CN113430124A - Culture medium for increasing number of soy sauce aspergillus spores and preparation method thereof - Google Patents
Culture medium for increasing number of soy sauce aspergillus spores and preparation method thereof Download PDFInfo
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- CN113430124A CN113430124A CN202110728834.4A CN202110728834A CN113430124A CN 113430124 A CN113430124 A CN 113430124A CN 202110728834 A CN202110728834 A CN 202110728834A CN 113430124 A CN113430124 A CN 113430124A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/50—Soya sauce
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N3/00—Spore forming or isolating processes
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Abstract
The invention provides a culture medium for increasing the number of soy sauce aspergillus spores, which comprises the following components in percentage by mass: 20-60% of bran, 20-50% of bean dregs, 10-30% of soybean protein and 2-6% of flour. According to the method, the soy protein is mixed with the bran, the bean dregs and the flour according to a certain proportion to prepare the koji culture medium, so that the problem of poor stability of the koji culture medium prepared from natural agricultural byproducts of the bran and the bean dregs is solved under the condition of realizing recycling of agricultural byproduct resources of the bean dregs and the bran, and the aspergillus oryzae can stably produce spores and keep high activity.
Description
Technical Field
The invention relates to the field of soy sauce brewing, in particular to a culture medium for increasing the number of soy sauce aspergillus spores and a preparation method thereof.
Background
The traditional production of the soy sauce koji generally takes bran, flour, rice bran and the like as raw materials, prepares the koji after a certain proportion of matching, mixing and pretreatment, has single raw material and insufficient nutrient substances, and cannot meet the full growth and metabolism of aspergillus oryzae. In addition, the agricultural and sideline products such as bran, rice bran and the like are used, so that the problem of large component fluctuation often exists, the quality of the koji batches is unstable, and the stable sporulation and survival capability of the aspergillus oryzae are influenced.
On the other hand, the bean dregs are byproducts in the bean product processing industry, most of the bean dregs are used as feed or fertilizer, even are treated as waste, and waste of raw materials is caused.
Disclosure of Invention
Based on the problems, the invention provides a culture medium capable of increasing the number of soy sauce aspergillus spores and a preparation method thereof. The soybean residue with a certain proportion is added into the culture medium of the soy sauce koji, so that the number of the soy sauce koji spores is increased, and the waste utilization of the soybean residue is realized.
The invention adopts the following technical scheme to realize the purpose of the invention:
in a first aspect: the invention provides a culture medium for increasing the number of soy sauce aspergillus spores, which comprises the following components in percentage by mass: 20-60% of bran, 20-50% of bean dregs, 10-30% of soybean protein and 2-6% of flour.
The invention adopts soybean protein, bran, bean dregs and flour which are mixed according to a certain proportion to prepare a starter culture medium with the total crude protein content of about 30 percent, the soybean protein has higher protein content, the bean dregs consist of organic matters, inorganic matters and water, wherein the crude fiber accounts for about 20 percent, the crude protein accounts for about 25 percent, but the soybean protein is unstable. The soybean protein can make up for the short unstable wheat bran, bean dregs and flour protein. The materials are combined according to a certain proportion, so that a synergistic effect can be achieved, and the number of the soy sauce aspergillus spores is improved and stabilized.
Within the scope of the present invention, the term "soy protein" is understood to mean a soy protein-rich product obtained by defatting, removing or partially removing carbohydrates from soy as a starting material, known to the person skilled in the art.
Within the scope of the present invention, the term "okara" is understood as a by-product of the process for producing soymilk or tofu known to the person skilled in the art.
Within the scope of the present invention, the term "bran" is understood to mean the seed coat of wheat known to the person skilled in the art after grinding the flour.
Within the scope of the present invention, the term "flour" is understood to mean a flour ground from wheat known to the person skilled in the art. According to the content of protein in the flour, the flour can be divided into high gluten flour, medium gluten flour, low gluten flour and non-gluten flour. High gluten flour, medium gluten flour, low gluten flour and non-gluten flour are included within the scope of the invention.
