CN104686813B - Feed probiotic microcapsule and its application - Google Patents
Feed probiotic microcapsule and its application Download PDFInfo
- Publication number
- CN104686813B CN104686813B CN201410674215.1A CN201410674215A CN104686813B CN 104686813 B CN104686813 B CN 104686813B CN 201410674215 A CN201410674215 A CN 201410674215A CN 104686813 B CN104686813 B CN 104686813B
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- China
- Prior art keywords
- enzymolysis
- hickory chick
- protein isolate
- wall material
- probiotic microcapsule
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K10/14—Pretreatment of feeding-stuffs with enzymes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
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Abstract
The invention provides a kind of probiotic microcapsule and its application.The probiotic microcapsule is made up of wall material, Lactobacillus plantarum, stachyose, hickory chick enzymolysis powder.It is as follows that coated method is carried out to probiotic microcapsule product：The stachyose that zymotic fluid quality 3 5% is added in probiotics fermention liquid after fermented tank fermented and cultured and 5 8% hickory chick are digested into powder, then zymotic fluid mixes with wall material solution, obtains mixed solution；The method being spray-dried using ultrasonic vacuum, one-level vacuum drying, two level inert protective gas add vibrations fluidized bed drying, and microcapsules coating is carried out to probiotics.Capsule of the present invention uses modified soybean protein isolate, has good enteric solubility, can be completely disintegrated in 1 1.5 hours after capsule arrival enteron aisle and discharge probiotics, and propagation turns into dominant microflora rapidly, suppresses the effect such as growth of pathogenic bacteria so as to reach.The present invention is made product have the nutritive peculiarity of hickory chick concurrently, has good palatability, improved animal feeding results using hickory chick enzymolysis powder.
Description
Technical field
The present invention relates to a kind of probiotics capsule product and its application, belong to additive for microbe feedstuff technical field.
Background technology
Lactic acid bacteria class microbial forage additive be antibacterials substitute in important one kind.Lactic acid bacteria be enteron aisle just
One of outstanding representative of beneficial bacterium, has important physiology and healthcare function in normal flora：It can be adjusted with antagonism pathogenic microorganism
Save animal gut microflora balance：Activated immune system, strengthen immunity, prevent the hair of a variety of diseases and adverse reaction
It is raw；Suppress the generation of tumour, protect animal health；Nutriment is synthesized, produces digestive enzymes, improves the work of animal digestion enzyme
Property, improve the metabolism of vitamin, neutralize enterotoxin, reduce generations of harmful substance such as amine, ammonia etc..However, lactic acid bacteria is growing
During do not formed bud embrace, resistance is poor, to external environment, such as oxygen, moisture, high temperature, mechanical presses, heat shock and hydrochloric acid in gastric juice all
It is very sensitive, it is difficult to ensure that effective viable bacteria amount of survival in the application of reality.Therefore, excellent lactic acid bacteria how is filtered out
Plant and extend its Commercial active, be always the Research Emphasis of lactic acid bacteria production firm all over the world.
Microcapsules technology is one of method for protecting thalline vigor maximally efficient and practical.Lactic acid bacteria is subjected to microcapsules
Change, thalline and extraneous poor environment can be separated, from trace element infringement in feed, slow down in pelletization temperature with
The influence of pressure；Solid particle is formed, beneficial to being uniformly distributed in premix, is also beneficial to store and transports：Using enteric
Property wall material after, moreover it is possible to ensure that thalline as much as possible reaches enteron aisle, the effect of really playing health care and treatment.
