CN104630182B - A kind of grower pigs compound enzyme and preparation method thereof - Google Patents
A kind of grower pigs compound enzyme and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of grower pigs compound enzyme and preparation method thereof, belong to enzymic preparation field；The grower pigs compound enzyme, is made up of the raw material of following parts by weight：Zytase 38, cellulase 8 15, beta glucan enzyme 5 10, medium temperature alpha amylase 35, galactosidase 12, pectase 12, Lactobacillus plantarum capsule 36,6 10 parts of galenical, 24 parts of Ginger P.E, 12 parts of radix glycyrrhizae.Using galenical, Ginger P.E, radix glycyrrhizae science compounding in product of the present invention, effectively slow down the moisture regain of enzyme preparation；Resistance to jelly, the heat resistance of complex enzyme can be strengthened, keep identical enzyme activity；Protective agent acts synergistically jointly with enzyme preparation, galenical and Ginger P.E; complex enzyme enzyme activity and effect is set to play to greatest extent; the utilization rate of feed and the growth rate of animal are improved, strengthens the appetite and resistance against diseases of animal, extends the shelf-life of product, protects environment.
Description
Technical field
The invention belongs to enzymic preparation field, and in particular to a kind of grower pigs compound enzyme and preparation method thereof.
Background technology
The enzyme preparation applied at present in feed industry mainly has 4 major classes：It is respectively intended to degraded cellulose, protein, shallow lake
Powder and phytic acid.
A kind of entitled store pig complex enzyme preparation for feeding, application number：201110026562.X the disclosure of the invention
A kind of store pig complex enzyme preparation for feeding, including following eight kinds of enzymes:Acid protease, amylase, 'beta '-mannase, wood
Dextranase, 1,4 beta-glucanase, cellulase, pectase and phytase；The enzyme activity ratio of eight kinds of enzymes is followed successively by 1: (0.8-
14.5)∶(0.19-0.3)∶(33.7-35)∶(25-30)∶(3.2-3.5)∶(0.19-2.0)∶(0-0.025).The invention is compound
Enzyme preparation can degrade various ANFs in feed, reduce enteron aisle chyme viscosity, improve feedstuff metabolisable energy power；Improve growth fertilizer
Educate digestibility of the pig to protein and starch；The nutrient content entered in rear intestinal chyme is reduced, effectively suppresses to be harmful in enteron aisle
The breeding of microorganism, intestinal microecology balance is reconciled, promote pig health；Growth, fattening pig average daily gain are improved, reduces material
Meat ratio, shorten the livestock on hand phase, increase economic efficiency.
A kind of complex enzyme preparation for feeding piglets, Application No. 201110026602.0, a kind of piglet of the disclosure of the invention are raised
Use complex enzyme formulation.The complex enzyme preparation for feeding piglets includes following seven kinds of enzymes:Acid protease, amylase, zytase, β-
Dextranase, cellulase, pectase and phytase；The enzyme activity ratio of seven kinds of enzymes is followed successively by 1: 1: (8-10): (3.3-4)
∶1∶(0.5-0.55)∶(0-0.02).Various ANFs in the degradable feed of invention complex enzyme formulation, reduce enteron aisle food
Rotten viscosity, improve feedstuff metabolisable energy power；Improve digestibility of the piglet to protein and starch；Reduce harmful microbe in enteron aisle
Excessive multiplication, the damage of intestinal wall is reduced, reconcile intestinal microecology balance；Piglet survival rate is improved, premunition, reduces diarrhea rate,
Improve the overall uniformity；Promote piglet weightening, reduce aquaculture cost.
A kind of pig's feed complex enzyme formulation added with activator, application number：201210500011.7 the invention provides
A kind of pig's feed complex enzyme formulation added with activator, the phytase in formula can resolve into the organophosphor in feed
The inorganic phosphate that can be absorbed, the protein-based nutriment in the degradable feed of protease, promote live pig to phosphorus in feed
With the absorption of nitrogen, while the discharge capacity of phosphorus and nitrogen in live pig fecaluria is also reduced, reduce its pollution on the environment.And activate
Agent can improve the reaction rate of enzyme, enzyme preparation is taken effect faster, and each ingredient combination in activator and enzyme preparation is used and may be used also
Make the shearing force of cold cuts reduce, improve pork tenderness.Pig's feed complex enzyme formulation produced by the present invention can improve pig to raising
The digestibility of nitrogen, phosphorus in material so that the excretion of phosphorus reduces more than 40% in live pig fecaluria, the excretion of nitrogen reduce 29% with
On；It is quick, there is significant effect within 2-3 days；Cold cuts shearing force reduces by more than 50%.
A kind of entitled pig's feed complex enzyme formulation and preparation method thereof application number：201410275557.6 the invention
A kind of pig's feed complex enzyme formulation is provided, belongs to feed additive field.The additive is made up of complex enzyme and reinforcing agent；
Complex enzyme be lipase, neutral proteinase, beta amylase, isoamylase, neutral phytase, 'beta '-mannase, zytase,
Alpha-galactosidase；Reinforcing agent by extract from pine needles, leaf of bamboo Thick many candies, caulis lonicerae powder, galangal powder, radix jurineae powder, lithospermum powder,
Vitamin C, sodium chloride, sodium butyrate, ferrous sulfate and calcium gluconate composition.The enzyme preparation can effectively improve the utilization of feed
Rate, feedstuff-meat ratio is reduced, promote the speed of growth of pig；The immunity of pig can also be played a part of strengthening, to improving pig
Health degree, reducing the pig incidence of disease has very big help, and market prospects are very wide.
A kind of entitled efficient pig's feed complex enzyme formulation added with enzymatic protective reagent, Application No.
201210381755.1, it is degradable the invention provides a kind of efficient pig's feed complex enzyme formulation added with enzymatic protective reagent
The material such as ANFs and large biological molecule such as beta glucan, phytic acid, pectin in feed, so as to promote pig in feed
The absorption of nutritional ingredient, improve efficiency of feed utilization；Protectant addition can reduce the inactivation rate of enzyme, extend effective time, make this
The effect of enzyme preparation is more significantly and lasting；High efficiency composition enzyme preparation feed conversion rate produced by the present invention is high, can make feedstuff-meat ratio
Decline more than 19%；The pig speed of growth improves more than 23%；The enzymatic activity retention time is grown, and in 18 months, enzymatic activity can be kept
More than 90%.
A kind of entitled efficient pig's feed complex enzyme formulation added with enzymatic protective reagent, application number：
201210360110.X, it is degradable the invention provides a kind of efficient pig's feed complex enzyme formulation added with enzymatic protective reagent
The material such as ANFs and large biological molecule such as beta glucan, phytic acid, pectin in feed, so as to promote pig in feed
The absorption of nutritional ingredient, improve efficiency of feed utilization；Protectant addition can reduce the inactivation rate of enzyme, extend effective time, make this
The effect of enzyme preparation is more significantly and lasting；High efficiency composition enzyme preparation feed conversion rate produced by the present invention is high, and feed can be used
Coefficient declines more than 19%；The pig speed of growth improves more than 23%；The enzymatic activity retention time is grown, and in 30 months, enzymatic activity can
It is maintained at more than 90%.
