CN104824379A - Biological composite enzyme for feed and preparation method thereof - Google Patents
Biological composite enzyme for feed and preparation method thereof Download PDFInfo
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- CN104824379A CN104824379A CN201510004750.0A CN201510004750A CN104824379A CN 104824379 A CN104824379 A CN 104824379A CN 201510004750 A CN201510004750 A CN 201510004750A CN 104824379 A CN104824379 A CN 104824379A
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Abstract
The invention discloses a biological composite enzyme for a feed and a preparation method thereof, belongs to the field of enzyme preparations and particularly discloses a composite enzyme and the preparation method thereof. The biological composite enzyme is composed of following raw materials, by weight: 10-20 parts of xylanase, 6-12 parts of cellulase, 3-5 parts of amylase, 2-5 parts of acidic protease, 1-2 parts of pectinase, 1-2 parts of mannase, 2-5 parts of capsules of lactobacillus plantarum, 1-3 parts of a plant preparation, 1-2 parts of a fresh ginger extract and 1-2 parts of radix glycyrrhizae. In the invention, by means of a synergistic effect of the protective agent, the enzyme preparation, the plant preparation, the fresh ginger extract and the enzyme preparation, the biological composite enzyme achieves best enzyme activity and efficacy, and can correspondingly improve the utilization rate of the feed and growth rate of animals, can enhance appetite and disease resistance of animals, and can prolong the shelf life of the feed and protect environment.
Description
Technical field
The invention belongs to enzymic preparation field, specifically complex enzyme and preparation method thereof.
Background technology
By adding enzyme preparation, acid protease and amylase can be supplemented, the albumen in auxiliary decomposition daily ration and starch, the ANFs in degraded daily ration, and the plant cell wall that can effectively decompose in corn/soybean meal based diets, intracellular nutriment such as starch, protein etc. are discharged.Decomposing soluble fiber simultaneously, reduces the water absorbing force of vegetable fibrous material and the viscosity of intestinal contents.
A kind of complex enzyme preparation for feeding piglets, its application number: 201110026602.0, a kind of complex enzyme preparation for feeding piglets of disclosure of the invention.This complex enzyme preparation for feeding piglets comprises following seven kinds of enzymes: acid acid protease, amylase, zytase, cellulase, pectase and phytase; The enzyme activity ratio of described 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 complex enzyme formulation degradable feed of the present invention, reduces enteron aisle chyme viscosity, improves feedstuff metabolisable energy; Improve the digestibility of piglet to protein and starch; Reduce harmful microbe excessive multiplication in enteron aisle, reduce the damage of intestines wall, reconcile intestinal microecology balance; Improve piglet survival rate, premunition, reduce diarrhea rate, improve the overall uniformity; Promote weight gain of piglets, reduce aquaculture cost.
A kind of pig's feed complex enzyme formulation being added with activator, its application number: 201210500011.7 inventions provide a kind of pig's feed complex enzyme formulation being added with activator, organophosphor in feed can be resolved into the inorganic phosphate that can absorb by the phytase in formula, protein-based nutriment in acid protease degradable feed, promote that live pig is to the absorption of Dietary phosphorus and nitrogen, also reduce the discharge capacity of phosphorus and nitrogen in live pig fecaluria simultaneously, reduce its pollution on the environment.And activator can improve the reaction rate of enzyme, enzyme preparation is taken effect faster, and each ingredient combination in activator and enzyme preparation use the shearing force of cold cuts also can be made to reduce, and improves pork tenderness.The pig's feed complex enzyme formulation that the present invention obtains can improve the digestibility of pig to nitrogen, phosphorus in feed, thus makes the excretion of phosphorus in live pig fecaluria reduce more than 40%, and the excretion of nitrogen reduces more than 29%; Instant effect, namely has significant effect in 2-3 days; Cold cuts shearing force reduces by more than 50%.
Name is called a kind of pig's feed complex enzyme formulation and preparation method thereof, its application number: 201410275557.6 inventions provide a kind of pig's feed complex enzyme formulation, belong to feed additive field.This additive is made up of complex enzyme and reinforcing agent; Complex enzyme is lipase, neutral acid protease, beta amylase, isoamylase, neutral phytase, 'beta '-mannase, zytase, alpha-acid protease; Reinforcing agent is made up of extract from pine needles, leaf of bamboo Thick many candies, caulis lonicerae powder, galangal powder, radix jurineae powder, Asian puccoon powder, vitamin C, sodium chloride, sodium butyrate, ferrous sulfate and calcium gluconae.This enzyme preparation effectively can improve the utilization rate of feed, reduces feedstuff-meat ratio, promotes the speed of growth of pig; Can also play the effect of the immunity strengthening pig, to the health degree improving pig, reduce the pig incidence of disease and have very large help, market prospects are very wide.