Such as：One kind is disclosed in patent CN1613455 and uses the coated probiotic microcapsule of three-layer protection layer；Patent
One kind is disclosed in CN1569043 and uses sodium alginate, calcium chloride as lactic acid bacteria made of wall material progress bed spray coating
Microcapsules.Above-mentioned patent improves the survival rate of probiotics or lactic acid bacteria to a certain extent, but inevitably,
Hydrogenated oil and fat temperature can cause to damage more than 55 DEG C to the lactic acid bacteria inside core in CN1613455, while the controlled release bag of outer layer
Clothing material, lactic acid bacteria is rapidly discharged in enteron aisle, a part of lactic acid bacteria can be caused to be excreted, reduce lactic acid bacteria
Utilization rate.Using sodium alginate as wall material in CN1569043, current microcapsules are commonly used in being coated with, but marine alga
Moisture holding capacity of sour sodium itself is poor, is easy to dehydration hardening rupture by the coated microcapsules gel of sodium alginate, in addition sodium alginate bag
The microcapsules of quilt not stomach juice-resistant, makes microcapsules cross stomach ability.A kind of entitled lactic acid bacteria containing Multi-layer microcapsule lactic acid bacteria
The preparation method of powder, number of patent application are：201310743621.4 invention belongs to nutritious health caring food technology field, especially relate to
A kind of and preparation method of the lactic acid bacteria powder containing collagen peptide and Multi-layer microcapsule lactic acid bacteria, it is characterised in that：By ocean fish-skin
20 parts~30 parts of the oligomeric Gly-His-Lys of collagen, 40 parts~50 parts with Multi-layer microcapsule lactic acid bacteria powder of 5 parts~20 parts of citric acid mix.
The invention has the advantages that probiotics can be made to be colonized growth and breeding in human intestines and stomach, it can improve gastroenteric environment, can kill
A variety of harmful levels of pathogens and harmful microorganism, it can produce various digestive ferments in human body, can help digest absorption, strong body
Body.A kind of lactobacillus micro-capsule and its production and use, application number：200810135259.1 the invention also discloses that one
Kind lactobacillus micro-capsule and its production and use.The lactobacillus micro-capsule is by outer layer wall material, freeze drying protectant and lactic acid
Bacterium forms.Purposes present invention also offers the preparation method of the lactobacillus micro-capsule and its as feed addictive.The present invention
Lactobacillus micro-capsule can effectively protect lactic acid bacteria in core, extend the time-to-live of lactic acid bacteria at ambient temperature, improve
Tolerance of the lactic acid bacteria to metal ion in feed；In addition, the lactobacillus micro-capsule stomach juice-resistant is good, can be collapsed rapidly in enteron aisle
Solution, discharges lactic acid bacteria, so as to really improve the utilization rate of lactic acid bacteria, plays balance intestinal micro-ecological environment and suppresses pathogen life
It is long, ensure the intestinal health of animal, reduce the effect such as incidence of disease of animal intestinal tract.
Great Bei agricultures company applies entitled《The production technology and its microcapsules of feeding lactobacillus microcapsules are with application and in advance
Batch mixing》Invention, application number：201110415081.8 a kind of production technology of feeding lactobacillus microcapsules of disclosure of the invention and its
Microcapsules and application and premix.The present invention by using can quickly be formed at low temperature vitrifying guard mode trehalose,
Lactose etc. and sodium glutamate, manganese sulfate, glycerine and polyvinylpyrrolidone are as main wall material composition, using special excusing from death ripple
Spray drying device, it is atomized using ultrasonic nozzle, is done at normal temperatures using vacuum and protective gas nitrogen etc.
It is dry, influence of the high temperature of the lyophilized low temperature of low temperature and high temperature spray-drying to lactic bacteria activity is avoided, bacterium survival rate can reach
To more than 90%.Technique realizes lactic acid bacteria and is dried at normal temperatures, and preparation technology is easy to control, the storage of obtained product
Phase is grown, and can significantly extend the storage time of product.It is directed to one kind and is related to ultrasonic atomization drying equipment in patent
Disclosed in 2011205033111.
In summary, existing microcapsules coating technique or lactic acid bacteria can be produced during its implementation larger broken
Bad effect, or manufactured microcapsules are poor to the tolerance of stomach environment, and these are all unfavorable for the industrialization of microcapsule product
And application.
The content of the invention
It is an object of the invention to provide a kind of probiotic microcapsule product, by wall material, Lactobacillus plantarum, stachyose, sheep tripe
Bacterium enzymolysis powder composition.