Preferably, the culture medium for increasing the number of the soy sauce aspergillus spores comprises the following components in percentage by mass: 27-50% of bran, 20-50% of bean dregs, 10-30% of soybean protein and 2-6% of flour.
Preferably, the culture medium for increasing the number of the soy sauce aspergillus spores comprises the following components in percentage by mass: 45% of bran, 35% of bean dregs, 15% of soybean protein and 5% of flour.
Preferably, the content of crude fiber in the bean dregs is more than or equal to 20 wt%, and the content of crude protein is more than or equal to 25 wt%.
In a second aspect: the invention provides a preparation method of a culture medium for increasing the number of soy sauce aspergillus spores, which comprises the following steps:
the method comprises the following steps: providing bran and bean dregs;
step two: uniformly mixing bran, bean dregs, soybean protein and flour according to a proportion, and performing sterilization treatment to obtain a culture medium for increasing the number of soy sauce aspergillus spores;
wherein the culture medium comprises the following components in percentage by weight: 20-60% of bran, 20-50% of bean dregs, 10-30% of soybean protein and 2-6% of flour.
Preferably, in the first step, the bran has a particle size of 20 meshes and the sieved residue is 20-60 wt%; the particle size of the bean dregs is 60 meshes, and the residue is 10-40%.
Preferably, the second step includes:
a. mixing bran, bean dregs, soybean protein and flour to obtain a dry material; adding water into the dry materials, and uniformly mixing to obtain a mixture; wherein the weight ratio of the dry materials to the water is 1: (1.2-1.5);
b. and (3) sterilizing the mixture at 110-130 ℃ for 30-40 min to obtain the culture medium for increasing the number of the soy sauce aspergillus spores.
In a third aspect: the invention provides a preparation method of soy sauce koji, which comprises the following steps:
1) inoculating Aspergillus oryzae to a culture medium containing soybean and/or wheat, and culturing to obtain intermediate strain;
2) inoculating the intermediate seed to the culture medium for increasing the number of the soy sauce koji spores, ventilating and culturing, and discharging the koji after maturation.
Preferably, in the step 1), the culture medium is prepared by the following method: soaking semen glycines in water, decocting, filtering, collecting filtrate to obtain bean juice, adding magnesium sulfate, potassium dihydrogen phosphate, soluble starch and agar powder, dissolving, packaging into test tube, and sterilizing.
Preferably, in the step 2), the intermediate seeds are cultured at a constant temperature of 20-30 ℃ for 1-5 days.
The invention has the beneficial effects that: according to the method, the soy protein is mixed with the bran, the bean dregs and the flour according to a certain proportion to prepare the koji culture medium, so that the problem of poor stability of the koji culture medium prepared from natural agricultural byproducts of the bran and the bean dregs is solved under the condition of realizing recycling of agricultural byproduct resources of the bean dregs and the bran, and the aspergillus oryzae can stably produce spores and keep high activity.
Detailed Description
In order to show technical solutions, purposes and advantages of the present invention more concisely and clearly, the technical solutions of the present invention are described in detail below with reference to specific embodiments. Unless otherwise specified, the reagents involved in the examples of the present invention are all commercially available products, and all of them are commercially available.
Example 1
The embodiment provides a culture medium for increasing the number of soy sauce aspergillus spores and a preparation method thereof, wherein the culture medium comprises the following components in percentage by mass: 60% of bran, 20% of bean dregs, 18% of soybean protein and 2% of flour. The preparation method of the culture medium comprises the following specific steps:
step one, raw material pretreatment:
the fineness of the bran is controlled to be 30 percent under a 20-mesh sieve, the proportion of the bean dregs is controlled to be 30 percent under a 60-mesh sieve, and the detection result of the crude protein of the bean dregs is 19 percent.