A kind of efficient pig's feed complex enzyme formulation being added with enzymatic protective reagent, its application number: 201210381755.1 inventions provide a kind of efficient pig's feed complex enzyme formulation being added with enzymatic protective reagent, the materials such as ANFs and large biological molecule such as the beta glucan in degradable feed, phytic acid, pectin, thus promote that pig is to the absorption of feed Middle nutrition composition, improves efficiency of feed utilization; Protectantly add the inactivation rate that can reduce enzyme, extend effective time, make the effect of this enzyme preparation more significantly and lasting; The high efficiency composition enzyme preparation feed conversion rate that the present invention obtains is high, and feedstuff-meat ratio can be used to decline more than 19%; The pig speed of growth improves more than 23%; The enzymatic activity retention time is long, and in 18 months, enzymatic activity can remain on more than 90%.A kind of efficient pig's feed complex enzyme formulation being added with enzymatic protective reagent, application number: 201210360110.X, invention provides a kind of efficient pig's feed complex enzyme formulation being added with enzymatic protective reagent, the materials such as ANFs and large biological molecule such as the beta glucan in degradable feed, phytic acid, pectin, thus promote that pig is to the absorption of feed Middle nutrition composition, improves efficiency of feed utilization; Protectantly add the inactivation rate that can reduce enzyme, extend effective time, make the effect of this enzyme preparation more significantly and lasting; The high efficiency composition enzyme preparation feed conversion rate that the present invention obtains is high, and feed coefficient can be used to decline more than 19%; The pig speed of growth improves more than 23%; The enzymatic activity retention time is long, and in 30 months, enzymatic activity can remain on more than 90%.
A kind of the feed for piglet complex enzyme formulation and production method thereof and application, application number: the 201410150709.X applying date: a kind of the feed for piglet complex enzyme formulation of 2014-04-15 disclosure of the invention and production method thereof and application.The enzyme of this complex enzyme formulation is lived and is consisted of: glucose oxidase 80-100U/g, zytase 3000-4000U/g, 'beta '-mannase 500-600U/g, acid protease 2000-3000U/g, neutral proteinase 800-1000U/g, amylase 8 00-1000U/g, fungal alpha-amylase 400-500U/g, glucoamylase 3000-4000U/g.This complex enzyme formulation adopts cornstarch and potassium dihydrogen phosphate, dipotassium hydrogen phosphate, mixture of calcium sulfate as diluent.This complex enzyme formulation using method is: the addition in newborn piglet perfect compound feed per ton is 500-750 gram.Name is called a kind of the feed for piglet complex enzyme formulation and production method thereof and application, application number: 201410150709.X, a kind of the feed for piglet complex enzyme formulation of disclosure of the invention and production method thereof and application.The enzyme of this complex enzyme formulation is lived and is consisted of: glucose oxidase 80-100U/g, zytase 3000-4000U/g, 'beta '-mannase 500-600U/g, amylase 8 00-1000U/g, fungal alpha-amylase 400-500U/g, glucoamylase 3000-4000U/g.This complex enzyme formulation adopts cornstarch and potassium dihydrogen phosphate, dipotassium hydrogen phosphate, mixture of calcium sulfate as diluent.This complex enzyme formulation using method is: the addition in newborn piglet perfect compound feed per ton is 500-750 gram.
The bark of eucommia is rich in phenylpropanoids, iridoids isoreactivity composition, has rubbish in purged body, strengthens cellular material metabolism, decomposer inner cholesterol, reduces body fat, broad-spectrum antiseptic, improves white blood cell count, the remarkable efficacy such as develop immunitypty.The result of pure eucommia bark powder in zoopery shows: can promote cholesterol and lipid-metabolism; Add the eel that pure eucommia bark powder is raised, there is the taste of natural eel; Pure eucommia bark powder is added to the quality that can improve broiler chicken in feed, make it that there is tempting fragrance and quality.
In sum, the market space that the application of feed biological complex enzyme has it wide and huge economic worth, but the heat endurance of feed biological complex enzyme, security, composite comprehensive and action effect give full play to the major issue being still enzyme preparation manufacturer and numerous raisers and jointly paying close attention to, prepare safer, more comprehensively, the better feed biological complex enzyme of enzyme action effect is corporation responsibility and the pursuit of industry technical staff.
Summary of the invention
A kind of feed biological complex enzyme of technical problem solved by the invention, is made up of the raw material of following parts by weight:
Zytase 10-20, cellulase 6-12, amylase 3-5, acid protease 2-5, pectase 1-2, mannase 1-2, Lactobacillus plantarum capsule 2-5, galenical 1-3 part, Ginger P.E 1-2 part, Radix Glycyrrhizae 1-2 part.
Described zytase, cellulase, amylase, acid protease are food-grade enzyme preparation;
The preparation method of described galenical is as follows:
Take cassia seed 10-18 part; Bark of eucommia 5-15 part; Poria cocos 10-15 part; Respectively said herbal medicine being crushed to particle diameter is less than 2 millimeters, then Homogeneous phase mixing add the water of 3-6 times of weight in container, add the acidic cellulase of said mixture constituent mass 1-2%, control temperature 45-55 DEG C, pH:3.5-4, keeps 1-2h, adds the mixture of mixed material 2-3 times of w ethanol and propyl alcohol subsequently, control temperature to 60 DEG C-78 DEG C of maintenance 3-4h, filter; Filter vacuum concentrates postlyophilization and obtains galenical.
The mass ratio of described ethanol and propyl alcohol is 1:1-1.5; Concentration of alcohol is 95%; Propanol concentration is 100%;
The preparation method of described Ginger P.E is as follows:
To be crushed to particle diameter is that less than 2 millimeters gingers are placed in container, adds the water of 3-6 times of weight, and control temperature 50 DEG C-60 DEG C keeps 2-3h, adds the mixture of mixed material 2-3 times of w ethanol and methyl alcohol, and control temperature to 30 DEG C-40 DEG C of maintenance 3-8h, filter; Filter vacuum concentrates postlyophilization and obtains Ginger P.E.
