CN104824337A - Preparation method of fermented soybean meal for feeding - Google Patents
Preparation method of fermented soybean meal for feeding Download PDFInfo
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- CN104824337A CN104824337A CN201510306993.XA CN201510306993A CN104824337A CN 104824337 A CN104824337 A CN 104824337A CN 201510306993 A CN201510306993 A CN 201510306993A CN 104824337 A CN104824337 A CN 104824337A
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Abstract
The invention discloses a preparation method of fermented soybean meal for feeding. The preparation method includes inoculating mixed bacteria solution into fermented soybean meal substrate and uniformly mixing; fermenting for 72 to 96 hours at 27-37 degrees Centigrade; drying and smashing to obtain fermented soybean meal for feeding. The mixed bacteria solution is composed of aspergillus niger liquid, saccharomycetes liquid, bacillus subtilis liquid and lactic acid bacteria liquid. The weight ratio of the mixed bacteria solution to the fermented soybean meal substrate is 6-18:1000, and the weight ratio of aspergillus niger liquid, saccharomycetes liquid, bacillus subtilis liquid and lactic acid bacteria liquid is 1-4:0.5-2:0.5-2:0.5-2. By the preparation method, polysaccharide in the fermented soybean meal can be effectively degraded, nutrient substance in the soybean meal is released, and crude protein of the fermented soybean meal is concentrated. Meanwhile, antinutritional factors such as glycinin, beta-conglycinin, oligosaccharide and phytic acid are effectively degraded, digestive utilization rate of nutrients is increased, and the key problems that glycan content in the fermented soybean meal is high and crude protein is hard to improve and poor in nutritive value are solved.
Description
Technical field
The invention belongs to fermentation technical field, particularly a kind of preparation method of feeding fermented dregs of beans.
Background technology
Dregs of beans, because of its good nutritive value, is the most widely used vegetable protein raw material of current feedstuff industry, but wherein there is more ANFs, limit its further development and application.The quality adopting the method for fermentable to promote dregs of beans at present more, mainly utilize bacterial fermentation process dregs of beans, dregs of beans after process eliminates part glycan and the multiple ANFs such as glycinin, β-companion's globulin, oligosaccharides, phytic acid, crude protein content is slightly promoted, and be rich in protease, amylase, organic acid and active probiotic, make dregs of beans change into the higher functional feed of nutritive value.
But fermented bean dregs prepared by traditional fermentation methods still contains glycan and the ANFs of larger proportion.Glycan is plant cell wall constituent, is the summation of the SNSPs such as cellulose, xylan, galactolipin, mannosan and pectin, has anti-trophism, not only can reduce the digestive utilization ratio of nutrient, and suppresses the healthy growth of livestock and poultry.ANFs refers to the material that in feed, some hinders nutritional labeling digestion, absorbs and utilize, and mainly comprises glycinin, β-companion's globulin, oligosaccharides, phytic acid, stachyose, gossypose etc.The high-load of high glycan and ANFs limits the further lifting of fermented bean dregs protein quality, and in the fermented bean dregs causing traditional fermentation methods to be prepared, crude protein limiting value only has about 50%.
Therefore, improve the preparation method of fermented bean dregs, solve glycan and ANFs content in fermented bean dregs higher, the problem that crude protein quality is difficult to promote is extremely important.
Summary of the invention
The object of the invention is to overcome above-mentioned deficiency existing in prior art, a kind of preparation method of feeding fermented dregs of beans is provided.Preparation method of the present invention adopts black-koji mould, saccharomycete, bacillus subtilis and lactic acid bacteria mixed culture fermentation technology, zytase is produced by fermentation, dextranase, mannase, cellulase and pectase etc., farthest destroy dregs of beans cell membrane, effectively degrade the polysaccharide in fermented bean dregs, nutriment in release dregs of beans, and then the crude protein of concentrated fermented bean dregs, can also effectively to degrade glycinin and β-companion's globulin simultaneously, oligosaccharides, the ANFs such as phytic acid, improve the digestive utilization ratio of nutrient, final acquisition compound sugar, high protein content, the feeding fermented dregs of beans of high nutrition.
In order to realize foregoing invention object, the invention provides following technical scheme:
The preparation method of feeding fermented dregs of beans of the present invention is: be inoculated into by mixed bacteria liquid in bean pulp fermentation substrate, mix, and at 27 ~ 37 DEG C of bottom fermentation 72 ~ 96h, then dries, and pulverizes, obtains feeding fermented dregs of beans; Described mixed bacteria liquid is made up of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension; The weight ratio of described mixed bacteria liquid and bean pulp fermentation substrate is 6 ~ 18:1000, and in described mixed bacteria liquid, the weight ratio of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension is 1 ~ 4:0.5 ~ 2:0.5 ~ 2:0.5 ~ 2; Black-koji mould viable count in described black-koji mould liquid is 2.5 × 10
7~ 4.6 × 10
7cFU/g; In described yeast liquid, saccharomycete viable count is 1.3 × 10
9~ 3.6 × 10
9cFU/g; Bacillus subtilis viable count in described bacillus subtilis liquid is 1.5 × 10
8~ 2.3 × 10
8cFU/g; In described lactobacillus suspension, viable count of lactobacillus is 2.3 × 10
8~ 4.0 × 10
8cFU/g.
Black-koji mould secretion can produce amylase, carbohydrase, citric acid, gluconic acid, gallic acid etc.Saccharomycete produces great amount of carbon dioxide gas in dregs of beans, and due to the formation of fermented bean dregs network organization, and is left in desmachyme, makes that fermented bean dregs is loose porous, volume increases; In addition, during the fermentation, the biochemical reaction of experience series of complex, creates the distinctive fermenting aroma of fermented bean dregs.The enzymes such as bacillus subtilis thalline self synthesis AMS, protease, lipase, cellulase.Conversion of glucose is lactic acid by lactic acid bacteria under the catalytic action of enzyme, and has biodegradable effect.The present invention utilizes black-koji mould, saccharomycete, bacillus subtilis and lactic acid bacteria mixed culture fermentation technology, zytase is produced by fermentation, dextranase, mannase, cellulase and pectase etc., farthest destroy dregs of beans cell membrane, effectively degrade the polysaccharide in fermented bean dregs, nutriment in release dregs of beans, and then the crude protein of concentrated fermented bean dregs, can also effectively to degrade glycinin and β-companion's globulin simultaneously, oligosaccharides, the ANFs such as phytic acid, improve the digestive utilization ratio of nutrient, dregs of beans is made to change into the higher functional feed of nutritive value, be rich in protease simultaneously, amylase, organic acid and active probiotic.
Applicant finds through test of many times, is 2.5 × 10 by viable count
7~ 4.6 × 10
7the black-koji mould liquid of CFU/g, viable count is 1.3 × 10
9~ 3.6 × 10
9the yeast liquid of CFU/g, viable count is 1.5 × 10
8~ 2.3 × 10
8the bacillus subtilis liquid of CFU/g, viable count is 2.3 × 10
8~ 4.0 × 10
8the lactobacillus suspension of CFU/g is that 1 ~ 4:0.5 ~ 2:0.5 ~ 2:0.5 ~ 2 join in bean pulp fermentation substrate according to weight ratio, zytase is produced by fermentation, dextranase, mannase, cellulase and pectase etc., farthest destroy dregs of beans cell membrane, effectively degrade the polysaccharide in fermented bean dregs, nutriment in release dregs of beans, and then the crude protein of concentrated fermented bean dregs, this preparation method can also effectively degrade glycinin and β-companion's globulin simultaneously, oligosaccharides, the ANFs such as phytic acid, improve the digestive utilization ratio of nutrient, final acquisition compound sugar, high protein content, the feeding fermented dregs of beans of high nutrition.
Preferably, described bean pulp fermentation substrate is made up of the raw material of following weight: dregs of beans 30 ~ 70 parts, 1 ~ 4 part, wheat bran, 1 ~ 4 part, ammonium sulfate, 0.5 ~ 2 part, molasses, 30 ~ 70 parts, water.The preparation method of bean pulp fermentation substrate is specially: take dregs of beans, wheat bran, ammonium sulfate and molasses by weight ratio, mix and blend, then inject running water, continues mixing and stirs, the bacterium liquid to be seeded such as then.This bean pulp fermentation substrate is after above-mentioned preparation method's fermentation, glycan in dregs of beans and the ANFs such as glycinin and β-companion's globulin, oligosaccharides, phytic acid are effectively degraded, crude protein content is high, can promote the healthy growth of livestock and poultry, is suitable for the livestocks such as wean pig, piglet, chicken of feeding.