A kind of entitled the feed for piglet complex enzyme formulation and its production method and application, application number：
201410150709.X, the disclosure of the invention a kind of the feed for piglet complex enzyme formulation and its production method and application.This is compound
The enzyme activity of enzyme preparation forms：Glucose oxidase 80-100U/g, alpha-galactosidase 20-25U/g, zytase 3000-
4000U/g, 1,4 beta-glucanase 500-600U/g, 'beta '-mannase 500-600U/g, acid protease 2000-3000U/g, in
Property proteinase 8 00-1000U/g, mesophilicα-diastase 800-1000U/g, fungal alpha-amylase 400-500U/g, glucoamylase
Enzyme 3000-4000U/g.The complex enzyme formulation is made using cornstarch and potassium dihydrogen phosphate, dipotassium hydrogen phosphate, mixture of calcium sulfate
For diluent.The complex enzyme formulation application method is：Addition in suckling piglet perfect compound feed per ton is 500-750
Gram.
The bark of eucommia, rich in phenylpropanoids, iridoids isoreactivity composition, have and remove internal rubbish, strengthen thin
Born of the same parents' metabolism, decomposer inner cholesterol, body fat is reduced, broad-spectrum antiseptic, it is aobvious to improve white blood cell count, strengthen immunity etc.
Write effect.Result of the pure eucommia bark powder in zoopery shows:Cholesterol and lipid-metabolism can be promoted；Add pure eucommia bark powder raising
Eel, have natural eel taste；Pure eucommia bark powder, which is added to, can improve the quality of broiler chicken in feed, with tempting
Fragrance and quality.
Poria cocos has clearing damp and promoting diuresis, and strengthening the spleen and reducing phlegm, antitoxic heart-soothing and sedative, the effect of internal heat anticance, can relax gastrointestinal smooth muscle,
Gastric acid secretion inhibiting, prevent necrosis of liver cells, antibacterial and other effects；Contained pachymic acid has strengthen immunity, antitumor and town
Quiet, hypoglycemic etc. effect；Pachymaran can reduce malaber reefcod serum A/G values, improve serum complement C3 content, improve blood
Liquid leukocyte count and leukocytes phagocytic rate, improve its non-specific immunity.
In summary, the application of grower pigs specific enzyme has its wide market space and huge economic value, but raw
The heat endurance of long pig specific enzyme, security, comprehensive and action effect give full play to of compounding are still enzyme preparation manufacturer
The major issue paid close attention to jointly with numerous raisers, prepares safer, and more comprehensively, the more preferable grower pigs of enzyme effect effect are special
It is corporation responsibility and the pursuit of industry technical staff with enzyme.
The content of the invention
Technical problem solved by the invention provides a kind of grower pigs compound enzyme；
The grower pigs compound enzyme, is made up of the raw material of following parts by weight：Zytase 3-8, cellulase 8-15,
1,4 beta-glucanase 5-10, mesophilicα-diastase 3-5, galactosidase 1-2, pectase 1-2, Lactobacillus plantarum capsule 3-6, plant
Preparation 6-10 parts, Ginger P.E 2-4 parts, radix glycyrrhizae 1-2 parts.
The zytase, 1,4 beta-glucanase, cellulase, mesophilicα-diastase, galactosidase are food grade enzyme
Preparation；
The preparation method of the galenical is as follows：
Weigh cassia seed 10-18 parts；Bark of eucommia 5-15 parts；Poria cocos 10-15 parts；Said herbal medicine is crushed into particle diameter respectively is
Less than 2 millimeters, then uniformly mixed in container and add the water of 3-6 times of weight, addition said mixture material constituent mass 1-
2% acidic cellulase, control 45-55 DEG C of temperature, pH：3.5-4,1-2h is kept, then adds 2-3 times of weight of mixed material
The mixture of ethanol and propyl alcohol, control temperature to 60 DEG C of -78 DEG C of holding 3-4h, filtering；It is freeze-dried and obtains after filter vacuum concentration
Obtain galenical.
The mass ratio of the ethanol and propyl alcohol is 1：1-1.5；Concentration of alcohol is 95%；Propanol concentration is 100%；
The preparation method of the Ginger P.E is as follows：
Particle diameter will be crushed to be placed in container for less than 2 millimeters gingers, add the water of 3-6 times of weight of ginger, control temperature
50 DEG C of -60 DEG C of holding 2-3h, add the mixture of 2-3 times of w ethanol of ginger and methanol, control temperature to 30 DEG C of -40 DEG C of holdings
3-8h, filtering；Freeze-drying obtains Ginger P.E after filter vacuum concentration.
The mass ratio of the ethanol and methanol is 1：2, concentration of alcohol 85-95%；Methanol concentration 100%.
The preparation method of the Lactobacillus plantarum capsule is as follows：
Lactobacillus plantarum capsule product digests powder group by wall material, Lactobacillus plantarum, freeze drying protectant, stachyose and hickory chick
Into.
Described wall material is made up of enzymatic hydrolysis of soybean protein isolate, chitosan, xanthans, carragheen, glycerine and trehalose；On
Stating concentration when each material is made into mixed solution is respectively：Enzymatic hydrolysis of soybean protein isolate 7-10%, chitosan 1-2%, xanthans
1-3%, carragheen 0.1-0.5%, glycerine 0.3-1%, trehalose 0.5-2%.
The preparation method of enzymatic hydrolysis of soybean protein isolate is：Compound concentration is that 10-13% soybean protein isolate solution is heated to
30-45 DEG C, pH to 3-5 is adjusted, the acid protease insulation enzymolysis 0.5-1.5 for adding soybean protein isolate weight 0.1-1% is small
When, after enzymolysis solution spray drying obtain enzymatic hydrolysis of soybean protein isolate.
Hickory chick enzymolysis powder, preparation method thereof is as follows：
(1) Morchella esculenta (L.) Pers sporophore drying and crushing；
(2) water-soluble homogeneous：The Morchella esculenta (L.) Pers sporophore of crushing is added in stainless steel cylinder, adds 3-6 times of weight of fructification
Water, 3-5 hours are soaked, are then by colloid mill, colloid mill operation condition by this Morchella esculenta (L.) Pers sporophore liquid：Adjustment colloid mill is determined
The gap of son and rotor is 0.5-1 microns, and colloid mill flow is 0.4-1 ton hours；
(3) heating enzymolysis：Morchella esculenta (L.) Pers sporophore liquid mixed liquor Jing Guo milling treatment of colloid is transferred to stainless steel enzymatic vessel
In be heated to 50-60 DEG C, adjust pH to 4.5-6.0, addition Morchella esculenta (L.) Pers sporophore weight 0.05-0.1% cellulase,
0.01-0.1% 1,4 beta-glucanase, 0.01-0.1% protease, enzymolysis 0.5-1.5 hours are incubated, in enzymolysis process constantly
Stirring.