The mass ratio of described ethanol and methyl alcohol is 1:2, concentration of alcohol 85-95%.Methanol concentration 100%.
The preparation method of described Lactobacillus plantarum capsule is as follows:
Lactobacillus plantarum capsule product is made up of wall material, Lactobacillus plantarum, freeze drying protectant, stachyose, hickory chick enzymolysis powder.
Described wall material is made up of enzymatic soybean protein isolate, shitosan, xanthans, carragheen, glycerine and trehalose; Concentration when above-mentioned each material is made into mixed solution is respectively: enzymatic soybean protein isolate 7-10%, shitosan 1-2%, xanthans 1-3%, carragheen 0.1-0.5%, glycerine 0.3-1%, trehalose 0.5-2%.
The preparation method of enzymatic soybean protein isolate is: compound concentration is that 10-13% soybean protein isolate solution is heated to 30-45 DEG C, adjustment pH to 3-5, add the protease insulation enzymolysis 0.5-1.5 hour of soybean protein isolate weight 0.1-1%, after enzymolysis, solution spray drying obtains enzymatic 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 pulverizing is added in stainless steel cylinder, add the water of fructification 3-6 times of weight, soak 3-5 hour, then this Morchella esculenta (L.) Pers sporophore liquid is passed through colloid mill, colloid mill operation condition is: adjustment colloid mill stator and the gap of rotor are 0.5-1 micron, colloid mill flow be 0.4-1 ton/hour;
(3) intensification enzymolysis: the Morchella esculenta (L.) Pers sporophore liquid mixed liquor through milling treatment of colloid is transferred in stainless steel enzymatic vessel and is heated to 50-60 DEG C, adjustment pH to 4.5-6.0, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.05-0.1%, the protease of 0.01-0.1%, insulation enzymolysis 0.5-1.5 hour, constantly stirs in enzymolysis process.
(4) dry: dry acquisition hickory chick enzymolysis powder after the mash filtrations after enzymolysis.
Described drying can adopt the dried forms such as conventional spraying dry, freeze drying.
Described freeze drying protectant is made up of skimmed milk power, trehalose, maltodextrin.
The preferred CGMCC NO.9405 of the Lactobacillus plantarum that capsule product of the present invention comprises.
Above-mentioned Lactobacillus plantarum capsule product preparation method is as follows:
1) core is prepared: in the Lactobacillus plantarum zymotic fluid cultivated through ferment tank, add the stachyose of zymotic fluid quality 3-5% and the hickory chick enzymolysis powder of 5-8%, then mix with frozen-dried protective agent solution, pre-freeze 0.5h at-50 DEG C, then freeze-drying 10-18h in vacuum freeze drier, grinds after freeze-drying and makes core;
The ratio of described zymotic fluid and frozen-dried protective agent solution is 1:1.4-0.8; Described frozen-dried protective agent solution is made up of skimmed milk power, trehalose, maltodextrin solution; The volume ratio of three kinds of solution is skimmed milk power: trehalose: maltodextrin=3:1:0.5;
The concentration of described skimmed milk power, trehalose, maltodextrin solution is respectively: skimmed milk power 5%, trehalose 1%, maltodextrin 2%.
2) bag quilt: core is suspended in fluid bed, wall material spraying bag quilt, the mixed liquor of shitosan, glycerine and trehalose is sprayed into from a shower nozzle, the mixed liquor of xanthans, carragheen and enzymatic soybean protein isolate is sprayed into from another shower nozzle, spray velocity controls identical, bag between 25-38 DEG C, is namely formed capsule after 30 minutes by temperature in fluid bed in process.This drying means adopts device processes described in patent 201120503311.1.
The preparation method of feed biological complex enzyme of the present invention is as follows:
By described Radix Glycyrrhizae, galenical and Ginger P.E ultramicro grinding respectively, then with zytase, cellulase, amylase, acid protease, pectase, mannase, packs the feed biological complex enzyme that gets product after Lactobacillus plantarum capsule Homogeneous phase mixing.
Described feed biological complex enzyme is applicable to feed-processing plant and plant's autogamy feed, should mix, can the present invention be mixed with a small amount of feed in advance, then be mixed in large quantities of feed, Direct-fed during use with other raw material in feed.Advise that complete diet pellet addition per ton is: 100-150g.
Beneficial effect:
Adopt galenical, Ginger P.E, Radix Glycyrrhizae science composite in product of the present invention, effectively slow down the moisture regain of enzyme preparation; Can strengthen complex enzyme simultaneously resistance toly to freeze, heat resistance, keep identical enzyme activity, its heat resisting temperature can improve 10-15 DEG C, resistance to cryogenic temperature can reduce 5-10 DEG C, effectively prevent the loss of complex enzyme enzyme activity in transport, preservation and use procedure, extend the shelf-life of complex enzyme, reach same enzyme activity, the like product shelf-life can extend 3-5 month.
The galenical that product of the present invention adds both can extend the shelf-life of complex enzyme formulation, can improve again the immunity of raising livestock and poultry, effectively prevent the generation of livestock and poultry epidemic disease.
