CN105410337A - Biological feed additive rich in bioactive peptides and probiotics as well as preparation method of additive - Google Patents
Biological feed additive rich in bioactive peptides and probiotics as well as preparation method of additive Download PDFInfo
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
Abstract
The invention discloses a biological feed additive rich in bioactive peptides and probiotics as well as a preparation method of the additive. The biological feed additive prepared with the method has the peptide content higher than 20%, the active probiotic content up to 1.0*10<9> cfu/g and contains fermentation metabolites such as bacteriocin and the like; meanwhile, the additive has the advantages that the additive is purely natural, non-toxic, harmless and free of pollution and residues.
Description
Technical field
The invention belongs to biological enzymolysis and fermentation technical field, particularly relate to and be a kind ofly rich in biology feed additive of active peptide and beneficial bacterium and preparation method thereof.
Background technology
Cottonseed Meal, rapeseed dregs, Soybean Meal, fish meal, wheat bran, corn, rice bran etc. are conventional feed industry raw materials.Cottonseed Meal is mainly raw material with cottonseed, products obtained therefrom after using prepressing extraction or direct leaching to deoil.Cottonseed Meal contains the crude protein of about 42.5%, the crude fibre of about 10.1%, the crude fat of about 0.7%, the coarse ash of about 6.5%.Rapeseed dregs is the accessory substance after rapeseed oil.It contains the crude protein of about 38.6%, the crude fibre of about 11.8%, the crude fat of about 1.4%, the coarse ash of about 7.3%.Soybean Meal is the byproduct obtained after soybean extracting bean oil.It contains the crude protein of about 43.0%, the crude fibre of about 5.1%, the crude fat of about 1.9%, the coarse ash of about 6.0%.Fish meal is raw material with one or more fish, and the high-protein feed raw material after processing of deoiling, dewater, pulverize, it contains the crude protein of about 62.5%, the crude fibre of about 0.5%, the crude fat of about 4.0%, the coarse ash of about 12.3%.Wheat bran is the byproduct obtained after wheat processing flour.It contains the crude protein of about 15.7%, the crude fibre of about 8.9%, the crude fat of about 3.9%, the coarse ash of about 4.9%.Corn contains the crude protein of about 8.7%, the crude fibre of about 1.6%, the crude fat of about 3.6%, the coarse ash of about 1.4%.Rice bran is the principal by product of paddy processing.It contains the crude protein of about 12.8%, the crude fibre of about 5.7%, the crude fat of about 16.5%, the coarse ash of about 7.5%.
Amylase, cellulase, hemicellulase, pectase, lipase, phytase, protease, keratinase, alpha-galactosidase, 1,4 beta-glucanase, 'beta '-mannase, zytase etc. are conventional feed fermentation or interpolation enzyme preparation.Amylase energy hydrolyzed starch, glycogen and the enzyme about the O-glucose key in polysaccharide, comprise AMS, beta amylase, gamma amylase, isoamylase etc.Cellulase is the glucogenic one group of enzyme of degraded cellulose, primarily of compositions such as circumscribed 1,4 beta-glucanase, Endo-β-glucanase, beta-glucosidase, zytases.Hemicellulase is a complicated enzyme system, and a lot of hemicellulose take wood sugar as main chain, and side chain has the monose such as mannose, glucose, arabinose, and that decomposes hemicellulose has zytase, galactosidase, mannase etc.Pectase refers to the enzyme of decompose pectin matter, comprises pectinesterase, polygalacturonase and pectin lyase.Lipase is under the jurisdiction of carboxylic ester hydrolase class, progressively triglyceride hydrolysis can be become glycerine and aliphatic acid, comprise phosphate, sterol enzyme and carboxy-lesterase.Phytase can take off phosphate group by catalysis phytic acid, comprises phytic acid 3-phosphohydrolase, phytic acid 6-phosphohydrolase and the cruel hydrolase of nonspecific orthophosphoric acid list.Protease is the class of enzymes of aminosal peptide chain, by the mode of its hydrolyzed peptide, is divided into endopeptidase and exopeptidase two class; By its activated centre, be divided into serine protease, thiol protease, metalloproteinases and aspartic protease; By the optimum pH of its reaction, be divided into acid protease, neutral proteinase and alkali protease.Keratinase is that one can exclusively be degraded keratic protease, and its degraded substrate is except keratin, and it goes back hydrolyzable multiple proteins substrate, comprises solvable and insoluble protein.Alpha-galactosidase refers to the enzyme that catalysis α-D-galactoside (as galactooligosaccharicomposition, galactomannan) end irreducibility α-D-galactose residue is hydrolyzed.1,4 beta-glucanase refers to the multiple enzyme of energy catalyzing hydrolysis beta glucan, comprises endo-type β-Isosorbide-5-Nitrae-dextranase, endo-type β-1,3-dextranase, circumscribed-type β-Isosorbide-5-Nitrae-dextranase, circumscribed-type β-1,3-dextranase.The manna oligosacchride that 'beta '-mannase is hydrolysis is main chain with β-Isosorbide-5-Nitrae-D-mannopyranose, the endo hydrolysis enzyme of mannocarolose (mannosan, glucomannan, galactomannans), belong to a kind of hemicellulose enzyme.Zytase is the enzyme system of a class degradation of xylan, comprises β-Isosorbide-5-Nitrae-endo-xylanase, xylobiase, α-Arabinose glycosides enzyme, α-D-glucuronidase, acetyl group zytase and phenolic acid esterase, degradable xylan class hemicellulose.
