CN104531575A - Bacillus coagulans solid-state fermentation production method - Google Patents
Bacillus coagulans solid-state fermentation production method Download PDFInfo
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Abstract
The invention relates to the field of microbial fermentation, and in particular relates to a bacillus coagulans solid-state fermentation production method. The method comprises the following steps: (1) first-stage seed preparation; (2) second-stage solid-state seed inoculation; (3) third-stage solid-state seed inoculation; and (4) solid-state fermentation inoculation, wherein a formula of a solid-state fermentation material comprises the following components in percentage by mass: 70-99% of wheat bran, 1-30% of soybean meal, 0.1-10% of glucose, 0.1-10% of urea, 0.1-10% of calcium carbonate and 0.1-5% of manganese sulfate, and accessory materials are added corresponding to the mass of the wheat bran and the soybean meal. According to the method disclosed by the invention, the solid-state fermentation inoculation quantity can be reduced by 50-95% compared with that of the prior art, and a pH value is kept between 7 and 8 in a fermentation process; moreover, a high bacterium number can be achieved, dominant populations can be formed for fermentation, bacterial contamination cannot be easily caused, relatively high spore numbers and spore rates can be achieved, the spore number is 1*10<9>CFU/g to 1*10<11>CFU/g, and the maximum spore rate can reach 99%; and the method is easy in production and promotion.
Description
Technical field
The present invention relates to field of microbial fermentation, be specifically related to a kind of Bacillus coagulans state fermentation production method.
Background technology
Bacillus coagulans (Bacillus coagulans) has been put into new feedstuff in the protection period and new feed additive in No. 1126th, The Ministry of Agriculture of the People's Republic of China, MOA bulletin " fodder additives kind catalogue " (2008); it ratifies duration of service is in May, 2004; the scope of application is broiler and growing-finishing pig; illustrate that its security, validity, economy and the impact on environment have passed the evaluation of national feed evaluation committee, and obtain Ministry of Agriculture's approval.U.S. FDA has also approved that it is the bacillus class of " generally believing safety ", and the applied research of this genus bacillus on the mankind and animal productiong becomes focus, and Be very effective.
Abuse of antibiotics can produce a series of drawback, as destroyed intestinal microecology balance, producing Resistant strain, causing livestock product drug residue, the health of the serious threat mankind and livestock and poultry.Therefore, the country such as European Union has completely forbidden the use of antibiotic feed additive in feed.Recent study shows, probiotic bacterium (Prob-iotics) has maintenance microbial population of animal intestinal tract balance, improves animal intestinal function, improves the action effect such as immunity of organisms and production performance, and the various drawbacks that use microbiotic can be avoided to bring, be widely used in the fields such as food, medical treatment, livestock and poultry and aquaculture.Bacillus coagulans (Bacillus coagulans) is a kind of probiotic bacterium that can produce lactic acid, have the advantage of genus bacillus and milk-acid bacteria concurrently, and strong stress resistance, animal intestinal micro-ecology can be regulated to balance, promote animal digestion and improve animal immunizing power etc., because being applied in fodder industry.
Though Bacillus coagulans is improved the plurality of advantages such as breeding performonce fo animals, but its action effect is not lumped together, the stability etc. of different manufacturer, different viable bacteria content, different feeding animal, different operational phase and preparation all affects its action effect, the stability wherein improving gemma number, gemma rate and preparation is the gordian technique of fermentative production Bacillus coagulans, is also the important factor playing its biological action.The coagulated bacillus living number reported at present can reach 10
9~ 10
11cfu/mL, but its spore forming rate is not high.The people such as people and Liu Xinlei such as Qi Wei use tubular fibre membrane filter method and High-density cultivation of Bacillus coagulans using fed-batch method respectively, and gemma rate is respectively 75% and 26.7%.