Lactobacillus micro-capsule provided by the invention, modified soybean protein isolate is with the addition of in its wall material, modified soybean protein isolate improves 15-25% than soybean protein isolate emulsifying capacity raising 25%, gelling ability, modified soybean protein isolate xanthans and carragheen and shitosan with the use of, gel strength improves more than 10%, enhances the stomach juice-resistant of capsule;
Capsule of the present invention adopts modified soybean protein isolate, has good enteric solubility, complete disintegration can discharge Lactobacillus plantarum, and propagation becomes dominant microflora rapidly after capsule arrives enteron aisle in 1-1.5 hour, thus reaches effects such as suppressing growth of pathogenic bacteria.
The common synergy of protective agent and enzyme preparation, galenical, Ginger P.E and enzyme preparation in product of the present invention; the enzyme activity of complex enzyme and effect are played to greatest extent; and improve the utilization rate of feed and the growth rate of animal accordingly; enhance appetite and the resistance against diseases of animal, extend the shelf-life of product and protect environment.
Detailed description of the invention
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, but not limits the scope of the invention, and the spirit and scope of the invention only limited by claims.To those skilled in the art, under the prerequisite not deviating from essence of the present invention and scope, the various change carry out the material component in these embodiments and consumption or change also belong to protection scope of the present invention.
Embodiment 1
The Breeding Process of the present invention's Lactobacillus plantarum used CGMCC No.9405 is as follows:
The original bacterial classification that sets out → test tube activation → dithyl sulfate (DES) mutagenesis → nitrosoguanidine (NTG) mutagenesis → plasma mutagenesis → dull and stereotyped primary dcreening operation → shaking flask sieves → mitotic stability test again.
Starting strain of the present invention is in MRS dextrose culture-medium, and the throughput rate of its lactic acid is 1.5g/L/d, almost stops growing when medium pH is 3.5, is 0.34mg/h/kg Chinese cabbage to the decomposition rate of natrium nitrosum.Starting strain by
li Zhengbe collected in the greenfeed of Fattening Sheep field, Yanchi county Ningxia, acquisition time on September 15th, 2013.
In order to improve the decomposition rate of its production of lactic acid speed, acid-fast ability and nitrite, DES and NTG technology is adopted to carry out mutagenesis to this bacterial classification successively, after mutagenesis, bacterial strain adopts MRS calcium carbonate flat board to carry out primary dcreening operation, then 500mL shake flask fermentation is adopted, biosensor analysis instrument carries out multiple sieve to Producing Strain, the lactobacillus plantarum strain that seed selection is excellent, then does passage assays, evaluates its genetic stability.
Lactobacillus plantarum tlj-2014 genetic stability result shows: through continuous passage ten times, property indices is all more stable, and heredity is better, and proterties is not replied, therefore using the object bacterial strain that Lactobacillus plantarum tlj-2014 obtains as seed selection.
Empirical tests finds: the production of lactic acid speed of this mutagenic strain can reach 35g/L/d, and this bacterial strain lactic acid concn after 71 hours fermentation reaches 95g/L; Can survive under pH is the condition of 1.80.Degrading nitrite speed is fast, and capacity of decomposition reaches 9.8mg/h/kg (speed of spontaneous fermentation process nitrite accumulation is approximately 1.1mg/h/kg), can resistance to 1% cholate.
1) DES mutagenic and breeding
A. on super-clean bench, get Lactobacillus plantarum L mono-ring (set out bacterium) on test tube slant, access is equipped with in the 250mL triangular flask of 50mL culture medium MRS (without agar, glucose 20g/L), 200rpm, cultivate about 12h for 37 DEG C, make thalline be in logarithmic growth in earlier stage.
B. get 5mL bacterium liquid, the centrifugal 10min of 5000rpm collects thalline, with brine 2 times.
C. 10 are diluted to pH7.0 phosphate buffer
7individual/mL bacteria suspension.
D. get the kaliumphosphate buffer of 32mL pH7.0,8mL bacteria suspension, 150mL triangular flask that 0.4mL DES to put into rotor in advance fully mix, make DES ultimate density be 1% (v/v).
E. in 37 DEG C of shaking tables, 150rpm reacts 30min, gets 1mL mixed liquor, adds 0.5mL 25%Na
2s
2o
3solution stopped reaction.
F. suitably dilute, get last dilution bacterium liquid 0.2mL, coat in calcium carbonate screening and culturing base (the calcium carbonate MRS culture medium containing 100g/L glucose) plate.Cultivate after 2 ~ 3 days at 37 DEG C, adopting photolithography the bacterial strain of this screening flat board to be transferred to pH is on the upper and natrium nitrosum screening and culturing base (single nitrogenous source is the modification MRS screening and culturing base of 2g/L natrium nitrosum) of LPHMRS culture medium (low ph value modification MRS culture medium) of 1.5,1.8 and 2.0.
G. cultivate after 2 ~ 3 days at 37 DEG C, choosing colony is comparatively large, can grow respectively and on LPHMRS culture medium, natrium nitrosum screening and culturing base on calcium carbonate screening and culturing base.Through Preliminary screening, the bacterium colony called after Lactobacillus plantarum L1 that picking goes out.
2) nitrosoguanidine mutagenesis
A. on super-clean bench, get Lactobacillus plantarum L1 mono-ring on test tube slant, access is equipped with in the 250mL triangular flask of 50mL culture medium MRS (without agar) (concentration of glucose is 60g/L), 200rpm, cultivates about 12h for 37 DEG C, makes thalline be in logarithmic growth in earlier stage.
