CN109576202B - Cultivation method for increasing viable count of probiotics by using traditional Chinese medicine additive - Google Patents

Cultivation method for increasing viable count of probiotics by using traditional Chinese medicine additive Download PDF

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CN109576202B
CN109576202B CN201811492949.2A CN201811492949A CN109576202B CN 109576202 B CN109576202 B CN 109576202B CN 201811492949 A CN201811492949 A CN 201811492949A CN 109576202 B CN109576202 B CN 109576202B
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于大海
李源博
刘红新
杨吉雨
谢里会
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Pu Sheng Tai Biotechnology Co ltd Of Jilin Province
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Abstract

The invention discloses a cultivation method for increasing the number of live bacteria of probiotics by using a traditional Chinese medicine additive, which comprises the following steps: step one, preparing grain culture medium raw materials using traditional Chinese medicine additives respectively according to parts by weight: 0.5-1.5 parts of a traditional Chinese medicine additive, 12-25 parts of corncobs, 10-25 parts of rice bran, 3-8 parts of bran, 5-10 parts of oat, 10-15 parts of buckwheat, 2-5 parts of sorghum rice, 2-5 parts of millet, 5-8 parts of coix seeds, 10-15 parts of soybeans, 10-15 parts of mung beans, 10-15 parts of black beans and 10-15 parts of soybeans; step two, putting the raw materials into a stirring pot in sequence, carrying out dry stirring, adding water with the temperature of 20-30 ℃ after all the raw materials are uniformly stirred, then carrying out stirring again, controlling the water content of the culture medium to be 88-93%, uniformly stirring, and then sterilizing at the temperature of 115 ℃ and under the pressure of 0.1MPa for 25-30 min to obtain a grain culture medium; step three, preparing seed liquid by reviving and expanding the probiotic strains to be cultured; and step four, inoculating the seed liquid into the grain culture medium for enrichment, adjusting the enrichment temperature to 37 ℃, and adjusting the enrichment culture time to 18-25 h.

Description

Cultivation method for increasing viable count of probiotics by using traditional Chinese medicine additive
Technical Field
The invention relates to the field of microbial fermentation, in particular to a cultivation method for increasing the number of live bacteria of probiotics by using a traditional Chinese medicine additive.
Background
Probiotics are a general term for living microorganisms which are fixedly planted in the intestinal tract and the reproductive system of a human body, can generate definite health efficacy so as to improve the micro-ecological balance of a host and have important effects on the health of the human body, and are widely applied to the production of foods, health care products and medicines. Research finds that the quantity of probiotics in human intestinal tract plays a decisive role in the health care and treatment efficacy of the probiotics, and besides factors such as proliferation of the probiotics in vivo, survival guarantee and biological activity maintenance of the probiotics during processing and storage, the sufficient quantity of viable bacteria is a necessary prerequisite for the probiotic supplement product to exert the effectiveness of the probiotic supplement product.
The plant grain raw material is used as a culture medium and is used as a fermentation substrate, and the grains can provide necessary carbon source and nitrogen source for the growth of probiotics and reduce the fermentation cost; the fermented grain can obviously improve the nutritive value and functional characteristics, and the aromatic components such as lactic acid, ethyl acetate, acetaldehyde and the like generated after fermentation can improve the flavor and aroma of grain products. The grains as the probiotic fermentation substrate can provide essential nutrient substances for growth of the probiotics and effectively proliferate the probiotics, but the grain culture mediums need to be subjected to complex pretreatment before fermentation culture, and the complex treatment processes comprise seed coat removal, cooking, enzymolysis and the like. The cereal puffing powder is prepared by puffing at high temperature and high pressure, and can reduce nutrition loss to the maximum extent in the curing process. The high temperature, high pressure and high shear force generated by the expansion can expand and essence starch particles in the grains, the starch molecule connection is opened, the surface area of the chyme particles is increased, and the digestibility of the starch is improved; meanwhile, the protein peptide chain is cracked into peptide and amino acid by puffing, so that the absorption rate of the protein is greatly improved. The puffed grains can effectively simplify the fermentation process and shorten the fermentation period, thereby reducing the fermentation cost.
Meanwhile, a plurality of foods, namely medicines, have no absolute boundary among the foods, a plurality of medicinal and edible traditional Chinese medicinal materials can well promote the number of viable bacteria of probiotics, and the properties of the probiotics can be better improved by taking plant grain raw materials as fermentation substrates of a culture medium and properly adding traditional Chinese medicine additives.
Disclosure of Invention
The invention designs and develops a cultivation method for improving the viable count of probiotics by using a traditional Chinese medicine additive, and aims to improve the maximum viable count remarkably by adding the traditional Chinese medicine additive into a culture medium in the cultivation process, reasonably controlling the addition amount of the traditional Chinese medicine additive, and accurately controlling the dry stirring time, the re-stirring time, the seed inoculation liquid and the enriched cultivation time according to the mass parts of raw materials in the preparation process.
The invention also aims to improve the maximum viable count after treating the particle size of the traditional Chinese medicine additive and the particle size of the corncobs.
The technical scheme provided by the invention is as follows:
a cultivation method for increasing the number of live bacteria of probiotics by using a traditional Chinese medicine additive comprises the following steps:
step one, preparing grain culture medium raw materials using traditional Chinese medicine additives respectively according to parts by weight:
0.5-1.5 parts of a traditional Chinese medicine additive, 12-25 parts of corncobs, 10-25 parts of rice bran, 3-8 parts of bran, 5-10 parts of oat, 10-15 parts of buckwheat, 2-5 parts of sorghum rice, 2-5 parts of millet, 5-8 parts of coix seeds, 10-15 parts of soybeans, 10-15 parts of mung beans, 10-15 parts of black beans and 10-15 parts of soybeans;
step two, putting the raw materials into a stirring pot in sequence, carrying out dry stirring, adding water with the temperature of 20-30 ℃ after all the raw materials are uniformly stirred, then carrying out stirring again, controlling the water content of the culture medium to be 88-93%, uniformly stirring, and then sterilizing at the temperature of 115 ℃ and under the pressure of 0.1MPa for 25-30 min to obtain a grain culture medium;
step three, preparing seed liquid by reviving and expanding the probiotic strains to be cultured;
inoculating the seed liquid into the grain culture medium for enrichment, adjusting the enrichment temperature to 37 ℃, and allowing the enrichment culture time to be 18-25 h;
the traditional Chinese medicine additive comprises the following components in parts by weight:
2-4 parts of pseudo-ginseng, 1-1.5 parts of salvia miltiorrhiza, 5-8 parts of Chinese yam, 10-13 parts of astragalus membranaceus, 15-25 parts of radix puerariae, 1-3 parts of radix paeoniae rubra, 1-3 parts of bighead atractylodes rhizome, 2-8 parts of cowberry fruit, 15-25 parts of honeysuckle and 10-18 parts of wolfberry fruit.