(4) dry：Dried after mash filtrations after enzymolysis and obtain hickory chick enzymolysis powder.
The drying can use the dried forms such as conventional spray drying, freeze-drying.
The freeze drying protectant is made up of skimmed milk power, trehalose and maltodextrin.
The preferred CGMCC NO.9405 of Lactobacillus plantarum that capsule product of the present invention includes.
Above-mentioned Lactobacillus plantarum capsule product preparation method is as follows：
1) core is prepared：Zymotic fluid quality 3-5% is added into the Lactobacillus plantarum zymotic fluid of fermented tank fermented and cultured
Stachyose and 5-8% hickory chick enzymolysis powder, then mixed with frozen-dried protective agent solution, pre-freeze 0.5h, Ran Hou at -50 DEG C
10-18h is freezed in vacuum freeze drier, is ground after lyophilized and core is made；
The ratio of zymotic fluid and the frozen-dried protective agent solution is 1:1.4-0.8；The frozen-dried protective agent solution is by degreasing
Milk powder, trehalose and maltodextrin solution composition；The volume ratio of three kinds of solution is skimmed milk power：Trehalose：Maltodextrin=3:
1:0.5；
The concentration of the skimmed milk power, trehalose and maltodextrin solution is respectively：Skimmed milk power 5%, trehalose 1%,
Maltodextrin 2%.
2) it is coated with：Core is suspended in fluid bed, wall material spraying coating, chitosan, glycerine are sprayed into from a shower nozzle
With the mixed liquor of trehalose, the mixed liquor of xanthans, carragheen and enzymatic hydrolysis of soybean protein isolate is sprayed into from another shower nozzle, is sprayed
Mist speed control is identical, and between 25-38 DEG C, capsule is formed after 30 minutes for temperature in fluid bed during coating.This is dried
Method uses equipment in patent 201120503311.1 to handle.
The mesophilicα-diastase has following characteristic：
(1) the enzyme Acclimation temperature wider range, temperature activity between 55-75 DEG C is higher, and optimum temperature is 65 DEG C；
Heat endurance：The enzyme preserves 24h under the conditions of 65 DEG C still has 80-83% enzyme activity, and 12h is preserved still under the conditions of 70 DEG C
With more than 50-60% enzyme activity.
(2) the enzyme optimal reaction pH value is 5.0；There is high enzyme vigor between pH value 4.0-6.0；
Absolute acid stability：Still there is more than 80% enzyme activity after preservation 18h under pH4-6.
(3) enzymatic activity：Bacterial strain 304 is 7000-9600U/ml by the acid mesophilicα-diastase enzyme activity prepared that ferments,
It is particularly suitable for liquefaction process and Mashing process and the industrialization demand deposited.
The amylase is mesophilicα-diastase, is obtained using solution fermentation culture.
The bacterial strain of production mesophilicα-diastase provided by the invention is specially bacillus subtilis 304
(Bacillussubtilis)304.The bacterial strain has been preserved in China typical culture collection center (letter on November 24th, 2013
Claim CCTCC, address is：Chinese Wuhan Wuhan Universitys), preserving number is CCTCC NO:M 2013600.
The bacillus subtilis 304 is by one plant of bacillus subtilis for producing mesophilicα-diastase for being isolated from acid soil
Starting strain obtains through UV-LiCl-dithyl sulfate Mutation screening, and bacterial strain feature is as follows：The bacterial strain is solid
Colony colour is milky on body flat board, and dry tack free fold, color is secretly opaque, neat in edge, and microscopy is elongated rod shape, gram
Stained positive, peritrichous, gemma are oval.
The preparation method of grower pigs compound enzyme of the present invention is as follows：
By the radix glycyrrhizae, galenical and Ginger P.E distinguish ultramicro grinding, then with zytase, cellulase,
1,4 beta-glucanase, amylase, galactosidase, pectase, Lactobacillus plantarum capsule are packed after uniformly mixing and got product
Grower pigs compound enzyme.
The enzyme is applied to feed-processing plant and plant's autogamy feed, should be mixed with other raw materials in feed during use
Uniformly, it can be well mixed, remix in large quantities of feeds, Direct-fed with a small amount of feed by of the invention in advance；It is it is recommended that per ton complete
Valency material addition is：100-150g.
Beneficial effect：
The present invention uses the zymotechnique of gradient cooling and gradient increased temperature with mesophilicα-diastase, while is added inoculation
With in good time feed supplement so that the present invention produce mesophilicα-diastase vigor high, tolerable temperature height, stability by force, suitable for industrial metaplasia
Production.Enzyme activity is 9500U/ml.Medium temperature alphalise starch crude enzyme liquid sample determines enzyme activity after the different time is preserved at 40-90 DEG C,
As a result show, sample preserves 24h under the conditions of 65 DEG C still has 83% enzyme activity, preserved under the conditions of 70 DEG C 12h still have 60% with
Upper enzyme activity.
Using galenical, Ginger P.E, radix glycyrrhizae science compounding in product of the present invention, returning for enzyme preparation effectively slow down
Tide；Resistance to jelly, the heat resistance of complex enzyme can be strengthened simultaneously, keep identical enzyme activity, its heat resisting temperature can improve 10-15 DEG C,
Freeze-resistant temperature can reduce 5-10 DEG C, effectively prevent the loss of complex enzyme enzyme activity during transport, preservation and use, prolong
The shelf-life of complex enzyme has been grown, has reached same enzyme activity, the like product shelf-life can extend 3-5 months.
The galenical of product addition of the present invention can both extend the shelf-life of complex enzyme formulation, can improve raising livestock and poultry again
Immunity, effectively prevent the generation of livestock and poultry epidemic disease.
Lactobacillus micro-capsule provided by the invention, modified soybean protein isolate, modified soybean separation are with the addition of in its wall material
Albumen improves 25%, gelling ability than soybean protein isolate emulsifying capacity and improves 15-25%, modified soybean protein isolate xanthans
With being used cooperatively for carragheen and chitosan, gel strength improves more than 10%, enhances the stomach juice-resistant of capsule；The present invention
Capsule uses modified soybean protein isolate, has good enteric solubility, capsule can be complete within 1-1.5 hours after reaching enteron aisle
Disintegration discharges Lactobacillus plantarum, and propagation turns into dominant microflora rapidly, suppresses the effect such as growth of pathogenic bacteria so as to reach.
