyme and application thereof |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101857833A (en) * | 2010-06-11 | 2010-10-13 | 山西三盟实业发展有限公司 | Standardized and industrialized production process for Shanxi mature vinegar |
| CN103087896A (en) * | 2013-01-15 | 2013-05-08 | 山西梁汾醋业有限公司 | Method for making aged vinegar by using extrusion of raw materials |
| CN103756918A (en) * | 2014-01-16 | 2014-04-30 | 山西梁汾醋业有限公司 | Protease generating strain amplification method and method for increasing content of amino-acid-state nitrogen in vinegar |
| CN104357306A (en) * | 2014-11-28 | 2015-02-18 | 山西梁汾醋业有限公司 | Method for brewing soybean sauce vinegar |
| CN104357307A (en) * | 2014-12-02 | 2015-02-18 | 山西梁汾醋业有限公司 | Brewage method of wire drawing vinegar |
| CN105907597A (en) * | 2016-06-08 | 2016-08-31 | 山西省农业科学院农产品加工研究所 | Preparation method of high-lactic-acid edible vinegar |
-
2017
- 2017-06-16 CN CN201710457502.0A patent/CN107022467B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101857833A (en) * | 2010-06-11 | 2010-10-13 | 山西三盟实业发展有限公司 | Standardized and industrialized production process for Shanxi mature vinegar |
| CN103087896A (en) * | 2013-01-15 | 2013-05-08 | 山西梁汾醋业有限公司 | Method for making aged vinegar by using extrusion of raw materials |
| CN103756918A (en) * | 2014-01-16 | 2014-04-30 | 山西梁汾醋业有限公司 | Protease generating strain amplification method and method for increasing content of amino-acid-state nitrogen in vinegar |
| CN104357306A (en) * | 2014-11-28 | 2015-02-18 | 山西梁汾醋业有限公司 | Method for brewing soybean sauce vinegar |
| CN104357307A (en) * | 2014-12-02 | 2015-02-18 | 山西梁汾醋业有限公司 | Brewage method of wire drawing vinegar |
| CN105907597A (en) * | 2016-06-08 | 2016-08-31 | 山西省农业科学院农产品加工研究所 | Preparation method of high-lactic-acid edible vinegar |
Non-Patent Citations (2)
| Title |
|---|
| 高产酸性蛋白酶黑曲霉菌种选育;刘鑫等;《四川大学学报(自然科学版)》;20050228;第42卷(第01期);第158-161页 * |
| 黑曲霉酸性蛋白酶在食醋酿造中的催化效应;刘鑫等;《化学研究与应用》;20040828;第16卷(第04期);第482-484页 * |
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