In above-mentioned preparation method, mixed bacteria liquid is inoculated in bean pulp fermentation substrate, mixes, at 27 ~ 37 DEG C of bottom fermentation 72 ~ 96h.Be shorter than 72h when fermenting, fermentation is abundant not, and glycan and ANFs are fully degraded; When being longer than 96h when fermenting, excessively, nutritive value reduces on the contrary for glycan and ANFs degraded.Mixed bacteria liquid preferably, to be describedly inoculated in bean pulp fermentation substrate by the best, mixes, at 27 ~ 37 DEG C of bottom fermentation 80h.
In above-mentioned preparation method, the weight ratio of described mixed bacteria liquid and bean pulp fermentation substrate is 6 ~ 18:1000.When weight ratio is less than 6:1000, obviously fermented bean dregs quality can be reduced; When inoculum concentration is greater than 18:1000, bacterial reproduction can be caused excessive, fermented bean dregs conversion ratio reduces, and causes unnecessary waste.Further preferably, the weight ratio of described mixed bacteria liquid and bean pulp fermentation substrate is 10 ~ 15:1000.Best preferred, the weight ratio of described mixed bacteria liquid and bean pulp fermentation substrate is 12:1000.
Preferably, in described mixed bacteria liquid, the weight ratio of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension is 2:1:1:1.By preferably above-mentioned, the degraded of glycan and ANFs can be promoted further, improve the quality of fermented bean dregs.
Preferably, described black-koji mould liquid is made up of following methods:
(1) black-koji mould is inoculated in black-koji mould slant medium, at 25 ~ 30 DEG C, cultivates 68 ~ 76h; From black-koji mould slant medium picking one ring black-koji mould, be seeded in black-koji mould fluid nutrient medium, cultivate 72 ~ 96h at 23 ~ 28 DEG C, obtain black-koji mould enrichment liquid;
(2) getting black-koji mould enrichment liquid prepared by step (1), is that the inoculum concentration of 0.5 ~ 5:100 is seeded in black-koji mould fluid nutrient medium by volume, regulates pH to 6.2 ~ 6.6, ventilate cavity 72 ~ 96h at 23 ~ 28 DEG C; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in black-koji mould fluid nutrient medium, regulate pH to 6.2 ~ 6.6, ventilate cavity 72 ~ 96h at 23 ~ 28 DEG C, until be extended to 5L, now in black-koji mould fluid nutrient medium, black-koji mould viable count is 2.5 × 10
7~ 4.6 × 10
7cFU/g;
Described black-koji mould slant medium is that raw material is made by glucose 8 ~ 12g, peptone 4 ~ 6g, potassium dihydrogen phosphate 0.5 ~ 2g, magnesium sulfate 0.1 ~ 1g, agar 10 ~ 30g, rose-bengal 0.03 ~ 0.04g, chloramphenicol 0.05 ~ 0.2g, distilled water 900 ~ 1100mL;
Described black-koji mould fluid nutrient medium is that raw material is formulated by peptone 4 ~ 6g, yeast extract powder 1 ~ 3g, magnesium sulfate 0.1 ~ 1g, glucose 10 ~ 30g, dipotassium hydrogen phosphate 0.5 ~ 2g, distilled water 900 ~ 1000mL.
Inventor finds through test of many times, adopt the black-koji mould liquid that said method is cultivated, effectively can secrete and produce amylase, carbohydrase, citric acid, gluconic acid, gallic acid etc., and then better and yeast liquid, bacillus subtilis liquid, lactobacillus suspension work in coordination with compatibility, glycan in degraded dregs of beans and ANFs, promote the crude protein quality of fermented bean dregs.
Preferably, the preparation method of described black-koji mould slant medium is as follows: get glucose 10g, peptone 5g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, agar 20g, rose-bengal 0.033g, chloramphenicol 0.1g, distilled water 1000mL prepare, 121 DEG C of sterilizing 20min.
Preferably, the preparation method of described black-koji mould fluid nutrient medium is as follows: get peptone 5g, yeast extract powder 2g, magnesium sulfate 0.5g, glucose 20g, dipotassium hydrogen phosphate 1g, distilled water 1000mL prepare, 121 DEG C of sterilizing 20min.
Preferably, described yeast liquid is made up of following methods:
(1) saccharomycete is inoculated in saccharomycete slant medium, cultivate 68 ~ 76h at 25 ~ 30 DEG C, from saccharomycete slant medium, picking one ring saccharomycete, is seeded in saccharomycete fluid nutrient medium, cultivate 36 ~ 72h at 35 ~ 37 DEG C, obtain saccharomycete enrichment liquid;
(2) get saccharomycete enrichment liquid prepared by step (1), the inoculum concentration being 0.5 ~ 5:100 according to volume ratio is seeded in saccharomycete fluid nutrient medium, regulates pH to 6.5 ~ 7.5, ventilate cavity 36 ~ 72h at 25 ~ 30 DEG C; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in saccharomycete fluid nutrient medium, and regulate pH to 6.5 ~ 7.5, ventilate cavity 36 ~ 72h at 25 ~ 30 DEG C, until be extended to 5L; Now in saccharomycete fluid nutrient medium, saccharomycete viable count is 1.3 × 10
9~ 3.6 × 10
9cFU/g;
Described saccharomycete slant medium is by glucose 3 ~ 7g, peptone 1 ~ 3g, yeast extract 0.5 ~ 2g, sodium chloride 0.1 ~ 1g, and agar 10 ~ 20g is that raw material is formulated;
Described saccharomycete fluid nutrient medium is that raw material is formulated by glucose 3 ~ 7g, peptone 1 ~ 3g, yeast extract 0.5 ~ 2g, sodium chloride 0.1 ~ 1g and distilled water 900 ~ 1100ml.
Inventor finds through test of many times, the yeast liquid adopting said method to cultivate, effectively can produce great amount of carbon dioxide gas in dregs of beans, and due to the formation of fermented bean dregs network organization, and be left in desmachyme, make that fermented bean dregs is loose porous, volume increases; In addition, during the fermentation, the biochemical reaction of experience series of complex, creates the distinctive fermenting aroma of fermented bean dregs; And then work in coordination with compatibility with black-koji mould liquid, bacillus subtilis liquid, lactobacillus suspension better, the glycan better in degraded dregs of beans and ANFs, promote the crude protein quality of fermented bean dregs.
Preferably, the preparation method of described saccharomycete slant medium is as follows: get glucose 5g, peptone 2g, yeast extract 1g, sodium chloride 0.5g, agar 15g prepare, saccharomycete slant medium pH is 6.5 ~ 7.5,121 DEG C of sterilizing 20min.
Preferably, the preparation method of described saccharomycete fluid nutrient medium is as follows: get glucose 5g, peptone 2g, yeast extract 1g, sodium chloride 0.5g, distilled water 1000ml prepare, 121 DEG C of sterilizing 20min.
Preferably, described bacillus subtilis liquid is made up of following methods:
(1) bacillus subtilis is inoculated in bacillus subtilis slant medium, 40 ~ 60h is cultivated at 35 ~ 38 DEG C, picking one ring bacillus subtilis from bacillus subtilis slant medium, be seeded in bacillus subtilis fluid nutrient medium, cultivate 36 ~ 72h at 35 ~ 38 DEG C, obtain bacillus subtilis enrichment liquid;
(2) get bacillus subtilis enrichment liquid prepared by step (1), the inoculum concentration being 0.5 ~ 5:100 according to volume ratio is seeded in bacillus subtilis fluid nutrient medium; Regulate pH to 7.0 ~ 7.2, at 35 ~ 38 DEG C, cultivate 36 ~ 72h; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in bacillus subtilis fluid nutrient medium, regulates pH to 7.0 ~ 7.2, cultivates 36 ~ 72h, until be extended to 5L at 35 ~ 38 DEG C; Now in fluid nutrient medium, bacillus subtilis viable count is 1.5 × 10
8~ 2.3 × 10
8cFU/g;
Described bacillus subtilis slant medium is that raw material is formulated by glucose 3 ~ 7g, peptone 1 ~ 4g, yeast extract 0.5 ~ 2g, sodium chloride 0.1 ~ 1g, agar 10 ~ 20g;
Described bacillus subtilis fluid nutrient medium is that raw material is formulated by tryptone 5 ~ 15g, yeast extract powder 1 ~ 10g, sodium chloride 5 ~ 20g, distilled water 900 ~ 1100ml.