In recent years, probiotics flourish, a collection of Bacillus coagulans manufacturer and research worker are emerged in large numbers, but the current mode of production both domestic and external mostly adopts liquid fermentation process, but there is shortcomings in liquid fermenting, as long in fermentation period, spore forming rate is low, aftertreatment sewage discharge is serious, high in cost of production problem, but state fermentation production method, gemma number and gemma rate higher, technological process is simple, less investment, processing ease, the features such as aftertreatment is simple, are accepted widely and are studied.Be directed to the industrialization of Bacillus coagulans, Chinese scholars is furtherd investigate Bacillus coagulans liquid fermenting high-density culture, think that condensation bud pole bacterium accumulates will pH value be caused to decline and produce strong restraining effect to the formation of thalli growth and gemma through the lactic acid that homofermentation generates in fermented liquid, for solid state fermentation, the problem that same existence is such, how more effective raising solid state fermentation Bacillus coagulans spore number and gemma rate.
Summary of the invention
The object of the invention is the state fermentation production method putting forward a kind of high yield Bacillus coagulans spore number and gemma rate.
Method according to the present invention comprises the following steps:
(1) first order seed preparation: by the inoculum size of 1 ~ 10%, cultivates in access YPD liquid nutrient medium, is primary seed solution.
(2) the solid-state seed inoculation method of secondary: primary seed solution is adjusted pH to 6.0 ~ 9.0, the inoculum size by 10% ~ 90% mixes to mix thoroughly with solid-state material cultivates.
(3) three grades of solid-state seed inoculation methods: by the inoculum size of 1% ~ 50%, get the solid-state seed of cultured secondary in sterilized water, stir, then pour in solid-state material to mix thoroughly and cultivate.
(4) solid state fermentation inoculation method: by the inoculum size of 1% ~ 50%, gets cultured three grades of solid-state seeds in sterilized water, stirs, then pour in solid-state material to mix thoroughly and cultivate,
The formula of wherein said solid-state fermentation material comprises in mass ratio: (1) major ingredient, 70 ~ 99% wheat brans, 1 ~ 30% dregs of beans; (2) auxiliary material, the quality based on major ingredient comprises 0.1 ~ 10% glucose, 0.1 ~ 10% urea, 0.1 ~ 10% calcium carbonate, 0.1 ~ 5% manganous sulfate.
According to the specific embodiment of the present invention, the state fermentation production method of high yield Bacillus coagulans spore number of the present invention and gemma rate comprises step:
(1) first order seed preparation: glycerine pipe preservation of bacteria strain one, by the inoculum size of 1 ~ 10%, gets 0.1 ~ 1ml bacterium liquid, in access YPD liquid nutrient medium, 40 DEG C, 150r/min cultivates 15 ~ 24h, is primary seed solution.
(2) the solid-state seed culture medium preparation of secondary: take wheat bran, dregs of beans in proportion, by 0.1 ~ 10% glucose, 0.1 ~ 10% urea, 0.1 ~ 10% calcium carbonate, be dissolved in 20 ~ 50% (to solid-state siccative meter) 50 ~ 60 DEG C of hot water, pour in the solid-state material weighed up and mix thoroughly, 121 DEG C, sterilizing 50min.
(3) secondary solid-state seed inoculation method: sterilized solid-state material is naturally cooled to about 50 DEG C, simultaneously by plastics film, the sterilizing of solid state fermentation sterilizing containers, pours in container after solid-state material cooling; Use sterilizing 1 ~ 50% sodium carbonate, primary seed solution is adjusted pH to 6.0 ~ 9.0, the inoculum size by 10% ~ 90% mixes with solid-state material mixes thoroughly, is placed between 40 ~ 50 DEG C of cultivations and cultivates 15 ~ 24h.
(4) three grades of solid-state seed culture medium preparations: take wheat bran, dregs of beans in proportion, by 0.1 ~ 10% glucose, 0.1 ~ 10% urea, 0.1 ~ 10% calcium carbonate, be dissolved in 20 ~ 80% (to solid-state siccative meter) 50 ~ 60 DEG C of hot water, pour in the solid-state material weighed up and mix thoroughly, 121 DEG C, sterilizing 50min.
(5) three grades of solid-state seed inoculation methods: sterilized solid-state material is naturally cooled to about 50 DEG C, simultaneously by plastics film, the sterilizing of solid state fermentation sterilizing containers, pour in container after solid-state material cooling; Inoculation by 1% ~ 50% measures the solid-state seed of cultured secondary, pour in sterilizing bucket, 20 ~ 80% (to solid-state siccative meters) add sterilized water in proportion, stir, pour in solid-state material and mix thoroughly, be placed between 40 ~ 50 DEG C of cultivations and cultivate 15 ~ 24h.
