CN105475988A - Hybrid intestinal micro-ecologic preparation and preparation method thereof - Google Patents
Hybrid intestinal micro-ecologic preparation and preparation method thereof Download PDFInfo
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- CN105475988A CN105475988A CN201510832927.6A CN201510832927A CN105475988A CN 105475988 A CN105475988 A CN 105475988A CN 201510832927 A CN201510832927 A CN 201510832927A CN 105475988 A CN105475988 A CN 105475988A
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/113—Acidophilus
Abstract
The invention provides a hybrid intestinal micro-ecologic preparation and a preparation method thereof. The preparation method comprises the following steps: carrying out third-stage fermentation culture on probiotics butyric acid bacteria, lactobacillus acidophilus, Bacillus coagulans and Bacillus licheniformis, adding a freeze drying protection gent, freeze-drying the obtained culture product, carrying out microcapsule embedding, freeze-drying the embedded product, and mixing the embedded product with isomaltooligosacharide, fructooligosaccharide and vitamins to obtain the micro-ecologic preparation. Butyric acid bacteria and other probiotics can be directly supplemented to the human body intestinal tract, so all the probiotics are propagated and intergrow in the intestinal tract and have synergism, and the intestine controlling, cancer preventing, immunity improving, digestion helping and harmful bacterium and corrupted group growth breeding inhibiting effects of the composite bacterium preparation are effectively performed, thereby the beneficial physiologic effects of the probiotics on human bodies are greatly improved; and the preparation method can effectively guarantee the quantity of live bacteria of the probiotics, the activity of the probiotics and the preservation time of the probiotics.
Description
Technical field
Patent of the present invention belongs to the category of probiotics, is specifically related to a kind of mixed type enteral microecological formulation and preparation method thereof.
Background technology
Live in the enteron aisle of Healthy People kind more than 400, tens of trillion bacterium, these microorganisms take part in physiology, the growth of human body, whole vital movement processes of biophysics and pharmacology and pathology.According to the effect to human body, we are divided into beneficial bacterium, harmful bacteria and two-way bacterium these bacteriums.These bacterium interdependences, mutually to restrict, be in relative balance state, micro-ecological environment maximum in composition body, becomes the natural defence line of safeguarding health, jointly completes the effect of nutrient absorption and toxin excretion.Human consumption is absorbed and plays an important role, its normal or imbalance, have very important impact to the health of human body and life-span.
Along with the increase of modern food kind and pattern, and environmental pollution, abuse of antibiotics, hormone, chemicotherapy, various factors stress be waited to cause in human body intestinal canal micro-ecological environment disorderly, other harmful bacteria will be taken advantage of the occasion and enter, cause flora imbalance, metabolic disorder, various toxin, corrupt substance, carcinogen increase, and produce multiple pathology, cause human body inner product poison to cause constipation, diarrhoea, hepatitis and other disease of digestive tracts, serious even can life-threatening.Except the health that is careful in one's diet, balanced diet, strengthen physical training except, people start to notice that some active bacteria formulations are improving the effect in health, and adopting probiotics, to rebuild healthy micro-ecological environment in enteron aisle be a kind of desirable method.
Probio (Probiotics) be a class by colonized action change host a certain position flora composition, produce and be conducive to the single of host health effect or form clear and definite mixed microorganism, the active bacteria formulation be made up of single or multiple-microorganism.The probio of current extensive use has Bacillus acidi lactici class, Bifidobacterium class, bacillus category and saccharomycete etc., Miyarisan and lactobacillus acidophilus (lactic acid bacteria), bacillus coagulans, bacillus licheniformis are beneficial bacteriums intrinsic in human body, they influence each other, mutually promote, collaborative suppression harmful bacteria, jointly plays very important effect in regulating intestinal canal dysfunction, maintenance intestinal flora balance and raising immunity of organism level etc.