Protective agent and the common synergy of enzyme preparation, galenical, Ginger P.E and enzyme preparation in product of the present invention,
So that the enzyme activity and effect of complex enzyme play to greatest extent, and the utilization rate of feed and the growth of animal have been correspondingly improved it
Rate, the appetite and resistance against diseases of animal are enhanced, extend the shelf-life of product and protect environment.
Embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention
It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention
Scope, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
The various changes carried out on the premise of invention spirit and scope to the material component in these embodiments and dosage or change
Belong to protection scope of the present invention.
Lactobacillus plantarum (Lactobacillus plantarum) tlj-2014 provided by the present invention, the bacterial strain preservation
In China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.9405, preservation
Location：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode 100101, preservation date
On July 2nd, 2014.
Embodiment 1
Lactobacillus plantarum CGMCC No.9405 Breeding Process is as follows used in the present invention：
The original strain that sets out → test tube activation → dithyl sulfate (DES) mutagenesis → nitrosoguanidine (NTG) mutagenesis → wait from
Daughter mutagenesis → flat board primary dcreening operation → shaking flask secondary screening → mitotic stability experiment.
For starting strain of the present invention in MRS dextrose culture-mediums, the throughput rate of its lactic acid is 1.5g/L/d,
Almost stopped growing when medium pH is 3.5, the decomposition rate to natrium nitrosum is 0.34mg/h/kg Chinese cabbages.Starting strain
It is collected in the greenfeed of Yanchi county Ningxia Fattening Sheep field by Li Zheng, acquisition time September in 2013 15 days.
In order to improve the decomposition rate of its production of lactic acid speed, acid-fast ability and nitrite, successively using DES and NTG
Technology carries out mutagenesis to the strain, and bacterial strain carries out primary dcreening operation using MRS calcium carbonate flat board after mutagenesis, is then sent out using 500mL shaking flasks
Ferment, biosensor analysis instrument carry out secondary screening to Producing Strain, the excellent lactobacillus plantarum strain of seed selection, then do passage assays,
Evaluate its genetic stability.
Lactobacillus plantarum tlj-2014 genetic stability results show：By continuous passage ten times, property indices are all
More stable, heredity is preferable, and character is not replied, therefore the purpose bacterium that Lactobacillus plantarum tlj-2014 is obtained as seed selection
Strain.
Empirical tests are found：The production of lactic acid speed of the mutagenic strain can reach 35g/L/d, and the bacterial strain was sent out by 71 hours
Lactic acid concn reaches 95g/L after ferment；Survived under conditions of being 1.80 in pH.Degrading nitrite speed is fast, capacity of decomposition
Reach 9.8mg/h/kg (speed of spontaneous fermentation process nitrite accumulation is about 1.1mg/h/kg), can be resistant to 1% cholate.
1) DES mutagenic and breedings
A. the rings of Lactobacillus plantarum L mono- (going out bacterium germination) on test tube slant are taken on super-clean bench, access is equipped with 50mL culture mediums
In MRS (no agar, glucose 20g/L) 250mL triangular flasks, 200rpm, 37 DEG C are cultivated 12h or so, thalline is in logarithm
Grow early stage.
B. 5mL bacterium solutions are taken, 5000rpm centrifugations 10min collects thalline, with brine 2 times.
C. 10 are diluted to pH7.0 phosphate buffers7Individual/mL bacteria suspensions.
D. 32mL pH7.0 kaliumphosphate buffer, 8mL bacteria suspensions, 0.4mL DES is taken to be placed in advance in the 150mL of rotor
It is sufficiently mixed in triangular flask, it is 1% (v/v) to make DES ultimate densities.
E. 150rpm reacts 30min in 37 DEG C of shaking tables, takes 1mL mixed liquors, adds 0.5mL 25%Na2S2O3In solution
Only react.
F. appropriate dilution, takes the bacterium solution 0.2mL of last dilution factor, is coated on calcium carbonate screening and culturing medium (Portugal containing 100g/L
The calcium carbonate MRS culture mediums of grape sugar) in plate.After 37 DEG C of cultures 2~3 days, using photolithography by the bacterial strain of the screening flat board
It is transferred on the LPHMRS culture mediums (low ph value is modified MRS culture mediums) that pH is 1.5,1.8 and 2.0 and natrium nitrosum screening and culturing
On base (single nitrogen source is the modification MRS screening and culturing mediums of 2g/L natrium nitrosums).
G. after 37 DEG C are cultivated 2~3 days, choosing colony is larger, respectively can be in LPHMRS culture mediums, natrium nitrosum screening training
Support and grown and in calcium carbonate screening and culturing medium on base.Through preliminary screening, the bacterium colony that picking goes out is named as Lactobacillus plantarum L1.
2) nitrosoguanidine mutagenesis
A. the rings of Lactobacillus plantarum L1 mono- on test tube slant are taken on super-clean bench, access is equipped with 50mL culture mediums MRS (no fine jades
Fat) (concentration of glucose 60g/L) 250mL triangular flasks in, 200rpm, 37 DEG C of culture 12h or so, thalline be in logarithm and give birth to
Long early stage.
B. 5mL bacterium solutions 5000rpm centrifugations 10min is taken to collect thalline, with brine 2 times.
C. 10 are diluted to pH6.0 phosphate buffers7Individual/mL bacteria suspensions.
D. take 10mL bacteria suspensions to be transferred in 100mL triangular flasks, add 10mg NTG, be configured to final concentration of 10mg/mL
NTG solution, and 4-5 drop acetone is added, so that NTG dissolves.
E. the 200rpm oscillating reactions 30min at 37 DEG C, 5000rpm centrifugation 10min collect thalline, use sterile saline
Wash for several times, stopped reaction.
F. appropriate dilution, takes the bacterium solution 0.2mL of last dilution factor, is coated on calcium carbonate screening and culturing medium (Portugal containing 100g/L
The calcium carbonate MRS culture mediums of grape sugar) in plate.After 37 DEG C of cultures 2~3 days, using photolithography by the bacterial strain of the screening flat board
It is transferred on the LPHMRS culture mediums (low ph value is modified MRS culture mediums) that pH is 1.5,1.8 and 2.0 and natrium nitrosum screening and culturing
On base (single nitrogen source is the modification MRS screening and culturing mediums of 2g/L natrium nitrosums).
G. bacterial strain method is selected：Choosing colony is larger, and difference can be in LPHMRS culture mediums, natrium nitrosum screening and culturing medium
Grow and in calcium carbonate screening and culturing medium.Through preliminary screening, 100 bacterium colonies for meeting conditions above of picking.
3) shaking flask secondary screening
A. the ring of Lactobacillus plantarum one on each test tube slant is taken respectively on super-clean bench, access is equipped with 50mL culture mediums MRS
In the 250mL triangular flasks of (no agar) (concentration of glucose 100g/L), 200rpm, 37 DEG C are cultivated 15h or so, are in thalline
Mid log phase.