Inventor finds through test of many times, adopt the bacillus subtilis liquid that said method is cultivated, self can effectively synthesize the enzymes such as AMS, protease, lipase, cellulase, and then work in coordination with compatibility with yeast liquid, black-koji mould liquid, lactobacillus suspension better, glycan better in degraded dregs of beans and ANFs, promote the crude protein quality of fermented bean dregs.
Preferably, the preparation method of described bacillus subtilis slant medium is as follows: get glucose 5g, peptone 2g, yeast extract 1g, sodium chloride 0.5g, agar 15g prepare, saccharomycete slant medium pH 6.5 ~ 7.5,121 DEG C of sterilizing 20min.
Preferably, the preparation method of described bacillus subtilis fluid nutrient medium is as follows: get tryptone 10g, yeast extract powder 5g, sodium chloride 10g, distilled water 1000mL prepare, 121 DEG C of sterilizing 20min.
Preferably, described lactobacillus suspension is made up of following methods:
(1) by lactobacillus inoculum in lactic acid bacteria slant medium, at 35 ~ 38 DEG C cultivate 40 ~ 60h; From lactic acid bacteria slant medium, picking one ring lactic acid bacteria, is seeded in lactic acid bacteria fluid nutrient medium, cultivates 36 ~ 72h, obtain lactic acid bacteria enrichment liquid at 28 ~ 32 DEG C;
(2) get lactic acid bacteria enrichment liquid prepared by step (1), the inoculum concentration being 0.5 ~ 5:100 according to volume ratio is seeded in lactic acid bacteria fluid nutrient medium, regulates pH to 5.5 ~ 6.0, cultivates 36 ~ 72h at 35 ~ 38 DEG C; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in lactic acid bacteria fluid nutrient medium, regulates pH to 5.5 ~ 6.0, cultivates 36 ~ 72h, until be extended to 5L at 35 ~ 38 DEG C; Now in lactic acid bacteria fluid nutrient medium, viable count of lactobacillus is 2.3 × 10
8~ 4.0 × 10
8cFU/g;
Described lactic acid bacteria liquid slant medium is that raw material is formulated by skimmed milk power 50 ~ 200g, dusty yeast 0.5 ~ 2g, dipotassium hydrogen phosphate 1 ~ 10g, agar 10 ~ 20g;
Described lactic acid bacteria fluid nutrient medium is that raw material is formulated by peptone 5 ~ 20g, powdered beef 1 ~ 10g, yeast extract powder 2 ~ 8g, beef peptone 5 ~ 14g, glucose 10 ~ 30g, Tween-80 0.5 ~ 2mL, dipotassium hydrogen phosphate 1 ~ 4g, sodium acetate 1 ~ 10g, Triammonium citrate 0.5 ~ 4g, magnesium sulfate 0.1 ~ 0.3g, manganese sulfate 0.01 ~ 0.1g, distilled water 900 ~ 1000ml.
Inventor finds through test of many times, and the lactobacillus suspension adopting said method to cultivate, effectively can be converted into lactic acid by glucose, and have biodegradable effect under the catalytic action of enzyme.And then work in coordination with compatibility with yeast liquid, bacillus subtilis liquid, black-koji mould liquid better, the glycan better in degraded dregs of beans and ANFs, promote the crude protein quality of fermented bean dregs.
Preferably, the preparation method of described lactic acid slant medium is as follows: extracting degreasing milk powder 100g, dusty yeast 1g, dipotassium hydrogen phosphate 5g, agar 15g prepare, 121 DEG C of sterilizing 20min.
Preferably, the preparation method of described lactic acid bacteria fluid nutrient medium is as follows: get peptone 10g, powdered beef 5g, yeast extract powder 4g, beef peptone 10g, glucose 20g, Tween-80 1mL, dipotassium hydrogen phosphate 2g, sodium acetate 5g, Triammonium citrate 2g, magnesium sulfate 0.2g, manganese sulfate 0.05g, distilled water 1000mL prepare, 121 DEG C of sterilizing 20min.
Compared with prior art, beneficial effect of the present invention:
(1) preparation method of the present invention is with black-koji mould, saccharomycete, the mode of bacillus subtilis and lactic acid bacteria mixed culture fermentation, zytase is produced by fermentation, dextranase, mannase, cellulase and pectase etc., farthest destroy dregs of beans cell membrane, effectively degrade the polysaccharide in fermented bean dregs, nutriment in release dregs of beans, and then the crude protein of concentrated fermented bean dregs, also effectively degrade glycinin and β-companion's globulin simultaneously, oligosaccharides, the ANFs such as phytic acid, improve the digestive utilization ratio of nutrient, solve glycan content in fermented bean dregs pointedly higher, crude protein is difficult to promote, the key issue that nutritive value is not high.
(2) through fermented bean dregs prepared by the inventive method, xylan, glucan, mannosan degradation rate are all more than 50%, particularly pectin degrading rate is up to more than 70%, and the fermented bean dregs glycan total amount after degraded is only 1/3rd of traditional zymotic dregs of beans.
(3) the inventive method eliminates the fermented bean dregs after glycan, and dry composition is correspondingly concentrated, and make the crude protein limiting value of traditional zymotic technique be promoted to 60% by 50%, fermented bean dregs quality gets a promotion further.
(4) the inventive method also degrades the ANFs such as glycinin and β-companion's globulin, stachyose, gossypose well, and ANFs degradation rate, all more than 90%, improves the digestive utilization ratio of nutrient in fermented bean dregs.
Detailed description of the invention
Below in conjunction with test example and detailed description of the invention, the present invention is described in further detail.But this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following embodiment, all technology realized based on content of the present invention all belong to scope of the present invention.
(1) preparation of black-koji mould liquid
A, black-koji mould is inoculated in black-koji mould slant medium, at 25 ~ 30 DEG C, cultivates 68 ~ 76h; From black-koji mould slant medium picking one ring black-koji mould, be seeded in black-koji mould fluid nutrient medium, cultivate 72 ~ 96h at 23 ~ 28 DEG C, obtain black-koji mould enrichment liquid;
B, get black-koji mould enrichment liquid prepared by steps A, be that the inoculum concentration of 0.5 ~ 5:100 is seeded in black-koji mould fluid nutrient medium by volume, regulate pH to 6.2 ~ 6.6, ventilate at 23 ~ 28 DEG C (air after bacteriological filtration process) cultivates 72 ~ 96h; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in black-koji mould fluid nutrient medium, regulate pH to 6.2 ~ 6.6, at 23 ~ 28 DEG C, 72 ~ 96h is cultivated in ventilation (air after bacteriological filtration process), until be extended to 5L, now in black-koji mould fluid nutrient medium, black-koji mould viable count is 2.5 × 10
7~ 4.6 × 10
7cFU/g;
Wherein, black-koji mould slant medium is that raw material is made by glucose 8 ~ 12g, peptone 4 ~ 6g, potassium dihydrogen phosphate 0.5 ~ 2g, magnesium sulfate 0.1 ~ 1g, agar 10 ~ 30g, rose-bengal 0.03 ~ 0.04g, chloramphenicol 0.05 ~ 0.2g, distilled water 900 ~ 1100mL;
Black-koji mould fluid nutrient medium is that raw material is formulated by peptone 4 ~ 6g, yeast extract powder 1 ~ 3g, magnesium sulfate 0.1 ~ 1g, glucose 10 ~ 30g, dipotassium hydrogen phosphate 0.5 ~ 2g, distilled water 900 ~ 1000mL.