(1) Miyarisan.Miyarisan is one of Body normal strain, research finds, Miyarisan have whole intestines, anticancer, improve the effect such as immunity, the growth and breeding of harmful bacteria and corrupt group can be suppressed in enteron aisle, the most significant biological nature of Miyarisan be its can with the intestinal beneficial bacterium symbiosis such as lactobacillus acidophilus, and promote that it grows propagation; Near in half a century, Miyarisan preparation is widely used in peptic ulcer that enteric flora disturbance, acute and chronic diarrhea, intestinal irritable syndrome, Antibiotic-associated Colitis, chronic hepatitis and supplemental treatment helicobacter pylori that a variety of causes such as digestion, children's, surgery, tumour, gynemetrics cause cause and pseudomembranous enteritis etc. in Asias such as Japan, Korea S, Taiwan;
(2) lactobacillus acidophilus.Last century end is found, belong to gram-positive cocci, obligate anaerobic, has adjustment stomach and intestine to human body equally, helps digest, improves immunity, reduces blood fat, reduces different physiological roles such as spoilage organisms generation amine, indoles harmful substance etc. in intestines, lactobacillus acidophilus is just contained in many Yoghourts in market;
(3) bacillus coagulans.As a kind of novel gemma probio of uniqueness, many research shows, it had both had the lactic acid producing characteristic of lactic acid bacteria, has again the abundant enzyme system of bacillus.Bacillus coagulans as the up-and-coming youngster of probio family, then can adjust intestinal microecology balance equally, improves immunity of organism, produces antibacterial substance and promote growth of animal etc., become the focus studied in probio;
(4) bacillus licheniformis.Its active bacteria formulation made makes it enter people's enteron aisle with the form of viable bacteria, not only to human body itself without any injury, and can produce multiple Antagonism material, these antagonistic substances can suppress malignant bacteria in enteron aisle; Multiple enzyme can also be produced and improve human consumption's enzymatic activity, promoting human body digesting and assimilating nutriment; And the immunologic function of human body can be strengthened, resist infection.Lichens bud pole bacterium is a kind of facultative anaerobe, can also play the effect that its biology takes oxygen by force, makes probio obtain better growing environment, maintains the ecological balance in enteron aisle, the symptoms such as diarrhoea can be made to alleviate or disappear.
At present, independent Miyarisan preparation, Pro-Bionate, bacillus coagulans preparation and bacillus licheniformis preparation are widely used at every country and area, but Miyarisan, lactobacillus acidophilus, bacillus coagulans and bacillus licheniformis probiotic combination are got up to make the probiotics that viable bacteria mixes, makes its product playing multi-strain-combined Action advantage there is not yet reported in literature.
Probiotics is a kind of oral non-pathogenic active bacteria formulation, refers to and uses microecology principle, utilize the growth promoting substance to the useful and harmless probio of host or probio, through the preparation that special process is made.It can improve the balance of enteric bacteria, brings about a wholesome effect to human or animal.The research of enteral microecological formulation has become our times microbiological art research emphasis, and the research and development of probiotic products can bring positive promotion meaning to the health of the mankind, therefore has important economic worth and social benefit.But these probioticses have a fatal weakness, these active microorganisms not adding protective layer are in transport, store and application process in, need to stand a series of poor environment (as aerobic state, high temperature, extruding, low ph value, high cholate etc.) impact, field planting in enteron aisle could be arrived, in this process, due to special thalline oxytolerant such as some anaerobic bacteria or facultative anaerobes etc., acidproof, resistance to elevated temperatures is poor, resistance is poor, not stomach juice-resistant and cholate, be subject to the impact of extraneous various biochemistry and physical factor, activity is caused greatly to reduce even dead, difficulty has enough quantity reach enteron aisle or settle down enteron aisle and play a role, have a strong impact on tiring of active bacteria formulation.The number of viable of probio few (all lower than 300,000,000/gram) in current probiotics, and the viable count of probio reduces (1.5 years viable counts be only preparation at the beginning of 50%-55%) after preparation with the increase of memory time, in addition, the bacterial classification that current probiotics product uses is more single, thus its treatment and health-care effect limited.
Summary of the invention
The object of the present invention is to provide a kind of mixed type enteral microecological formulation, provide its corresponding preparation method to be another goal of the invention of the present invention simultaneously.
Based on above-mentioned purpose, the present invention takes following technical scheme: a kind of mixed type enteral microecological formulation, containing probiotic microcapsule 5 ~ 60g, oligoisomaltose 30 ~ 60g, FOS 9.9 ~ 34.75g, vitamin 0.1 ~ 0.5g in every 100g preparation; Described probiotic microcapsule is made up of the raw material of following weight portion: Miyarisan 1 ~ 25 part, lactobacillus acidophilus 1 ~ 25 part, bacillus coagulans 1 ~ 25 part, bacillus licheniformis 1 ~ 25 part.
Described vitamin is one or more the mixing in vitamin B1, vitamin B2, vitamin B6, vitamin C and folic acid.