Step two, uniformly mixing and sterilizing the raw materials:
mixing the pretreated bran, bean dregs, soybean protein and flour according to the mass percentage of 60%, 20%, 18% and 2%, and then mixing the dry materials according to the following ratio: uniformly stirring water in a weight ratio of 1:1.3, and sterilizing in a starter culture machine at 121 ℃ for 30 min. The culture medium for increasing the number of the soy sauce koji spores is obtained.
The invention also provides examples 2-6 and comparative examples 1-12, wherein the dosage of each component of each example and comparative example is shown in Table 1:
table 1: the amounts of the respective components of examples 1 to 6 and comparative examples 1 to 12
The culture media prepared in the above examples 1 to 6 and comparative examples 1 to 12 were subjected to starter propagation, and the spore-increasing effect thereof was examined, specifically as follows:
1. preparing strains:
1) preparing a slant culture medium:
soaking 1kg of soybean in 2.0L of water for 8h, cooking the soybean, filtering, collecting filtrate to obtain soybean juice, adding 0.8g of magnesium sulfate, 1.5g of potassium dihydrogen phosphate, 15g of soluble starch and 20g of agar powder into 1L of soybean juice, dissolving, packaging into test tubes, and placing in a sterilization pot at 121 ℃ for 20min to obtain the slant culture medium.
2) Preparing a triangular flask culture medium:
mixing the pretreated bran, bean dregs, bran coat and prepared flour according to the weight ratio of 50 percent, 22 percent, 20 percent and 8 percent, and then mixing the materials according to the dry material ratio: stirring uniformly with water at a ratio of 1:1.3, subpackaging into triangular flasks, and placing in a sterilizing pot at 121 deg.C for 20min to obtain triangular flask culture medium.
3) Inoculating and culturing: inoculating Aspergillus oryzae Shanghai brewing 3.042 into slant culture medium of test tube, and culturing at 29 deg.C for 3 days to obtain slant strain. Inoculating the slant seeds into a triangular flask culture medium, and culturing at the constant temperature of 29 ℃ for 2 days to obtain triangular flask seeds.
2. Inoculating and culturing:
the seeds of the triangular flask are respectively inoculated into a koji machine filled with the koji culture media prepared in examples 1-6 and comparative examples 1-12, ventilated culture is carried out for 3 days at 30 ℃, and koji is produced after maturation.
3. Detecting the spore number and the enzyme activity of the yeast:
(1) the protease activity is detected by adopting a welfare method in an SB/T10317-1999 protease activity assay;
(2) the detection method of the number of the spores is GB 4789.15-2010/national food safety standard/food microbiology inspection/detection of mold and yeast count.
(3) The moisture detection method adopts GBT 6284-2006/general method for measuring moisture in chemical products/drying reduction method for detection.
(4) The spore germination rate detection method adopts SB/T10316-1999 method for measuring the spore germination rate
The test results are shown in the following table 2:
table 2: various detection results of yeast
As can be seen from Table 2, comparative examples 1 to 4 lack one of the components, comparative examples 5 to 8 lack two of the components, and comparative examples 9 to 12 contain only one of the components. Compared with the comparative example, the data of the number of spores, moisture, neutral protease activity and spore germination rate of the seed koji in the example are all higher than those of the comparative example. Therefore, the components of the invention generate a synergistic effect, and the spore number, the moisture content, the neutral protease activity and the spore germination rate of the koji in the examples 1 to 6 are all improved, which shows that the bran, the bean dregs, the soybean protein and the flour selected by the invention generate unexpected interaction, and the interaction can improve the activity of the aspergillus in the koji making process. Although the effects of the number of spores, moisture, neutral protease activity and spore germination rate of the koji mold in examples 1 to 6 are good, it is obvious that the component proportion effect of example 3 is the best, because the inventor adjusts the dosage proportion of the bran, the bean dregs, the soybean protein and the flour, and finds that when the weight ratio of the bran, the bean dregs, the soybean protein and the flour is 9:7:3:1, the aspergillus fermentation activity can be promoted to the maximum extent, and the germination rate of the spores is increased by 98%.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (10)
1. The culture medium for increasing the number of the soy sauce aspergillus spores is characterized by comprising the following components in percentage by mass: 20-60% of bran, 20-50% of bean dregs, 10-30% of soybean protein and 2-6% of flour.