(2) preparation of yeast liquid
A, be inoculated in saccharomycete slant medium by saccharomycete, cultivate 68 ~ 76h at 25 ~ 30 DEG C, from saccharomycete slant medium, picking one ring saccharomycete, is seeded in saccharomycete fluid nutrient medium, cultivates 36 ~ 72h, obtain saccharomycete enrichment liquid at 35 ~ 37 DEG C;
B, get saccharomycete enrichment liquid prepared by steps A, the inoculum concentration being 0.5 ~ 5:100 according to volume ratio is seeded in saccharomycete fluid nutrient medium, regulates pH to 6.5 ~ 7.5, and ventilate at 25 ~ 30 DEG C (air after bacteriological filtration process) cultivates 36 ~ 72h; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in saccharomycete fluid nutrient medium, regulates pH to 6.5 ~ 7.5, and ventilate at 25 ~ 30 DEG C (air after bacteriological filtration process) cultivates 36 ~ 72h, until be extended to 5L; Now in saccharomycete fluid nutrient medium, saccharomycete viable count is 1.3 × 10
9~ 3.6 × 10
9cFU/g;
Wherein, saccharomycete fluid nutrient medium I is by glucose 3 ~ 7g, peptone 1 ~ 3g, yeast extract 0.5 ~ 2g, sodium chloride 0.1 ~ 1g, and agar 10 ~ 20g is that raw material is formulated;
Saccharomycete fluid nutrient medium is that raw material is formulated by glucose 3 ~ 7g, peptone 1 ~ 3g, yeast extract 0.5 ~ 2g, sodium chloride 0.1 ~ 1g and distilled water 900 ~ 1100ml.
(3) preparation of bacillus subtilis liquid
A, bacillus subtilis is inoculated in bacillus subtilis slant medium, 40 ~ 60h is cultivated at 35 ~ 38 DEG C, picking one ring bacillus subtilis from bacillus subtilis slant medium, be seeded in bacillus subtilis fluid nutrient medium, cultivate 36 ~ 72h at 35 ~ 38 DEG C, obtain bacillus subtilis enrichment liquid;
B, get bacillus subtilis enrichment liquid prepared by steps A, the inoculum concentration being 0.5 ~ 5:100 according to volume ratio is seeded in bacillus subtilis fluid nutrient medium; Regulate pH to 7.0 ~ 7.2, at 35 ~ 38 DEG C, cultivate 36 ~ 72h; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in bacillus subtilis fluid nutrient medium, regulates pH to 7.0 ~ 7.2, cultivates 36 ~ 72h, until be extended to 5L at 35 ~ 38 DEG C; Now in fluid nutrient medium, bacillus subtilis viable count is 1.5 × 10
8~ 2.3 × 10
8cFU/g; Wherein, bacillus subtilis fluid nutrient medium is that raw material is formulated by glucose 3 ~ 7g, peptone 1 ~ 4g, yeast extract 0.5 ~ 2g, sodium chloride 0.1 ~ 1g, agar 10 ~ 20g;
Bacillus subtilis fluid nutrient medium is that raw material is formulated by tryptone 5 ~ 15g, yeast extract powder 1 ~ 10g, sodium chloride 5 ~ 20g, distilled water 900 ~ 1100ml.
(4) preparation of lactobacillus suspension
A, by lactobacillus inoculum in lactic acid bacteria slant medium, at 35 ~ 38 DEG C cultivate 40 ~ 60h; From lactic acid bacteria slant medium, picking one ring lactic acid bacteria, is seeded in lactic acid bacteria fluid nutrient medium, cultivates 36 ~ 72h, obtain lactic acid bacteria enrichment liquid at 28 ~ 32 DEG C;
B, get lactic acid bacteria enrichment liquid prepared by steps A, the inoculum concentration being 0.5 ~ 5:100 according to volume ratio is seeded in lactic acid bacteria fluid nutrient medium, regulates pH to 5.5 ~ 6.0, cultivates 36 ~ 72h at 35 ~ 38 DEG C; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in lactic acid bacteria fluid nutrient medium, regulates pH to 5.5 ~ 6.0, cultivates 36 ~ 72h, until be extended to 5L at 35 ~ 38 DEG C; Now in lactic acid bacteria fluid nutrient medium, viable count of lactobacillus is 2.3 × 10
8~ 4.0 × 10
8cFU/g;
Wherein, lactic acid bacteria fluid nutrient medium is that raw material is formulated by skimmed milk power 50 ~ 200g, dusty yeast 0.5 ~ 2g, dipotassium hydrogen phosphate 1 ~ 10g, agar 10 ~ 20g;
Lactic acid bacteria fluid nutrient medium is that raw material is formulated by peptone 5 ~ 20g, powdered beef 1 ~ 10g, yeast extract powder 2 ~ 8g, beef peptone 5 ~ 14g, glucose 10 ~ 30g, Tween-80 0.5 ~ 2mL, dipotassium hydrogen phosphate 1 ~ 4g, sodium acetate 1 ~ 10g, Triammonium citrate 0.5 ~ 4g, magnesium sulfate 0.1 ~ 0.3g, manganese sulfate 0.01 ~ 0.1g, distilled water 900 ~ 1000ml.
(5) preparation of fermented bean dregs
Take raw material dregs of beans 300 ~ 700kg, wheat bran 10 ~ 40kg, ammonium sulfate 10 ~ 40kg, molasses 5 ~ 20kg add in mixer successively, inject the running water of 300 ~ 700kg after mix and blend 5 ~ 20min, continue mixing stirring 5 ~ 20min, make fermentation substrate; In fermentation substrate, inject 6 ~ 18kg mixed bacteria liquid, wherein mixed bacteria liquid is that 1 ~ 4:0.5 ~ 2:0.5 ~ 2:0.5 ~ 2 form by black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension according to weight ratio; Be uniformly mixed 10 ~ 20min, then heap fermentation 72 ~ 96h at 27 DEG C ~ 37 DEG C; Finally fermented bean dregs is transferred to drying plant, drying to moisture content at 45 ~ 65 DEG C is about 12%, pulverizes, and packing, obtains fermented bean dregs product.
The nutritive index of fermented bean dregs is analyzed, mainly comprises: xylan, glucan, mannosan, pectin, crude protein, acid-soluble albumen, amino acid, glycinin, β accompany globulin, stachyose, raffinose, lactic acid, little peptide, urase.Wherein, respectively refer to that object detection method is as follows:
A, crude protein, GB/T 6432;
B, acid-soluble albumen, QB/T 2653;
C, stachyose, raffinose, NY/T 2218-2012;
D, lactic acid, GB/T 23877-2009;
E, little peptide, GB/T 22492-2008;
F, glycinin, β accompany globulin, ELISA kit, enterprise's mark;
G, xylan, glucan, mannosan, pectin, gas chromatography, enterprise's mark;
The testing result of the fermented bean dregs that preparation method of the present invention obtains is as follows: xylan degrading rate >=50%, glucan degradation rate >=50%, mannosan degradation rate >=50%, pectin degrading rate >=80%, crude protein content >=60%, acid-soluble albumen >=85%, glycinin antigen degradation rate >=90%, β accompany immunoglobulin antigen degradation rate >=90%, stachyose degradation rate >=99%, gossypose >=99%, lactic acid >=5.5%, little peptide >=15%, urease to suppress degradation rate >=90%.
Embodiment 1
(1) preparation of black-koji mould liquid
A, black-koji mould is inoculated in black-koji mould slant medium, at 28 DEG C, cultivates 72h; From black-koji mould slant medium picking one ring black-koji mould, be seeded in black-koji mould fluid nutrient medium, cultivate 72 ~ 96h at 23 ~ 28 DEG C, obtain black-koji mould enrichment liquid;
B, get black-koji mould enrichment liquid prepared by steps A, be that the inoculum concentration of 0.5 ~ 5:100 is seeded in black-koji mould fluid nutrient medium by volume, regulate pH to 6.2 ~ 6.6, ventilate cavity 72 ~ 96h at 23 ~ 28 DEG C; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in black-koji mould fluid nutrient medium, regulate pH to 6.2 ~ 6.6, ventilate cavity 72 ~ 96h at 23 ~ 28 DEG C, until be extended to 5L, now in black-koji mould fluid nutrient medium, black-koji mould viable count is 2.5 × 10
7~ 4.6 × 10
7cFU/g;
Wherein, the preparation method of black-koji mould slant medium is as follows: get glucose 10g, peptone 5g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, agar 20g, rose-bengal 0.033g, chloramphenicol 0.1g, distilled water 1000mL prepare, 121 DEG C of sterilizing 20min.
The preparation method of black-koji mould fluid nutrient medium is as follows: get peptone 5g, yeast extract powder 2g, magnesium sulfate 0.5g, glucose 20g, dipotassium hydrogen phosphate 1g, distilled water 1000mL prepare, 121 DEG C of sterilizing 20min.