B. 5mL bacterium solutions are taken respectively, and access is equipped with the 50mL calcium carbonate screening fluid nutrient medium (carbonic acid of the glucose containing 250g/L
Calcium MRS culture mediums) in plate, pH 1.5,1.8 and 2.0 LPHMRS fluid nutrient mediums (low ph value is modified MRS culture mediums) and
(note on natrium nitrosum liquid screening medium (single nitrogen source is the modification MRS screening and culturing mediums of 2g/L natrium nitrosums)：Using
250mL triangular flasks).200rpm, 37 DEG C are cultivated 3-4 days, are detected Pfansteihl in calcium carbonate screening fluid nutrient medium respectively daily and are produced
The consumption speed of raw speed, the biomass in LPHMRS fluid nutrient mediums and natrium nitrosum liquid screening medium nitrite
Rate.After fermentation ends, compare Pfansteihl in the calcium carbonate screening fluid nutrient medium of 100 plants of strains and produce speed, LPHMRS liquid
The wear rate of biomass and natrium nitrosum liquid screening medium nitrite in culture medium.
C. selection has high Pfansteihl generation speed concurrently, (strain is only capable of in minimum pH1.8 culture medium the low pH of tolerance
Growth) and nitrite the high bacterial strain of wear rate, be named as L2 bacterium.
4) genetic stability is tested
Continuous ten passages on inclined-plane by L2 bacterium, and detect the fermentation feelings after passage every time with the method for shaking flask secondary screening
Condition.Experiment finds that continuous ten passages, the strain character do not have significant change on inclined-plane, and property indices are all normal, say
The genetic stability of the bright strain is stronger.Strain Designation is Lactobacillus plantarum (Lactobacillus plantarum) tlj-
2014。
5) 5L fermentation tanks are tested
A. the rings of Lactobacillus plantarum L2 mono- on inclined-plane are taken, access is equipped with 50mL culture mediums MRS (no agar) (concentration of glucose
For 150g/L) 250mL triangular flasks in, 200rpm, 37 DEG C of culture 12h or so, thalline is in mid log phase.
B. 5L of the strain access equipped with 3L MRS fluid nutrient mediums (initial glucose 150g/L) of logarithmic phase is fermented
In tank.Inoculum concentration is 10%, and 100rpm is cultivated 8 hours at 37 DEG C, logarithm dissolved oxygen early stage control 10% (ventilation 0.5L/min), after
Phase Anaerobic culturel 63 hours.After fermentation ends, Lactobacillus plantarum L2 lactic acid production reaches 95g/L.
C. by the strain access of logarithmic phase equipped with LPHMRS fluid nutrient mediums (the initial glucose 50g/ that 3L pH are 1.8
L in 5L fermentation tanks).Inoculum concentration is 10%, and 100rpm is cultivated 8 hours at 37 DEG C, (the ventilation of logarithm dissolved oxygen early stage control 10%
0.5L/min), later stage anaerobism, whole process are controlled zymotic fluid pH 1.8 with 0.5mol/L sodium hydroxide, total incubation time
For 48 hours.After fermentation ends, detection Lactobacillus plantarum L2 biomass is 2.5g/L, illustrates that Lactobacillus plantarum L2 can be
Survived in pH1.8 environment.
D. by the strain access of logarithmic phase, equipped with 3L natrium nitrosums liquid screening medium, (single nitrogen source is 2g/L nitrous acid
The modification MRS screening and culturing mediums of sodium) 5L fermentation tanks in.Inoculum concentration is 10%, and 100rpm is cultivated 8 hours at 37 DEG C, before logarithm
Phase dissolved oxygen control 10% (ventilation 0.5L/min), later stage anaerobism, fermentation process adds 20g/L according to the wear rate stream of nitrite
Sodium nitrite solution, cultivate 2-3 days.After fermentation ends, degradeds of the fermentation process Lactobacillus plantarum L2 to natrium nitrosum is calculated
Speed.As a result find：Under this condition, L2 can reach 563mg/h/L to the degradation rate of natrium nitrosum.
Embodiment 2
A kind of preparation method of mesophilicα-diastase, comprises the following steps：
(1) actication of culture
The slant strains of intact bacillus subtilis 304 are inoculated in slant medium, 31 DEG C of culture 24h are carried out
Actication of culture, so activation 2 times；
The slant medium forms：Beef extract 3g, sodium chloride 5g, peptone 10g, glucose 2g, agar 15g, steam
Distilled water l000mL, 6,121 DEG C of sterilizing 20min of pH value；
(2) liquid seeds expand culture
1. first order seed culture：The ring of slant strains 2 after step (1) is activated is accessed in 500 milliliters of shaking flasks, culture medium dress
100 milliliters, rotary shaker 250rpm of amount, 31 DEG C of cultivation temperature, incubation time 10h；
2. secondary seed culture：First order seed is accessed in 500 milliliters of secondary seed shaking flasks according to 10% inoculum concentration, training
The condition of supporting is identical with first order seed；
3. three-level seed culture：Secondary seed is accessed in 5000 milliliters of three-level seed flasks with 10% inoculum concentration, culture
1000 milliliters, rotary shaker 250rpm of base loading amount, 38 DEG C of cultivation temperature, incubation time 10h；
4. first class seed pot culture：Three-level seed is accessed into first class seed pot of the total measurement (volume) as 150L using 10% inoculum concentration,
Fermentation medium loading amount 100L, 31 DEG C, mixing speed 200rpm of cultivation temperature, ventilation (V/V) 1:1, tank pressure 0.05Mpa, training
Support time 10h；
Institute's one-level, two level, three-level seed culture medium weight composition are：
Dusty yeast 0.3%, glucose 1%, peptone 0.3%, beef extract 0.5%, dipotassium hydrogen phosphate 0.8%, Chinese herbal medicine
Agent powder 1.5%, trehalose 1%, calcium sulfate 1g, magnesium chloride 2g, sodium citrate 1g, insufficient section pure water are supplied, pH value 6,121
DEG C sterilizing 30min.
The seed tank culture base weight forms：
Maltodextrin 5%, dusty yeast 0.4%, herbal mediciment powder 1.5%, trehalose 1%, peptone 0.1%, corn steep liquor
0.1%, dipotassium hydrogen phosphate 0.8%, magnesium sulfate 0.05%, sodium citrate 0.1%, insufficient section pure water supplies, pH value 6,121
DEG C sterilizing 30min.