Lactic acid bacteria, saccharomycete, bacillus, mould etc. are that bacterial classification produced by conventional fermented feed, are again feeding beneficial bacteriums.Wherein lactic acid bacteria culturers comprises animal bifidobacteria, bifidobacterium bifidum, bifidobacterium infantis, bifidobacterium longum, bifidobacterium breve, bifidobacterium adolescentis, enterococcus faecalis, VREF, lactoenterococcus, lactobacillus acidophilus, Lactobacillus casei, Lactobacillus delbrueckii subsp. lactis, Lactobacillus plantarum, Pediococcus acidilactici, streptococcus thermophilus, lactobacillus reuteri, lactobacillus cellobiosas, lactobacillus fermenti, lactobacillus delbruockii subspecies bulgaricus, Bu Shi lactobacillus, lactobacillus paraceasi etc., yeast seeds comprises candida utili, Candida lipolytica, candida tropicalis, saccharomyces cerevisiae, Issatchenkia orientalis, Pichia pastoris etc., Bacillus comprises bacillus licheniformis, bacillus subtilis, bacillus lentus, bacillus pumilus, bacillus megaterium, bacillus coagulans, Brevibacillus laterosporus, clostridium butyricum etc., mold species comprises aspergillus niger, aspergillus oryzae, Rhizopus oryzae, Mucor etc.The feedstuffs such as Cottonseed Meal, rapeseed dregs, Soybean Meal, fish meal, wheat bran, corn, rice bran, to grade nutritional labeling containing abundant crude protein, crude fibre, crude fat, cinder, Direct-fed animal digestion absorption rate is low.Like product in the market greatly mainly with enzymolysis as enzymolysis dregs of beans, ferment such as fermented bean dregs etc. be main.Enzymolysis dregs of beans peptide content mostly below 10%, non-activity beneficial bacterium content; Fermented bean dregs peptide content is generally below 10%, and active beneficial bacterium content is 1.0 × 10
7cfu/g or following.
Summary of the invention
The present invention solves the aforementioned problems in the prior a kind of of proposition to be rich in biology feed additive of active peptide and beneficial bacterium and preparation method thereof, and make obtained additive peptide content more than 20%, active beneficial bacterium content is up to 1.0 × 10
9more than cfu/g, and containing fermentating metabolism products such as bacteriocins (also claiming antibacterial peptide).
In order to solve the problems of the technologies described above, the technical measures that the present invention takes are:
Be rich in a preparation method for the biology feed additive of active peptide and beneficial bacterium, comprise the following steps:
Step one, prepares beneficial bacterium seed liquor, adopts the method for mixing or the mixed culture fermentation simultaneously of multiple bacterial classification after single strain fermentation, under 20-40 DEG C of condition of culture, carries out aerobic or anaerobic fermentation 8-48h, obtain beneficial bacterium seed liquor;
Step 2, prepares nutrient solution, nutritional labeling is fully mixed, and add after water fully dissolves, sterilization treatment 10-30 minute under 80-121 DEG C of condition, obtains nutrient solution;
Step 3, solid state fermentation, by nutrient solution prepared in beneficial bacterium seed liquor prepared in step one, step 2 together with enzyme preparation, add in solid state fermentation raw material and be fully uniformly mixed, under 5-40 DEG C of ambient temperature conditions, solid state fermentation 12-96 hour, obtains solid state fermentation thing;
Step 4, post processing, the solid state fermentation thing obtained by step 3 solid state fermentation, under 30-300 DEG C of temperature conditions, carries out drying process, obtains the biology feed additive being rich in active peptide and beneficial bacterium.
In order to optimize technique scheme, the technical measures that the present invention takes also comprise:
Preferably, the beneficial bacterium in beneficial bacterium seed liquor described in above-mentioned steps one is selected from one or more in lactic acid bacteria, saccharomycete, bacillus, mould.