Preferably, the seed liquid inoculation ratio is
Figure BDA0001896125830000021
In the formula, WDIs the mass portion of the traditional Chinese medicine additive, WCIs the mass fraction of corncobs, WBIs the mass fraction of the bean product, WRIs the sum of the mass parts of sorghum rice and millet.
Preferably, in the fourth step, the enrichment culture time is
Figure BDA0001896125830000031
In the formula, WDIs the mass portion of the traditional Chinese medicine additive, WOTo removeThe other components of the traditional Chinese medicine additive are mixed in parts by weight, WBIs the mass fraction of the bean product, WRThe sum of the mass parts of sorghum rice and millet, t is the enrichment culture time, tmaxFor maximum time of enrichment culture, tminMinimum time for enrichment culture, teIn order to increase the time of the experience of the culture,
Figure BDA0001896125830000032
the correction coefficient is the time of enrichment culture, eta is the empirical time correction coefficient of enrichment culture, and e is the base number of natural logarithm.
Preferably, in the third step, the preparation process of the seed liquid is as follows:
adding the probiotic strains into an MRS culture medium according to the volume ratio of 1%, and performing static culture at 37 ℃ for 12 hours to recover the strains; inoculating resuscitation bacteria liquid into the MRS culture medium according to the volume proportion of 10%, and performing static culture at 37 ℃ for 12 hours to prepare probiotic bacteria seed liquid;
the MRS culture medium is sterilized for 30min at 115 ℃ and 0.1Mpa, and is naturally cooled for use after sterilization.
Preferably, the MRS culture medium comprises the following components in parts by weight:
20 parts of peptone, 10 parts of beef extract, 5 parts of yeast extract, 4 parts of diammonium hydrogen citrate, 20 parts of glucose, 803.2 parts of tween, 5 parts of sodium acetate, 2 parts of dipotassium hydrogen phosphate, 0.25 part of magnesium sulfate, 0.05 part of manganese sulfate, 18 parts of agar and 1000 parts of distilled water.
Preferably, in the second step, the dry stirring time is 30 to 50 minutes, and the re-stirring time is 80 to 100 minutes.
Preferably, in the second step, the dry stirring time is
Figure BDA0001896125830000033
In the formula, WCIs the mass fraction of corncobs, WBIs the mass fraction of the bean product, WRIs sorghum rice and milletThe sum of the parts by mass of rice, T1For dry stirring time, T1_maxFor maximum time of dry stirring, T1_minFor a minimum time of dry stirring, T1_eFor dry mixing empirical time, delta1Correction of the coefficient for dry stirring time, gamma1The coefficients were corrected for dry stirring empirical time.
Preferably, the re-stirring time is
Figure BDA0001896125830000041
In the formula, WCIs the mass fraction of corncobs, WBIs the mass fraction of the bean product, WRIs the sum of the mass fractions of sorghum rice and millet, T2For a further stirring time, T2_maxFor maximum time of restirring, T2_minFor minimum time of re-stirring, T2_eFor the empirical time of stirring, delta2To correct the coefficient for remixing time, gamma2The coefficients were corrected for empirical time for re-stirring.
Preferably, δ1The value is 1.43-1.55, gamma1The value is 0.23, delta2The value is 2.35-2.47, gamma2The value is 1.13.
It is preferable that the first and second liquid crystal layers are formed of,
Figure BDA0001896125830000042
the value of eta is 1.3-1.5, and the value of eta is 2.3.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention provides a culture method for improving the number of viable bacteria of probiotics by using a traditional Chinese medicine additive, wherein the culture medium provided by the invention is used for culturing the probiotics, and the maximum number of viable bacteria can be obviously improved by adding the traditional Chinese medicine additive, reasonably controlling the addition amount of the traditional Chinese medicine additive, and accurately controlling the dry stirring time, the re-stirring time, the seed inoculation liquid and the enriched culture time according to the mass parts of the raw materials;
2. the invention can improve the maximum viable count after processing the particle size of the traditional Chinese medicine additive and the particle size of the corncobs.
Detailed Description
The present invention is described in further detail below to enable those skilled in the art to practice the invention with reference to the description.
The invention provides a grain culture medium for increasing the number of live bacteria of probiotics by using a traditional Chinese medicine additive, which comprises the following components in parts by weight:
the traditional Chinese medicine composition comprises 12-25 parts of corncobs, 10-25 parts of rice bran, 3-8 parts of bran, 5-10 parts of oat, 10-15 parts of buckwheat, 2-5 parts of sorghum rice, 2-5 parts of millet, 5-8 parts of coix seeds, 10-15 parts of soybeans, 10-15 parts of mung beans, 10-15 parts of black beans, 10-15 parts of soybeans and 88-93% of water;
wherein the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000051
In the formula, WDIs the mass portion of the traditional Chinese medicine additive, WCIs the mass fraction of corncobs, WBIs the mass fraction of the bean product, WRThe sum of the mass parts of sorghum rice and millet, e is the base number of natural logarithm, and lambda is an empirical adjustment coefficient;
the traditional Chinese medicine additive comprises the following components in parts by weight:
2-4 parts of pseudo-ginseng, 1-1.5 parts of salvia miltiorrhiza, 5-8 parts of Chinese yam, 10-13 parts of astragalus membranaceus, 15-25 parts of radix puerariae, 1-3 parts of radix paeoniae rubra, 1-3 parts of bighead atractylodes rhizome, 2-8 parts of cowberry fruit, 15-25 parts of honeysuckle and 10-18 parts of wolfberry fruit.
The invention also provides a preparation method of the grain culture medium for improving the viable count of probiotics by using the traditional Chinese medicine additive, which comprises the following steps:
the method comprises the following steps: respectively preparing the raw materials of the grain culture medium according to the parts by weight:
the traditional Chinese medicine composition comprises, by weight, 10-25 parts of rice bran, 3-8 parts of bran, 5-10 parts of oat, 10-15 parts of buckwheat, 2-5 parts of sorghum rice, 2-5 parts of millet, 5-8 parts of coix seed, 10-15 parts of soybean, 10-15 parts of mung bean, 10-15 parts of black bean and 10-15 parts of soybean;
wherein the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000052
In the formula, WDIs the mass portion of the traditional Chinese medicine additive, WCIs the mass fraction of corncobs, WBIs the mass fraction of the bean product, WRThe sum of the mass parts of sorghum rice and millet, e is the base number of natural logarithm, and lambda is an empirical adjustment coefficient;
the traditional Chinese medicine additive comprises the following components in parts by weight:
2-4 parts of pseudo-ginseng, 1-1.5 parts of salvia miltiorrhiza, 5-8 parts of Chinese yam, 10-13 parts of astragalus membranaceus, 15-25 parts of radix puerariae, 1-3 parts of radix paeoniae rubra, 1-3 parts of bighead atractylodes rhizome, 2-8 parts of cowberry fruit, 15-25 parts of honeysuckle and 10-18 parts of wolfberry fruit.