B. get the centrifugal 10min of 5mL bacterium liquid 5000rpm and collect thalline, with brine 2 times.
C. 10 are diluted to pH6.0 phosphate buffer
7individual/mL bacteria suspension.
D. get 10mL bacteria suspension to be transferred in 100mL triangular flask, add the NTG of 10mg, be mixed with the NTG solution that final concentration is 10mg/mL, and add 4-5 and drip acetone, be beneficial to NTG and dissolve.
E. at 37 DEG C, the centrifugal 10min of 200rpm oscillating reactions 30min, 5000rpm collects thalline, with SPSS washing several, and stopped reaction.
F. suitably dilute, get last dilution bacterium liquid 0.2mL, coat in calcium carbonate screening and culturing base (the calcium carbonate MRS culture medium containing 100g/L glucose) plate.Cultivate after 2 ~ 3 days at 37 DEG C, adopting photolithography the bacterial strain of this screening flat board to be transferred to pH is on the upper and natrium nitrosum screening and culturing base (single nitrogenous source is the modification MRS screening and culturing base of 2g/L natrium nitrosum) of LPHMRS culture medium (low ph value modification MRS culture medium) of 1.5,1.8 and 2.0.
G. select bacterial strain method: choosing colony is comparatively large, to grow on LPHMRS culture medium, natrium nitrosum screening and culturing base respectively and on calcium carbonate screening and culturing base.Through Preliminary screening, picking 100 meets the bacterium colony of above condition.
3) shaking flask is sieved again
A. on super-clean bench, get Lactobacillus plantarum one ring on each test tube slant respectively, access is equipped with in the 250mL triangular flask of 50mL culture medium MRS (without agar) (concentration of glucose is 100g/L), 200rpm, cultivates about 15h, makes thalline be in mid log phase for 37 DEG C.
B. get 5mL bacterium liquid respectively, LPHMRS fluid nutrient medium (low ph value modification MRS culture medium) and the natrium nitrosum liquid screening medium (single nitrogenous source is the modification MRS screening and culturing base of 2g/L natrium nitrosum) upper (note: adopt 250mL triangular flask) that 50mL calcium carbonate screens in fluid nutrient medium (the calcium carbonate MRS culture medium containing 250g/L glucose) plate, pH is 1.5,1.8 and 2.0 is equipped with in access.200rpm, cultivates 3-4 days for 37 DEG C, detects the wear rate that Pfansteihl in calcium carbonate screening fluid nutrient medium produces speed, biomass in LPHMRS fluid nutrient medium and natrium nitrosum liquid screening medium nitrite every day respectively.After fermentation ends, compare the wear rate that Pfansteihl in the calcium carbonate screening fluid nutrient medium of 100 strain bacterial classifications produces speed, biomass in LPHMRS fluid nutrient medium and natrium nitrosum liquid screening medium nitrite.
C. select to have concurrently the bacterial strain that high Pfansteihl produces speed, the wear rate of tolerate low pH (this bacterial classification only can grow in the minimum culture medium for pH1.8) and nitrite is high, by its called after L2 bacterium.
4) genetic stability test
L2 bacterium is gone down to posterity for continuous ten times on inclined-plane, and detects the fermentation situation after at every turn going down to posterity by the method that shaking flask is sieved again.Experiment finds, inclined-plane goes down to posterity for continuous ten times, and this bacterial classification proterties does not have significant change, and property indices is all normal, illustrates that the genetic stability of this bacterial classification is stronger.Strain Designation is Lactobacillus plantarum (Lactobacillus plantarum) tlj-2014.
5) 5L fermentation tank test
A. get Lactobacillus plantarum L2 mono-ring on inclined-plane, access is equipped with in the 250mL triangular flask of 50mL culture medium MRS (without agar) (concentration of glucose is 150g/L), 200rpm, cultivates about 12h, makes thalline be in mid log phase for 37 DEG C.
B. the access of the bacterial classification of logarithmic phase is equipped with in the 5L fermentation tank of 3L MRS fluid nutrient medium (initial glucose is 150g/L).Inoculum concentration is that at 10%, 37 DEG C, 100rpm cultivates 8 hours, and logarithm dissolved oxygen in early stage controls 10% (ventilation 0.5L/min), later stage Anaerobic culturel 63 hours.After fermentation ends, the lactic acid production of Lactobacillus plantarum L2 reaches 95g/L.
C. 3L pH being equipped with in the access of the bacterial classification of logarithmic phase is in the 5L fermentation tank of LPHMRS fluid nutrient medium (initial glucose is 50g/L) of 1.8.Inoculum concentration is that at 10%, 37 DEG C, 100rpm cultivates 8 hours, and logarithm dissolved oxygen in early stage controls 10% (ventilation 0.5L/min), and later stage anaerobism, zymotic fluid pH controls 1.8 by the NaOH of whole process 0.5mol/L, and total incubation time is 48 hours.After fermentation ends, the biomass detecting Lactobacillus plantarum L2 is 2.5g/L, illustrates that Lactobacillus plantarum L2 can survive in the environment of pH1.8.
