CN104642746A - Preparation method of feeding lactic acid bacteria micro pills - Google Patents
Preparation method of feeding lactic acid bacteria micro pills Download PDFInfo
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Abstract
The invention provides a preparation method of feeding lactic acid bacteria micro pills. The preparation method comprises the following steps: culturing lactic acid bacteria until the metaphase of a logarithmic growth phase appears; adding sodium chloride into a culture solution to perform continuous culture until the prophase of a stable growth phase appears; centrifuging; cleaning bacterial sludge by using normal saline, and collecting the bacterial sludge; re-suspending the bacterial sludge in a fresh MRS culture medium, slowly heating to 45-55 DEG C, keeping the temperature for 30 minutes, then quickly cooling to 0-4 DEG C, taking out the bacterial sludge, and uniformly mixing the bacterial sludge with an appropriate amount of a filling agent, sodium alginate, lactalbumin, xylooligosaccharide, fructo-oligosaccharide, resistant starch and an antioxidant to obtain a mixture; and then putting the mixture into an extrusion pill-rolling machine for pelletizing, and drying after pelletizing to obtain the feeding lactic acid bacteria micro pills. The preparation method can be used for effectively improving the stability and tolerance of the lactic acid bacteria in the micro pills, so that the survival rate of the lactic acid bacteria during storage, transportation and processing of the lactic acid bacteria micro pills is increased and the living bacterium rate of a feed is increased; and meanwhile, the preparation method is simple and easy to control, and is suitable for industrial large-scale production.
Description
Technical field
The present invention relates to a kind of lactobacillus preparation technical field, particularly a kind of preparation method of feeding lactobacillus micropill.
Background technology
Lactic acid bacteria is the normal flora in animal intestinal, amphimicrobian lactic acid bacteria can produce the materials such as lactic acid, biacetyl, hydrogen peroxide, bacteriocin, has and suppresses pathogen and spoilage organisms, maintenance intestinal microecology balance, control to suffer from diarrhoea, improve immunity, improve livestock products quality physiological function.Therefore, it has become the active demand of feed industry.Most lactic acid bacteria has good storage stability under cryogenic, but most Feed Enterprise does not have cold chain condition, so lactic acid bacteria usually can mortality in storage and transport process; On the other hand, feed companies is when making particle or puffing material, and all along with heat treatment process, lactic acid bacteria non-refractory, in the process of granulating, loss late is very high.At present, in raising lactic acid bacteria temperature tolerance, the study hotspot of feed industry has been become.Probio is made the feasible way that pulvis microcapsules are regarded as addressing this problem.
Microcapsules to be wrapped in by the molecule of solid, liquid or even gas by macromolecular material that is natural or synthesis in microencapsulation that diameter is the semi permeability of 1-500 μm or sealing cyst membrane, material in microcapsules is due to isolated with external environment, can from the impact of environment, thus keep stable, and under proper condition, encapsulated material can discharge again.Probio is made microcapsules and achieve good effect at protection bioactive molecule, tissue and cell to resist in adverse environment.Adopt microcapsules technology to be embedded by lactic acid bacteria thalline, the resistivity of thalline environmental factor to external world can be strengthened, significantly improve thalline in Cord blood phase and the survival rate after arriving enteron aisle, make lactic acid bacteria play wholesome effect better.But this microcapsules are pulvis shape after making, and there is following problem in pulvis microcapsules, first be that thalline storage-stable is good not, again, to thermo-responsive, and pulvis microcapsules as feed addictive time, feed granulating process can be subject to high temperature, high humidity again, the multiple destruction such as Thief zone, cause the survival rate of lactic acid bacteria in feed extremely low, therefore pulvis probiotic microcapsule is unworkable in feed industry.
Authorization Notice No. is that the Chinese patent of CN102178238B discloses name and is called a kind of heat resistant type microcapsule life tonifying bacterium and its production and use, the heat resistant type microcapsule life tonifying bacterium particle diameter of this disclosure of the invention is between 20 ~ 400 μm, embedding rate >=90%, viable count is 109 ~ 1010cfu/g, instant in cold water, at the temperature of 80 ~ 90 DEG C, through 3 minutes, its viable count only declines 1 ~ 1.5 order of magnitude.This invention also discloses the preparation method of this heat resistant type microcapsule life tonifying bacterium.Although the product heat resistant type microcapsule life tonifying bacterium resistance that this invention provides and stability improve, but heat resistant type microcapsule life tonifying bacterium is equally in pulvis, there is the problem same with the pulvis probiotic microcapsule of foregoing description, in addition, the processing method of this invention is the method for emulsified gel, cultured thalline is needed to join in solution, carry out secondary embedding, and embedding conditions is harsh, need high speed homogenization process, also to experience freeze drying and fluidized bed drying after embedding, last longer, process operation is very loaded down with trivial details, cost is higher.