2. The culture medium according to claim 1, comprising the following components in percentage by mass: 27-50% of bran, 20-50% of bean dregs, 10-30% of soybean protein and 2-6% of flour.
3. The culture medium according to claim 2, wherein the culture medium comprises the following components in percentage by mass: 45% of bran, 35% of bean dregs, 15% of soybean protein and 5% of flour.
4. The culture medium according to any one of claims 1 to 3, wherein the content of crude fiber in the bean dregs is not less than 20 wt%, and the content of crude protein is not less than 25 wt%.
5. A preparation method of a culture medium for increasing the number of soy sauce aspergillus spores is characterized by comprising the following steps:
the method comprises the following steps: providing bran and bean dregs;
step two: uniformly mixing bran, bean dregs, soybean protein and flour according to a proportion, and performing sterilization treatment to obtain a culture medium for increasing the number of soy sauce aspergillus spores;
wherein the culture medium comprises the following components in percentage by weight: 20-60% of bran, 20-50% of bean dregs, 10-30% of soybean protein and 2-6% of flour.
6. The preparation method of claim 5, wherein in the first step, the bran has a particle size of 20 meshes and a screen residue of 20-60 wt%; the particle size of the bean dregs is 60 meshes, and the residue is 10-40%.
7. The method of claim 5, wherein step two comprises:
a. mixing bran, bean dregs, soybean protein and flour to obtain a dry material; adding water into the dry materials, and uniformly mixing to obtain a mixture; wherein the weight ratio of the dry materials to the water is 1: (1.2-1.5);
b. and (3) sterilizing the mixture at 110-130 ℃ for 30-40 min to obtain the culture medium for increasing the number of the soy sauce aspergillus spores.
8. A preparation method of soy sauce koji is characterized by comprising the following steps:
1) inoculating Aspergillus oryzae to a culture medium containing soybean and/or wheat, and culturing to obtain intermediate strain;
2) inoculating the intermediate seed to the culture medium for increasing the number of the soy sauce koji spores as claimed in any one of claims 1 to 3, culturing in a ventilation manner, and maturing to obtain the koji.
9. The method according to claim 8, wherein the culture medium is prepared by the following method in step 1): soaking semen glycines in water, decocting, filtering, collecting filtrate to obtain bean juice, adding magnesium sulfate, potassium dihydrogen phosphate, soluble starch and agar powder, dissolving, packaging into test tube, and sterilizing.
10. The method according to claim 8, wherein the intermediate species is cultured at a constant temperature of 20 to 30 ℃ for 1 to 5 days in step 2).
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Cited By (2)
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| CN114317287A (en) * | 2021-12-31 | 2022-04-12 | 佛山市海天(高明)调味食品有限公司 | Aspergillus oryzae fermentation solid material, aspergillus oryzae strain culture medium and preparation method and application thereof |
| CN115354011A (en) * | 2022-08-23 | 2022-11-18 | 山东巧媳妇食品集团有限公司 | Preparation method of soy sauce koji with high protease yield and stable enzyme activity |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114317287A (en) * | 2021-12-31 | 2022-04-12 | 佛山市海天(高明)调味食品有限公司 | Aspergillus oryzae fermentation solid material, aspergillus oryzae strain culture medium and preparation method and application thereof |
| CN114317287B (en) * | 2021-12-31 | 2023-09-15 | 佛山市海天(高明)调味食品有限公司 | Aspergillus oryzae fermentation solid material, aspergillus oryzae strain culture medium, and preparation method and application thereof |
| CN115354011A (en) * | 2022-08-23 | 2022-11-18 | 山东巧媳妇食品集团有限公司 | Preparation method of soy sauce koji with high protease yield and stable enzyme activity |
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