D. the access of the bacterial classification of logarithmic phase is equipped with in the 5L fermentation tank of 3L natrium nitrosum liquid screening medium (single nitrogenous source is the modification MRS screening and culturing base of 2g/L natrium nitrosum).Inoculum concentration is that at 10%, 37 DEG C, 100rpm cultivates 8 hours, and logarithm dissolved oxygen in early stage controls 10% (ventilation 0.5L/min), and later stage anaerobism, sweat adds the sodium nitrite solution of 20g/L according to the wear rate stream of nitrite, cultivates 2-3 days.After fermentation ends, calculate sweat Lactobacillus plantarum L2 to the degradation rate of natrium nitrosum.Found that: under this condition, L2 can reach 563mg/h/L to the degradation rate of natrium nitrosum.
Embodiment 2
Lactobacillus plantarum capsule product is made up of wall material, Lactobacillus plantarum, freeze drying protectant, stachyose, hickory chick enzymolysis powder.
Described wall material is made up of enzymatic soybean protein isolate, shitosan, xanthans, carragheen, glycerine and trehalose; Concentration when above-mentioned each material is made into mixed solution is respectively: enzymatic soybean protein isolate 10%, shitosan 1%, xanthans 3%, carragheen 0.1%, glycerine 0.5%, trehalose 0.5%.The preparation method of enzymatic soybean protein isolate is: compound concentration is that 10-13% soybean protein isolate solution is heated to 30-33 DEG C, pH is to 3 in adjustment, the acid protease adding soybean protein isolate weight 0.2% is incubated enzymolysis 1.5 hours, and after enzymolysis, solution spray drying obtains enzymatic soybean protein isolate.
Lactobacillus plantarum is
preservation is compilednumber CGMCC NO.9405 bacterial classification.
The preparation method of described 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 pulverizing is added in stainless steel cylinder, add the water of fructification 3 times of weight, soak 5 hours, then this Morchella esculenta (L.) Pers sporophore liquid is passed through colloid mill, colloid mill operation condition is: the gap of adjustment colloid mill stator and rotor is 0.6 ± 1 micron, and colloid mill flow is 0.5 ton/hour;
(3) intensification enzymolysis: the Morchella esculenta (L.) Pers sporophore liquid mixed liquor through milling treatment of colloid is transferred in stainless steel enzymatic vessel and is heated to 50 DEG C, pH is to 6.0 in adjustment, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.05%, the acid protease of 0.1%, insulation enzymolysis 1.5 hours, constantly stirs in enzymolysis process.
(4) dry: after the mash filtrations after enzymolysis, spraying dry obtains hickory chick enzymolysis powder.
The preparation process of above-mentioned Lactobacillus plantarum capsule product is as follows:
1) core is prepared: in the Lactobacillus plantarum zymotic fluid cultivated through ferment tank, add the stachyose of zymotic fluid quality 3% and the hickory chick enzymolysis powder of 8%, then mix with frozen-dried protective agent solution, pre-freeze 0.5h at-50 DEG C, then freeze-drying 10h in vacuum freeze drier, grinds after freeze-drying and makes core; 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, maltodextrin solution; The volume ratio of three kinds of solution is skimmed milk power: trehalose: maltodextrin=3:1:0.5;
The concentration of described skimmed milk power, trehalose, maltodextrin solution is respectively: skimmed milk power 5%, trehalose 1%, maltodextrin 2%.
2) bag quilt: core is suspended in fluid bed, wall material spraying bag quilt, the mixed liquor of shitosan, glycerine and trehalose is sprayed into from a shower nozzle, the mixed liquor of xanthans, carragheen and enzymatic soybean protein isolate is sprayed into from another shower nozzle, spray velocity controls identical, bag at 28 DEG C, is namely formed capsule after 30 minutes by temperature in fluid bed in process.Adopt a kind of ultrasonic atomization drying device preparation as described in patent 201120503311.1.
Lactobacillus plantarum capsule product bile tolerance is tested.
Capsule product in embodiment 2-3 be placed in pig cholate solution (solution concentration 3.0%) insulation of 37 DEG C and constantly stir, take out after 2h, with sterile saline washing, with opening one's purse, liquid dissolves, measure Lactobacillus Survival, and compare with the bacterium liquid do not embedded.Test result as
following table instituteshow.
| Metamorphosis | Survival rate | |
| Embodiment 1 | Not disintegration | 87.5％ |
| Embodiment 2 | Not disintegration | 89.3％ |
| Do not wrap and contrasted | -- | 0.65％ |
Embodiment 3
A kind of feed biological complex enzyme of technical problem solved by the invention, is made up of the raw material of following parts by weight:
Zytase 15, cellulase 10, amylase 4, acid protease 4, pectase 1, mannase 2, Lactobacillus plantarum capsule 3, galenical 2 parts, Ginger P.E 1 part, 1.5 parts, Radix Glycyrrhizae.
The preparation method of described galenical is as follows:
Take cassia seed 15 parts; The bark of eucommia 10 parts; 12 parts, Poria cocos; Respectively said herbal medicine being crushed to particle diameter is less than 2 millimeters, then Homogeneous phase mixing add the water of 5 times of weight in container, add the acidic cellulase of said mixture constituent mass 2%, control temperature 50 DEG C, pH:3.8, keeps 1h, adds the mixture of mixed material 3 times of w ethanol and propyl alcohol subsequently, control temperature to 70 DEG C keeps 3h, filters; Filter vacuum concentrates postlyophilization and obtains galenical.