A preparation method for mixed type enteral microecological formulation, comprises the following steps:
(1) preparation of active bacteria mud: take each raw material by following parts by weight: Miyarisan 1 ~ 25 part, lactobacillus acidophilus 1 ~ 25 part, bacillus coagulans 1 ~ 25 part, bacillus licheniformis 1 ~ 25 part, as seed bacterial strain after mixing, add in culture medium and carry out three grade fermemtation, every grade of fermentation time is 24 ~ 48h, and the bacterial classification amount that fermentation at different levels is added is 3 ~ 8% of culture volume at different levels; After cultivation terminates, centrifugal and with sterilized water washing 2 ~ 3 times, obtain active bacteria mud;
(2) freeze drying protectant freeze-drying is added: after the weight ratio of active bacteria mud and freeze drying protectant being pressed 100:35 ~ 60 mixes, freeze-drying obtains freeze-dried powder;
(3) embedding of embedding liquid is added: to be mixed by the weight ratio of 100:35 ~ 60 with embedding liquid by freeze-dried powder and make embedding mixed liquor;
(4) preparation of microcapsules: embedding mixed liquor is poured in embedding injector, high pressure injection embedding consolidation liquid, constantly stirs simultaneously, after the embedding solidification process of 2 ~ 10h, filtration, sterilized water washing 2 ~ 3 times, freeze-drying obtains probiotic microcapsule; Described embedding mixed liquor is 1:1 ~ 1:6 with the volume ratio of embedding consolidation liquid;
(5) preparation of active microbiological preparations: mixed with oligoisomaltose, FOS, vitamin by above-mentioned probiotic microcapsule and obtain active microbiological preparations, containing probiotic microcapsule 5 ~ 60g, oligoisomaltose 30 ~ 60g, FOS 10 ~ 34.5g, vitamin 0.1 ~ 0.5g in every 100g preparation.
The percentage by weight of described culture medium at different levels consists of: weight hundred score of glucose, tryptone, yeast extract is 1 ~ 5%, beef extract, sodium chloride, ammonium sulfate, dipotassium hydrogen phosphate, ferrous sulfate, magnesium sulfate, sodium acid carbonate, calcium carbonate, Tween.80 weight hundred score are 0.01 ~ 0.5%, and all the other are water.
Described freeze drying protectant is made by the raw material of following weight portion: skimmed milk power 30 ~ 55 parts, sodium glutamate 5 ~ 25 parts, lactose 5 ~ 35 parts, starch 20 ~ 50 parts, vitamine C sodium 0.5 ~ 5 part, add water and be settled to 100mL, adds thermal agitation and make it dissolve, sterilizing; Sterilizing 15min at sterilizing is specially 105 DEG C, is cooled to 10 ~ 35 DEG C.
Described embedding liquid is made by the raw material of following weight portion: sodium alginate 10 ~ 50 parts, 20 ~ 40 parts, gelatin, 30 ~ 50 parts, aliphatic acid, add water and be settled to 100mL, stirs 10min, leave standstill 2 ~ 6h, sterilizing, sterilizing 15min at sterilizing is specially 105 DEG C, is cooled to 10 ~ 35 DEG C.
Described embedding consolidation liquid is prepared by the following method: take calcium chloride 0.5 ~ 20.0g, potassium chloride 0.5 ~ 15.0g, add water and be settled to 100mL, sterilizing; Sterilizing 15min at sterilizing is specially 105 DEG C, is cooled to 10 ~ 35 DEG C.
Step (2) and the middle freeze-drying of step (4) are carried out under the following conditions: vacuum 0.5 ~ 1.3Pa, temperature-30 ~-50 DEG C, the time is 7 ~ 10h.
The pressure injecting embedding consolidation liquid in step (2) is 0.1 ~ 0.4MPa.
The present invention is by probios such as Miyarisans in direct supplementary human body intestinal canal, can reach and make various bacterium in enteron aisle internal breeding symbiosis, synergy, more effectively play composite bacteria preparation whole intestines, anticancer, improve immunity, help digest, suppress the effect such as harmful bacteria and corrupt group's growth and breeding, the physiological action making probio useful to human body improves greatly, and the preservation time of this preparation method more effective quantity of guarantee probiotics viable bacteria of energy and the activity of probio and probio.