Publication number is that the China of CN101338288 application discloses that name is called a kind of method improving plant bacterium lacticum stability, this Lactobacillus plantarum is plant lactobacillus LP-Onlly, add NaCl in the fermentation logarithmic phase later stage or carry out high-temperature process or carry out K cryogenic treatment, to improve the stability of Lactobacillus plantarum cell.It is by adopting microbial metabolism engineering strategy, mainly by utilizing infiltration and temperature two kinds of pressure stress treatment technology, and raising plant bacterium lacticum stability energy.This will improve the method that stability provides a tool realistic meaning for lactic acid bacteria, thus to improve product quality, provides normal temperature probiotic products Machinability Evaluation, extends the product shelf life-span.But in actual applications, the method adopts single high salt or single high temperature or low temperature to carry out short time Stress treatment to Lactobacillus plantarum in the logarithmic phase later stage, not obvious to the raising of lactic acid bacteria tolerance, lactic acid bacteria can not resist the various destructions of post-production, makes the survival rate of lactic acid bacteria lower.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of feeding lactobacillus micropill, this preparation method effectively can improve the stability of lactic acid bacteria in lactic acid bacteria micropill, tolerance, thus improve lactic acid bacteria micropill storing, the survival rate of transport and the lactic acid bacteria in process, improve the viable bacteria rate in feed, this preparation method is simple and easy to control simultaneously, is applicable to industrialization large-scale production.
For solving the problems of the technologies described above, the present invention is achieved through the following technical solutions.
A preparation method for feeding lactobacillus micropill, is prepared by following step;
(1) lactic acid bacteria bacterium mud is prepared: when lactic acid bacteria being cultured to exponential phase mid-term, about 10 ~ 16h, adds a certain amount of sodium chloride and make the final concentration of sodium chloride in nutrient solution control at 0.4 ~ 0.7mol/L in nutrient solution; Lactic acid bacteria is continued to be cultured to early stage stable growth phase under this concentration, about 6 ~ 10h, centrifugal 10min under centrifugal acceleration is 4600g condition, with the physiological saline cleaning bacterium mud twice of 0.85 ~ 0.9%, collect bacterium mud, then by resuspended for bacterium mud with fresh MRS medium, slowly be warming up to 45 ~ 55 DEG C, and after keeping 30min at such a temperature, be cooled to rapidly 0 ~ 4 DEG C, for subsequent use;
(2) 280 ~ 400 parts, the bacterium mud of preparation in filler 200 ~ 250 parts, sodium alginate 25 ~ 50 parts, lactalbumin 25 ~ 50 parts, xylo-oligosaccharide 30 ~ 50 parts, FOS 30 ~ 50 parts, resistant starch 30 ~ 50 parts, antioxidant 1 ~ 3 part and step (1) is taken, then load weighted bacterium mud and other compositions are mixed in agitator, make softwood
(3) by softwood obtained for step (2), put into and extrude finisher,pill and granulate, after granulation, be drying to obtain feeding lactobacillus micropill.
Preparation process (1) is cultured to exponential phase lactic acid bacteria and adds sodium chloride mid-term, and now lactic acid bacterium number is more, flushes, and adds sodium chloride, is conducive to the lactic acid bacteria obtaining better resistance; And now add sodium chloride, the enough Stress treatment time can be ensured, bring good effect, sodium chloride addition controls to be as the criterion at 0.4 ~ 0.7 mol/L to make the final concentration of sodium chloride in nutrient solution, through experiment, we find under this concentration range, less for most lactic acid bacteria destruction, its growth rate can not be subject to too large suppression substantially, effectively can improve again the resistance of lactic acid bacteria simultaneously.
Granulation conditions described in step (3) processes 10 ~ 15 minutes under 800 ~ 1000 revs/min.
Drying described in step (3) is by moisture drying to 4% ~ 7% at 40 ~ 48 DEG C.
Described filler is prefabricated gelatinized starch, microcrystalline cellulose, lactose, one or more the mixing in sweet mellow wine.
Described antioxidant is vitamin C, one or both in vitamin E.
Described lactic acid bacteria bacterium mud be selected from Lactobacillus plantarum, Lactobacillus casei, lactobacillus acidophilus, streptococcus thermophilus one or more.