It is as follows that coated method is carried out to probiotic microcapsule product：By the probiotics fermention after fermented tank fermented and cultured
Zymotic fluid quality 3-5% stachyose and 5-8% hickory chick enzymolysis powder are added in liquid, then zymotic fluid mixes with wall material solution
Close, obtain mixed solution；The method being spray-dried using ultrasonic vacuum, one-level vacuum drying, two level inert protective gas add shake
Fluidized bed drying, microcapsules coating is carried out to probiotics.
The composition of the wall material solution is as follows：Enzymatic hydrolysis of soybean protein isolate 4-10%, chitosan 0.5-1%, Propiram are more
Sugared 0.2-0.5%, xanthans 0.2-1%, carragheen 0.1-0.5%, glycerine 0.5-2%, trehalose 0.2-0.5%, surplus are
Distilled water；Above-mentioned is percent weight in volume.
Various components are independently weighed by above-mentioned composition, is mixed in distilled water and dissolves by heating, pH is naturally, be completely dissolved rear cold
But to room temperature, the wall material solution is produced.
The preparation method of the enzymatic hydrolysis of soybean protein isolate is as follows：Compound concentration is that 10-13% soybean protein isolate is molten
Liquid, 30-45 DEG C is heated to, adjusts pH to 3-5, add soybean protein isolate weight 0.1-1% acid protease, insulation enzymolysis
0.5-1.5 hours, after enzymolysis solution spray drying obtain enzymatic hydrolysis of soybean protein isolate.
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：Morchella esculenta (L.) Pers sporophore after crushing is added in stainless steel cylinder, adds 3-6 times of fructification quality
Water, soak 3-5 hours, be then by colloid mill, colloid mill operation condition by this Morchella esculenta (L.) Pers sporophore liquid：Adjust colloid mill
The gap of stator 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 Jing Guo milling treatment of colloid is transferred in stainless steel enzymatic vessel and heated
To 50-60 DEG C, pH to 4.5-6.0 is adjusted, adds Morchella esculenta (L.) Pers sporophore weight 0.05-0.1% cellulase, 0.01-0.1%
1,4 beta-glucanase, 0.01-0.1% protease, insulation enzymolysis 0.5-1.5 hours, be stirred continuously in enzymolysis process；
(4) dry：Dried after mash filtrations after enzymolysis and obtain hickory chick enzymolysis powder.
The Lactobacillus plantarum preferred strain tlj-2014, the bacterial strain are preserved in China Microbiological on July 2nd, 2014
(abbreviation CGMCC, address are culture presevation administration committee common micro-organisms center：City of BeiJing, China Chaoyang District North Star West Road 1
Institute 3, postcode：100101), deposit number is CGMCC NO.9405, and Classification And Nomenclature is Lactobacillus plantarum (Lactobacillus
plantarum)。
Lactobacillus plantarum tlj-2014 bacterial strain features are as follows in the present invention：To observe under the microscope, the bacterial strain is rod-short,
Gram's staining is positive, and atrichia, does not produce gemma；On solid medium, the bacterium bacterium colony is white, and surface is smooth, fine and close,
Form is circle, and edge is more neat.
Physicochemical characteristicses are：Catalase (-), gelatin liquefaction (-), indoles experiment (+), motility (-), fermentation gas
(-), nitrate reductase (-), fermentation gas (-), hydrogen sulfide gas (-) is produced, grows (+) in pH4.0MRS culture mediums.
Lactobacillus plantarum tlj-2014 of the present invention carries out seed selection using following flows：
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.Starting strain is the blue or green storage feeding that Li Zheng is collected in Yanchi county Ningxia Fattening Sheep field
Material, 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, can be resistant to 1% cholate.
Another object of the present invention is to provide application of the capsule as feed addictive.The capsule of the present invention adds as feed
Agent is added to can be widely used in animal feed, but present invention is preferably used in the feed of poultry.Capsule product is in feed
Addition is 3-5%, or uses 10-20 grams daily per boss fowl.
Beneficial effect：
Probiotics capsule provided by the invention, modified soybean protein isolate, modified soybean separation egg are with the addition of in its wall material
25%, gelling ability is improved than soybean protein isolate emulsifying capacity improve 15-25%, modified soybean protein isolate and xanthan in vain
Glue, carragheen and chitosan are used cooperatively, and gel strength improves more than 10%, enhances the stomach juice-resistant of capsule；This hair
' Yanming ' capsules for clearing uses modified soybean protein isolate, has good enteric solubility, capsule can be complete within 1-1.5 hours after reaching enteron aisle
Full disintegration discharges probiotics, and propagation turns into dominant microflora rapidly, suppresses the effect such as growth of pathogenic bacteria so as to reach.