Step two: the raw materials are sequentially put into a stirring pot after being puffed, dry stirring is carried out, all the raw materials are stirred uniformly, water with the temperature of 20-30 ℃ is added for stirring, the water content of the culture medium is controlled to 88-93% after the water is added, the culture medium is stirred uniformly to obtain a grain culture medium, and the grain culture medium is sterilized for 25-30 min at the temperature of 115 ℃ and the pressure of 0.1 MPa.
The invention also provides a cultivation method for improving the viable count of probiotics by using the traditional Chinese medicine additive, which comprises the following steps:
step 1, respectively preparing grain culture medium raw materials using traditional Chinese medicine additives in parts by weight:
the traditional Chinese medicine composition comprises 12-25 parts of corncobs, 10-25 parts of rice bran, 3-8 parts of bran, 5-10 parts of oat, 10-15 parts of buckwheat, 2-5 parts of sorghum rice, 2-5 parts of millet, 5-8 parts of coix seeds, 10-15 parts of soybeans, 10-15 parts of mung beans, 10-15 parts of black beans and 10-15 parts of soybeans;
wherein the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000061
In the formula, WDIs the mass portion of the traditional Chinese medicine additive, WCIs the mass fraction of corncobs, WBIs the mass fraction of the bean product, WRThe sum of the mass parts of sorghum rice and millet, e is the base number of natural logarithm, and lambda is an empirical adjustment coefficient;
the traditional Chinese medicine additive comprises the following components in parts by weight:
2-4 parts of pseudo-ginseng, 1-1.5 parts of salvia miltiorrhiza, 5-8 parts of Chinese yam, 10-13 parts of astragalus membranaceus, 15-25 parts of radix puerariae, 1-3 parts of radix paeoniae rubra, 1-3 parts of bighead atractylodes rhizome, 2-8 parts of cowberry fruit, 15-25 parts of honeysuckle and 10-18 parts of wolfberry fruit.
Step 2, putting the raw materials into a stirring pot in sequence after puffing treatment, carrying out dry stirring, adding water with the temperature of 20-30 ℃ after all the raw materials are uniformly stirred, then stirring, controlling the water content of the culture medium to 88-93% after the water is added, then uniformly stirring to obtain a grain culture medium, and sterilizing the grain culture medium for 25-30 min at the temperature of 115 ℃ and under the pressure of 0.1 MPa;
step 3, preparation of probiotic seed liquid: preparing a seed solution from the preserved probiotic strains after recovery and expansion, adding probiotic strains into an improved MRS culture medium in a proportion of 1% (v/v), and performing static culture at 37 ℃ for 12 hours to recover the strains; inoculating the resuscitating bacteria liquid into a modified MRS culture medium according to the proportion of 10% (v/v), performing static culture at 37 ℃ for 12 hours to prepare probiotic bacteria seed liquid, sterilizing the prepared modified lactic acid bacteria culture medium (modified MRS) at 115 ℃ and 0.1Mpa for 30min, and naturally cooling after sterilization, wherein the formula is shown in Table 1;
TABLE 1 modified MRS media
Figure BDA0001896125830000062
Figure BDA0001896125830000071
Step 4, enrichment culture of probiotics: inoculating the probiotic seed liquid obtained in the step 3 into the enrichment culture medium obtained in the step 2 for enrichment, wherein the enrichment culture temperature is 37 ℃.
In another embodiment, the probiotic bacterial strain is Lactobacillus plantarum, Lactobacillus bulgaricus, Lactobacillus casei, enterococcus faecalis, Lactobacillus salivarius, Lactobacillus rhamnosus, Lactobacillus fermentum
In another embodiment, the Chinese medicinal additive is processed to make the Chinese medicinal additive granules meet the following conditions: the proportion of particles with the mesh size of more than 120 meshes is 5-10%, the proportion of particles with the mesh size of 100-120 meshes is 30-35%, the proportion of particles with the mesh size of 80-100 meshes is 40-45%, and the balance is particles with the mesh size of less than 80 meshes.
In another embodiment, the corn cob particles are processed such that the corn cob particles satisfy the following conditions: the proportion of the particles is 1-3% when the particles are 7-9 mm, the proportion of the particles is 75-85% when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 5-8%.
In another embodiment, the dry stirring time is 30-50 minutes, the stirring time is 80-100 minutes, the inoculation proportion of the seed liquid to the enrichment culture medium is 3% (volume ratio, v/v), and the enrichment culture time is 18-25 hours.
In another embodiment, the dry stirring time is
Figure BDA0001896125830000081
The re-stirring time is
Figure BDA0001896125830000082
In the formula, WCIs the mass fraction of corncobs, WBIs the mass fraction of the bean product, WRIs the sum of the mass fractions of sorghum rice and millet, T1For dry stirring time, T1_maxFor maximum time of dry stirring, T1_minFor a minimum time of dry stirring, T1_eFor dry mixing empirical time, delta1Correction of the coefficient for dry stirring time, gamma1For dry-stirring empirical time correction factor, T2For a further stirring time, T2_maxFor maximum time of restirring, T2_minFor minimum time of re-stirring, T2_eFor the empirical time of stirring, delta2To correct the coefficient for remixing time, gamma2The coefficients were corrected for empirical time for re-stirring.
In another embodiment, the seed liquid inoculation ratio is
Figure BDA0001896125830000083
In the formula, WDIs the mass portion of the traditional Chinese medicine additive, WCIs the mass fraction of corncobs, WBIs the mass fraction of the bean product, WRIs the sum of the mass parts of sorghum rice and millet.
In another embodiment, the enrichment culture time is
Figure BDA0001896125830000084
In the formula, WDIs the mass portion of the traditional Chinese medicine additive, WOIn order to remove the mass parts of other components of the traditional Chinese medicine additive, WBIs the mass fraction of the bean product, WRThe sum of the mass parts of sorghum rice and millet, t is the enrichment culture time, tmaxFor maximum time of enrichment culture, tminMinimum time for enrichment culture, teIn order to increase the time of the experience of the culture,
Figure BDA0001896125830000085
the correction coefficient is the time of enrichment culture, eta is the empirical time correction coefficient of enrichment culture, and e is the base number of natural logarithm.
In another embodiment, λ is 0.38-0.45, δ1The value is 1.43-1.55, gamma1The value is 0.23, delta2The value is 2.35-2.47, gamma2The value of the additive is 1.13,
Figure BDA0001896125830000091
the value of eta is 1.3-1.5, and the value of eta is 2.3.