Preferably, above-mentioned lactic acid bacteria is selected from one or more in animal bifidobacteria, bifidobacterium bifidum, bifidobacterium infantis, bifidobacterium longum, bifidobacterium breve, bifidobacterium adolescentis, enterococcus faecalis, VREF, lactoenterococcus, lactobacillus acidophilus, Lactobacillus casei, Lactobacillus delbrueckii subsp. lactis, Lactobacillus plantarum, Pediococcus acidilactici, streptococcus thermophilus, lactobacillus reuteri, lactobacillus cellobiosas, lactobacillus fermenti, lactobacillus delbruockii subspecies bulgaricus, Bu Shi lactobacillus or lactobacillus paraceasi.
Preferably, above-mentioned saccharomycete is selected from one or more in candida utili, Candida lipolytica, candida tropicalis, saccharomyces cerevisiae, Issatchenkia orientalis or Pichia pastoris.
Preferably, above-mentioned bacillus is selected from one or more in bacillus licheniformis, bacillus subtilis, bacillus lentus, bacillus pumilus, bacillus megaterium, bacillus coagulans, Brevibacillus laterosporus or clostridium butyricum.
Preferably, above-mentioned mould is selected from one or more in aspergillus niger, aspergillus oryzae, Rhizopus oryzae or Mucor.
Preferably, the medium component of beneficial bacterium seed liquor of fermenting in above-mentioned steps one be selected from rice bran, wheat bran, corn flour, soybean meal powder, soybean-cake flour, tomato powder, cornstarch, Dried Corn Steep Liquor Powder, corn steep liquor, glucose, sucrose, lactose, ox bone peptone, fish peptone, soy peptone, tryptone, casein peptone,
any three kinds or more mixed preparing in the leaching of peptone, lactoalbumin hydrolysate, yeast extract, yeast powder, dusty yeast, beef extract, beef extract powder, sodium acetate, diammonium hydrogen citrate, ammonium sulfate, ammonium chloride, sodium chloride, ferric sesquichloride, calcium chloride, potassium chloride, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium sulfate, manganese sulfate, zinc sulfate, ferrous sulfate, sodium nitrate, L-cysteine hydrochloride or Tween-80.
Preferably, the culture medium pH value before sterilization of beneficial bacterium seed liquor of fermenting in above-mentioned steps one is 4.0-9.0, and beneficial bacterium seed liquor pH prepared by step one is 3.0-9.0, and beneficial bacterium total viable count is 1.0 × 10
8more than cfu/ml.
Preferably, when preparing nutrient solution in above-mentioned steps two, described nutritional labeling Cottonseed Meal hydrolysate, rapeseed dregs hydrolysate, Soybean Meal hydrolysate, Fish meal hydrolysates, glucose, sucrose, lactose, ox bone peptone, the molten slurry of fish, the molten starch of fish, fish peptone, soy peptone, tryptone, casein peptone,
two or more mixed preparing in the leaching of peptone, lactoalbumin hydrolysate, yeast extract, yeast powder, dusty yeast, beef extract, beef extract powder, ammonium sulfate, ammonium chloride, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium sulfate, manganese sulfate, zinc sulfate or Tween-80.
Preferably, described in above-mentioned steps three, enzyme preparation is selected from one or more in amylase, cellulase, hemicellulase, pectase, lipase, phytase, protease, keratinase, alpha-galactosidase, 1,4 beta-glucanase, 'beta '-mannase or zytase.
Preferably, described in above-mentioned steps three, solid state fermentation raw material is selected from one or more in Cottonseed Meal, rapeseed dregs, Soybean Meal, fish meal, wheat bran, corn or rice bran.
Preferably, in above-mentioned steps four, dry process adopts vacuum drying, expansion drying, fluidized bed drying or pneumatic conveying drying.
Preferably, the step of described solid state fermentation raw material being carried out pulverization process by pulverizer is also comprised in above-mentioned steps three.
On the other hand, the present invention also provides a kind of as the biology feed additive being rich in active peptide and beneficial bacterium prepared by above-mentioned preparation method.
Further, the above-mentioned biology feed additive water content being rich in active peptide and beneficial bacterium is below 12%, and peptide content is more than 20%, and beneficial bacterium total viable count is 1.0 × 10
9more than cfu/g.
Meanwhile, the present invention also provides a kind of biological feedstuff being rich in active peptide and beneficial bacterium, is added with above-mentioned additive.
The present invention adopts technique scheme, compared with prior art, has following technique effect: additive active peptide content prepared by preparation method of the present invention is high, and beneficial bacterium viable count content is high; Simultaneously containing fermentating metabolism products such as bacteriocins (also claiming antibacterial peptide); Have pure natural concurrently, the advantage of nontoxic, harmless, pollution-free, noresidue simultaneously.