Beneficial effect:
1, compare in prior art, the present invention has the following advantages:
The present invention adopts the high-temperature process lactic acid bacteria of 45 ~ 55 DEG C again after first adopting the high Ficus caricaL lactic acid bacteria of 0.4 ~ 0.7mol/L sodium chloride, this combined treatment has synergy, cross-protection can be induced, improve lactic acid bacteria tolerance and stability, better to the protected effect of lactic acid bacteria in follow-up pelletization, simultaneously the present invention adds appropriate sodium chloride when lactic acid bacteria being cultured to exponential phase mid-term and continuing to cultivate in nutrient solution, achieve the salt stress process of long period in incubation, next step operation is carried out without the need to collected by centrifugation, be conducive to continuous print operation, be applicable to industrialization large-scale production.
The lactic acid bacteria micropill prepared by method of the present invention has closed, structure closely, be conducive to the cryogenic inner environment that maintenance one is stable, thus reduce the destruction of ambient temperature fluctuation to lactobacillus cell, improve the survival rate of lactic acid bacteria in storage, transport and pelletization, especially during feed granulating high temperature, high humidity to the product of the destruction of lactic acid bacteria far below powdery microencapsulation form, in addition, micropill production capacity is large, cost of equipment is lower, and size distribution is good, mobility is high, is easy to large-scale application in feed industry.
The proportioning of the filler of micropill and other composition is very important for the mouldability of micropill, under the effect of adhesive water, if proportioning is low, difference spherical in shape, if proportioning is high, the proportioning of other auxiliary materials just reduces, be unfavorable for keeping feeding lactobacillus micropill in processing, growth promoting function in survival rate in utilization and utilization process, therefore inventor passes through long-term practice, the specific composition composition of feeding lactobacillus micropill of the present invention and the proportioning of each composition are drawn, micropill is made to have good mouldability, the stability of lactic acid bacteria is kept in process, improve the survival rate of lactic acid bacteria in process and utilization process.
The present invention adopts sodium alginate and lactalbumin as protective agent; sodium alginate is a kind of food-grade wall material of enteric solubility; it can make micropill in enteron aisle disintegration release again; avoid lactic acid bacteria disintegration release and the problem of being killed by hydrochloric acid in gastric juice under one's belt; improve the survival rate of lactic acid bacteria in enteron aisle; thus effectively playing the beneficial function of lactic acid bacteria, lactalbumin can cushion the coercion of hydrochloric acid in gastric juice to lactic acid bacteria, with sodium alginate coordinating protection lactic acid bacteria.
The present invention adopts xylo-oligosaccharide, FOS and resistant starch as growth promoter, and they belong to prebiotics, can not be utilized in enteron aisle by harmful bacteria, can promote that lactic acid bacteria breeds on the contrary; In the process of the granulation of feed, they can play synergistic protective effect with sodium alginate and lactalbumin, improve the survival rate of lactic acid bacteria in pelletization; Another itself is exactly a kind of nutriment, to human body and animal all beneficial.
2, the present invention adopts prefabricated gelatinized starch, microcrystalline cellulose, lactose, and sweet mellow wine, as filler, makes feeding lactobacillus micropill have good mouldability, keeps well spherical.
3, lactic acid bacteria is a kind of anaerobism or micro-aerobic microorganism, and can be exposed to air the process lactic acid bacteria of granulating, therefore the present invention adopts vitamin C and vitamin E as antioxidant, is conducive to the survival improving lactic acid bacteria.
Detailed description of the invention
The present invention is described in detail below in conjunction with specific embodiment.
Embodiment 1
A preparation method for feeding lactobacillus micropill, is prepared by following step;
(1) lactic acid bacteria bacterium mud is prepared: when Lactobacillus plantarum being cultured in MRS nutrient solution exponential phase mid-term, about 10 ~ 16h, adds a certain amount of sodium chloride and make the final concentration of sodium chloride in nutrient solution control at 0.4mol/L in respective nutrient solution; Lactobacillus plantarum is continued to be cultured to early stage stable growth phase under this concentration, about 6 ~ 10h, centrifugal 10min under centrifugal acceleration is 4600g condition, with 0.85% physiological saline cleaning bacterium mud twice, collect bacterium mud, then bacterium mud is resuspended in fresh MRS medium, slowly be warming up to 45 DEG C, and after keeping 30min at such a temperature, be cooled to rapidly 0 ~ 4 DEG C, for subsequent use;
(2) 280 parts, the Lactobacillus plantarum bacterium mud of preparation in prefabricated gelatinized starch 200 parts, sodium alginate 25 parts, lactalbumin 25 parts, xylo-oligosaccharide 30 parts, FOS 30 parts, resistant starch 30 parts, vitamin C 1 part and step (1) is taken, then load weighted bacterium mud and other compositions are mixed in agitator, make softwood, now the water content of softwood is about 35 ~ 42%;
(3) by softwood obtained for step (2), put into and extrude finisher,pill under 800 revs/min, process granulation in 10 minutes, after granulation, at 40 DEG C, namely moisture drying to 4% is obtained feeding lactobacillus micropill.