The present invention digests powder using hickory chick, its nutritional ingredient is effectively discharged into product by enzymolysis processing,
Also cause that the peat-reek of hickory chick is effectively discharged by enzymolysis simultaneously, product is had the nutritive peculiarity of hickory chick concurrently, have
There is good palatability, improve animal feeding results, compared with similar approximate commercially available prod, livestock and poultry can be effectively improved to this product
Preference degree 10-15% (by the comparison of feed intake).
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.
Embodiment 1
Lactobacillus plantarum tlj-2014 of the present invention specific Breeding Process is as follows：
1.DES mutagenic and breedings
1) rings of Lactobacillus plantarum L mono- on test tube slant are taken on super-clean bench, access is equipped with 50mL culture mediums MRS (no fine jades
Fat, glucose 20g/L) 250mL triangular flasks in, 200rpm, 37 DEG C of culture 12h or so, thalline is in logarithmic growth early stage.
2) 5mL bacterium solutions are taken, 5000rpm centrifugations 10min collects thalline, with brine 2 times.
3) 107/mL bacteria suspensions are diluted to pH7.0 phosphate buffers.
4) 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.
5) 150rpm reacts 30min in 37 DEG C of shaking tables, takes 1mL mixed liquors, adds in 0.5mL 25%Na2S2O3 solution
Only react.
6) 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).
7) 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
1) 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.
2) 5mL bacterium solutions 5000rpm centrifugations 10min is taken to collect thalline, with brine 2 times.
3) 107/mL bacteria suspensions are diluted to pH6.0 phosphate buffers.
4) 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.
5) the 200rpm oscillating reactions 30min at 37 DEG C, 5000rpm centrifugation 10min collect thalline, use sterile saline
Wash for several times, stopped reaction.
6) 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).
7) 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
1) 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.
2) 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.
3) 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. fermentation tank is tested
1) 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.
2) 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.
3) 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.
4) 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 probiotic microcapsule product, it is made up of wall material, Lactobacillus plantarum, stachyose, hickory chick enzymolysis powder.
It is as follows that coated method is carried out to probiotic microcapsule product：By the Lactobacillus plantarum after fermented tank fermented and cultured
The stachyose and 8% hickory chick enzymolysis powder of zymotic fluid quality 3% are added in CGMCCNO.9405 zymotic fluids, it is then molten with wall material
Liquid mixes, and obtains mixed solution；The method being spray-dried using ultrasonic vacuum, one-level vacuum drying, two level inert protective gas
Add vibrations fluidized bed drying, microcapsules coating is carried out to probiotics.
The method that the ultrasonic atomization is dried uses method disclosed in application for a patent for invention 201110415081.8.Specifically
To open ultrasonic atomization drying equipment, after house vacuum pressure to be dried maintains 2.5-3.0kPa, adjustment flow velocity is 0-
20mL/min, the temperature in hothouse is at 20-30 DEG C or so；Protective gas carbon dioxide, temperature are filled in secondary drying fluid bed
Maintain 10-20 DEG C.The frequency of described ultrasonic nozzle is 20kHz-30kHz.The mixing of zymotic fluid and wall material solution composition
Suspension is dried into vacuum drying cabinet immediately after ultrasonic nozzle is atomized, drop just shape in a short time
Into capsule, then by the drying time of 3-25 seconds, the bottom of hothouse is fallen on；Subsequently into vibrations fluid bed, that is, carry out two
Level vibrations fluidized bed drying, through 20-40min, collects in discharging opening, that is, obtains probiotics capsule.
The water content of capsule is 8%, bacterium survival rate 93%.
The composition of the wall material solution is as follows：Enzymatic hydrolysis of soybean protein isolate 4%, chitosan 1%, pulullan polysaccharide 0.2%,
Xanthans 1%, carragheen 0.1%, glycerine 0.5%, trehalose 0.5%, surplus are distilled water；Above-mentioned is bulking value percentage
Than.