(2) preparation of yeast liquid
A, be inoculated in saccharomycete slant medium by saccharomycete, cultivate 72h at 28 DEG C, from saccharomycete slant medium, picking one ring saccharomycete, is seeded in saccharomycete fluid nutrient medium, cultivates 36 ~ 72h, obtain saccharomycete enrichment liquid at 35 ~ 37 DEG C;
B, get saccharomycete enrichment liquid prepared by steps A, the inoculum concentration being 0.5 ~ 5:100 according to volume ratio is seeded in saccharomycete fluid nutrient medium, regulates pH to 6.5 ~ 7.5, ventilate cavity 36 ~ 72h at 28 DEG C; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in saccharomycete fluid nutrient medium, and regulate pH to 6.5 ~ 7.5, ventilate cavity 36 ~ 72h at 28 DEG C, until be extended to 5L; Now in saccharomycete fluid nutrient medium, saccharomycete viable count is 1.3 × 10
9~ 3.6 × 10
9cFU/g;
Wherein, the preparation method of saccharomycete slant medium is as follows: get glucose 5g, peptone 2g, yeast extract 1g, sodium chloride 0.5g, agar 15g prepare, saccharomycete slant medium pH is 6.5 ~ 7.5,121 DEG C of sterilizing 20min.
The preparation method of saccharomycete fluid nutrient medium is as follows: get glucose 5g, peptone 2g, yeast extract 1g, sodium chloride 0.5g, distilled water 1000ml prepare, 121 DEG C of sterilizing 20min.
(3) preparation of bacillus subtilis liquid
A, bacillus subtilis is inoculated in bacillus subtilis slant medium, 48h is cultivated at 37 DEG C, picking one ring bacillus subtilis from bacillus subtilis slant medium, be seeded in bacillus subtilis fluid nutrient medium, cultivate 36 ~ 72h at 37 DEG C, obtain bacillus subtilis enrichment liquid;
B, get bacillus subtilis enrichment liquid prepared by steps A, the inoculum concentration being 0.5 ~ 5:100 according to volume ratio is seeded in bacillus subtilis fluid nutrient medium; Regulate pH to 7.0 ~ 7.2, at 37 DEG C, cultivate 36 ~ 72h; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in bacillus subtilis fluid nutrient medium, regulates pH to 7.0 ~ 7.2, cultivates 36 ~ 72h, until be extended to 5L at 37 DEG C; Now in fluid nutrient medium, bacillus subtilis viable count is 1.5 × 10
8~ 2.3 × 10
8cFU/g;
Wherein, the preparation method of bacillus subtilis slant medium is as follows: get glucose 5g, peptone 2g, yeast extract 1g, sodium chloride 0.5g, agar 15g prepare, saccharomycete slant medium pH 6.5 ~ 7.5,121 DEG C of sterilizing 20min.
The preparation method of bacillus subtilis fluid nutrient medium is as follows: get tryptone 10g, yeast extract powder 5g, sodium chloride 10g, distilled water 1000mL prepare, 121 DEG C of sterilizing 20min.
(4) preparation of lactobacillus suspension
A, by lactobacillus inoculum in lactic acid bacteria slant medium, cultivate 48h at 37 DEG C; From lactic acid bacteria slant medium, picking one ring lactic acid bacteria, is seeded in lactic acid bacteria fluid nutrient medium, cultivates 36 ~ 72h, obtain lactic acid bacteria enrichment liquid at 28 ~ 32 DEG C;
B, get lactic acid bacteria enrichment liquid prepared by steps A, the inoculum concentration being 0.5 ~ 5:100 according to volume ratio is seeded in lactic acid bacteria fluid nutrient medium, regulates pH to 5.5 ~ 6.0, cultivates 36 ~ 72h at 37 DEG C; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in lactic acid bacteria fluid nutrient medium, regulates pH to 5.5 ~ 6.0, cultivates 36 ~ 72h, until be extended to 5L at 37 DEG C; Now in lactic acid bacteria fluid nutrient medium, viable count of lactobacillus is 2.3 × 10
8~ 4.0 × 10
8cFU/g;
Wherein, the preparation method of lactic acid bacteria slant medium is as follows: extracting degreasing milk powder 100g, dusty yeast 1g, dipotassium hydrogen phosphate 5g, agar 15g prepare, 121 DEG C of sterilizing 20min.
The preparation method of lactic acid bacteria fluid nutrient medium is as follows: get peptone 10g, powdered beef 5g, yeast extract powder 4g, beef peptone 10g, glucose 20g, Tween-80 1mL, dipotassium hydrogen phosphate 2g, sodium acetate 5g, Triammonium citrate 2g, magnesium sulfate 0.2g, manganese sulfate 0.05g, distilled water 1000mL prepare, 121 DEG C of sterilizing 20min.
(5) preparation of fermented bean dregs
Take raw material dregs of beans 500kg, wheat bran 20kg, ammonium sulfate 20kg, molasses 10kg add in mixer successively, inject the running water of 450kg after mix and blend 10min, continue mixing and stir 10min, make fermentation substrate; In fermentation substrate, inject 12kg mixed bacteria liquid, wherein mixed bacteria liquid is that 2:1:1:1 forms by black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension according to weight ratio; Be uniformly mixed 10min, then heap fermentation 80h at 27 DEG C ~ 37 DEG C; Finally fermented bean dregs is transferred to drying plant, drying to moisture content at 45 ~ 65 DEG C is about 12%, pulverizes, and packing, obtains fermented bean dregs product.
Comparative example 1
Investigate fermentation time to the impact of soybean products quality.
Comparative example 1-1: the time except heap fermentation is except 68h, all the other are all with embodiment 1.
Comparative example 1-2: the time except heap fermentation is except 72h, all the other are all with embodiment 1.
Comparative example 1-3: the time except heap fermentation is except 76h, all the other are all with embodiment 1.
Comparative example 1-4: the time except heap fermentation is except 88h, all the other are all with embodiment 1.
Comparative example 1-5: the time except heap fermentation is except 96h, all the other are all with embodiment 1.
Comparative example 1-6: the time except heap fermentation is except 100h, all the other are all with embodiment 1.
Analyze the Polyose degradation rate of each soybean products, oligosaccharides degradation rate, antigen degradation rate, crude protein, little peptide, lactic acid, analyze the transformation efficiency of fermented bean dregs simultaneously, its result is as shown in table 1 below.
The Quality Detection result of each fermented bean dregs of table 1
From upper table 1, when being less than 72h when fermenting, because fermentation time is inadequate, its Polyose degradation rate is only 48.03%, oligosaccharides degradation rate is only 95.01%, antigen degradation rate is 82.32%, crude protein (percentage composition) is 50.03%, little peptide is (percentage composition) 9.25%, lactic acid is (percentage composition) 4.51%, transformation efficiency is low to moderate 86.03%, has had a strong impact on the quality of fermented bean dregs.Be 72h ~ 96h when fermenting, fermentation time is suitable for, and the quality of each fermented bean dregs obtains obvious raising.When wherein fermentation time is 80h, Polyose degradation rate is 51.30%, oligosaccharides degradation rate is 99.21%, antigen degradation rate is 90.61%, crude protein (percentage composition) is 60.87%, little peptide is (percentage composition) 15.12%, lactic acid is (percentage composition) 5.46%, transformation efficiency is 91.37%, is in preferably effect with less advantage.After this, along with the continuation of fermentation time extends, the testing result change of each index is mild, shows that fermentation time is longer than 96h, can cause excessive fermentation, cause unnecessary waste, nutritional benefit composition also can be caused simultaneously to a certain extent to be also decomposed.
Comparative example 2
Investigate the weight ratio of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension in mixed bacteria liquid to the impact of fermented bean dregs quality.
Comparative example 2-1: except in mixed bacteria liquid, the weight ratio of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension is 1:2:0.5:0.5, all the other are all with embodiment 1.
Comparative example 2-2: except in mixed bacteria liquid, the weight ratio of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension is 4:1:2:2, all the other are all with embodiment 1.
Comparative example 2-3: except in mixed bacteria liquid, the weight ratio of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension is 3:0.5:1:1, all the other are all with embodiment 1.
Comparative example 2-4: except in mixed bacteria liquid, the weight ratio of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension is 0.5:3:3:3, all the other are all with embodiment 1.
Comparative example 2-5: except in mixed bacteria liquid, the weight ratio of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension is 5:0.1:0.5:3, all the other are all with embodiment 1.
Analyze the Polyose degradation rate of each soybean products, oligosaccharides degradation rate, antigen degradation rate, crude protein, little peptide, lactic acid, analyze the transformation efficiency of fermented bean dregs simultaneously, its result is as shown in table 2 below.