Wherein, solid-state fermentation material is prepared: take wheat bran, dregs of beans in proportion, by 0.1 ~ 10% glucose, 0.1 ~ 10% urea, 0.1 ~ 10% calcium carbonate, 0.1 ~ 5% manganous sulfate, the quality interpolation of more than preparing burden relative to wheat bran and dregs of beans, be dissolved in 20 ~ 80% (to solid-state siccative meter) 50 ~ 60 DEG C of hot water, pour in the solid-state material weighed up and mix thoroughly, 121 DEG C, sterilizing 50 ~ 120min.
According to method of the present invention, wherein, solid state fermentation inoculation method: sterilized solid-state material is naturally cooled to about 50 DEG C, by the sterilizing of solid state fermentation sterilizing containers, pours in container after solid-state material cooling; Inoculation by 1% ~ 50% measures cultured three grades of solid-state seeds, pour in sterilized water, sterilized water in proportion 20 ~ 80% (to solid-state siccative meter) adds, and stirs, then pour in solid-state material and mix thoroughly, be placed in 40 ~ 50 DEG C cultivate between cultivation 48 ~ 72h after oven drying at low temperature.
The state fermentation production method technique of high yield Bacillus coagulans spore number of the present invention and gemma rate is simple, eliminate the investment of the equipment such as liquid seeds tank, save cost, adopted solid expanding production step by step, compared traditional solid state fermentation inoculum size and reduce 50%-95%, between yeast phase, pH value maintains between 7-8, and bacterium number is high, formation advantage group bacterium is fermented, and is not easy microbiological contamination, gemma number and gemma rate higher, gemma number is 1 × 10
9~ 1 × 10
11cFU/g, gemma rate is the highest can reach 99%, is easy to produce and promote.
Embodiment
Embodiment 1
(1) first order seed preparation: glycerine pipe preservation of bacteria strain one, gets 0.5ml, accesses in one bottle of YPD liquid nutrient medium (300ml/1000ml), prepares 5 bottles of 1500ml seed liquor altogether; 40 DEG C, 150r/min cultivates 17h, is primary seed solution.
(2) secondary solid-state seed culture medium preparation: the solid-state seed culture medium of preparation 2.5kg: take wheat bran 2250g, dregs of beans 250g mixing stand-by, take glucose 25g, urea 50g, calcium carbonate 50g, be dissolved in 1250ml 50 ~ 60 DEG C of hot water, pouring in the solid-state material weighed up mixes thoroughly until all soak, load in cloth bag, fasten, 121 DEG C, laboratory Autoclave sterilizing 50min.
(3) secondary solid-state seed inoculation method: sterilized solid-state material is naturally cooled to about 50 DEG C, is placed in sterilizing 30min under ultraviolet lamp simultaneously, pours in white basin after solid-state material cooling by plastics film, the white basin of 35cm*45cm*15cm; Use sterilizing 20% sodium carbonate, primary seed solution 1500ml is adjusted pH to 8.0, and mix with solid-state material and mix thoroughly, until all soak, nothing is obviously lumpd, and covers sterilized plastics film and shuts, do not leave space, put into 45 DEG C of incubators and cultivate 20-24h.
(4) three grades of solid-state seed culture medium preparations: the solid-state material of preparation 50kg, totally 10 basins, each basin 5kg distribution: take wheat bran 4500g, dregs of beans 500g mixing stand-by, take glucose 50g, urea 100g, calcium carbonate 100g, be dissolved in 2L 50 ~ 60 DEG C of hot water, pour in the solid-state material weighed up and mix thoroughly until all soak, load in cloth bag, fasten, 121 DEG C, horizontal Autoclave sterilizing 120min.