Example 1
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
12 parts of corncobs, 10 parts of rice bran, 3 parts of bran, 5 parts of oat, 10 parts of buckwheat, 2 parts of sorghum rice, 2 parts of millet, 5 parts of pearl barley, 10 parts of soybean, 10 parts of mung bean, 10 parts of black bean and 10 parts of soybean;
the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000092
Wherein, the value of lambda is 0.38;
the traditional Chinese medicine additive comprises the following components in parts by weight:
2 parts of pseudo-ginseng, 1 part of salvia miltiorrhiza, 5 parts of Chinese yam, 10 parts of astragalus membranaceus, 15 parts of kudzu root, 1 part of red paeony root, 1 part of bighead atractylodes rhizome, 2 parts of cowberry fruit, 15 parts of honeysuckle flower and 10 parts of medlar;
the traditional Chinese medicine additive is processed to ensure that the traditional Chinese medicine additive granules meet the following conditions: the proportion of particles with the mesh size of more than 120 meshes is 5 percent, the proportion of particles with the mesh size of 100-120 meshes is 30 percent, the proportion of particles with the mesh size of 80-100 meshes is 40 percent, and the balance is particles with the mesh size of less than 80 meshes;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 1 percent of 7-9 mm, the proportion of the particles is 75 percent of 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 5 percent;
the dry stirring time is
Figure BDA0001896125830000093
The re-stirring time is
Figure BDA0001896125830000094
In the formula, delta1The value is 1.43, gamma1The value is 0.23, delta2The value is 2.35, gamma2A value of 1.13, T1_minIs 30min, T1_maxIs 50min, T1_eIs 45min, T2_minIs 80min, T2_maxIs 100min, T2_eIs 95 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000101
The time of enrichment culture is
Figure BDA0001896125830000102
In the formula, tminIs 20h, tmaxIs 25h, teThe reaction time is 22.5h,
Figure BDA0001896125830000103
the value of eta is 1.3, and the value of eta is 2.3.
Example 2
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
25 parts of corncobs, 25 parts of rice bran, 8 parts of bran, 10 parts of oat, 15 parts of buckwheat, 5 parts of sorghum rice, 5 parts of millet, 8 parts of coix seeds, 15 parts of soybeans, 15 parts of mung beans, 15 parts of black beans and 15 parts of soybeans;
the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000104
In the formula, the value of lambda is 0.45;
the traditional Chinese medicine additive comprises the following components in parts by weight:
4 parts of pseudo-ginseng, 1.5 parts of salvia miltiorrhiza, 8 parts of Chinese yam, 13 parts of astragalus membranaceus, 15 parts of kudzu root, 3 parts of red paeony root, 3 parts of bighead atractylodes rhizome, 8 parts of cowberry fruit, 25 parts of honeysuckle flower and 18 parts of medlar;
the traditional Chinese medicine additive is processed to ensure that the traditional Chinese medicine additive granules meet the following conditions: the proportion of particles with the mesh size of more than 120 meshes is 10 percent, the proportion of particles with the mesh size of 100-120 meshes is 35 percent, the proportion of particles with the mesh size of 80-100 meshes is 45 percent, and the balance is particles with the mesh size of less than 80 meshes;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 3 percent when the particles are 7-9 mm, the proportion of the particles is 85 percent when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 8 percent;
the dry stirring time is
Figure BDA0001896125830000105
The re-stirring time is
Figure BDA0001896125830000111
In the formula, delta1Value of 1.55, gamma1The value is 0.23, delta2The value is 2.47, gamma2A value of 1.13, T1_minIs 30min, T1_maxIs 50min, T1_eIs 45min, T2_minIs 80min, T2_maxIs 100min, T2_eIs 95 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000112
The time of enrichment culture is
Figure BDA0001896125830000113
In the formula, tminIs 20h, tmaxIs 25h, teThe reaction time is 22.5h,
Figure BDA0001896125830000114
the value of eta is 1.5, and the value of eta is 2.3.
Example 3
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
18 parts of corncobs, 15 parts of rice bran, 5 parts of bran, 8 parts of oat, 13 parts of buckwheat, 3 parts of sorghum rice, 4 parts of millet, 7 parts of coix seeds, 13 parts of soybeans, 12 parts of mung beans, 13 parts of black beans and 12 parts of soybeans;
the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000115
In the formula, lambda is 0.4;
the traditional Chinese medicine additive comprises the following components in parts by weight:
3 parts of pseudo-ginseng, 1.3 parts of salvia miltiorrhiza, 6 parts of Chinese yam, 11 parts of astragalus membranaceus, 18 parts of kudzu root, 2 parts of red paeony root, 2 parts of bighead atractylodes rhizome, 5 parts of cowberry fruit, 21 parts of honeysuckle flower and 13 parts of wolfberry fruit;
the traditional Chinese medicine additive is processed to ensure that the traditional Chinese medicine additive granules meet the following conditions: 8 percent of particles with the mesh size of more than 120 meshes, 33 percent of particles with the mesh size of 100-120 meshes, 43 percent of particles with the mesh size of 80-100 meshes and the balance of particles with the mesh size of less than 80 meshes;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 2 percent when the particles are 7-9 mm, the proportion of the particles is 80 percent when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 7 percent;
the dry stirring time is
Figure BDA0001896125830000121
The re-stirring time is
Figure BDA0001896125830000122
In the formula, delta1Value of 1.5, gamma1The value is 0.23, delta2Value of 2.4, gamma2A value of 1.13, T1_minIs 30min, T1_maxIs 50min, T1_eIs 45min, T2_minIs 80min, T2_maxIs 100min, T2_eIs 95 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000123
The time of enrichment culture is
Figure BDA0001896125830000124
In the formula, tminIs 20h, tmaxIs 25h, teThe reaction time is 24 hours,
Figure BDA0001896125830000125
the value of eta is 1.45 and the value of eta is 2.3.
Comparative example 1
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
25 parts of corncobs, 25 parts of rice bran, 8 parts of bran, 10 parts of oat, 15 parts of buckwheat, 5 parts of sorghum rice, 5 parts of millet, 8 parts of coix seeds, 15 parts of soybeans, 15 parts of mung beans, 15 parts of black beans and 15 parts of soybeans;
the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000126
In the formula, the value of lambda is 0.45;
the traditional Chinese medicine additive comprises the following components in parts by weight:
4 parts of pseudo-ginseng, 1.5 parts of salvia miltiorrhiza, 8 parts of Chinese yam, 13 parts of astragalus membranaceus, 15 parts of kudzu root, 3 parts of red paeony root, 3 parts of bighead atractylodes rhizome, 8 parts of cowberry fruit, 25 parts of honeysuckle flower and 18 parts of medlar;
processing the traditional Chinese medicine additive to enable particles of the traditional Chinese medicine additive to meet the requirement of 80-100 meshes;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 3 percent when the particles are 7-9 mm, the proportion of the particles is 85 percent when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 8 percent;
the dry stirring time is
Figure BDA0001896125830000131
The re-stirring time is
Figure BDA0001896125830000132
In the formula, delta1Value of 1.55, gamma1The value is 0.23, delta2The value is 2.47, gamma2A value of 1.13, T1_minIs 30min, T1_maxIs 50min, T1_eIs 45min, T2_minIs 80min, T2_maxIs 100min, T2_eIs 95 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000133
The time of enrichment culture is
Figure BDA0001896125830000134
In the formula, tminIs 20h, tmaxIs 25h, teThe reaction time is 22.5h,
Figure BDA0001896125830000135
the value of eta is 1.5, and the value of eta is 2.3.