Embodiment 2
A preparation method for feeding lactobacillus micropill, is prepared by following step;
(1) lactic acid bacteria bacterium mud is prepared: when Lactobacillus casei being cultured in MRS nutrient solution exponential phase mid-term (about 10 ~ 16h), in respective nutrient solution, adding a certain amount of sodium chloride make the final concentration of sodium chloride in nutrient solution control at 0.5mol/L; Lactic acid bacteria is continued to be cultured to the early stage stable growth phase (about 6 ~ 10h) under this concentration, centrifugal 10min under centrifugal acceleration is 4600g condition, twice, physiological saline cleaning bacterium mud with 0.85%, collect bacterium mud, then by resuspended for bacterium mud with fresh MRS medium, be slowly warming up to 47 DEG C, and keep at such a temperature after 30min, be cooled to rapidly 0 ~ 4 DEG C, for subsequent use;
(2) microcrystalline cellulose 220 parts, sodium alginate 30 parts, lactalbumin 30 parts, xylo-oligosaccharide 35 parts, FOS 35 parts, resistant starch 35 parts, the vitamin E of 2 parts and 290 parts, the Lactobacillus casei bacterium mud of the middle preparation of step (1) is taken, then load weighted bacterium mud and other compositions are mixed in agitator, make softwood, now the water content of softwood is about 35 ~ 42%;
(3) by softwood obtained for step (2), put into and extrude finisher,pill under 850 revs/min, process granulation in 12 minutes, after granulation, at 42 DEG C, namely moisture drying to 5% is obtained feeding lactobacillus micropill.
Embodiment 3
A preparation method for feeding lactobacillus micropill, is prepared by following step;
(1) lactic acid bacteria bacterium mud is prepared: when lactobacillus acidophilus being cultured in MRS nutrient solution exponential phase mid-term, about 10 ~ 16h, adds a certain amount of sodium chloride and make the final concentration of sodium chloride in nutrient solution control at 0.6Mmol/L in respective nutrient solution; Lactic acid bacteria is continued to be cultured to early stage stable growth phase under this concentration, about 6 ~ 10h, centrifugal 10min under centrifugal acceleration is 4600g condition, with the physiological saline cleaning bacterium mud twice of 0.9%, collect bacterium mud, then by resuspended for bacterium mud with fresh MRS medium, slowly be warming up to 50 DEG C, and after keeping 30min at such a temperature, be cooled to rapidly 0 ~ 4 DEG C, for subsequent use;
(2) 300 parts, the lactobacillus acidophilus bacterium mud of preparation in lactose 230 parts, sodium alginate 40 parts, lactalbumin 40 parts, xylo-oligosaccharide 45 parts, FOS 45 parts, resistant starch 40 parts, the vitamin E of 1 part, the vitamin C of 1 part and step (1) is taken, then load weighted bacterium mud and other compositions are mixed in agitator, make softwood, now the water content of softwood is about 35 ~ 42%;
(3) by softwood obtained for step (2), put into and extrude finisher,pill under 900 revs/min, process granulation in 13 minutes, after granulation, at 45 DEG C, namely moisture drying to 6% is obtained feeding lactobacillus micropill.
Embodiment 4
A preparation method for feeding lactobacillus micropill, is prepared by following step;
(1) lactic acid bacteria bacterium mud is prepared: when streptococcus thermophilus being cultured in MRS nutrient solution exponential phase mid-term, about 10 ~ 16h, adds a certain amount of sodium chloride and make the final concentration of sodium chloride in nutrient solution control at 0.7mol/L in respective nutrient solution; Lactic acid bacteria is continued to be cultured to early stage stable growth phase under this concentration, about 6 ~ 10h, centrifugal 10min under centrifugal acceleration is 4600g condition, with the physiological saline cleaning bacterium mud twice of 0.9%, collect bacterium mud, then by resuspended for bacterium mud with fresh MRS medium, slowly be warming up to 52 DEG C, and after keeping 30min at such a temperature, be cooled to rapidly 0 ~ 4 DEG C, for subsequent use;
(2) 320 parts, the streptococcus thermophilus bacterium mud of preparation in 240 parts, sweet mellow wine, sodium alginate 45 parts, lactalbumin 45 parts, xylo-oligosaccharide 45 parts, FOS 45 parts, resistant starch 45 parts, the vitamin E of 1 part, the vitamin C of 1.5 parts and step (1) is taken, then load weighted bacterium mud and other compositions are mixed in agitator, make softwood, now the water content of softwood is about 35 ~ 42%;
(3) by softwood obtained for step (2), put into and extrude finisher,pill under 950 revs/min, process granulation in 14 minutes, after granulation, at 48 DEG C, namely moisture drying to 7% is obtained feeding lactobacillus micropill.