(5) three grades of solid-state seed inoculation methods: sterilized solid-state material is naturally cooled to about 50 DEG C, 10 plastics films and 10 white basins of 47cm*68cm*37cm are placed in sterilizing 30min under ultraviolet lamp simultaneously, pour in white basin respectively after solid-state material cooling, every basin dress 5kg material (refer to solid siccative 5kg here, wet feed is about 10kg); Get the solid-state seed of cultured secondary (siccative is 2.5kg, wet feed 5kg), pour in bucket, add sterilized water 35L, stir, every basin is poured 3.5L seed liquor into and is mixed, and is placed between 45 DEG C of cultivations and cultivates 20-24h.
(6) 1 ton of solid-state material is prepared: take 0.9 ton, wheat bran, dregs of beans 0.1 ton mixing is stand-by, first getting manganous sulfate 1.4kg is dissolved in 400L 50 ~ 60 DEG C of hot water, then add glucose 10kg, urea 20kg to dissolve, finally add calcium carbonate 20kg, pouring in the solid-state material weighed up mixes thoroughly until all soak, be sub-packed in 200 cloth bags, every bag of 5kg, 121 DEG C, horizontal Autoclave sterilizing 120min.
(7) 1 tons of solid-state material inoculations: sterilized solid-state material is naturally cooled to about 50 DEG C, pour in fermentation vat after solid-state material cooling, three grades of solid-state seeds (siccative is 50kg, wet feed 100kg) in Example 1, pour in 700L sterilized water, stir, regulate the temperature to 45 DEG C between cultivating, until sporulation, cultivate 2-3 days, oven drying at low temperature, after testing gemma number 8.36 × 10
10cFU/g, gemma rate is 95%.
Embodiment 2
In order to better prove that this fermentating formula can stablize the pH value in fermenting process, doing the formula adding urea and do not add urea respectively and contrasting.
(1) technique of urea formula is added: take 900g wheat bran, the mixing of 100g dregs of beans is stand-by, first getting manganous sulfate 1.4g is dissolved in 400ml, then 10g glucose is added, 20g urea dissolves, finally add 20g calcium carbonate, pouring in the solid-state material weighed up mixes thoroughly until all soak, be loaded in cloth bag, 121 DEG C, sterilizing 50min, sterilized solid-state material is naturally cooled to about 50 DEG C, pour in fermenting container after solid-state material cooling, level liquid seed 600ml in Example 1, adjust ph to 8.0, mix with solid-state material, put into 45 DEG C of cultivations between cultivation, the change of pH value is monitored in culturing process, cultivate 3 days, oven drying at low temperature detects.
(2) technique of urea formula is not added: take 900g wheat bran, the mixing of 100g dregs of beans is stand-by, first getting manganous sulfate 1.4g is dissolved in 400ml, then 10g glucose is added, finally add 20g calcium carbonate, pouring in the solid-state material weighed up mixes thoroughly until all soak, be loaded in cloth bag, 121 DEG C, sterilizing 50min, sterilized solid-state material is naturally cooled to about 50 DEG C, pour in fermenting container after solid-state material cooling, level liquid seed 600ml in Example 1, adjust ph to 8.0, mix with solid-state material, put into 45 DEG C of cultivations between cultivation, the change of pH value is monitored in culturing process, cultivate 3 days, oven drying at low temperature detects.
(3) test-results: from data, acid is produced in Bacillus coagulans fermenting process, without the obvious decline from the pH value of fermentation the 8th hour solid-state material of urea formula, until 60 hours pH value are stabilized in about 5.0, reason is produce that hyper acid inhibits the Growth and metabolism of Bacillus coagulans, and produce urase in urea formula Bacillus coagulans fermenting process, the ammonia produced after decomposing urea and acid neutralize, make the pH value in fermenting process maintain partial neutral always, be conducive to the growth of Bacillus coagulans and the formation of gemma.After fermentation, carry out drying and processing to sample, detect gemma number and gemma rate, the gemma number without urea formula is 5.11 × 10
8cFU/g, gemma rate is 55%, and the gemma number of urea formula is 4 × 10
10cFU/g, gemma rate is 97%, and therefore urea formula has the effect of obviously stablizing pH value in Bacillus coagulans solid ferment process.