Comparative example 2
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
25 parts of corncobs, 25 parts of rice bran, 8 parts of bran, 10 parts of oat, 15 parts of buckwheat, 5 parts of sorghum rice, 5 parts of millet, 8 parts of coix seeds, 15 parts of soybeans, 15 parts of mung beans, 15 parts of black beans and 15 parts of soybeans;
the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000141
In the formula, the value of lambda is 0.45;
the traditional Chinese medicine additive comprises the following components in parts by weight:
4 parts of pseudo-ginseng, 1.5 parts of salvia miltiorrhiza, 8 parts of Chinese yam, 13 parts of astragalus membranaceus, 15 parts of kudzu root, 3 parts of red paeony root, 3 parts of bighead atractylodes rhizome, 8 parts of cowberry fruit, 25 parts of honeysuckle flower and 18 parts of medlar;
the traditional Chinese medicine additive is processed to ensure that the traditional Chinese medicine additive granules meet the following conditions: the proportion of particles with the mesh size of more than 120 meshes is 10 percent, the proportion of particles with the mesh size of 100-120 meshes is 35 percent, the proportion of particles with the mesh size of 80-100 meshes is 45 percent, and the balance is particles with the mesh size of less than 80 meshes;
processing the corncobs to ensure that the corncob particles are less than 9 mm;
the dry stirring time is
Figure BDA0001896125830000142
The re-stirring time is
Figure BDA0001896125830000143
In the formula, delta1Value of 1.55, gamma1The value is 0.23, delta2The value is 2.47, gamma2A value of 1.13, T1_minIs 30min, T1_maxIs 50min, T1_eIs 45min, T2_minIs 80min, T2_maxIs 100min, T2_eIs 95 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000144
The time of enrichment culture is
Figure BDA0001896125830000145
In the formula, tminIs 20h, tmaxIs 25h, teThe reaction time is 22.5h,
Figure BDA0001896125830000151
the value of eta is 1.5, and the value of eta is 2.3.
Comparative example 3
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
1.5 parts of traditional Chinese medicine additive, 25 parts of corncob, 25 parts of rice bran, 8 parts of bran, 10 parts of oat, 15 parts of buckwheat, 5 parts of sorghum rice, 5 parts of millet, 8 parts of pearl barley, 15 parts of soybean, 15 parts of mung bean, 15 parts of black bean and 15 parts of soybean;
the traditional Chinese medicine additive comprises the following components in parts by weight:
4 parts of pseudo-ginseng, 1.5 parts of salvia miltiorrhiza, 8 parts of Chinese yam, 13 parts of astragalus membranaceus, 15 parts of kudzu root, 3 parts of red paeony root, 3 parts of bighead atractylodes rhizome, 8 parts of cowberry fruit, 25 parts of honeysuckle flower and 18 parts of medlar;
the traditional Chinese medicine additive is processed to ensure that the traditional Chinese medicine additive granules meet the following conditions: the proportion of particles with the mesh size of more than 120 meshes is 10 percent, the proportion of particles with the mesh size of 100-120 meshes is 35 percent, the proportion of particles with the mesh size of 80-100 meshes is 45 percent, and the balance is particles with the mesh size of less than 80 meshes;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 3 percent when the particles are 7-9 mm, the proportion of the particles is 85 percent when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 8 percent;
the dry stirring time is
Figure BDA0001896125830000152
The re-stirring time is
Figure BDA0001896125830000153
In the formula, delta1Value of 1.55, gamma1The value is 0.23, delta2The value is 2.47, gamma2A value of 1.13, T1_minIs 30min, T1_maxIs 50min, T1_eIs 45min, T2_minIs 80min, T2_maxIs 100min, T2_eIs 95 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000154
The time of enrichment culture is
Figure BDA0001896125830000161
In the formula, tminIs 20h, tmaxIs 25h, teThe reaction time is 22.5h,
Figure BDA0001896125830000162
the value of eta is 1.5, and the value of eta is 2.3.
Comparative example 4
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
25 parts of corncobs, 25 parts of rice bran, 8 parts of bran, 10 parts of oat, 15 parts of buckwheat, 5 parts of sorghum rice, 5 parts of millet, 8 parts of coix seeds, 15 parts of soybeans, 15 parts of mung beans, 15 parts of black beans and 15 parts of soybeans;
the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000163
In the formula, the value of lambda is 0.45;
the traditional Chinese medicine additive comprises the following components in parts by weight:
4 parts of pseudo-ginseng, 1.5 parts of salvia miltiorrhiza, 8 parts of Chinese yam, 13 parts of astragalus membranaceus, 15 parts of kudzu root, 3 parts of red paeony root, 3 parts of bighead atractylodes rhizome, 8 parts of cowberry fruit, 25 parts of honeysuckle flower and 18 parts of medlar;
the traditional Chinese medicine additive is processed to ensure that the traditional Chinese medicine additive granules meet the following conditions: the proportion of particles with the mesh size of more than 120 meshes is 10 percent, the proportion of particles with the mesh size of 100-120 meshes is 35 percent, the proportion of particles with the mesh size of 80-100 meshes is 45 percent, and the balance is particles with the mesh size of less than 80 meshes;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 3 percent when the particles are 7-9 mm, the proportion of the particles is 85 percent when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 8 percent;
the dry stirring time is 30 minutes;
the re-stirring time is
Figure BDA0001896125830000164
In the formula, delta2The value is 2.47, gamma2A value of 1.13, T2_minIs 80min, T2_maxIs 100min, T2_eIs 95 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000171
The time of enrichment culture is
Figure BDA0001896125830000172
In the formula, tminIs 20h, tmaxIs 25h, teThe reaction time is 22.5h,
Figure BDA0001896125830000173
the value of eta is 1.5, and the value of eta is 2.3.
Comparative example 5
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
25 parts of corncobs, 25 parts of rice bran, 8 parts of bran, 10 parts of oat, 15 parts of buckwheat, 5 parts of sorghum rice, 5 parts of millet, 8 parts of coix seeds, 15 parts of soybeans, 15 parts of mung beans, 15 parts of black beans and 15 parts of soybeans;
the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000174
In the formula, the value of lambda is 0.45;
the traditional Chinese medicine additive comprises the following components in parts by weight:
4 parts of pseudo-ginseng, 1.5 parts of salvia miltiorrhiza, 8 parts of Chinese yam, 13 parts of astragalus membranaceus, 15 parts of kudzu root, 3 parts of red paeony root, 3 parts of bighead atractylodes rhizome, 8 parts of cowberry fruit, 25 parts of honeysuckle flower and 18 parts of medlar;
the traditional Chinese medicine additive is processed to ensure that the traditional Chinese medicine additive granules meet the following conditions: the proportion of particles with the mesh size of more than 120 meshes is 10 percent, the proportion of particles with the mesh size of 100-120 meshes is 35 percent, the proportion of particles with the mesh size of 80-100 meshes is 45 percent, and the balance is particles with the mesh size of less than 80 meshes;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 3 percent when the particles are 7-9 mm, the proportion of the particles is 85 percent when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 8 percent;
dry stirring time was 50 minutes;
the re-stirring time is
Figure BDA0001896125830000181
In the formula, delta2The value is 2.47, gamma2A value of 1.13, T2_minIs 80min, T2_maxIs 100min, T2_eIs 95 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000182
The time of enrichment culture is
Figure BDA0001896125830000183
In the formula, tminIs 20h, tmaxIs 25h, teThe reaction time is 22.5h,
Figure BDA0001896125830000184
the value of eta is 1.5, and the value of eta is 2.3.