The mass ratio of described ethanol and propyl alcohol is 1:1; Concentration of alcohol is 95%; Propanol concentration is 100%;
The preparation method of described Ginger P.E is as follows:
To be crushed to particle diameter is that less than 2 millimeters gingers are placed in container, adds the water of 4 times of weight, and control temperature 55 DEG C keeps 2h, adds the mixture of mixed material 3 times of w ethanol and methyl alcohol, and control temperature to 35 DEG C keeps 5h, filters; Filter vacuum concentrates postlyophilization and obtains Ginger P.E.
The mass ratio of described ethanol and methyl alcohol is 1:2, concentration of alcohol 90%.Methanol concentration 100%.
Embodiment 4 is basic with routine 1-4
A kind of feed biological complex enzyme of technical problem solved by the invention, is made up of the raw material of following parts by weight:
Zytase 10, cellulase 12, amylase 5, acid protease 2, pectase 2, mannase 1, Lactobacillus plantarum capsule 5, galenical 3 parts, Ginger P.E 2 parts, 2 parts, Radix Glycyrrhizae.
Described acidic xylanase, phytase, cellulase, amylase, acid protease are food-grade enzyme preparation;
Embodiment 6
A kind of feed biological complex enzyme of technical problem solved by the invention, is made up of the raw material of following parts by weight:
Zytase 20, cellulase 6, amylase 5, acid protease 2, pectase 1, mannase 2, Lactobacillus plantarum capsule 3, galenical 3 parts, Ginger P.E 2 parts, 1 part, Radix Glycyrrhizae.
The preparation method of feed biological complex enzyme of the present invention is as follows:
By described Radix Glycyrrhizae, galenical and Ginger P.E ultramicro grinding respectively, then with zytase, cellulase, amylase, acid protease, pectase, mannase, packs the feed biological complex enzyme that gets product after Lactobacillus plantarum capsule Homogeneous phase mixing.
The result of use test of embodiment 5 embodiment of the present invention 1 feed biological complex enzyme in weanling pig
Test method:
The healthy weanling pig 120 of subjects to be a certain large-scale regular plant in Hunan average weight be (7.16 ± 0.15) kg, statistical analysis weight differences is not remarkable.Adopt single-factor Randomized Designs, by 120 healthy weanling pigs, by male and female half and half, be divided into 2 groups (control group and test group), often organize 6 repetitions, each repetition 10 pigs.The product 100g of the present invention that in test group feed, interpolation embodiment 1 per ton is obtained, control group does not add product of the present invention.Feed continuously 30 days, growth indexes
as table 1
table 1
| Project | Control group | Test group | Deviation |
| Starting weight (kg) | 7.09 | 7.23 | -- |
| End heavy (kg) | 18.34 | 21.58 | -- |
| Daily gain (g/d ﹒ head) | 375 | 478 | 27.47％ |
| Daily ingestion amount (g/d ﹒ head) | 607.5 | 598.5 | -1.48％ |
| Feedstuff-meat ratio | 1.62 | 1.25 | -22.84％ |
| Diarrhea rate (%) | 14 | 2 | -92.86％ |
Result of the test shows, in weanling pig daily ration, add this product, and the daily gain of piglet improves 27.47% than control group; Feedstuff-meat ratio reduces 22.84%; Diarrhea rate reduces 92.86%, and product of the present invention can improve piglet body immunity, increases cultivation quality and benefits.
This bacterial classification
preservationunit: Chinese microorganism strain
preservationmanagement
the committeecommon micro-organisms center
preservationunit address: in address
state northno. 3, No. 1, North Star West Road, Jing Shi Chaoyang District institute, postcode: 100101
preservationdate: on July 2nd, 2014
preservation is compilednumber: CGMCC NO.9405
Classification And Nomenclature: Lactobacillus plantarum (Lactobacillus plantarum) tlj-2014.
Claims (8)
1. a feed biological complex enzyme, be made up of the raw material of following parts by weight: zytase 10-20, cellulase 6-12, amylase 3-5, acid protease 2-5, pectase 1-2, mannase 1-2, Lactobacillus plantarum capsule 2-5, galenical 1-3 part, Ginger P.E 1-2 part, Radix Glycyrrhizae 1-2 part.
2. feed biological complex enzyme according to claim 1, the preparation method of described galenical is as follows:
Take cassia seed 10-18 part; Bark of eucommia 5-15 part; Poria cocos 10-15 part; Respectively said herbal medicine being crushed to particle diameter is less than 2 millimeters, then Homogeneous phase mixing add the water of 3-6 times of weight in container, add the acidic cellulase of said mixture constituent mass 1-2%, control temperature 45-55 DEG C, pH:3.5-4, keeps 1-2h, adds the mixture of mixed material 2-3 times of w ethanol and propyl alcohol subsequently, control temperature to 60 DEG C-78 DEG C of maintenance 3-4h, filter; Filter vacuum concentrates postlyophilization and obtains galenical.
3. feed biological complex enzyme according to claim 1, the preparation method of described Ginger P.E is as follows:
To be crushed to particle diameter is that less than 2 millimeters gingers are placed in container, adds the water of 3-6 times of weight, and control temperature 50 DEG C-60 DEG C keeps 2-3h, adds the mixture of mixed material 2-3 times of w ethanol and methyl alcohol, and control temperature to 30 DEG C-40 DEG C of maintenance 3-8h, filter; Filter vacuum concentrates postlyophilization and obtains Ginger P.E.