The seeding tank zymotic fluid cell concentration is 7.0x 108Individual/ml；
(3) ferment tank
Seeding tank zymotic fluid is accessed into fermentation tank with 6% inoculum concentration, 35 DEG C, mixing speed 250r/min of cultivation temperature, led to
Air quantity (V/V) 1:2, incubation time 13h；Then with 1.5 DEG C/h rate of temperature fall slow cooling to 15 DEG C, incubated 13h；Continue
With 1.5 DEG C/h rate of temperature fall slow cooling to 3 DEG C, now, first class seed pot zymotic fluid is added into access fermentation with 2% inoculum concentration
Tank, incubated 13h；35 DEG C finally are slowly increased to 1.5 DEG C/h heating rates, incubated 30h.
PH is controlled：By mending ammoniacal liquor or phosphoric acid,diluted, pH value in fermentation process is controlled to be maintained at 6.5；
The fermentation medium forms：Maltodextrin 100g, corn flour 55g, beancake powder 20g, herbal mediciment powder 40g,
Trehalose 35g, dusty yeast 6g, corn steep liquor 3g, ammonium sulfate 2g, dipotassium hydrogen phosphate 2g, potassium dihydrogen phosphate 2g, sodium citrate 3g, disappear
Infusion 0.5g, pure water l000mL, 6.5,121 DEG C of sterilizing 20min of pH value；
The preparation method of the Chinese herbal medicine powder is as follows：
Weigh 25 parts of the Radix Astragali, 15 parts of Radix Codonopsis, 15 parts of radix bupleuri, 10 parts of radix scutellariae；Said herbal medicine is crushed to particle diameter as 2 respectively
Millimeter is following, is then uniformly mixed in container and adds the water of 5 times of weight, and control temperature is to 33 DEG C, pH value 6.5, by mixing
Than adding 0.3% cellulase degradation 1.5 hours, 80 DEG C of control temperature is kept for 8 minutes quality of material, is then cooled to 50 DEG C,
PH is 6.0, by mixed material weight than be separately added into 0.20% papain and pectinase enzymatic hydrolysis 50 minutes, be cooled to 8
DEG C kept for 2 hours, increase the temperature to 90 DEG C and kept for 15 minutes, finally add the mixing of 2 times of w ethanols of mixed material and propyl alcohol
Thing, control temperature to 70 DEG C of holding 3.4h, filtering；Freeze-drying obtains Chinese herbal medicine powder after filter vacuum concentration.
The mass ratio of the ethanol and propyl alcohol is 1：1.2.
(1) zymotic fluid is filtered, concentration, allotment, refined filtration, dry mesophilicα-diastase.
Obtain zymotic fluid through above-mentioned preparation method is through the medium temperature alphalise starch crude enzyme liquid enzyme activity obtained by centrifuging and taking supernatant
9500U/ml.Medium temperature alphalise starch crude enzyme liquid sample determines enzyme activity after the different time is preserved at 40-90 DEG C, as a result shows, sample
Product preserve 24h under the conditions of 65 DEG C still has 83% enzyme activity, and preserving 12h under the conditions of 70 DEG C still has more than 60% enzyme activity；
Embodiment 3
Lactobacillus plantarum capsule product digests powder group by wall material, Lactobacillus plantarum, freeze drying protectant, stachyose and hickory chick
Into.
Described wall material is made up of enzymatic hydrolysis of soybean protein isolate, chitosan, xanthans, carragheen, glycerine and trehalose；On
Stating concentration when each material is made into mixed solution is respectively：Enzymatic hydrolysis of soybean protein isolate 10%, chitosan 1%, xanthans 3%,
Carragheen 0.1%, glycerine 0.5%, trehalose 0.5%.
The preparation method of enzymatic hydrolysis of soybean protein isolate is：The soybean protein isolate solution that compound concentration is 10-13% heats
To 30-33 DEG C, pH to 3 is adjusted, adds the acid protease insulation enzymolysis of soybean protein isolate weight 0.2% 1.5 hours, enzymolysis
Solution spray drying afterwards obtains enzymatic hydrolysis of soybean protein isolate.
Lactobacillus plantarum is deposit number CGMCC NO.9405 strains.
The preparation method of the hickory chick enzymolysis powder is as follows：
(1) Morchella esculenta (L.) Pers sporophore drying and crushing；
(2) water-soluble homogeneous：The Morchella esculenta (L.) Pers sporophore of crushing is added in stainless steel cylinder, adds the water of 3 times of weight of fructification,
Immersion 5 hours, then it is by colloid mill, colloid mill operation condition by this Morchella esculenta (L.) Pers sporophore liquid：Adjust colloid mill stator with
The gap of rotor is 0.6 ± 1 micron, and colloid mill flow is 0.5 ton hour；
(3) heating enzymolysis：Morchella esculenta (L.) Pers sporophore liquid mixed liquor Jing Guo milling treatment of colloid is transferred to stainless steel enzymatic vessel
In be heated to 50 DEG C, adjust pH to 6.0, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.05%, 0.01% β-Portugal gathers
Carbohydrase, 0.1% protease, insulation enzymolysis 1.5 hours, are stirred continuously in enzymolysis process.
(4) dry：Spray drying obtains hickory chick enzymolysis powder after mash filtrations after enzymolysis.
The preparation process of above-mentioned Lactobacillus plantarum capsule product is as follows：
1) core is prepared：Zymotic fluid quality 3% is added into the Lactobacillus plantarum zymotic fluid of fermented tank fermented and cultured
Stachyose and 8% hickory chick enzymolysis powder, are then mixed, pre-freeze 0.5h at -50 DEG C, then in vacuum with frozen-dried protective agent solution
10h is freezed in freeze drier, is ground after lyophilized and core is made；The ratio of zymotic fluid and frozen-dried protective agent solution is 1:
1.4；Frozen-dried protective agent solution is made up of skimmed milk power, trehalose and maltodextrin solution；The volume ratio of three kinds of solution is degreasing
Milk powder：Trehalose：Maltodextrin=3:1:0.5；
The concentration of the skimmed milk power, trehalose and maltodextrin solution is respectively：Skimmed milk power 5%, trehalose 1%,
Maltodextrin 2%.
2) it is coated with：Core is suspended in fluid bed, wall material spraying coating, chitosan, glycerine are sprayed into from a shower nozzle
With the mixed liquor of trehalose, the mixed liquor of xanthans, carragheen and enzymatic hydrolysis of soybean protein isolate is sprayed into from another shower nozzle, is sprayed
Mist speed control is identical, and temperature forms capsule after 28 DEG C, 30 minutes in fluid bed during coating.Using such as patent
201120503311.1 prepared by described device.
Lactobacillus plantarum capsule product bile tolerance is tested.