Comparative example 6
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
25 parts of corncobs, 25 parts of rice bran, 8 parts of bran, 10 parts of oat, 15 parts of buckwheat, 5 parts of sorghum rice, 5 parts of millet, 8 parts of coix seeds, 15 parts of soybeans, 15 parts of mung beans, 15 parts of black beans and 15 parts of soybeans;
the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000185
In the formula, the value of lambda is 0.45;
the traditional Chinese medicine additive comprises the following components in parts by weight:
4 parts of pseudo-ginseng, 1.5 parts of salvia miltiorrhiza, 8 parts of Chinese yam, 13 parts of astragalus membranaceus, 15 parts of kudzu root, 3 parts of red paeony root, 3 parts of bighead atractylodes rhizome, 8 parts of cowberry fruit, 25 parts of honeysuckle flower and 18 parts of medlar;
the traditional Chinese medicine additive is processed to ensure that the traditional Chinese medicine additive granules meet the following conditions: the proportion of particles with the mesh size of more than 120 meshes is 10 percent, the proportion of particles with the mesh size of 100-120 meshes is 35 percent, the proportion of particles with the mesh size of 80-100 meshes is 45 percent, and the balance is particles with the mesh size of less than 80 meshes;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 3 percent when the particles are 7-9 mm, the proportion of the particles is 85 percent when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 8 percent;
the dry stirring time is
Figure BDA0001896125830000191
Stirring for 80 minutes;
in the formula, delta1Value of 1.55, gamma1Value of 0.23, T1_minIs 30min, T1_maxIs 50min, T1_eIs 45 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000192
The time of enrichment culture is
Figure BDA0001896125830000193
In the formula, tminIs 20h, tmaxIs 25h, teThe reaction time is 22.5h,
Figure BDA0001896125830000194
the value of eta is 1.5, and the value of eta is 2.3.
Comparative example 7
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
25 parts of corncobs, 25 parts of rice bran, 8 parts of bran, 10 parts of oat, 15 parts of buckwheat, 5 parts of sorghum rice, 5 parts of millet, 8 parts of coix seeds, 15 parts of soybeans, 15 parts of mung beans, 15 parts of black beans and 15 parts of soybeans;
the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000195
In the formula, the value of lambda is 0.45;
the traditional Chinese medicine additive comprises the following components in parts by weight:
4 parts of pseudo-ginseng, 1.5 parts of salvia miltiorrhiza, 8 parts of Chinese yam, 13 parts of astragalus membranaceus, 15 parts of kudzu root, 3 parts of red paeony root, 3 parts of bighead atractylodes rhizome, 8 parts of cowberry fruit, 25 parts of honeysuckle flower and 18 parts of medlar;
the traditional Chinese medicine additive is processed to ensure that the traditional Chinese medicine additive granules meet the following conditions: the proportion of particles with the mesh size of more than 120 meshes is 10 percent, the proportion of particles with the mesh size of 100-120 meshes is 35 percent, the proportion of particles with the mesh size of 80-100 meshes is 45 percent, and the balance is particles with the mesh size of less than 80 meshes;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 3 percent when the particles are 7-9 mm, the proportion of the particles is 85 percent when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 8 percent;
the dry stirring time is
Figure BDA0001896125830000201
Stirring for 100 minutes;
in the formula, delta1Value of 1.55, gamma1Value of 0.23, T1_minIs 30min, T1_maxIs 50min, T1_eIs 45 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000202
The time of enrichment culture is
Figure BDA0001896125830000203
In the formula, tminIs 20h, tmaxIs 25h, teThe reaction time is 22.5h,
Figure BDA0001896125830000204
the value of eta is 1.5, and the value of eta is 2.3.
Comparative example 8
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
25 parts of corncobs, 25 parts of rice bran, 8 parts of bran, 10 parts of oat, 15 parts of buckwheat, 5 parts of sorghum rice, 5 parts of millet, 8 parts of coix seeds, 15 parts of soybeans, 15 parts of mung beans, 15 parts of black beans and 15 parts of soybeans;
the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000205
In the formula, the value of lambda is 0.45;
the traditional Chinese medicine additive comprises the following components in parts by weight:
4 parts of pseudo-ginseng, 1.5 parts of salvia miltiorrhiza, 8 parts of Chinese yam, 13 parts of astragalus membranaceus, 15 parts of kudzu root, 3 parts of red paeony root, 3 parts of bighead atractylodes rhizome, 8 parts of cowberry fruit, 25 parts of honeysuckle flower and 18 parts of medlar;
the traditional Chinese medicine additive is processed to ensure that the traditional Chinese medicine additive granules meet the following conditions: the proportion of particles with the mesh size of more than 120 meshes is 10 percent, the proportion of particles with the mesh size of 100-120 meshes is 35 percent, the proportion of particles with the mesh size of 80-100 meshes is 45 percent, and the balance is particles with the mesh size of less than 80 meshes;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 3 percent when the particles are 7-9 mm, the proportion of the particles is 85 percent when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 8 percent;
the dry stirring time is
Figure BDA0001896125830000211
The re-stirring time is
Figure BDA0001896125830000212
In the formula, delta1Value of 1.55, gamma1The value is 0.23, delta2The value is 2.47, gamma2A value of 1.13, T1_minIs 30min, T1_maxIs 50min, T1_eIs 45min, T2_minIs 80min, T2_maxIs 100min, T2_eIs 95 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation proportion of the seed liquid added to the enrichment medium is 3 percent (volume ratio, v/v);
the time of enrichment culture is
Figure BDA0001896125830000213
In the formula, tminIs 20h, tmaxIs 25h, teThe reaction time is 22.5h,
Figure BDA0001896125830000214
the value of eta is 1.5, and the value of eta is 2.3.