Embodiment 5
A preparation method for feeding lactobacillus micropill, is prepared by following step;
(1) lactic acid bacteria bacterium mud is prepared: when streptococcus thermophilus being cultured in MRS nutrient solution exponential phase mid-term, about 10 ~ 16h, adds a certain amount of sodium chloride and make the final concentration of sodium chloride in nutrient solution control at 0.7Mmol/L in respective nutrient solution; Lactic acid bacteria is continued to be cultured to early stage stable growth phase under this concentration, about 6 ~ 10h, centrifugal 10min under centrifugal acceleration is 4600g condition, with the physiological saline cleaning bacterium mud twice of 0.9%, collect bacterium mud, then by resuspended for bacterium mud with fresh MRS medium, slowly be warming up to 55 DEG C, and after keeping 30min at such a temperature, be cooled to rapidly 0 ~ 4 DEG C, for subsequent use;
(2) 340 parts, the streptococcus thermophilus bacterium mud of preparation in microcrystalline cellulose 250 parts, sodium alginate 50 parts, lactalbumin 50 parts, xylo-oligosaccharide 50 parts, FOS 50 parts, resistant starch 50 parts, the vitamin E of 1 part, the vitamin C of 2 parts and step (1) is taken, then load weighted bacterium mud and other compositions are mixed in agitator, make softwood, now the water content of softwood is about 35 ~ 42%;
(3) by softwood obtained for step (2), put into and extrude finisher,pill under 1000 revs/min, process granulation in 15 minutes, after granulation, at 48 DEG C, namely moisture drying to 7% is obtained feeding lactobacillus micropill.
Embodiment 6
A preparation method for feeding lactobacillus micropill, is prepared by following step;
(1) lactic acid bacteria bacterium mud is prepared: when Lactobacillus plantarum, Lactobacillus casei being cultured to respectively in MRS nutrient solution exponential phase mid-term, about 10 ~ 16h, adds a certain amount of sodium chloride and makes the final concentration of sodium chloride in nutrient solution control at 0.4mol/L in respective nutrient solution; Under this concentration, Lactobacillus plantarum, Lactobacillus casei are continued to be cultured to early stage stable growth phase, about 6 ~ 10h, centrifugal 10min under centrifugal acceleration is 4600g condition, with 0.85% physiological saline cleaning bacterium mud twice, collect bacterium mud, then bacterium mud is resuspended in fresh MRS medium, slowly be warming up to 45 DEG C, and after keeping 30min at such a temperature, be cooled to rapidly 0 ~ 4 DEG C, for subsequent use;
(2) Lactobacillus plantarum bacterium mud, totally 350 parts, the Lactobacillus casei bacterium mud of preparation in filler 200 parts, sodium alginate 25 parts, lactalbumin 25 parts, xylo-oligosaccharide 30 parts, FOS 30 parts, resistant starch 30 parts, vitamin C 1 part and step (1) is taken, then load weighted bacterium mud and other compositions are mixed in agitator, make softwood, now the water content of softwood is about 35 ~ 42%;
(3) by softwood obtained for step (2), put into and extrude finisher,pill under 800 revs/min, process granulation in 10 minutes, after granulation, at 40 DEG C, namely moisture drying to 4% is obtained feeding lactobacillus micropill.
The described filler of the present embodiment is the mixing of prefabricated gelatinized starch, microcrystalline cellulose.