Detailed description of the invention
The invention provides a kind of preparation method being rich in the biology feed additive of active peptide and beneficial bacterium, comprise the following steps:
Step one, prepares beneficial bacterium seed liquor, adopts the method for mixing or the mixed culture fermentation simultaneously of multiple bacterial classification after single strain fermentation, under 20-40 DEG C of condition of culture, carries out aerobic or anaerobic fermentation 8-48h;
Step 2, prepares nutrient solution, nutritional labeling is fully mixed, and adds after water fully dissolves, sterilization treatment 10-30 minute under 80-121 DEG C of condition;
Step 3, solid state fermentation, by nutrient solution prepared in beneficial bacterium seed liquor prepared in step one, step 2 together with enzyme preparation, adds in solid state fermentation raw material and is fully uniformly mixed, under 5-40 DEG C of ambient temperature conditions, solid state fermentation 12-96 hour;
Step 4, post processing, the solid state fermentation thing obtained by step 3 solid state fermentation, under 30-300 DEG C of temperature conditions, carries out drying process, obtains the biology feed additive being rich in active peptide and beneficial bacterium.
Carry out detailed and concrete introduction below by specific embodiment to the present invention, to make better to understand the present invention, but following embodiment does not limit the scope of the invention.
Embodiment one
The present embodiment is prepared by following concrete steps:
(1) seed liquor preparation.Triangular flask culture medium: the leaching of glucose 2.0%, ox bone peptone 1.0%, beef extract 1.0%, yeast powder 0.5%, anhydrous sodium acetate 0.5%, potassium dihydrogen phosphate 0.2%, diammonium hydrogen citrate 0.2%, epsom salt 0.058%, four water manganese sulfate 0.025%, Tween-80 0.1%.Seed and fermentation medium: the leaching of corn flour 3.0%, soybean meal powder 2.0%, tomato powder 0.5%, fish peptone 0.8%, yeast powder 0.5%, glucose 0.5%, potassium dihydrogen phosphate 0.2%, epsom salt 0.058%, Tween-80 0.1%.Regulate pH6.8 before sterilizing, 118 DEG C of sterilizings 20 minutes, inoculated and cultured Lactobacillus plantarum after being cooled to 37 DEG C.Triangular flask puts 37 DEG C of static gas wave refrigerator 16 hours, obtains triangular flask seed liquor; Seeding tank, fermentation tank tank pressure 0.01-0.06MPa, mixing speed 50rpm/min; Seeding tank 37 DEG C is cultivated 12 hours, obtains seeding tank seed liquor; Fermentation tank 37 DEG C is cultivated 18 hours, obtains fermentation seed liquid.
(2) nutrient solution preparation.Configuration nutrient solution: Fish meal hydrolysates 10%, glucose 5.0%, yeast extract 2.0%, Tween-80 0.1%, 118 DEG C of sterilizings 20 minutes, are cooled to 37 DEG C, obtain nutrient solution.
(3) enzyme preparation configuration.Configuration enzyme preparation: phytase 30%, acid protease 30%, amylase 2 0%, cellulase 10%, hemicellulase 10%, Homogeneous phase mixing, obtains complex enzyme formulation.
(4) solid state fermentation Feedstock treating.Get Cottonseed Meal 40%, Soybean Meal 30%, rapeseed dregs 30%, after mixing, pulverized 30 mesh sieves, obtain solid state fermentation raw material.
(5) solid state fermentation.By the solid state fermentation raw material in step (4), the seed liquor obtained with step (1), the nutrient solution that step (2) is obtained, enzyme preparation in step (3) fully stirs, obtain solid material after mixing, dress polybag carries out solid state fermentation, under 30 DEG C of ambient temperature conditions, ferment 36 hours, obtain solid state fermentation thing.
(6) dry process.By solid state fermentation thing, in 40 DEG C of fluidized bed at elevated drying process 3h, namely make described biology feed additive.
The biology feed additive that the present embodiment prepares, through detecting, peptide content 21.3%, total viable count of lactic acid bacteria is 8.3 × 10
9cfu/g, moisture 9.2%.