Capsule product in embodiment 2,3 is placed in 37 DEG C of pig gall salting liquid (solution concentration 3.0%) and be incubated not
Disconnected stirring, take out after 2h, washed with sterile saline, dissolved with solution cyst fluid, determine Lactobacillus Survival, and with not embedding
Bacterium solution is compared.Test result is as shown in the table.
| Metamorphosis | Survival rate | |
| Embodiment 1 | It is not disintegrated | 87.5% |
| Embodiment 2 | It is not disintegrated | 89.3% |
| Non- peridium pair is shone | -- | 0.65% |
Embodiment 4
A kind of grower pigs compound enzyme, is made up of the raw material of following parts by weight：Zytase 3, cellulase 10, β-Portugal
Dextranase 8, mesophilicα-diastase 4, galactosidase 1, pectase 1, Lactobacillus plantarum capsule 5,8 parts of galenical, ginger carries
Take 3 parts of thing, 1.5 parts of radix glycyrrhizae.
The preparation method of the galenical is as follows：
Weigh 15 parts of cassia seed；10 parts of the bark of eucommia；12 parts of Poria cocos；Respectively by said herbal medicine be crushed to particle diameter for 2 millimeters with
Under, then uniformly mixed in container and add the water of 5 times of weight, the acidity of addition said mixture material constituent mass 2% is fine
Plain enzyme is tieed up, controls temperature 50 C, pH：3.8,1h is kept, then adds the mixture of 3 times of w ethanols of mixed material and propyl alcohol,
Temperature is controlled to 70 DEG C of holding 3h, filtering；Freeze-drying obtains galenical after filter vacuum concentration.
The mass ratio of the ethanol and propyl alcohol is 1：1；Concentration of alcohol is 95%；Propanol concentration is 100%；
The preparation method of the Ginger P.E is as follows：
Particle diameter will be crushed to be placed in container for less than 2 millimeters gingers, add the water of 4 times of weight of ginger, control temperature 55
DEG C 2h is kept, add the mixture of 3 times of w ethanols of ginger and methanol, control temperature to 35 DEG C of holding 5h, filtering；Filter vacuum
Freeze-drying obtains Ginger P.E after concentration.
The mass ratio of the ethanol and methanol is 1：2, concentration of alcohol 90%；Methanol concentration 100%.
Embodiment 5
Substantially with example 1-4
A kind of grower pigs compound enzyme, is made up of the raw material of following parts by weight：Zytase 8, cellulase 15, β-Portugal
Dextranase 5, mesophilicα-diastase 5, galactosidase 2, pectase 2, Lactobacillus plantarum capsule 3,10 parts of galenical, ginger carries
Take 2 parts of thing, 2 parts of radix glycyrrhizae.
The zytase, 1,4 beta-glucanase, cellulase, mesophilicα-diastase, galactosidase are food grade enzyme
Preparation；
Embodiment 6
A kind of grower pigs compound enzyme, is made up of the raw material of following parts by weight：Zytase 5, cellulase 8, β-Portugal
Dextranase 10, mesophilicα-diastase 5, galactosidase 1, pectase 1, Lactobacillus plantarum capsule 3,6 parts of galenical, ginger carries
Take 2 parts of thing, 1 part of radix glycyrrhizae.
The preparation method of grower pigs compound enzyme of the present invention is as follows：
By the radix glycyrrhizae, galenical and Ginger P.E distinguish ultramicro grinding, then with zytase, cellulase,
1,4 beta-glucanase, amylase, galactosidase, pectase, Lactobacillus plantarum capsule are packed after uniformly mixing and got product
Grower pigs compound enzyme.
Feeding experiment：
Materials and methods
The selection and packet of test pig
On a certain large-scale regular pig farm in Hunan, the healthy DLY three way cross that 36 body weight are (30 ± 2) kg is chosen
Grower pigs, are randomly divided into 3 groups, and every group of 2 columns repeat, often repeatedly 6, galt and gilt half and half.Formal test advance behavior
The prerun of 1 week phase, and complete expelling parasite and routine immunization work.Experiment is divided to front and rear two phase to carry out, early stage (30kg-60kg), the later stage
(61kg~90kg).
Feeding management
Dry mash being fed, free choice feeding, being limited with cannot not have enough surplusly, automatic drinking bowl drinking-water.Feed day early stage three times, day in later stage
Feed it is secondary, feed consumption rate is recorded in units of column.Pig house is brick structure, cement flooring, single-column type, and examination pig is raised in 9 columns respectively
Interior, the environmental condition of each column home is consistent, cleans daily morning and afternoon that colony house is each once, while observes the behavior of pig, appetite, excrement.
Disease and treatment are recorded during experiment.
Feed and formula
Based on full price national standard grows pig feed, feed per ton adds the growth of test group, control group 1 and control group 2
Pig specific enzyme 100g；
Test index：Before the test at the end of (30kg-60kg), mid-term and later stage (61kg~90kg), claim respectively
Individual weight, weigh and carry out on an empty stomach in the morning.Stage by stage, it is recorded in units of column per daily material consumption, the incidence of disease, is calculated simultaneously
Daily ingestion amount and food utilization efficiency, and above-mentioned data are carried out with statistical analysis, growth pig growth performance is shown in Table 2.
Table 2
Above-mentioned experiment full period test result is analyzed such as table 3：
Table 3
| Project | Test group | Control group 1 | Deviation (%) | Control group 2 | Deviation (%) |
| Daily gain (g) | 877 | 746 | 131(17.56) | 657 | 220(33.49) |
| Feed intake (kg) | 189.73 | 193.46 | -3.73(-1.92) | 210.17 | -20.44(-9.7) |
| Feed-weight ratio | 2.80 | 3.36 | -0.56(-16.67) | 4.15 | -1.35(-32.53) |
| Diarrhea rate (%) | 0 | 4 | -4(-100) | 17 | -17(-100) |
| Hair color scores | 9 | 6.5 | 2.5(38.46) | 4 | 5(125) |
Claims (9)
1. a kind of grower pigs compound enzyme, is made up of the raw material of following parts by weight：Zytase 3-8, cellulase 8-15, β-
Dextranase 5-10, mesophilicα-diastase 3-5, galactosidase 1-2, pectase 1-2, Lactobacillus plantarum capsule 3-6, plant system
Agent 6-10 parts, Ginger P.E 2-4 parts, radix glycyrrhizae 1-2 parts；The preparation method of the galenical is as follows：
Weigh cassia seed 10-18 parts；Bark of eucommia 5-15 parts；Poria cocos 10-15 parts；Said herbal medicine is crushed to particle diameter as 2 millis respectively
Rice is following, is then uniformly mixed in container and adds the water of 3-6 times of weight, addition said mixture material constituent mass 1-2%
Acidic cellulase, control 45-55 DEG C of temperature, pH：3.5-4,1-2h is kept, then adds 2-3 times of weight second of mixed material
The mixture of alcohol and propyl alcohol, control temperature to 60 DEG C of -78 DEG C of holding 3-4h, filtering；It is freeze-dried and obtains after filter vacuum concentration
Galenical；
The mesophilicα-diastase has following characteristic：
(1) the enzyme Acclimation temperature wider range, temperature activity between 55-75 DEG C is higher, and optimum temperature is 65 DEG C；
Heat endurance：The enzyme preserves 24h under the conditions of 65 DEG C still has 80-83% enzyme activity, and preserving 12h under the conditions of 70 DEG C still has
More than 50-60% enzyme activity；
(2) the enzyme optimal reaction pH value is 5.0, there is high enzyme vigor between pH value 4.0-6.0；Absolute acid stability：In pH4-6
Still there is more than 80% enzyme activity after lower preservation 18h；
(3) enzymatic activity：Bacterial strain 304 is 7000-9600U/ml by the acid mesophilicα-diastase enzyme activity prepared that ferments；Especially
It is adapted to liquefaction process and Mashing process and the industrialization demand deposited；The bacterial strain for producing mesophilicα-diastase is specially bacillus subtilis
304Bacillus subtilis304, preserving number are CCTCC NO:M 2013600.