The Quality Detection result of each fermented bean dregs of table 2
From the indices testing result of embodiment 1 and comparative example 2-1 ~ 2-3 in above-mentioned table 2, in fermentation process of the present invention, when in mixed bacteria liquid, the weight ratio of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension is 1 ~ 4:0.5 ~ 2:0.5 ~ 2:0.5 ~ 2, the quality of fermented bean dregs can be significantly improved, its Polyose degradation rate is greater than 51%, oligosaccharides degradation rate is greater than 99%, and antigen degradation rate is greater than 90%, crude protein is greater than 60%, little peptide is greater than 15%, lactic acid is greater than 5%, transformation efficiency is greater than 90%.Wherein, when black-koji mould liquid in mixed bacteria liquid, yeast liquid, the weight ratio of bacillus subtilis liquid and lactobacillus suspension is 2:1:1:1, its Polyose degradation rate is 51.30%, oligosaccharides degradation rate is 99.21%, antigen degradation rate is 90.61%, crude protein (percentage composition) is 60.87%, little peptide is (percentage composition) 15.12%, lactic acid is (percentage composition) 5.46%, transformation efficiency is 91.37%, a little more than comparative example 2-1, 2-2 and 2-3, preferably effect is in less advantage, i.e. black-koji mould liquid in mixed bacteria liquid, yeast liquid, the weight ratio of bacillus subtilis liquid and lactobacillus suspension is 2:1:1:1 is optimum implementation of the present invention.
And the fermented bean dregs quality of comparative example 2-4 and comparative example 2-5 is compared with comparative example 2-1 ~ 2-3, be decreased significantly, its Polyose degradation rate is less than 50%, oligosaccharides degradation rate is less than 90%, and antigen degradation rate is less than 90%, crude protein is less than 55%, little peptide is less than 15%, lactic acid is less than 5%, transformation efficiency is less than 70%.Show, when the weight ratio of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension in mixed bacteria liquid is not in the protection domain of the claims in the present invention, significantly to reduce the quality of fermented bean dregs.
Comparative example 3
Step (1) ~ step (4) is all with embodiment 1.
Step (5) is as follows:
A, take raw material dregs of beans 500kg, wheat bran 20kg, ammonium sulfate 20kg, molasses 10kg add in mixer successively, inject the running water of 450kg after mix and blend 10min, continue mixing and stir 10min, make fermentation substrate; Black-koji mould liquid prepared by 12kg step (1) is injected in fermentation substrate; Be uniformly mixed 10min, then heap fermentation 80h at 27 DEG C ~ 37 DEG C; Finally fermented bean dregs is transferred to drying plant, drying to moisture content at 45 ~ 65 DEG C is about 12%, pulverizes, and packing, obtains fermented bean dregs A.
B, take raw material dregs of beans 500kg, wheat bran 20kg, ammonium sulfate 20kg, molasses 10kg add in mixer successively, inject the running water of 450kg after mix and blend 10min, continue mixing and stir 10min, make fermentation substrate; Yeast liquid prepared by 12kg step (2) is injected in fermentation substrate; Be uniformly mixed 10min, then heap fermentation 80h at 27 DEG C ~ 37 DEG C; Finally fermented bean dregs is transferred to drying plant, drying to moisture content at 45 ~ 65 DEG C is about 12%, pulverizes, and packing, obtains fermented bean dregs B.
C, take raw material dregs of beans 500kg, wheat bran 20kg, ammonium sulfate 20kg, molasses 10kg add in mixer successively, inject the running water of 450kg after mix and blend 10min, continue mixing and stir 10min, make fermentation substrate; Bacillus subtilis liquid prepared by 12kg step (3) is injected in fermentation substrate; Be uniformly mixed 10min, then heap fermentation 80h at 27 DEG C ~ 37 DEG C; Finally fermented bean dregs is transferred to drying plant, drying to moisture content at 45 ~ 65 DEG C is about 12%, pulverizes, and packing, obtains fermented bean dregs C.
D, take raw material dregs of beans 500kg, wheat bran 20kg, ammonium sulfate 20kg, molasses 10kg add in mixer successively, inject the running water of 450kg after mix and blend 10min, continue mixing and stir 10min, make fermentation substrate; Lactobacillus suspension prepared by 12kg step (4) is injected in fermentation substrate; Be uniformly mixed 10min, then heap fermentation 80h at 27 DEG C ~ 37 DEG C; Finally fermented bean dregs is transferred to drying plant, drying to moisture content at 45 ~ 65 DEG C is about 12%, pulverizes, and packing, obtains fermented bean dregs D.
E, take raw material dregs of beans 500kg, wheat bran 20kg, ammonium sulfate 20kg, molasses 10kg add in mixer successively, inject the running water of 450kg after mix and blend 10min, continue mixing and stir 10min, make fermentation substrate; In fermentation substrate, inject 12kg mixed bacteria liquid, wherein mixed bacteria liquid is that 1:1:1 forms by yeast liquid, bacillus subtilis liquid and lactobacillus suspension according to weight ratio; Be uniformly mixed 10min, then heap fermentation 80h at 27 DEG C ~ 37 DEG C; Finally fermented bean dregs is transferred to drying plant, drying to moisture content at 45 ~ 65 DEG C is about 12%, pulverizes, and packing, obtains fermented bean dregs E.
F, take raw material dregs of beans 500kg, wheat bran 20kg, ammonium sulfate 20kg, molasses 10kg add in mixer successively, inject the running water of 450kg after mix and blend 10min, continue mixing and stir 10min, make fermentation substrate; In fermentation substrate, inject 12kg mixed bacteria liquid, wherein mixed bacteria liquid is that 2:1:1 forms by black-koji mould liquid, bacillus subtilis liquid and lactobacillus suspension according to weight ratio; Be uniformly mixed 10min, then heap fermentation 80h at 27 DEG C ~ 37 DEG C; Finally fermented bean dregs is transferred to drying plant, drying to moisture content at 45 ~ 65 DEG C is about 12%, pulverizes, and packing, obtains fermented bean dregs F.
G, take raw material dregs of beans 500kg, wheat bran 20kg, ammonium sulfate 20kg, molasses 10kg add in mixer successively, inject the running water of 450kg after mix and blend 10min, continue mixing and stir 10min, make fermentation substrate; In fermentation substrate, inject 12kg mixed bacteria liquid, wherein mixed bacteria liquid is that 2:1 forms by black-koji mould liquid, bacillus subtilis liquid according to weight ratio; Be uniformly mixed 10min, then heap fermentation 80h at 27 DEG C ~ 37 DEG C; Finally fermented bean dregs is transferred to drying plant, drying to moisture content at 45 ~ 65 DEG C is about 12%, pulverizes, and packing, obtains fermented bean dregs G.
Grab sample detects, and its result is:
Glycan total content 7.64% in fermented bean dregs A, oligosaccharides total content 0.15%, glycinin 5.49%, β-companion globulin 4.53%, crude protein 63.27%.Correspondingly, Polyose degradation rate is 51.02%, and oligosaccharides degradation rate is 96.05%, and antigen degradation rate is 60.70%, and crude protein improves 37.84%.
In fermented bean dregs B, glycan total content is 14.93%, oligosaccharides total content 3.35%, glycinin 3.03%, β-companion globulin 2.82%, crude protein 49.01%; Correspondingly, Polyose degradation rate is 4.32%, and oligosaccharides degradation rate is 11.84%, and antigen degradation rate is 77.06%, and crude protein improves 14.18%.
In fermented bean dregs C, glycan total content is 11.92%, oligosaccharides total content 0.89%, glycinin 4.54%, β-companion globulin 4.19%, crude protein 51.26%; Correspondingly, Polyose degradation rate is 23.59%, and oligosaccharides degradation rate is 76.50%, and antigen degradation rate is 65.8%, and crude protein improves 19.21%.
In fermented bean dregs D, glycan total content is 13.31%, oligosaccharides total content 0.38%, glycinin 9.33%, β-companion globulin 8.97%, crude protein 50.59%; Correspondingly, Polyose degradation rate is 14.68%, and oligosaccharides degradation rate is 90.0%, and antigen degradation rate is 28.23%, and crude protein improves 17.65%.
In fermented bean dregs E, glycan total content is 11.04%, oligosaccharides total content 0.35%, glycinin 2.32%, β-companion globulin 2.19%, crude protein 52.31%; Correspondingly, Polyose degradation rate is 29.23%, and oligosaccharides degradation rate is 90.79%, and antigen degradation rate is 82.31%, and crude protein improves 21.65%.