Embodiment two
The present embodiment is prepared by following concrete steps:
(1) seed liquor preparation.Triangular flask culture medium: lactoalbumin hydrolysate 2.0%, soy peptone 1.2%, yeast extract 0.5%, sucrose 3.0%, sodium chloride 0.5%, potassium dihydrogen phosphate 0.2%, dipotassium hydrogen phosphate 0.2%, epsom salt 0.058%, Tween-80 0.1%.Seed and fermentation medium: the leaching of corn flour 3.0%, glucose 0.5%, tryptone 0.8%, yeast powder 0.5%, potassium dihydrogen phosphate 0.2%, epsom salt 0.058%, Tween-80 0.1%.Regulate pH5.8 before sterilizing, 115 DEG C of sterilizings 30 minutes, inoculation fermentation lactobacillus, saccharomyces cerevisiae after being cooled to 30 DEG C.Triangular flask puts 30 DEG C, 80rpm/min shaken cultivation 18 hours, obtains triangular flask seed liquor; Seeding tank, fermentation tank tank pressure 0.01-0.04MPa, mixing speed 100rpm/min; Seeding tank 30 DEG C is cultivated 16 hours, obtains seeding tank seed liquor; Fermentation tank 30 DEG C is cultivated 24 hours, obtains fermentation seed liquid.
(2) nutrient solution preparation.Configuration nutrient solution: sucrose 4.0%, Cottonseed Meal hydrolysate 8%, casein peptone 0.5%, yeast extract 2.0%, zinc sulfate 0.05%, Tween-80 0.1%, 115 DEG C of sterilizings 30 minutes, be cooled to 30 DEG C, obtain nutrient solution.
(3) enzyme preparation configuration.Configuration enzyme preparation: acid protease 20%, neutral proteinase 20%, amylase 2 0%, cellulase 20%, alpha-galactosidase 20%, Homogeneous phase mixing, obtains complex enzyme formulation.
(4) solid state fermentation Feedstock treating.Get Cottonseed Meal 30%, rapeseed dregs 20%, fish meal 20%, wheat bran 20%, corn 10%, after mixing, pulverized 60 mesh sieves, obtain solid state fermentation raw material.
(5) solid state fermentation.By the solid state fermentation raw material in step (4), the seed liquor obtained with step (1), the nutrient solution that step (2) is obtained, enzyme preparation in step (3) fully stirs, and obtains solid material after mixing, and dress fermentation tank carries out solid state fermentation, stirring 1 time is opened every 6 hours, under 30 DEG C of ambient temperature conditions, ferment 22 hours, obtain solid state fermentation thing.
(6) dry process.By solid state fermentation thing, vacuum drying treatment 4h at 50 DEG C, namely makes described biology feed additive.
The biology feed additive that the present embodiment prepares, peptide content 22.5%, lactic acid bacteria and saccharomycete total viable count are 2.1 × 10
9cfu/g, moisture 9.8%.
Embodiment three
The present embodiment is prepared by following concrete steps:
(1) seed liquor preparation.Triangular flask culture medium: the leaching of glucose 2.0%, ox bone peptone 1.0%, beef extract 1.0%, yeast powder 0.5%, anhydrous sodium acetate 0.5%, potassium dihydrogen phosphate 0.2%, diammonium hydrogen citrate 0.2%, epsom salt 0.058%, four water manganese sulfate 0.025%, Tween-80 0.1%.Seed and fermentation medium: the leaching of glucose 3.0%, yeast powder 0.8%, tryptone 0.4%, calcium chloride 0.5%, potassium dihydrogen phosphate 0.2%, epsom salt 0.058%, Tween-80 0.1%.Regulate pH6.4 before sterilizing, 121 DEG C of sterilizings 20 minutes, inoculating lactic acid intestines bacterium, Pediococcus acidilactici, candida utili, bacillus coagulans respectively after being cooled to 30 DEG C.Lactic acid intestines bacterium, Pediococcus acidilactici triangular flask put 30 DEG C of static gas wave refrigerator 24 hours, and candida utili, bacillus coagulans put 30 DEG C, 150rpm/min shaken cultivation 24 hours, obtain triangular flask seed liquor; Lactic acid intestines bacterium, Pediococcus acidilactici seeding tank, fermentation tank tank pressure 0.01-0.04MPa; Candida utili, bacillus coagulans seeding tank, fermentation tank tank pressure 0.02-0.08MPa, mixing speed 150rpm/min; Seeding tank 30 DEG C is cultivated 16 hours, obtains seeding tank seed liquor; Fermentation tank 30 DEG C is cultivated 30 hours, obtains fermentation seed liquid.
(2) nutrient solution preparation.Configuration nutrient solution: lactose 8.0%, Soybean Meal hydrolysate 12%, casein peptone 2.0%, yeast extract 2.0%, ammonium chloride 0.3%, manganese sulfate 0.2%, 121 DEG C of sterilizings 20 minutes, be cooled to 30 DEG C, obtain nutrient solution.
(3) enzyme preparation configuration.Configuration enzyme preparation: phytase 30%, acid protease 20%, neutral starch enzyme 20%, cellulase 10%, pectase 10%, zytase 10% Homogeneous phase mixing, obtains complex enzyme formulation.