2. grower pigs compound enzyme according to claim 1, it is characterised in that the preparation method of the Ginger P.E is such as
Under：
Particle diameter will be crushed to be placed in container for less than 2 millimeters gingers, the water of addition 3-6 times of weight of ginger, control temperature 50 C-
The mixture of 60 DEG C of holding 2-3h, addition 2-3 times of w ethanol of ginger and methanol, control temperature to 30 DEG C of -40 DEG C of holding 3-8h,
Filtering；Freeze-drying obtains Ginger P.E after filter vacuum concentration.
3. according to grower pigs compound enzyme described in claim 1, it is characterised in that the preparation method of the Lactobacillus plantarum capsule is such as
Under：
Lactobacillus plantarum capsule product is made up of wall material, Lactobacillus plantarum, freeze drying protectant, stachyose and hickory chick enzymolysis powder；
Described wall material is made up of enzymatic hydrolysis of soybean protein isolate, chitosan, xanthans, carragheen, glycerine and trehalose；It is above-mentioned each
Concentration when material is made into mixed solution is respectively：Enzymatic hydrolysis of soybean protein isolate 7-10%, chitosan 1-2%, xanthans 1-
3%th, carragheen 0.1-0.5%, glycerine 0.3-1%, trehalose 0.5-2%；The preparation method of enzymatic hydrolysis of soybean protein isolate is：Match somebody with somebody
Concentration processed is that 10-13% soybean protein isolate solution is heated to 30-45 DEG C, adjusts pH to 3-5, adds soybean protein isolate weight
0.1-1% acid protease insulation digests 0.5-1.5 hours, and solution spray drying acquisition enzymatic hydrolysis of soybean separates egg after enzymolysis
In vain.
4. according to grower pigs compound enzyme described in claim 3, it is characterised in that Lactobacillus plantarum is CGMCC NO.9405.
5. according to grower pigs compound enzyme described in claim 1-3, it is characterised in that Lactobacillus plantarum capsule product preparation method is such as
Under：
1) core is prepared：Zymotic fluid quality 3-5% water is added into the Lactobacillus plantarum zymotic fluid of fermented tank fermented and cultured
Threose and 5-8% hickory chick digest powder, are then mixed with frozen-dried protective agent solution, pre-freeze 0.5h at -50 DEG C, then in vacuum
10-18h is freezed in freeze drier, is ground after lyophilized and core is made；
The ratio of zymotic fluid and the frozen-dried protective agent solution is 1:1.4-0.8；The frozen-dried protective agent solution by skimmed milk power,
Trehalose and maltodextrin solution composition；The volume ratio of three kinds of solution is skimmed milk power：Trehalose：Maltodextrin=3:1:0.5；
The concentration of the skimmed milk power, trehalose and maltodextrin solution is respectively：Skimmed milk power 5%, trehalose 1%, malt
Dextrin 2%；
2) it is coated with：Core is suspended in fluid bed, wall material spraying coating, chitosan, glycerine and sea are sprayed into from a shower nozzle
The mixed liquor of algae sugar, the mixed liquor of xanthans, carragheen and enzymatic hydrolysis of soybean protein isolate, spraying speed are sprayed into from another shower nozzle
Degree control is identical, and between 25-38 DEG C, capsule is formed after 30 minutes for temperature in fluid bed during coating.
6. according to grower pigs compound enzyme described in claim 1, it is characterised in that be made up of the raw material of following parts by weight：Wood is poly-
Carbohydrase 3, cellulase 10,1,4 beta-glucanase 8, mesophilicα-diastase 4, galactosidase 1, pectase 1, Lactobacillus plantarum capsule
5,8 parts of galenical, 3 parts of Ginger P.E, 1.5 parts of radix glycyrrhizae.
7. according to grower pigs compound enzyme described in claim 1, it is characterised in that be made up of the raw material of following parts by weight：Wood is poly-
Carbohydrase 8, cellulase 15,1,4 beta-glucanase 5, mesophilicα-diastase 5, galactosidase 2, pectase 2, Lactobacillus plantarum capsule
3,10 parts of galenical, 2 parts of Ginger P.E, 2 parts of radix glycyrrhizae.
8. according to grower pigs compound enzyme described in claim 1, it is characterised in that be made up of the raw material of following parts by weight：Wood is poly-
Carbohydrase 5, cellulase 8,1,4 beta-glucanase 10, mesophilicα-diastase 5, galactosidase 1, pectase 1, Lactobacillus plantarum capsule
3,6 parts of galenical, 2 parts of Ginger P.E, 1 part of radix glycyrrhizae.
It is 9. as follows according to the preparation method of grower pigs compound enzyme described in claim 1：
By the radix glycyrrhizae, galenical and Ginger P.E distinguish ultramicro grinding, then with zytase, cellulase, β-Portugal
Dextranase, amylase, galactosidase, pectase, Lactobacillus plantarum capsule pack after uniformly mixing and get product growth
Pig compound enzyme.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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| CN105087523A (en) * | 2015-09-01 | 2015-11-25 | 沈阳农业大学 | Complex enzyme for animal feed and application of complex enzyme |
| CN106234795A (en) * | 2016-08-10 | 2016-12-21 | 河南大陆农牧技术股份有限公司 | A kind of for sick pig rehabilitation or the health-nutrition tonic of children's pig in age |
| CN106721009A (en) * | 2016-12-23 | 2017-05-31 | 山西大禹生物工程股份有限公司 | Growth promotion puies forward quality feed addictive and preparation method |
| CN108925759A (en) * | 2017-05-25 | 2018-12-04 | 南宁市磲合生物科技有限公司 | A kind of feeding Chinese herbal medicine complex enzyme and preparation method thereof |
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| CN101874547A (en) * | 2009-11-27 | 2010-11-03 | 济南天天香有限公司 | Complex enzyme for feed |
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