Various components are independently weighed by above-mentioned composition, is mixed in distilled water and dissolves by heating, pH is naturally, be completely dissolved rear cold
But to room temperature, the wall material solution is produced.
The preparation method of the enzymatic hydrolysis of soybean protein isolate is as follows：Compound concentration is 13% soybean protein isolate solution,
45 DEG C are heated to, adjusts pH to 3, adds the acid protease of soybean protein isolate weight 1%, insulation enzymolysis 0.5 hour, enzymolysis
Solution spray drying afterwards obtains enzymatic hydrolysis of soybean protein isolate.
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：Morchella esculenta (L.) Pers sporophore after crushing is added in stainless steel cylinder, adds 3 times of fructification quality
Water, soak 5 hours, be then by colloid mill, colloid mill operation condition by this Morchella esculenta (L.) Pers sporophore liquid：Adjust colloid mill stator
Gap with rotor is 0.5-1 microns, and colloid mill flow is 0.4 ton hour；
(3) heating enzymolysis：Morchella esculenta (L.) Pers sporophore liquid Jing Guo milling treatment of colloid is transferred in stainless steel enzymatic vessel and heated
To 50 DEG C, adjust pH to 4.5, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.05%, 0.01% 1,4 beta-glucanase,
0.01% protease, insulation enzymolysis 1.5 hours, is stirred continuously in enzymolysis process；
(4) dry：Dried after mash filtrations after enzymolysis and obtain hickory chick enzymolysis powder.
Embodiment 3
A kind of probiotic microcapsule product, it is made up of wall material, Lactobacillus plantarum, stachyose, hickory chick enzymolysis powder.
It is as follows that coated method is carried out to probiotic microcapsule product：By the Lactobacillus plantarum after fermented tank fermented and cultured
The stachyose and 5% hickory chick enzymolysis powder of zymotic fluid quality 5% are added in CGMCCNO.9405 zymotic fluids, it is then molten with wall material
Liquid mixes, and obtains mixed solution；The method being spray-dried using ultrasonic vacuum, one-level vacuum drying, two level inert protective gas
Add vibrations fluidized bed drying, microcapsules coating is carried out to probiotics.
The composition of the wall material solution is as follows：Enzymatic hydrolysis of soybean protein isolate 10%, chitosan 0.5%, pulullan polysaccharide
0.5%th, xanthans 0.2%, carragheen 0.5%, glycerine 2%, trehalose 0.2%, surplus are distilled water；Above-mentioned is weighing body
Product percentage.
Various components are independently weighed by above-mentioned composition, is mixed in distilled water and dissolves by heating, pH is naturally, be completely dissolved rear cold
But to room temperature, the wall material solution is produced.
The preparation method of the enzymatic hydrolysis of soybean protein isolate is as follows：Compound concentration is 10% soybean protein isolate solution,
35 DEG C are heated to, adjusts pH to 5, adds the acid protease of soybean protein isolate weight 0.1%, insulation enzymolysis 1.5 hours, enzyme
Solution spray drying obtains enzymatic hydrolysis of soybean protein isolate after solution.
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：Morchella esculenta (L.) Pers sporophore after crushing is added in stainless steel cylinder, adds 6 times of fructification quality
Water, soak 3 hours, be then by colloid mill, colloid mill operation condition by this Morchella esculenta (L.) Pers sporophore liquid：Adjust colloid mill stator
Gap with rotor is 0.5-1 microns, and colloid mill flow is 1 ton hour；
(3) heating enzymolysis：Morchella esculenta (L.) Pers sporophore liquid Jing Guo milling treatment of colloid is transferred in stainless steel enzymatic vessel and heated
To 60 DEG C, adjust pH to 6.0, add the cellulase of Morchella esculenta (L.) Pers sporophore weight 0.05%, 0.1% 1,4 beta-glucanase,
0.1% protease, insulation enzymolysis 0.5 hour, is stirred continuously in enzymolysis process；
(4) dry：Dried after mash filtrations after enzymolysis and obtain hickory chick enzymolysis powder.