In fermented bean dregs F, glycan total content is 7.96%, oligosaccharides total content 0.22%, glycinin 4.23%, β-companion globulin 3.91%, crude protein 60.04%; Correspondingly, Polyose degradation rate is 51.02%, and oligosaccharides degradation rate is 94.21%, and antigen degradation rate is 68.08%, and crude protein improves 39.63%.
In fermented bean dregs G, glycan total content is 7.76%, oligosaccharides total content 0.13%, glycinin 4.37%, β-companion globulin 4.03%, crude protein 60.12%; Correspondingly, Polyose degradation rate is 50.30%, and oligosaccharides degradation rate is 96.58%, and antigen degradation rate is 67.05%, and crude protein improves 25.47%.
The above results is listed in the table below in 3.
Table 3 bacterial classification is on the impact of fermented bean dregs quality
| Group | Polyose degradation rate | Oligosaccharides degradation rate | Antigen degradation rate | Crude protein |
| Embodiment 1 | 51.30％ | 99.21％ | 90.61％ | 60.87％ |
| Fermented bean dregs A | 51.02％ | 96.05％ | 60.70％ | 59.27％ |
| Fermented bean dregs B | 4.32％ | 11.84％ | 77.06％ | 49.01％ |
| Fermented bean dregs C | 23.59％ | 76.50％ | 65.80％ | 51.26％ |
| Fermented bean dregs D | 14.68％ | 90.00％ | 28.23％ | 50.59％ |
| Fermented bean dregs E | 29.23％ | 90.79％ | 82.31％ | 52.31％ |
| Fermented bean dregs F | 51.02％ | 94.21％ | 68.08％ | 60.04％ |
| Fermented bean dregs G | 50.30％ | 96.58％ | 67.05％ | 60.12％ |
From fermented bean dregs A and embodiment 1, when only adding black-koji mould liquid, its oligosaccharides degradation rate reduces 3.16%, and antigen degradation rate reduces 29.91%, and crude protein content is substantially suitable with embodiment 1 with Polyose degradation rate.
From fermented bean dregs B and embodiment 1, when only adding yeast liquid, its Polyose degradation rate reduces 46.98%, and oligosaccharides degradation rate reduces 82.41%, and antigen degradation rate reduces 13.55%, and crude protein content reduces 11.86%.
From fermented bean dregs C and embodiment 1, when only adding bacillus subtilis liquid, its Polyose degradation rate reduces 27.71%, and oligosaccharides degradation rate reduces 22.71%, and antigen degradation rate reduces 24.81%, and crude protein content reduces 9.61%.
From fermented bean dregs D and embodiment 1, when only adding lactobacillus suspension, its Polyose degradation rate reduces 36.62%, and oligosaccharides degradation rate reduces 9.21%, and antigen degradation rate reduces 62.38%, and crude protein content reduces 10.28%.
From fermented bean dregs E and embodiment 1, when only adding yeast liquid, bacillus subtilis liquid and lactobacillus suspension, its Polyose degradation rate reduces 22.07%, and oligosaccharides degradation rate reduces 8.42%, antigen degradation rate reduces 8.3%, and crude protein content reduces 8.56%.
From fermented bean dregs F and embodiment 1, when only adding black-koji mould liquid, bacillus subtilis liquid and lactobacillus suspension, oligosaccharides degradation rate reduces 5.00%, and antigen degradation rate reduces 22.53%, and crude protein is substantially suitable with embodiment 1 with Polyose degradation rate.
From fermented bean dregs G and embodiment 1, when only adding black-koji mould liquid and bacillus subtilis liquid, oligosaccharides degradation rate reduces 2.63%, and antigen degradation rate reduces 23.56%, and crude protein is substantially suitable with embodiment 1 with Polyose degradation rate.
The above results shows, the present invention adopts black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension to be that mixed bacteria liquid carries out bean pulp fermentation, black-koji mould, saccharomycete, bacillus subtilis and lactic acid bacteria co-fermentation create the effect of Synergistic, significantly improve the content of Polyose degradation rate, oligosaccharides degradation rate and antigen degradation rate and crude protein.Lack wherein any one bacterial classification and all do not reach technique effect of the present invention.
Comparative example 4
Following methods is adopted to prepare fermented bean dregs respectively.
(1) preparation of traditional zymotic dregs of beans 1: preparation dregs of beans solid fermentation base-material, be mixed into nutritious supplementary pharmaceutical, make solid fermentation culture medium, to solid fermentation inoculation of medium ocean rhodotorula and lactic acid bacteria culturers, after solid fermentation, obtain Rhodotorula benthica fermentation feed.Fermented bean dregs finished product is put in shady and cool place after weighing, packing.
(2) preparation of traditional zymotic dregs of beans 2: first class inoculum is cultivated to obtain in inclined-plane preservation or frozen bafillus natto bacterial classification KMB culture media shaking vase; Bean cake powder, cornstarch, corn steep liquor, NaCl is water-soluble, adjust ph, sterilizing, access first class inoculum Shaking culture obtains second class inoculum; Then be soaked in water dregs of beans, then add NaCl, carbon source glucose, sucrose or cornstarch, nitrogenous source gelatin, aerobic fermentation 48 hours at 40 DEG C, then dry sample preparation at 60 DEG C.Fermented bean dregs finished product is put in shady and cool place after weighing, packing.
(3) preparation of traditional zymotic dregs of beans 3: by expanding cultured bacillus subtilis 3 strain, the fermentation culture of saccharomyces cerevisiae 1 strain and lactic acid bacteria 1 strain is to mix at 1.8: 1: 1 by weight, it is in the Cottonseed Meal (crossing 40 mesh sieves) of 50% that inoculum concentration with 5% is inoculated into water content, add the molasses of 5% simultaneously, and mix (initial pH value is 7.0), aerobic fermentation 48 hours at 40 DEG C, then dry sample preparation at 60 DEG C.Fermented bean dregs finished product is put in shady and cool place after weighing, packing.
(4) preparation of traditional zymotic dregs of beans 4: add water and the moisture soybean meal slurry that stirs to obtain in dregs of beans raw material, moisture soybean meal slurry is carried out sterilization treatment, add urea and ammonium and stir, obtain dregs of beans compost, then in dregs of beans compost, access can produce the microorganism fungus kind of lysine, temperature controlled fermentation, obtains fermented bean dregs, dry finished product.
The testing result of each fermented bean dregs is as shown in table 4 below.
The testing result of each fermented bean dregs quality of table 4
From above-mentioned table 4, the indices of fermented bean dregs embodiment 1 prepared is compared with the indices of traditional zymotic dregs of beans 1 ~ 4, and the Polyose degradation rate of embodiment 1 improves 37.15%, 39.21%, 41.48%, 42.55% respectively; Oligosaccharides degradation rate improves 12.43%, 9.06%, 29.38%, 26.4% respectively; Antigen degradation rate improves 38.16%, 36.91%, 34.48%, 12.02% respectively; Crude protein improves 10.42%, 13.17%, 11.69%, 11.82% respectively; Little peptide improves 5.8%, 6.26%, 3.57%, 1.04% respectively; Lactic acid improves 3.12%, 1.73%, 0.43%, 2.04% respectively.Show, preparation method of the present invention, compared with traditional preparation method, significantly improves Polyose degradation rate and the antigen degradation rate of fermented bean dregs, significantly improves protein quality.
Claims (10)
1. a preparation method for feeding fermented dregs of beans, is characterized in that: be inoculated into by mixed bacteria liquid in bean pulp fermentation substrate, mix, and at 27 ~ 37 DEG C of bottom fermentation 72 ~ 96h, then dries, and pulverizes, obtains feeding fermented dregs of beans; Described mixed bacteria liquid is made up of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension; The weight ratio of described mixed bacteria liquid and bean pulp fermentation substrate is 6 ~ 18:1000, and in described mixed bacteria liquid, the weight ratio of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension is 1 ~ 4:0.5 ~ 2:0.5 ~ 2:0.5 ~ 2; Black-koji mould viable count in described black-koji mould liquid is 2.5 × 10
7~ 4.6 × 10
7cFU/g; In described yeast liquid, saccharomycete viable count is 1.3 × 10
9~ 3.6 × 10
9cFU/g; Bacillus subtilis viable count in described bacillus subtilis liquid is 1.5 × 10
8~ 2.3 × 10
8cFU/g; In described lactobacillus suspension, viable count of lactobacillus is 2.3 × 10
8~ 4.0 × 10
8cFU/g.