(4) solid state fermentation Feedstock treating.Get Soybean Meal 30%, rapeseed dregs 30%, wheat bran 20%, corn 20%, after mixing, pulverized 80 mesh sieves, obtain solid state fermentation raw material.
(5) solid state fermentation.By the solid state fermentation raw material in step (4), the seed liquor obtained with step (1), the nutrient solution that step (2) is obtained, enzyme preparation in step (3) fully stirs, obtain solid material after mixing, dress polybag carries out solid state fermentation, under 25 DEG C of ambient temperature conditions, ferment 48 hours, obtain solid state fermentation thing.
(6) dry process.By solid state fermentation thing, in 55 DEG C of downstream drying process 2h, namely make described biology feed additive.
The biology feed additive that the present embodiment is obtained, peptide content 23.3%, lactic acid bacteria, saccharomycete, bacillus Mixed Microbes total viable count are 5.3 × 10
9cfu/g, moisture 9.5%.
Embodiment four
The present embodiment is prepared by following concrete steps:
(1) seed liquor preparation.Triangular flask culture medium: the leaching of tryptone 1.0%, sodium chloride 1.0%, yeast powder 0.5%, glucose 0.5%.Seed and fermentation medium: soybean-cake flour 5.0%, cornstarch 2.5%, yeast extract 1.5%, potassium dihydrogen phosphate 0.2%, epsom salt 0.15%, calcium chloride 0.01%, manganese sulfate 0.01%.Regulate pH7.2 before sterilizing, 121 DEG C of sterilizings 30 minutes, inoculate bacillus subtilis, bacillus licheniformis, bacillus lentus, bacillus pumilus after being cooled to 37 DEG C respectively.Triangular flask puts 37 DEG C, 180rpm/min shaken cultivation 24 hours, obtains triangular flask seed liquor; Seeding tank, fermentation tank tank pressure 0.02-0.08MPa, mixing speed 180rpm/min; Seeding tank 37 DEG C is cultivated 20 hours, obtains seeding tank seed liquor; Fermentation tank 37 DEG C is cultivated 24 hours, obtains fermentation seed liquid.
(2) nutrient solution preparation.Configuration nutrient solution: glucose 6.0%, wheat bran hydrolysate 8%, fish peptone 2.0%, ammonium sulfate 1.3%, magnesium sulfate 0.2%, 121 DEG C of sterilizings 30 minutes, be cooled to 37 DEG C, obtain nutrient solution.
(3) enzyme preparation configuration.Configuration enzyme preparation: alkali protease 40%, neutral starch enzyme 20%, cellulase 20%, amylase 10%, 1,4 beta-glucanase 10%, Homogeneous phase mixing, obtains complex enzyme formulation.
(4) solid state fermentation Feedstock treating.Get Soybean Meal 50%, rapeseed dregs 30%, rice bran 10%, fish meal 10%, after mixing, pulverized 80 mesh sieves, obtain solid state fermentation raw material.
(5) solid state fermentation.By the solid state fermentation raw material in step (4), the seed liquor obtained with step (1), the nutrient solution that step (2) is obtained, enzyme preparation in step (3) fully stirs, obtain solid material after mixing, dress chain-belt type installation for fermenting carries out solid state fermentation, under 35 DEG C of ambient temperature conditions, ferment 48 hours, obtain solid state fermentation thing.
(6) dry process.By solid state fermentation thing, at 240 DEG C, expansion drying process 1 hour, namely makes described biology feed additive.
The biology feed additive that the present embodiment is obtained, peptide content 25.0%, bacillus living adds up to 1.0 × 10
10cfu/g, moisture 8.9%.
Embodiment five
The present embodiment is prepared by following concrete steps:
(1) seed liquor preparation.Triangular flask culture medium: fish peptone 1.0%, glucose 0.5%, sodium chloride 0.5%, beef extract 0.3%.Seed and fermentation medium: tryptone 2.0%, beef extract 1.0%, yeast extract 0.6%, glucose 0.4%, dipotassium hydrogen phosphate 0.2%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.04%, calcium chloride 0.02%, ferrous sulfate 0.01%, L-cysteine hydrochloride 0.05%.Regulate pH7.2 before sterilizing, 118 DEG C of sterilizings 20 minutes, inoculated and cultured Bu Shi lactobacillus, animal bifidobacteria, clostridium butyricum after being cooled to 37 DEG C.Triangular flask puts 34 DEG C of static Anaerobic culturel 24 hours, obtains triangular flask seed liquor; Seeding tank, fermentation tank tank pressure 0.01-0.06MPa, mixing speed 50rpm/min; Seeding tank 34 DEG C of Anaerobic culturel 12 hours, obtain seeding tank seed liquor; Fermentation tank 34 DEG C of Anaerobic culturel 20 hours, obtain fermentation seed liquid.