Embodiment 7
A preparation method for feeding lactobacillus micropill, is prepared by following step;
(1) lactic acid bacteria bacterium mud is prepared: when Lactobacillus plantarum, Lactobacillus casei, lactobacillus acidophilus being cultured to respectively in MRS nutrient solution exponential phase mid-term, about 10 ~ 16h, adds a certain amount of sodium chloride and makes the final concentration of sodium chloride in nutrient solution control at 0.5mol/L in respective nutrient solution; Under this concentration, Lactobacillus plantarum, Lactobacillus casei are continued to be cultured to early stage stable growth phase, about 6 ~ 10h, centrifugal 10min under centrifugal acceleration is 4600g condition, with the physiological saline cleaning bacterium mud twice of 0.85%, collect bacterium mud, then by resuspended for bacterium mud with fresh MRS medium, slowly be warming up to 47 DEG C, and after keeping 30min at such a temperature, be cooled to rapidly 0 ~ 4 DEG C, for subsequent use;
(2) filler 220 parts, sodium alginate 30 parts, lactalbumin 30 parts, xylo-oligosaccharide 35 parts, FOS 35 parts, resistant starch 35 parts, the vitamin E of 2 parts and Lactobacillus plantarum bacterium mud, Lactobacillus casei bacterium mud, totally 360 parts, the lactobacillus acidophilus bacterium mud of the middle preparation of step (1) is taken, then load weighted bacterium mud and other compositions are mixed in agitator, make softwood, now the water content of softwood is about 35 ~ 42%;
(3) by softwood obtained for step (2), put into and extrude finisher,pill under 850 revs/min, process granulation in 12 minutes, after granulation, at 42 DEG C, namely moisture drying to 5% is obtained feeding lactobacillus micropill.
The described filler of the present embodiment is the mixing of lactose and sweet mellow wine.
Embodiment 8
A preparation method for feeding lactobacillus micropill, is prepared by following step;
(1) lactic acid bacteria bacterium mud is prepared: when lactobacillus acidophilus, streptococcus thermophilus being cultured to respectively in MRS nutrient solution exponential phase mid-term, about 10 ~ 16h, adds a certain amount of sodium chloride and makes the final concentration of sodium chloride in nutrient solution control at 0.6mol/L in respective nutrient solution; Under this concentration, lactobacillus acidophilus, streptococcus thermophilus are continued to be cultured to early stage stable growth phase, about 6 ~ 10h, centrifugal 10min under centrifugal acceleration is 4600g condition, with the physiological saline cleaning bacterium mud twice of 0.9%, collect bacterium mud, then by resuspended for bacterium mud with fresh MRS medium, slowly be warming up to 50 DEG C, and after keeping 30min at such a temperature, be cooled to rapidly 0 ~ 4 DEG C, for subsequent use;
(2) lactobacillus acidophilus bacterium mud, totally 370 parts, the streptococcus thermophilus bacterium mud of preparation in lactose 230 parts, sodium alginate 40 parts, lactalbumin 40 parts, xylo-oligosaccharide 45 parts, FOS 45 parts, resistant starch 40 parts, the vitamin E of 1 part, the vitamin C of 1 part and step (1) is taken, then load weighted bacterium mud and other compositions are mixed in agitator, make softwood, now the water content of softwood is about 35 ~ 42%;
(3) by softwood obtained for step (2), put into and extrude finisher,pill under 900 revs/min, process granulation in 13 minutes, after granulation, at 45 DEG C, namely moisture drying to 6% is obtained feeding lactobacillus micropill.
Filler described in the present embodiment is the mixing of prefabricated gelatinized starch, microcrystalline cellulose and lactose
Embodiment 9
A preparation method for feeding lactobacillus micropill, is prepared by following step;
(1) lactic acid bacteria bacterium mud is prepared: when Lactobacillus casei, lactobacillus acidophilus, streptococcus thermophilus being cultured to respectively in MRS nutrient solution exponential phase mid-term, about 10 ~ 16h, adds a certain amount of sodium chloride and makes the final concentration of sodium chloride in nutrient solution control at 0.7mol/L in respective nutrient solution; Under this concentration, Lactobacillus casei, lactobacillus acidophilus, streptococcus thermophilus are continued to be cultured to early stage stable growth phase, about 6 ~ 10h, centrifugal 10min under centrifugal acceleration is 4600g condition, with the physiological saline cleaning bacterium mud twice of 0.9%, collect bacterium mud, then by resuspended for bacterium mud with fresh MRS medium, slowly be warming up to 52 DEG C, and after keeping 30min at such a temperature, be cooled to rapidly 0 ~ 4 DEG C, for subsequent use;
(2) Lactobacillus casei bacterium mud, lactobacillus acidophilus bacterium mud, 380 parts, the streptococcus thermophilus bacterium mud of preparation in filler 240 parts, sodium alginate 45 parts, lactalbumin 45 parts, xylo-oligosaccharide 45 parts, FOS 45 parts, resistant starch 45 parts, the vitamin E of 1 part, the vitamin C of 1.5 parts and step (1) is taken, then load weighted bacterium mud and other compositions are mixed in agitator, make softwood, now the water content of softwood is about 35 ~ 42%;
(3) by softwood obtained for step (2), put into and extrude finisher,pill under 950 revs/min, process granulation in 14 minutes, after granulation, at 48 DEG C, namely moisture drying to 7% is obtained feeding lactobacillus micropill.