4., according to feed biological complex enzyme described in claim 1, the preparation method of described Lactobacillus plantarum capsule is as follows:
Lactobacillus plantarum capsule product is made up of wall material, Lactobacillus plantarum, freeze drying protectant, stachyose, hickory chick enzymolysis powder.
Described wall material is made up of enzymatic soybean protein isolate, shitosan, xanthans, carragheen, glycerine and trehalose; Concentration when above-mentioned each material is made into mixed solution is respectively: enzymatic soybean protein isolate 7-10%, shitosan 1-2%, xanthans 1-3%, carragheen 0.1-0.5%, glycerine 0.3-1%, trehalose 0.5-2%; The preparation method of enzymatic soybean protein isolate is: compound concentration is that 10-13% soybean protein isolate solution is heated to 30-45 DEG C, adjustment pH to 3-5, add the acidity acid acid protease insulation enzymolysis 0.5-1.5 hour of soybean protein isolate weight 0.1-1%, after enzymolysis, solution spray drying obtains enzymatic soybean protein isolate.
5. according to feed biological complex enzyme described in claim 4, the preferred CGMCC NO.9405 of Lactobacillus plantarum.
6., according to feed biological complex enzyme described in claim 1-4, preparation method is as follows for Lactobacillus plantarum capsule product:
1) core is prepared: in the Lactobacillus plantarum zymotic fluid cultivated through ferment tank, add the stachyose of zymotic fluid quality 3-5% and the hickory chick enzymolysis powder of 5-8%, then mix with frozen-dried protective agent solution, pre-freeze 0.5h at-50 DEG C, then freeze-drying 10-18h in vacuum freeze drier, grinds after freeze-drying and makes core;
The ratio of described zymotic fluid and frozen-dried protective agent solution is 1:1.4-0.8; Described frozen-dried protective agent solution is made up of skimmed milk power, trehalose, maltodextrin solution; The volume ratio of three kinds of solution is skimmed milk power: trehalose: maltodextrin=3:1:0.5;
The concentration of described skimmed milk power, trehalose, maltodextrin solution is respectively: skimmed milk power 5%, trehalose 1%, maltodextrin 2%.
2) bag quilt: core is suspended in fluid bed, wall material spraying bag quilt, the mixed liquor of shitosan, glycerine and trehalose is sprayed into from a shower nozzle, the mixed liquor of xanthans, carragheen and enzymatic soybean protein isolate is sprayed into from another shower nozzle, spray velocity controls identical, bag between 25-38 DEG C, is namely formed capsule after 30 minutes by temperature in fluid bed in process.
7., according to feed biological complex enzyme described in claim 1, be made up of the raw material of following parts by weight:
Zytase 15, cellulase 10, amylase 4, acid protease 4, pectase 1, mannase 2, Lactobacillus plantarum capsule 3, galenical 2 parts, Ginger P.E 1 part, 1.5 parts, Radix Glycyrrhizae;
Be made up of the raw material of following parts by weight:
Zytase 10, cellulase 12, amylase 5, acid protease 2, pectase 2, mannase 1, Lactobacillus plantarum capsule 5, galenical 3 parts, Ginger P.E 2 parts, 2 parts, Radix Glycyrrhizae;
Zytase 20, cellulase 6, amylase 5, acid protease 2, pectase 1, mannase 2, Lactobacillus plantarum capsule 3, galenical 3 parts, Ginger P.E 2 parts, 1 part, Radix Glycyrrhizae.
8. according to the preparation method of feed biological complex enzyme described in claim 1, as follows:
By described Radix Glycyrrhizae, galenical and Ginger P.E ultramicro grinding respectively, then with zytase, cellulase, amylase, acid protease, pectase, mannase, packs the feed biological complex enzyme that gets product after Lactobacillus plantarum capsule Homogeneous phase mixing.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106615691A (en) * | 2016-10-11 | 2017-05-10 | 上海高龙生物科技有限公司 | Anti-infection enzyme complexing agent and application thereof in feed |
| CN114601027A (en) * | 2022-04-26 | 2022-06-10 | 驻马店华中正大有限公司 | Acid, essential oil, enzyme and probiotic compound preparation and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103805582A (en) * | 2013-12-27 | 2014-05-21 | 湖南鸿鹰生物科技有限公司 | Beta-glucanase containing special enzyme for growing pigs and preparation method thereof |
| CN104605168A (en) * | 2014-12-04 | 2015-05-13 | 邵素英 | Biological composite enzyme for forages and preparation method thereof |
-
2015
- 2015-01-06 CN CN201510004750.0A patent/CN104824379A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103805582A (en) * | 2013-12-27 | 2014-05-21 | 湖南鸿鹰生物科技有限公司 | Beta-glucanase containing special enzyme for growing pigs and preparation method thereof |
| CN104605168A (en) * | 2014-12-04 | 2015-05-13 | 邵素英 | Biological composite enzyme for forages and preparation method thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106615691A (en) * | 2016-10-11 | 2017-05-10 | 上海高龙生物科技有限公司 | Anti-infection enzyme complexing agent and application thereof in feed |
| CN106615691B (en) * | 2016-10-11 | 2019-07-02 | 上海国龙生物技术集团有限公司 | A kind of anti-infective Enzyme reactors and its application in feed |
| CN114601027A (en) * | 2022-04-26 | 2022-06-10 | 驻马店华中正大有限公司 | Acid, essential oil, enzyme and probiotic compound preparation and preparation method thereof |
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