(2) nutrient solution preparation.Configuration nutrient solution: Fish meal hydrolysates 10%, glucose 5.0%, yeast extract 2.0%, ammonium sulfate 0.5%, 118 DEG C of sterilizings 20 minutes, be cooled to 34 DEG C, obtain nutrient solution.
(3) enzyme preparation configuration.Configuration enzyme preparation: acid protease 30%, keratinase 20%, amylase 2 0%, pectase 10%, cellulase 10%, 'beta '-mannase 10%, Homogeneous phase mixing, obtains complex enzyme formulation.
(4) solid state fermentation Feedstock treating.Get Cottonseed Meal 40%, rapeseed dregs 30%, fish meal 20%, wheat bran 10%, pulverized 120 mesh sieves after mixing, obtain solid state fermentation raw material.
(5) solid state fermentation.By the solid state fermentation raw material in step (4), the seed liquor obtained with step (1), the nutrient solution that step (2) is obtained, enzyme preparation in step (3) fully stirs, obtain solid material after mixing, dress polybag carries out solid state fermentation, under 34 DEG C of ambient temperature conditions, ferment 38 hours, obtain solid state fermentation thing.
(6) dry process.By solid state fermentation thing, in 40 DEG C of fluidized bed at elevated drying process 2h, namely make described biology feed additive.
The biology feed additive that the present embodiment is obtained, peptide content 24.8%, lactic acid bacteria and bacillus living add up to 1.3 × 10
9cfu/g, moisture 9.9%.
Embodiment six
The present embodiment is prepared by following concrete steps:
(1) seed liquor preparation.Triangular flask culture medium: sucrose 3.0%, wheat bran 1.0%, sodium nitrate 0.2%, dipotassium hydrogen phosphate 0.1%, epsom salt 0.05%, potassium chloride 0.05%, ferrous sulfate heptahydrate 0.001%.Seed and fermentation medium: corn flour 5.0%, soybean meal powder 2.0%, sodium nitrate 0.4%, dipotassium hydrogen phosphate 0.3%, epsom salt 0.1%, ferrous sulfate heptahydrate 0.01%.Regulate pH6.5 before sterilizing, 121 DEG C of sterilizings 30 minutes, inoculate aspergillus oryzae, Issatchenkia orientalis, bacillus subtilis after being cooled to 30 DEG C respectively.Triangular flask puts 30 DEG C, 250rpm/min shaken cultivation 22 hours, obtains triangular flask seed liquor; Seeding tank, fermentation tank tank pressure 0.02-0.08MPa, mixing speed 250rpm/min; Seeding tank 30 DEG C is cultivated 18 hours, obtains seeding tank seed liquor; Fermentation tank 30 DEG C is cultivated 25 hours, obtains fermentation seed liquid.
(2) nutrient solution preparation.Configuration nutrient solution: Fish meal hydrolysates 6%, glucose 4.0%, casein peptone 0.5%, ammonium sulfate 0.3%, potassium chloride 0.2%, 121 DEG C of sterilizings 30 minutes, be cooled to 30 DEG C, obtain nutrient solution.
(3) enzyme preparation configuration.Configuration enzyme preparation: alkali protease 20%, neutral starch enzyme 20%, acid protease 10%, cellulase 20%, amylase 2 0%, lipase 10%, Homogeneous phase mixing, obtains complex enzyme formulation.
(4) solid state fermentation Feedstock treating.Get Soybean Meal 40%, corn 40%, rice bran 10%, fish meal 10%, after mixing, pulverized 100 mesh sieves, obtain solid state fermentation raw material.
(5) solid state fermentation.By the solid state fermentation raw material in step (4), the seed liquor obtained with step (1), the nutrient solution that step (2) is obtained, enzyme preparation in step (3) fully stirs, obtain solid material after mixing, dress tray carries out solid state fermentation, under 30 DEG C of ambient temperature conditions, ferment 86 hours, obtain solid state fermentation thing.
(6) dry process.By solid state fermentation thing, at 80 DEG C, vacuum drying treatment 2 hours, namely makes described biology feed additive.
The biology feed additive that the present embodiment prepares, peptide content 20.9%, mould, saccharomycete, bacillus mixed live add up to 2.3 × 10
9cfu/g, moisture 9.5%.
Can be found out by each above-mentioned embodiment, biology feed additive prepared by preparation method of the present invention, peptide content is all more than 20%, and active beneficial bacterium content is up to 1.0 × 10
9more than cfu/g, and containing fermentating metabolism products such as bacteriocins (also claiming antibacterial peptide); Meanwhile, in the feed administering transgenic that the biology feed additive prepared by the above embodiments prepares, good effect is also achieved.