Table 1 ferment the solid-state material of 72h pH value change
| Fermentation period | Urea formula pH value | Without urea formula pH value |
| 0 | 7.50 | 7.50 |
| 4 | 7.48 | 7.45 |
| 8 | 7.33 | 6.93 |
| 12 | 7.40 | 6.87 |
| 16 | 7.21 | 6.55 |
| 20 | 7.15 | 6.49 |
| 24 | 7.19 | 6.05 |
| 28 | 7.20 | 5.87 |
| 32 | 6.98 | 5.91 |
| 36 | 6.98 | 5.48 |
| 40 | 7.11 | 5.55 |
| 44 | 7.19 | 5.63 |
| 48 | 7.34 | 5.34 |
| 52 | 7.33 | 5.36 |
| 56 | 7.29 | 5.24 |
| 60 | 7.33 | 4.89 |
| 64 | 7.28 | 5.01 |
| 68 | 7.45 | 4.99 |
| 72 | 7.49 | 4.89 |
Embodiment 3
(1) first order seed preparation: glycerine pipe preservation of bacteria strain one, gets 0.5ml, accesses in one bottle of YPD liquid nutrient medium (300ml/1000ml), prepares 5 bottles of 1500ml seed liquor altogether; 40 DEG C, 150r/min cultivates 17h, is primary seed solution.
(2) secondary solid-state seed culture medium preparation: the solid-state seed culture medium of preparation 2.5kg: take wheat bran 2250g, dregs of beans 250g mixing stand-by, take glucose 2.5g, urea 2.5g, calcium carbonate 2.5g, be dissolved in 1250ml 50 ~ 60 DEG C of hot water, pouring in the solid-state material weighed up mixes thoroughly until all soak, load in cloth bag, fasten, 121 DEG C, laboratory Autoclave sterilizing 50min.
(3) secondary solid-state seed inoculation method: sterilized solid-state material is naturally cooled to about 50 DEG C, is placed in sterilizing 30min under ultraviolet lamp simultaneously, pours in white basin after solid-state material cooling by plastics film, the white basin of 35cm*45cm*15cm; Use sterilizing 20% sodium carbonate, primary seed solution 1500ml is adjusted pH to 8.0, and mix with solid-state material and mix thoroughly, until all soak, nothing is obviously lumpd, and covers sterilized plastics film and shuts, do not leave space, put into 45 DEG C of incubators and cultivate 20-24h.
(4) three grades of solid-state seed culture medium preparations: the solid-state material of preparation 50kg, totally 10 basins, each basin 5kg distribution: take wheat bran 4500g, dregs of beans 500g mixing stand-by, take glucose 50g, urea 50g, calcium carbonate 50g, be dissolved in 2L 50 ~ 60 DEG C of hot water, pour in the solid-state material weighed up and mix thoroughly until all soak, load in cloth bag, fasten, 121 DEG C, horizontal Autoclave sterilizing 120min.
(5) three grades of solid-state seed inoculation methods: sterilized solid-state material is naturally cooled to about 50 DEG C, 10 plastics films and 10 white basins of 47cm*68cm*37cm are placed in sterilizing 30min under ultraviolet lamp simultaneously, pour in white basin respectively after solid-state material cooling, every basin dress 5kg material (refer to solid siccative 5kg here, wet feed is about 10kg); Get the solid-state seed of cultured secondary (siccative is 2.5kg, wet feed 5kg), pour in bucket, add sterilized water 35L, stir, every basin is poured 3.5L seed liquor into and is mixed, and is placed between 45 DEG C of cultivations and cultivates 20-24h.
(6) 1 ton of solid-state material is prepared: take 0.9 ton, wheat bran, dregs of beans 0.1 ton mixing is stand-by, first getting manganous sulfate 1kg is dissolved in 400L 50 ~ 60 DEG C of hot water, then add glucose 1kg, urea 1kg to dissolve, finally add calcium carbonate 1kg, pouring in the solid-state material weighed up mixes thoroughly until all soak, be sub-packed in 200 cloth bags, every bag of 5kg, 121 DEG C, horizontal Autoclave sterilizing 120min.