Comparative example 9
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
25 parts of corncobs, 25 parts of rice bran, 8 parts of bran, 10 parts of oat, 15 parts of buckwheat, 5 parts of sorghum rice, 5 parts of millet, 8 parts of coix seeds, 15 parts of soybeans, 15 parts of mung beans, 15 parts of black beans and 15 parts of soybeans;
the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000221
In the formula, the value of lambda is 0.45;
the traditional Chinese medicine additive comprises the following components in parts by weight:
4 parts of pseudo-ginseng, 1.5 parts of salvia miltiorrhiza, 8 parts of Chinese yam, 13 parts of astragalus membranaceus, 15 parts of kudzu root, 3 parts of red paeony root, 3 parts of bighead atractylodes rhizome, 8 parts of cowberry fruit, 25 parts of honeysuckle flower and 18 parts of medlar;
the traditional Chinese medicine additive is processed to ensure that the traditional Chinese medicine additive granules meet the following conditions: the proportion of particles with the mesh size of more than 120 meshes is 10 percent, the proportion of particles with the mesh size of 100-120 meshes is 35 percent, the proportion of particles with the mesh size of 80-100 meshes is 45 percent, and the balance is particles with the mesh size of less than 80 meshes;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 3 percent when the particles are 7-9 mm, the proportion of the particles is 85 percent when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 8 percent;
the dry stirring time is
Figure BDA0001896125830000222
The re-stirring time is
Figure BDA0001896125830000223
In the formula, delta1Value of 1.55, gamma1The value is 0.23, delta2The value is 2.47, gamma2A value of 1.13, T1_minIs 30min, T1_maxIs 50min, T1_eIs 45min, T2_minIs 80min, T2_maxIs 100min, T2_eIs 95 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000224
The time for enrichment culture is 18 h.
Comparative example 10
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
25 parts of corncobs, 25 parts of rice bran, 8 parts of bran, 10 parts of oat, 15 parts of buckwheat, 5 parts of sorghum rice, 5 parts of millet, 8 parts of coix seeds, 15 parts of soybeans, 15 parts of mung beans, 15 parts of black beans and 15 parts of soybeans;
the traditional Chinese medicine additive comprises the following components in parts by weight
Figure BDA0001896125830000231
In the formula, the value of lambda is 0.45;
the traditional Chinese medicine additive comprises the following components in parts by weight:
4 parts of pseudo-ginseng, 1.5 parts of salvia miltiorrhiza, 8 parts of Chinese yam, 13 parts of astragalus membranaceus, 15 parts of kudzu root, 3 parts of red paeony root, 3 parts of bighead atractylodes rhizome, 8 parts of cowberry fruit, 25 parts of honeysuckle flower and 18 parts of medlar;
the traditional Chinese medicine additive is processed to ensure that the traditional Chinese medicine additive granules meet the following conditions: the proportion of particles with the mesh size of more than 120 meshes is 10 percent, the proportion of particles with the mesh size of 100-120 meshes is 35 percent, the proportion of particles with the mesh size of 80-100 meshes is 45 percent, and the balance is particles with the mesh size of less than 80 meshes;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 3 percent when the particles are 7-9 mm, the proportion of the particles is 85 percent when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 8 percent;
the dry stirring time is
Figure BDA0001896125830000232
The re-stirring time is
Figure BDA0001896125830000233
In the formula, delta1Value of 1.55, gamma1The value is 0.23, delta2The value is 2.47, gamma2A value of 1.13, T1_minIs 30min, T1_maxIs 50min, T1_eIs 45min, T2_minIs 80min, T2_maxIs 100min, T2_eIs 95 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000241
The time for enrichment culture is 25 h.
Comparative example 11
Preparing grain culture medium raw materials using traditional Chinese medicine additives according to parts by weight:
25 parts of corncobs, 25 parts of rice bran, 8 parts of bran, 10 parts of oat, 15 parts of buckwheat, 5 parts of sorghum rice, 5 parts of millet, 8 parts of coix seeds, 15 parts of soybeans, 15 parts of mung beans, 15 parts of black beans and 15 parts of soybeans;
processing the corncobs to enable the corncob particles to meet the following conditions: the proportion of the particles is 3 percent when the particles are 7-9 mm, the proportion of the particles is 85 percent when the particles are 3-7 mm, the balance is particles with the particle size less than 3mm, and the water content of the corncob raw material is 8 percent;
the dry stirring time is
Figure BDA0001896125830000242
The re-stirring time is
Figure BDA0001896125830000243
In the formula, delta1Value of 1.55, gamma1The value is 0.23, delta2The value is 2.47, gamma2A value of 1.13, T1_minIs 30min, T1_maxIs 50min, T1_eIs 45min, T2_minIs 80min, T2_maxIs 100min, T2_eIs 95 min;
preparing seed liquid by using lactobacillus plantarum;
the inoculation ratio of the seed liquid to the enrichment culture medium is
Figure BDA0001896125830000244
The time of enrichment culture is
Figure BDA0001896125830000251
In the formula, tminIs 20h, tmaxIs 25h, teThe reaction time is 22.5h,
Figure BDA0001896125830000252
the value of eta is 1.5, and the value of eta is 2.3.
Test examples
And (3) determining the viable count of the examples 1-3 and the comparative examples 1-11, and determining the number of the probiotics in the fermentation liquor by adopting a flow cytometry counting method to obtain the viable count and the dead count of the probiotics in the grain fermentation liquor. The flow cytometry was performed as follows:
step 1, sample preparation, namely properly diluting the taken fermentation liquor to ensure that the number range of viable bacteria in the to-be-detected bacteria liquid sample is 5.5 multiplied by 105～6.5×105CFU/mL. Filtering the diluted fermentation liquid sample by a 300-mesh gauze, adding 200 mu L of the diluted sample to be detected into a 1.5mL EP tube, and carrying out warm bath in a constant-temperature incubator at 37 ℃ for 3 minutes;
and 2, dyeing the sample, namely adding 2 mu L of 100 XSybr Green I and 2 mu L of 100 XPI into 200 mu L of the sample to be detected, and dyeing for 10 minutes at 37 ℃ in a dark place. Sybr Green I and pi (propidium iodide) are two common DNA fluorescent dyes. Sybr Green I penetrates the cell membrane of live and dead cells, whereas PI does not penetrate the cell membrane of live cells well. Using Sybr Green I and PI to stain bacteria, the bacteria can be divided into three groups: viable bacteria of Sybr Green I (+) PI (-), dead bacteria of Sybr Green I (+) PI (+), and damaging bacteria between the two populations;
step 3, C6Accuri absolute counting, analyzing samples by a C6Accuri flow cytometer, setting a threshold value to be FSC-H larger than 10000, selecting slow flow, setting the sampling amount to be 60 mu L, and carrying out absolute counting detection;
and 4, drawing a cell density map by taking FL1-H as an abscissa and FL2-A as an ordinate. Analyzing the density map of the water sample, the bacterial liquid and the heated bacterial liquid, grouping cells according to the samples, and counting by enclosing a gate;
as shown in Table 2, the culture medium treated by the present invention was tested for viable bacteria, and the maximum viable bacteria counts of examples 1 to 3 were 5.5X 108CFU/mL or more, the maximum viable count of 6.1X 10 was reached using the medium of example 28CFU/mL is obviously improved compared with a culture medium (comparative example 11) without adding traditional Chinese medicine additives, and is also obviously improved compared with a culture medium (comparative example 1) which is simply processed by traditional Chinese medicines, a culture medium (comparative example 2) which is not processed by corncobs in an untreated way, a culture medium (comparative example 3) which is not processed by traditional Chinese medicine additives in a precise way according to the mass parts of raw materials in a dry stirring time, a culture medium (comparative examples 4 and 5) which is not processed by the traditional Chinese medicine additives in a precise way according to the mass parts of the raw materials in a dry stirring time, a culture medium (comparative examples 6 and 7) which is not processed by the traditional Chinese medicine additives in a precise way after water is added in a stirring time, a culture medium (comparative example 8) which is not processed by the traditional Chinese medicine additives in a precise way according to the mass parts of the raw materials in.