Embodiment 4
Probiotics capsule product simulated gastric fluid experiment measure.
Capsule product in embodiment 1-2 is placed in 37 DEG C of simulated gastric fluid and is incubated and is stirred continuously, is taken out after 2h, is used
Sterile saline washs, and is dissolved with solution cyst fluid, determines bacterium survival rate, and compared with the bacterium solution with not embedding.Result of the test
It is shown in Table 1.
The capsule of table 1 is disintegrated and bacterium survival assays result
| Product | Metamorphosis | Survival rate |
| Embodiment 1 | It is not disintegrated | 91.5% |
| Embodiment 2 | It is not disintegrated | 93.3% |
| Non- peridium pair is shone | 0.68% |
Embodiment 5
Feeding effect experiment of the capsule product of the embodiment of the present invention 1 as feed addictive in milking sow.
The feeding experiment of 21 days by a definite date has been carried out on Zhongweiof Ningxia, northwest China selection pig farm.Experiment uses single factor test contrast design, with
Machine chooses 50 milking sows healthy, farrowing head number and birth counterpoise are similar, parity is 2 or 3 tires, is randomly divided into 2
Group (i.e. test group and control group), every group of 15 repetitions.Wherein：This product that the addition of test group daily ration is prepared into by embodiment 1,
Control group is addition market like product.The results such as weaned piglet weight, diarrhea rate, the death rate are recorded, are specifically shown in Table 2.
The milking sow feeding experiment result of table 2
| Project | Test group | Control group | Remarks |
| Number born alive (head number) | 10.9 | 11.0 | |
| Birth counterpoise (kg) | 1.46 | 1.48 | |
| 21 days small weaning pigs (head) | 10 | 8.30 | |
| 21 days weight of weaning litters (kg) | 66.4 | 44.4 | |
| 21 days wean counterpoises (kg) | 6.64 | 5.35 | |
| Head net gain (kg) | 5.18 | 3.87 | |
| Grice diarrhoea rate (%) | 6.0 | 11.4 | |
| The death rate (%) | 1.8 | 4.1 |
Result of the test shows：Product of the present invention can improve sow and piglet body immunity, reduce antibiotic dosage, increase
Add cultivation quality and benefits.
Claims (4)
1. a kind of probiotic microcapsule, it is made up of wall material, Lactobacillus plantarum, stachyose, hickory chick enzymolysis powder, the plant breast bar
Bacterium deposit number is CGMCC NO.9405；It is as follows that coated method is carried out to the probiotic microcapsule：Fermented tank is fermented
Zymotic fluid quality 3-5% stachyose and 5-8% hickory chick enzymolysis powder are added in Lactobacillus plantarum zymotic fluid after culture, so
After fermentation liquid mixes with wall material solution, obtains mixed solution, the method being spray-dried using ultrasonic vacuum, one-level vacuum drying,
Two level inert protective gas adds vibrations fluidized bed drying, and microcapsules coating is carried out to probiotics；The composition of the wall material solution is such as
Under：In terms of percent weight in volume, enzymatic hydrolysis of soybean protein isolate 4-10%, chitosan 0.5-1%, pulullan polysaccharide 0.2-
0.5%th, xanthans 0.2-1%, carragheen 0.1-0.5%, glycerine 0.5-2%, trehalose 0.2-0.5%, surplus are distilled water.
A kind of 2. probiotic microcapsule as claimed in claim 1, it is characterised in that the preparation of the enzymatic hydrolysis of soybean protein isolate
Method is as follows：Compound concentration is 10-13% soybean protein isolate solution, is heated to 30-45 DEG C, adjusts pH to 3-5, is added big
Beans protein isolate weight 0.1-1% acid protease, insulation digest 0.5-1.5 hours, and solution is spray-dried acquisition after enzymolysis
Enzymatic hydrolysis of soybean protein isolate.