Be described in detail specific embodiments of the invention above, but it is just as example, the present invention is not restricted to specific embodiment described above.To those skilled in the art, any equivalent modifications that the present invention is carried out and substituting also all among category of the present invention.Therefore, equalization conversion done without departing from the spirit and scope of the invention and amendment, all should contain within the scope of the invention.
Claims (10)
1. be rich in a preparation method for the biology feed additive of active peptide and beneficial bacterium, it is characterized in that, comprise the following steps:
Step one, prepares beneficial bacterium seed liquor, adopts the method for mixing or the mixed culture fermentation simultaneously of multiple bacterial classification after single strain fermentation, under 20-40 DEG C of condition of culture, carries out aerobic or anaerobic fermentation 8-48h, obtain beneficial bacterium seed liquor;
Step 2, prepares nutrient solution, nutritional labeling is fully mixed, and add after water fully dissolves, sterilization treatment 10-30 minute under 80-121 DEG C of condition, obtains nutrient solution;
Step 3, solid state fermentation, by nutrient solution prepared in beneficial bacterium seed liquor prepared in step one, step 2 together with enzyme preparation, add in solid state fermentation raw material and be fully uniformly mixed, under 5-40 DEG C of ambient temperature conditions, solid state fermentation 12-96 hour, obtains solid state fermentation thing;
Step 4, post processing, the solid state fermentation thing obtained by step 3 solid state fermentation, under 30-300 DEG C of temperature conditions, carries out drying process, obtains the biology feed additive being rich in active peptide and beneficial bacterium.
2. preparation method according to claim 1, is characterized in that, the beneficial bacterium in the seed liquor of beneficial bacterium described in step one is selected from one or more in lactic acid bacteria, saccharomycete, bacillus, mould.
3. preparation method according to claim 1, it is characterized in that, the medium component of beneficial bacterium seed liquor of fermenting in step one be selected from rice bran, wheat bran, corn flour, soybean meal powder, soybean-cake flour, tomato powder, cornstarch, Dried Corn Steep Liquor Powder, corn steep liquor, glucose, sucrose, lactose, ox bone peptone, fish peptone, soy peptone, tryptone, casein peptone,
any three kinds or more mixed preparing in the leaching of peptone, lactoalbumin hydrolysate, yeast extract, yeast powder, dusty yeast, beef extract, beef extract powder, sodium acetate, diammonium hydrogen citrate, ammonium sulfate, ammonium chloride, sodium chloride, ferric sesquichloride, calcium chloride, potassium chloride, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium sulfate, manganese sulfate, zinc sulfate, ferrous sulfate, sodium nitrate, L-cysteine hydrochloride or Tween-80.
4. preparation method according to claim 1, it is characterized in that, nutritional labeling described in step 2 comprise Cottonseed Meal hydrolysate, rapeseed dregs hydrolysate, Soybean Meal hydrolysate, Fish meal hydrolysates, glucose, sucrose, lactose, ox bone peptone, the molten slurry of fish, the molten starch of fish, fish peptone, soy peptone, tryptone, casein peptone,
any two or more than two kinds mixed preparing in the leaching of peptone, lactoalbumin hydrolysate, yeast extract, yeast powder, dusty yeast, beef extract, beef extract powder, ammonium sulfate, ammonium chloride, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium sulfate, manganese sulfate, zinc sulfate or Tween-80.
5. preparation method according to claim 1, it is characterized in that, enzyme preparation described in step 3 is selected from one or more in amylase, cellulase, hemicellulase, pectase, lipase, phytase, protease, keratinase, alpha-galactosidase, 1,4 beta-glucanase, 'beta '-mannase or zytase.
6. preparation method according to claim 1, is characterized in that, the raw material of solid state fermentation described in step 3 is selected from one or more in Cottonseed Meal, rapeseed dregs, Soybean Meal, fish meal, wheat bran, corn or rice bran.
7. preparation method according to claim 1, is characterized in that, in step 4, dry process adopts vacuum drying, expansion drying, fluidized bed drying or pneumatic conveying drying.
8. the biology feed additive being rich in active peptide and beneficial bacterium prepared by claim 1-7 any one preparation method.
9. the biology feed additive being rich in active peptide and beneficial bacterium according to claim 8, is characterized in that, water content is below 12%, and peptide content is more than 20%, and beneficial bacterium total viable count is 1.0 × 10
9more than cfu/g.
10. be rich in a biological feedstuff for active peptide and beneficial bacterium, it is characterized in that, be added with additive as claimed in claim 8 or 9.
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