TABLE 2 maximum viable count
Examples Maximum viable count (CFU/mL)
Example 1 5.5×108
Practice ofExample 2 6.1×108
Example 3 5.8×108
Comparative example 1 4.4×108
Comparative example 2 4.3×108
Comparative example 3 5.0×108
Comparative example 4 2.4×108
Comparative example 5 3.2×108
Comparative example 6 3.5×108
Comparative example 7 3.1×108
Comparative example 8 2.8×108
Comparative example 9 4.0×108
Comparative example 10 4.1×108
Comparative example 11 1.3×108
While embodiments of the invention have been disclosed above, it is not limited to the applications listed in the description and the embodiments, which are fully applicable in all kinds of fields of application of the invention, and further modifications may readily be effected by those skilled in the art, and the invention is thus not limited to the specific details without departing from the generic concept defined in the claims and the equivalent scope.

Claims (5)

1. A cultivation method for increasing the number of live bacteria of probiotics by using a traditional Chinese medicine additive is characterized by comprising the following steps:
step one, preparing grain culture medium raw materials using traditional Chinese medicine additives respectively according to parts by weight:
0.5-1.5 parts of a traditional Chinese medicine additive, 12-25 parts of corncobs, 10-25 parts of rice bran, 3-8 parts of bran, 5-10 parts of oat, 10-15 parts of buckwheat, 2-5 parts of sorghum rice, 2-5 parts of millet, 5-8 parts of coix seeds, 10-15 parts of soybeans, 10-15 parts of mung beans, 10-15 parts of black beans and 10-15 parts of soybeans;
step two, putting the raw materials into a stirring pot in sequence, carrying out dry stirring, adding water with the temperature of 20-30 ℃ after all the raw materials are uniformly stirred, then carrying out stirring again, controlling the water content of the culture medium to be 88-93%, uniformly stirring, and then sterilizing at the temperature of 115 ℃ and under the pressure of 0.1MPa for 25-30 min to obtain a grain culture medium;
step three, preparing seed liquid by reviving and expanding the probiotic strains to be cultured;
inoculating the seed liquid into the grain culture medium for enrichment, adjusting the enrichment temperature to 37 ℃, and adjusting the enrichment culture time to 18-25 h;
the traditional Chinese medicine additive comprises the following components in parts by weight:
2-4 parts of pseudo-ginseng, 1-1.5 parts of salvia miltiorrhiza, 5-8 parts of Chinese yam, 10-13 parts of astragalus membranaceus, 15-25 parts of kudzu root, 1-3 parts of red paeony root, 1-3 parts of bighead atractylodes rhizome, 2-8 parts of cowberry fruit, 15-25 parts of honeysuckle and 10-18 parts of wolfberry fruit;
the seed liquid is inoculated according to the proportion of
Figure FDA0002708563970000011
In the formula, WDIs the mass portion of the traditional Chinese medicine additive, WCIs the mass fraction of corncobs, WBIs the mass fraction of the bean product, WRThe sum of the mass parts of sorghum rice and millet;
in the fourth step, the enriched culture time is
Figure FDA0002708563970000012
In the formula, WDIs the mass portion of the traditional Chinese medicine additive, WOIn order to remove the mass parts of other components of the traditional Chinese medicine additive, WBIs the mass fraction of the bean product, WRThe sum of the mass parts of sorghum rice and millet, t is the enrichment culture time, tmaxFor maximum time of enrichment culture, tminMinimum time for enrichment culture, teIn order to increase the time of the experience of the culture,
Figure FDA0002708563970000021
the correction coefficient is the enrichment culture time, eta is the enrichment culture experience time correction coefficient, and e is the base number of the natural logarithm;
in the second step, the dry stirring time is
Figure FDA0002708563970000022
In the formula, WCThe weight portion of the corncob is,WBis the mass fraction of the bean product, WRIs the sum of the mass fractions of sorghum rice and millet, T1For dry stirring time, T1_maxFor maximum time of dry stirring, T1_minFor a minimum time of dry stirring, T1_eFor dry mixing empirical time, delta1Correction of the coefficient for dry stirring time, gamma1The time correction factor is a dry stirring empirical time;
the re-stirring time is
Figure FDA0002708563970000023
In the formula, WCIs the mass fraction of corncobs, WBIs the mass fraction of the bean product, WRIs the sum of the mass fractions of sorghum rice and millet, T2For a further stirring time, T2_maxFor maximum time of restirring, T2_minFor minimum time of re-stirring, T2_eFor the empirical time of stirring, delta2To correct the coefficient for remixing time, gamma2The coefficients were corrected for empirical time for re-stirring.
2. The cultivation method for increasing the viable count of probiotics by using the traditional Chinese medicine additive according to claim 1, wherein in the third step, the preparation process of the seed liquid comprises the following steps:
adding the probiotic strains into an MRS culture medium according to the volume proportion of 1%, and performing static culture at 37 ℃ for 12 hours to recover the strains; inoculating resuscitation bacteria liquid into the MRS culture medium according to the volume proportion of 10%, and performing static culture at 37 ℃ for 12 hours to prepare probiotic bacteria seed liquid;
the MRS culture medium is sterilized for 30min at 115 ℃ and 0.1Mpa, and is naturally cooled for use after sterilization.
3. The cultivation method for increasing the viable count of probiotics by using the traditional Chinese medicine additive according to claim 2, wherein the MRS culture medium comprises the following components in parts by weight:
20 parts of peptone, 10 parts of beef extract, 5 parts of yeast extract, 4 parts of diammonium hydrogen citrate, 20 parts of glucose, 803.2 parts of tween, 5 parts of sodium acetate, 2 parts of dipotassium hydrogen phosphate, 0.25 part of magnesium sulfate, 0.05 part of manganese sulfate, 18 parts of agar and 1000 parts of distilled water.
4. A cultivation method using Chinese herbal medicine additive to increase the viable count of probiotics as claimed in claim 3, wherein δ1The value is 1.43-1.55, gamma1The value is 0.23, delta2The value is 2.35-2.47, gamma2The value is 1.13.
5. The cultivation method for increasing the viable count of probiotics by using the Chinese medicinal additive according to claim 4,
Figure FDA0002708563970000031
the value of eta is 1.3-1.5, and the value of eta is 2.3.
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