(7) 1 tons of solid-state material inoculations: sterilized solid-state material is naturally cooled to about 50 DEG C, pour in fermentation vat after solid-state material cooling, three grades of solid-state seeds (siccative is 50kg, wet feed 100kg) in Example 1, pour in 700L sterilized water, stir, regulate the temperature to 45 DEG C between cultivating, until sporulation, cultivate 2-3 days, oven drying at low temperature, after testing gemma number 5 × 10
9cFU/g, gemma rate is 87%.
Embodiment 4
(1) first order seed preparation: glycerine pipe preservation of bacteria strain one, gets 0.5ml, accesses in one bottle of YPD liquid nutrient medium (300ml/1000ml), prepares 5 bottles of 1500ml seed liquor altogether; 40 DEG C, 150r/min cultivates 17h, is primary seed solution.
(2) secondary solid-state seed culture medium preparation: the solid-state seed culture medium of preparation 2.5kg: take wheat bran 2250g, dregs of beans 250g mixing stand-by, take glucose 250g, urea 250g, calcium carbonate 250g, be dissolved in 1250ml 50 ~ 60 DEG C of hot water, pouring in the solid-state material weighed up mixes thoroughly until all soak, load in cloth bag, fasten, 121 DEG C, laboratory Autoclave sterilizing 50min.
(3) secondary solid-state seed inoculation method: sterilized solid-state material is naturally cooled to about 50 DEG C, is placed in sterilizing 30min under ultraviolet lamp simultaneously, pours in white basin after solid-state material cooling by plastics film, the white basin of 35cm*45cm*15cm; Use sterilizing 20% sodium carbonate, primary seed solution 1500ml is adjusted pH to 8.0, and mix with solid-state material and mix thoroughly, until all soak, nothing is obviously lumpd, and covers sterilized plastics film and shuts, do not leave space, put into 45 DEG C of incubators and cultivate 20-24h.
(4) three grades of solid-state seed culture medium preparations: the solid-state material of preparation 50kg, totally 10 basins, each basin 5kg distribution: take wheat bran 4500g, dregs of beans 500g mixing stand-by, take glucose 5000g, urea 5000g, calcium carbonate 5000g, be dissolved in 2L 50 ~ 60 DEG C of hot water, pour in the solid-state material weighed up and mix thoroughly until all soak, load in cloth bag, fasten, 121 DEG C, horizontal Autoclave sterilizing 120min.
(5) three grades of solid-state seed inoculation methods: sterilized solid-state material is naturally cooled to about 50 DEG C, 10 plastics films and 10 white basins of 47cm*68cm*37cm are placed in sterilizing 30min under ultraviolet lamp simultaneously, pour in white basin respectively after solid-state material cooling, every basin dress 5kg material (refer to solid siccative 5kg here, wet feed is about 10kg); Get the solid-state seed of cultured secondary (siccative is 2.5kg, wet feed 5kg), pour in bucket, add sterilized water 35L, stir, every basin is poured 3.5L seed liquor into and is mixed, and is placed between 45 DEG C of cultivations and cultivates 20-24h.
(6) 1 ton of solid-state material is prepared: take 0.9 ton, wheat bran, dregs of beans 0.1 ton mixing is stand-by, first getting manganous sulfate 50kg is dissolved in 400L 50 ~ 60 DEG C of hot water, then add glucose 100kg, urea 100kg to dissolve, finally add calcium carbonate 100kg, pouring in the solid-state material weighed up mixes thoroughly until all soak, be sub-packed in 200 cloth bags, every bag of 5kg, 121 DEG C, horizontal Autoclave sterilizing 120min.
(7) 1 tons of solid-state material inoculations: sterilized solid-state material is naturally cooled to about 50 DEG C, pour in fermentation vat after solid-state material cooling, three grades of solid-state seeds (siccative is 50kg, wet feed 100kg) in Example 1, pour in 700L sterilized water, stir, regulate the temperature to 45 DEG C between cultivating, until sporulation, cultivate 2-3 days, oven drying at low temperature, after testing gemma number 7.31 × 10
10cFU/g, gemma rate is 97%.