Filler described in the present embodiment is the mixing of microcrystalline cellulose, lactose and sweet mellow wine.
Embodiment 10
A preparation method for feeding lactobacillus micropill, is prepared by following step;
(1) lactic acid bacteria bacterium mud is prepared: when Lactobacillus plantarum, Lactobacillus casei, lactobacillus acidophilus, streptococcus thermophilus being cultured in MRS nutrient solution exponential phase mid-term, about 10 ~ 16h, adds a certain amount of sodium chloride and makes the final concentration of sodium chloride in nutrient solution control at 0.7Mmol/L in respective nutrient solution; Under this concentration, Lactobacillus plantarum, Lactobacillus casei, lactobacillus acidophilus, streptococcus thermophilus are continued to be cultured to early stage stable growth phase, about 6 ~ 10h, centrifugal 10min under centrifugal acceleration is 4600g condition, with the physiological saline cleaning bacterium mud twice of 0.9%, collect bacterium mud, then by resuspended for bacterium mud with fresh MRS medium, slowly be warming up to 55 DEG C, and after keeping 30min at such a temperature, be cooled to rapidly 0 ~ 4 DEG C, for subsequent use;
(2) Lactobacillus plantarum bacterium mud, Lactobacillus casei bacterium mud, lactobacillus acidophilus bacterium mud, totally 390 parts, the streptococcus thermophilus bacterium mud of preparation in microcrystalline cellulose 250 parts, sodium alginate 50 parts, lactalbumin 50 parts, xylo-oligosaccharide 50 parts, FOS 50 parts, resistant starch 50 parts, the vitamin E of 1 part, the vitamin C of 2 parts and step (1) is taken, then load weighted bacterium mud and other compositions are mixed in agitator, make softwood, now the water content of softwood is about 35 ~ 42%;
(3) by softwood obtained for step (2), put into and extrude finisher,pill under 1000 revs/min, process granulation in 15 minutes, after granulation, at 48 DEG C, namely moisture drying to 7% is obtained feeding lactobacillus micropill.
Filler described in the present embodiment is the mixing of prefabricated gelatinized starch, microcrystalline cellulose, lactose and sweet mellow wine.
Embodiment 11
The present embodiment provides the preparation method of two kinds of micropills, method 1 and method 2, and the difference of these two kinds of methods is the preparation of lactic acid bacteria bacterium mud, and blend step, the granulation step of two kinds of methods and wall material are identical, all adopt preparation process of the present invention.
Method 1:: adopt preparation process of the present invention.
Method 2: lactic acid bacteria is cultured in MRS nutrient solution early stage stable growth phase, then collects bacterium mud and is resuspended in fresh MRS nutrient solution, preserve under 0 ~ 4 DEG C of condition, then adopt the inventive method softwood, granulation.
Assay method: get respectively by method 2(contrast method) prepare sample, according to method 1(the inventive method) the feeding lactobacillus micropill sample 1g for preparing, under the condition of low temperature, wear into meal, join in 9g sterilized water, stir 10 ~ 20min, after all dissolving, gradient dilution, adopt the method for plate count to detect viable count, the culture medium of plate count is MRS culture medium; Viable lactic acid bacteria rate result is as following table 1
Table 1
Embodiment 12
By pulvis lactobacillus micro-capsule 1g, the feeding lactobacillus micropill 1g for preparing according to the inventive method, join (pH 2.0 in 9ml SGF respectively, 50U/mL, 2h) with high speed disperser to its homogeneous, after 12000 revs/min ~ 16000 revs/min process 60s, adopt plate count method respectively, measure feeding lactobacillus micropill, the viable bacteria rate of pulvis lactobacillus micro-capsule in SGF, result is as following table 2:
Table 2
From upper table 2, in human body simulation gastric juice (pH 2.0,50U/mL, 2h), micropill can be supplied to the good protected effect of lactic acid bacteria, and compared to common pulvis lactobacillus micro-capsule, the viable bacteria rate of micropill of the present invention is higher.