2. preparation method according to claim 1, is characterized in that, described bean pulp fermentation substrate is made up of the raw material of following weight: dregs of beans 30 ~ 70 parts, 1 ~ 4 part, wheat bran, 1 ~ 4 part, ammonium sulfate, 0.5 ~ 2 part, molasses, 30 ~ 70 parts, water.
3. preparation method according to claim 1, is characterized in that: be describedly inoculated in bean pulp fermentation substrate by mixed bacteria liquid, mixes, at 27 ~ 37 DEG C of bottom fermentation 80h.
4. preparation method according to claim 1, is characterized in that: the weight ratio of described mixed bacteria liquid and bean pulp fermentation substrate is 10 ~ 15:1000.
5. preparation method according to claim 4, is characterized in that: the weight ratio of described mixed bacteria liquid and bean pulp fermentation substrate is 12:1000.
6. preparation method according to claim 1, is characterized in that: in described mixed bacteria liquid, the weight ratio of black-koji mould liquid, yeast liquid, bacillus subtilis liquid and lactobacillus suspension is 2:1:1:1.
7. the preparation method according to any one of claim 1 ~ 6, is characterized in that, described black-koji mould liquid is made up of following methods:
(1) black-koji mould is inoculated in black-koji mould slant medium, at 25 ~ 30 DEG C, cultivates 68 ~ 76h; From black-koji mould slant medium picking one ring black-koji mould, be seeded in black-koji mould fluid nutrient medium, cultivate 72 ~ 96h at 23 ~ 28 DEG C, obtain black-koji mould enrichment liquid;
(2) getting black-koji mould enrichment liquid prepared by step (1), is that the inoculum concentration of 0.5 ~ 5:100 is seeded in black-koji mould fluid nutrient medium by volume, regulates pH to 6.2 ~ 6.6, ventilate cavity 72 ~ 96h at 23 ~ 28 DEG C; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in black-koji mould fluid nutrient medium, regulate pH to 6.2 ~ 6.6, ventilate cavity 72 ~ 96h at 23 ~ 28 DEG C, until be extended to 5L, now in black-koji mould fluid nutrient medium, black-koji mould viable count is 2.5 × 10
7~ 4.6 × 10
7cFU/g;
Described black-koji mould slant medium is that raw material is made by glucose 8 ~ 12g, peptone 4 ~ 6g, potassium dihydrogen phosphate 0.5 ~ 2g, magnesium sulfate 0.1 ~ 1g, agar 10 ~ 30g, rose-bengal 0.03 ~ 0.04g, chloramphenicol 0.05 ~ 0.2g, distilled water 900 ~ 1100mL;
Described black-koji mould fluid nutrient medium is that raw material is formulated by peptone 4 ~ 6g, yeast extract powder 1 ~ 3g, magnesium sulfate 0.1 ~ 1g, glucose 10 ~ 30g, dipotassium hydrogen phosphate 0.5 ~ 2g, distilled water 900 ~ 1000mL.
8. the preparation method according to any one of claim 1 ~ 6, is characterized in that, described yeast liquid is made up of following methods:
(1) saccharomycete is inoculated in saccharomycete slant medium, cultivate 68 ~ 76h at 25 ~ 30 DEG C, from saccharomycete slant medium, picking one ring saccharomycete, is seeded in saccharomycete fluid nutrient medium, cultivate 36 ~ 72h at 35 ~ 37 DEG C, obtain saccharomycete enrichment liquid;
(2) get saccharomycete enrichment liquid prepared by step (1), the inoculum concentration being 0.5 ~ 5:100 according to volume ratio is seeded in saccharomycete fluid nutrient medium, regulates pH to 6.5 ~ 7.5, ventilate cavity 36 ~ 72h at 25 ~ 30 DEG C; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in saccharomycete fluid nutrient medium, and regulate pH to 6.5 ~ 7.5, ventilate cavity 36 ~ 72h at 25 ~ 30 DEG C, until be extended to 5L; Now in saccharomycete fluid nutrient medium, saccharomycete viable count is 1.3 × 10
9~ 3.6 × 10
9cFU/g;
Described saccharomycete slant medium is by glucose 3 ~ 7g, peptone 1 ~ 3g, yeast extract 0.5 ~ 2g, sodium chloride 0.1 ~ 1g, and agar 10 ~ 20g is that raw material is formulated;
Described saccharomycete fluid nutrient medium is that raw material is formulated by glucose 3 ~ 7g, peptone 1 ~ 3g, yeast extract 0.5 ~ 2g, sodium chloride 0.1 ~ 1g and distilled water 900 ~ 1100ml.
9. the preparation method according to any one of claim 1 ~ 6, is characterized in that, described bacillus subtilis liquid is obtained by following methods:
(1) bacillus subtilis is inoculated in bacillus subtilis slant medium, 40 ~ 60h is cultivated at 35 ~ 38 DEG C, picking one ring bacillus subtilis from bacillus subtilis slant medium, be seeded in bacillus subtilis fluid nutrient medium, cultivate 36 ~ 72h at 35 ~ 38 DEG C, obtain bacillus subtilis enrichment liquid;
(2) get bacillus subtilis enrichment liquid prepared by step (1), the inoculum concentration being 0.5 ~ 5:100 according to volume ratio is seeded in bacillus subtilis fluid nutrient medium; Regulate pH to 7.0 ~ 7.2, at 35 ~ 38 DEG C, cultivate 36 ~ 72h; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in bacillus subtilis fluid nutrient medium, regulates pH to 7.0 ~ 7.2, cultivates 36 ~ 72h, until be extended to 5L at 35 ~ 38 DEG C; Now in fluid nutrient medium, bacillus subtilis viable count is 1.5 × 10
8~ 2.3 × 10
8cFU/g;
Described bacillus subtilis slant medium is that raw material is formulated by glucose 3 ~ 7g, peptone 1 ~ 4g, yeast extract 0.5 ~ 2g, sodium chloride 0.1 ~ 1g, agar 10 ~ 20g;
Described bacillus subtilis fluid nutrient medium is that raw material is formulated by tryptone 5 ~ 15g, yeast extract powder 1 ~ 10g, sodium chloride 5 ~ 20g, distilled water 900 ~ 1100ml.
10. the preparation method according to any one of claim 1 ~ 6, is characterized in that, described lactobacillus suspension is obtained by following methods:
(1) by lactobacillus inoculum in lactic acid bacteria slant medium, at 35 ~ 38 DEG C cultivate 40 ~ 60h; From lactic acid bacteria slant medium, picking one ring lactic acid bacteria, is seeded in lactic acid bacteria fluid nutrient medium, cultivates 36 ~ 72h, obtain lactic acid bacteria enrichment liquid at 28 ~ 32 DEG C;
(2) get lactic acid bacteria enrichment liquid prepared by step (1), the inoculum concentration being 0.5 ~ 5:100 according to volume ratio is seeded in lactic acid bacteria fluid nutrient medium, regulates pH to 5.5 ~ 6.0, cultivates 36 ~ 72h at 35 ~ 38 DEG C; After this inoculum concentration that each inoculation is all 0.5 ~ 5:100 according to volume ratio is seeded in lactic acid bacteria fluid nutrient medium, regulates pH to 5.5 ~ 6.0, cultivates 36 ~ 72h, until be extended to 5L at 35 ~ 38 DEG C; Now in lactic acid bacteria fluid nutrient medium, viable count of lactobacillus is 2.3 × 10
8~ 4.0 × 10
8cFU/g;
Described lactic acid bacteria liquid slant medium is that raw material is formulated by skimmed milk power 50 ~ 200g, dusty yeast 0.5 ~ 2g, dipotassium hydrogen phosphate 1 ~ 10g, agar 10 ~ 20g;
Described lactic acid bacteria fluid nutrient medium is that raw material is formulated by peptone 5 ~ 20g, powdered beef 1 ~ 10g, yeast extract powder 2 ~ 8g, beef peptone 5 ~ 14g, glucose 10 ~ 30g, Tween-80 0.5 ~ 2mL, dipotassium hydrogen phosphate 1 ~ 4g, sodium acetate 1 ~ 10g, Triammonium citrate 0.5 ~ 4g, magnesium sulfate 0.1 ~ 0.3g, manganese sulfate 0.01 ~ 0.1g, distilled water 900 ~ 1000ml.
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