A kind of 3. probiotic microcapsule as claimed in claim 1, it is characterised in that the preparation method of the hickory chick enzymolysis powder
It is as follows：
(1) Morchella esculenta (L.) Pers sporophore drying and crushing；
(2) water-soluble homogeneous：Morchella esculenta (L.) Pers sporophore after crushing is added in stainless steel cylinder, adds the water of 3-6 times of fructification quality,
3-5 hours are soaked, are then by colloid mill, colloid mill operation condition by this Morchella esculenta (L.) Pers sporophore liquid：Adjust colloid mill stator
Gap with 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 Jing Guo 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, addition Morchella esculenta (L.) Pers sporophore weight 0.05-0.1% cellulase, 0.01-0.1% β-
The protease of dextranase, 0.01-0.1%, insulation digest 0.5-1.5 hours, are stirred continuously in enzymolysis process；
(4) dry：Dried after mash filtrations after enzymolysis and obtain hickory chick enzymolysis powder.
4. application of any described probiotic microcapsules of claim 1-3 in feed addictive.
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| CN201711396070.3A CN107927326A (en) | 2014-11-21 | 2014-11-21 | A kind of feed probiotic microcapsule and its application |
| CN201711397186.9A CN107981030A (en) | 2014-11-21 | 2014-11-21 | A kind of feed probiotic microcapsule and its application |
| CN201410674215.1A CN104686813B (en) | 2014-11-21 | 2014-11-21 | Feed probiotic microcapsule and its application |
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| CN201711396070.3A Division CN107927326A (en) | 2014-11-21 | 2014-11-21 | A kind of feed probiotic microcapsule and its application |
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| CN105248842A (en) * | 2015-10-13 | 2016-01-20 | 武汉农信丰生物科技有限公司 | Preparation method for livestock and poultry feed additive |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1357407A (en) * | 2001-10-24 | 2002-07-10 | 中国科学院新疆化学研究所 | Conjugate linoleic acid or conjugate linoleate microcapsule |
| CN101496555A (en) * | 2008-01-31 | 2009-08-05 | 北京大北农科技集团股份有限公司 | Lactobacillus micro-capsule as well as preparation method and use |
| CN102511661A (en) * | 2011-12-28 | 2012-06-27 | 北京好友巡天生物技术有限责任公司 | Beneficial fungus feed additive for large-scale composite edible fungi and method for breeding flavored pigs |
| CN103156063A (en) * | 2011-12-13 | 2013-06-19 | 北京大北农科技集团股份有限公司 | Production process of feeding lactobacillus microcapsule and microcapsule, application and premix thereof |
| CN103859237A (en) * | 2014-04-02 | 2014-06-18 | 金陵科技学院 | Compound aquatic microcapsule initial baits, preparation method and application of initial baits |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102657287B (en) * | 2012-06-05 | 2013-05-08 | 荆州双胞胎饲料有限公司 | Compound feed addictive with bacteria and oxidation resisting functions |
-
2014
- 2014-11-21 CN CN201410674215.1A patent/CN104686813B/en active Active
- 2014-11-21 CN CN201711396070.3A patent/CN107927326A/en not_active Withdrawn
- 2014-11-21 CN CN201711397186.9A patent/CN107981030A/en not_active Withdrawn
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1357407A (en) * | 2001-10-24 | 2002-07-10 | 中国科学院新疆化学研究所 | Conjugate linoleic acid or conjugate linoleate microcapsule |
| CN101496555A (en) * | 2008-01-31 | 2009-08-05 | 北京大北农科技集团股份有限公司 | Lactobacillus micro-capsule as well as preparation method and use |
| CN103156063A (en) * | 2011-12-13 | 2013-06-19 | 北京大北农科技集团股份有限公司 | Production process of feeding lactobacillus microcapsule and microcapsule, application and premix thereof |
| CN102511661A (en) * | 2011-12-28 | 2012-06-27 | 北京好友巡天生物技术有限责任公司 | Beneficial fungus feed additive for large-scale composite edible fungi and method for breeding flavored pigs |
| CN103859237A (en) * | 2014-04-02 | 2014-06-18 | 金陵科技学院 | Compound aquatic microcapsule initial baits, preparation method and application of initial baits |
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| CN107927326A (en) | 2018-04-20 |
| CN107981030A (en) | 2018-05-04 |
| CN104686813A (en) | 2015-06-10 |
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