Embodiment 13
By the feeding lactobacillus micropill prepared by the inventive method, pulvis lactobacillus micro-capsule of the prior art is preserved 30 days respectively at normal temperature (about 20 DEG C), low temperature (4 DEG C) is preserved 30 days, then each 1g micropill of feeding lactobacillus micropill under two kinds of preservation conditions is got respectively, the each 1g of pulvis lactobacillus micro-capsule preserved under two kinds of preservation conditions, under the condition of low temperature, wear into meal, join in 9g sterilized water, stir 10 ~ 20min, after all dissolving, gradient dilution, the method of plate count is adopted to detect viable count, the culture medium of plate count is MRS culture medium.The survival outcome of viable bacteria is as following table 3:
Table 3
From upper table 3: the lactic acid bacteria be embedded in micropill has good storage stability, compared to the lactic acid bacteria bacterium powder that freeze drying and spraying dry obtain, the survival rate of lactic acid bacteria has been enhanced 60 ~ 80 times and 7 ~ 14 times respectively.
Embodiment 14
The present embodiment is by feeding lactobacillus micropill, and pulvis microcapsules measure its viable bacteria rate after feed granulating;
Granulating process: after micropill or pulvis microcapsules are mixed in proportion with other feedstuffs, adjusting screw(rod) rotating speed 21r/min, screw aperture 3.0mm; Regulate moisture 15 ~ 17%, pelleting temperature is 55 ~ 65 DEG C, regulating time 40s; Steam pressure 0.3 ~ 0.35Mpa, pellet draw ratio is 2.5:1, carries out the granulation of particulate material.
Condition determination: take containing feeding lactobacillus micropill Feed Sample and each 1g of feed containing pulvis microcapsules respectively, add in the centrifuge tube that 9mL sterilized water is housed, with high speed disperser to its homogeneous, after 12000 revs/min ~ 16000 revs/min process 60s, adopt plate count method respectively, in feed, the survival rate of lactic acid bacteria is as following table 4:
Table 4
From upper table 4, after feed granulating process, feeding lactobacillus micropill can better resist the various destructions in pelletization, has better protective effect to lactic acid bacteria, and viable bacteria rate improves more than 100 times than pulvis lactobacillus micro-capsule viable bacteria rate.
Claims (6)
1. a preparation method for feeding lactobacillus micropill, is characterized in that: prepared by following step;
(1) lactic acid bacteria bacterium mud is prepared: when lactic acid bacteria being cultured to exponential phase mid-term, in nutrient solution, adding a certain amount of sodium chloride make the final concentration of sodium chloride in nutrient solution control at 0.4 ~ 0.7mol/L; Lactic acid bacteria is continued to be cultured to early stage stable growth phase under this concentration, centrifugal, with physiological saline cleaning bacterium mud, collect bacterium mud, then bacterium mud is resuspended in fresh MRS medium, is slowly warming up to 45 ~ 55 DEG C, and after keeping 30min at such a temperature, be cooled to rapidly 0 ~ 4 DEG C, for subsequent use;
(2) 280 ~ 400 parts, the bacterium mud of preparation in filler 200 ~ 250 parts, sodium alginate 25 ~ 50 parts, lactalbumin 25 ~ 50 parts, xylo-oligosaccharide 30 ~ 50 parts, FOS 30-50 part, resistant starch 30 ~ 50 parts, antioxidant 1 ~ 3 part and step (1) is taken, then load weighted bacterium mud and other compositions are mixed in agitator, make softwood
(3) by above-mentioned obtained softwood, put into and extrude finisher,pill and granulate, after granulation, be drying to obtain feeding lactobacillus micropill.
2. the preparation method of a kind of feeding lactobacillus micropill according to claim 1, is characterized in that: the granulation conditions described in step (3) processes 10 ~ 15 minutes under 800 ~ 1000 revs/min.
3. the preparation method of a kind of feeding lactobacillus micropill according to claim 1 and 2, is characterized in that: the drying described in step (3) is by moisture drying to 4 ~ 7% at 40 ~ 48 DEG C.
4. the preparation method of a kind of feeding lactobacillus micropill according to claim 1, is characterized in that: described in step (2), filler is prefabricated gelatinized starch, microcrystalline cellulose, lactose, one or more the mixing in sweet mellow wine.
5. the preparation method of a kind of feeding lactobacillus micropill according to claim 1, is characterized in that: described in step (2), antioxidant is vitamin C, one or both in vitamin E.
6. the preparation method of a kind of feeding lactobacillus micropill according to claim 1, is characterized in that: described in step (1), lactic acid bacteria is selected from one or more in Lactobacillus plantarum, Lactobacillus casei, lactobacillus acidophilus, streptococcus thermophilus.
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