Claims (4)
1. a Bacillus coagulans state fermentation production method, is characterized in that, said method comprising the steps of:
(1) first order seed preparation: by the inoculum size of 1 ~ 10%, cultivates in access liquid nutrient medium, is primary seed solution;
(2) the solid-state seed inoculation of secondary: primary seed solution is adjusted pH to 6.0 ~ 9.0, and the inoculum size by 10% ~ 90% mixes to mix thoroughly with solid-state fermentation material cultivates;
(3) three grades of solid-state seed inoculations: by the inoculum size of 1% ~ 50%, get the solid-state seed of cultured secondary in sterilized water, stir, then pour in solid-state fermentation material to mix thoroughly and cultivate;
(4) solid state fermentation inoculation: by the inoculum size of 1% ~ 50%, get cultured three grades of solid-state seeds in sterilized water, stir, then pour in solid-state fermentation material to mix thoroughly and cultivate,
Wherein, the formula of described solid-state fermentation material comprises in mass ratio: (1) major ingredient, 70 ~ 99% wheat brans, 1 ~ 30% dregs of beans; (2) auxiliary material, the quality based on major ingredient comprises 0.1 ~ 10% glucose, 0.1 ~ 10% urea, 0.1 ~ 10% calcium carbonate, 0.1 ~ 5% manganous sulfate.
2. Bacillus coagulans state fermentation production method according to claim 1, it is characterized in that, in step (2), sterilized solid-state material is naturally cooled to 50 DEG C, simultaneously by plastics film, the sterilizing of solid state fermentation sterilizing containers, pour in container after solid-state material cooling, use sterilizing 1 ~ 50% sodium carbonate, primary seed solution is adjusted pH to 6.0 ~ 9.0, inoculum size by 10% ~ 90% mixes with solid-state material mixes thoroughly, is placed between 40 ~ 50 DEG C of cultivations and cultivates 15 ~ 24h.
3. Bacillus coagulans state fermentation production method according to claim 1, it is characterized in that, in step (3), sterilized solid-state material is naturally cooled to 50 DEG C, simultaneously by plastics film, the sterilizing of solid state fermentation sterilizing containers, pour in container after solid-state material cooling; Inoculation by 1% ~ 50% measures the solid-state seed of cultured secondary, pours in sterilizing bucket, in solid-state siccative, 20 ~ 80% add sterilized water in proportion, stir, pour in solid-state fermentation material and mix thoroughly, be placed between 40 ~ 50 DEG C of cultivations and cultivate 15 ~ 24h.
4. Bacillus coagulans state fermentation production method according to claim 1, it is characterized in that, in step (4), sterilized solid-state material is naturally cooled to about 50 DEG C, by the sterilizing of solid state fermentation sterilizing containers, pour in container after solid-state fermentation material cooling; Inoculation by 1% ~ 50% measures cultured three grades of solid-state seeds, pours in sterilized water, in solid-state siccative, sterilized water is 20 ~ 80% interpolations in proportion, stir, then pour in solid-state fermentation material and mix thoroughly, be placed in 40 ~ 50 DEG C cultivate between cultivation 48 ~ 72h after oven drying at low temperature.
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| CN106974063A (en) * | 2017-04-19 | 2017-07-25 | 广东轻工职业技术学院 | It is a kind of that the method that bacillus coagulans produces High efficiency protein feed is cooperateed with feeding enzyme |
| CN110669694A (en) * | 2019-10-25 | 2020-01-10 | 湖北华扬科技发展有限公司 | Bacillus histolyticus phosphorus reduction culture medium, fermentation method and application |
| CN111088188A (en) * | 2020-01-03 | 2020-05-01 | 江苏三仪生物工程有限公司 | Preparation of feed additive containing lactic acid and bacillus coagulans and detection culture medium thereof |
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| CN106974063B (en) * | 2017-04-19 | 2021-02-19 | 广东轻工职业技术学院 | Method for producing high-efficiency protein feed by using feed enzyme in cooperation with bacillus coagulans |
| CN110669694A (en) * | 2019-10-25 | 2020-01-10 | 湖北华扬科技发展有限公司 | Bacillus histolyticus phosphorus reduction culture medium, fermentation method and application |
| CN111088188A (en) * | 2020-01-03 | 2020-05-01 | 江苏三仪生物工程有限公司 | Preparation of feed additive containing lactic acid and bacillus coagulans and detection culture medium thereof |
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