Specifically, beneficial effect of the present invention is:
(1) the present invention utilizes microcapsules coating technique to make a kind of new mixed type enteral microecological formulation, effectively enhance the resistivity of microorganism to poor environment factors such as high temperature, drying, hydrochloric acid in gastric juice and bile, improve the stability of product quality concentration and product, be convenient to transport, storage and use, guaranteeing its tolerance granulation high temperature, extruding, stomach juice-resistant, bile tolerance and arrive intestinal colonisation smoothly;
(2) Miyarisan, lactobacillus acidophilus, bacillus coagulans and bacillus licheniformis probiotic combination are got up to make new mixed type probiotics by the present invention first, make it in enteron aisle internal breeding symbiosis, complement each other, more effectively play composite bacteria preparation and safeguard the beneficial effect of the microecological balance of enteron aisle, thus reach and regulate immunity of organisms, delay senility, Tumor suppression, adjusting blood lipid, improve the object of stomach;
(3) in probiotics of the present invention, beneficial bacterium total viable count is 5.0 × 10
10~ 9.9 × 10
11individual/gram, not only Board Lot is huge, keep sample at normal temperatures investigation after 2 years viable bacteria still higher than more than 90%, have good stability; And probiotics safety non-toxic of the present invention, be suitable for various crowd.
(4) the present invention add in probiotic microcapsule oligoisomaltose, FOS and vitamin be not only conducive to probio enter enteron aisle after can quick active breeding, and supplement necessary vitamin for human body, promote health.
Detailed description of the invention
Embodiment is only for further illustrating the present invention below, does not limit the present invention in any form.
Embodiment 1
A kind of mixed type enteral microecological formulation, containing probiotic microcapsule 20g, oligoisomaltose 50g, FOS 29.75g, VB12mg, VB22mg, VB62mg, VC200mg, folic acid 44mg in every 100g preparation.
Its preparation method, comprises the following steps:
(1) Miyarisan 15 parts, lactobacillus acidophilus 15 parts, bacillus coagulans 15 parts, bacillus licheniformis 15 parts, as seed bacterial strain after mixing, add in culture medium and carry out three grade fermemtation, every grade of fermentation time is 36h, and the bacterial classification amount of fermentation at different levels is 5% of culture volume at different levels; After cultivation terminates, centrifugal and wash 2 times with sterilized water, obtain active bacteria mud; Culture medium percentage at different levels consists of: glucose 1%, tryptone 1%, yeast extract 1%, beef extract 0.3%, sodium chloride 0.2%, ammonium sulfate 0.1%, dipotassium hydrogen phosphate 0.4%, ferrous sulfate 0.1%, magnesium sulfate 0.05%, calcium carbonate 0.1%, sodium acid carbonate 0.01%, Tween.800.02%, and all the other are water;
(2) freeze drying protectant freeze-drying is added: the weight ratio of active bacteria mud and freeze drying protectant being pressed 100:45 mixes, and at vacuum 0.5Pa, at-40 DEG C, freeze-drying obtains freeze-dried powder, and freeze-drying time is 8h; Freeze drying protectant is made by the raw material of following weight portion: skimmed milk power 40 parts, sodium glutamate 15 parts, lactose 35 parts, starch 20 parts, vitamine C sodium 3 parts, add water and be settled to 100mL, adding thermal agitation makes it dissolve, and then 105 DEG C of sterilizing 15min, are cooled to 10 ~ 35 DEG C;
(3) embedding of embedding liquid is added: to be mixed by the weight ratio of 100:45 with embedding liquid by freeze-dried powder and make embedding mixed liquor; Embedding liquid is made by the raw material of following weight portion: sodium alginate 35 parts, 25 parts, gelatin, 40 parts, aliphatic acid, add water and be settled to 100mL, stirs 10min, and leave standstill 2 ~ 6h, then 105 DEG C of sterilizing 15min, are cooled to 10 ~ 35 DEG C;
(4) preparation of microcapsules: embedding mixed liquor is poured in embedding injector, embedding consolidation liquid is injected under pressure 0.4MPa condition, constantly stir simultaneously, after the embedding solidification process of 6h, filtration, sterilized water wash 2 times, at vacuum 0.5Pa, at cryogenic temperature-40 DEG C, freeze-drying obtains probiotic microcapsule, and freeze-drying time is 8h; Embedding consolidation liquid is prepared by the following method: take calcium chloride 7.5g, potassium chloride 5g, adding water is settled to 100mL, thoroughly dissolve, then 105 DEG C of sterilizing 15min, are cooled to 10 ~ 35 DEG C; Embedding mixed liquor is 1:4 with the volume proportion of embedding consolidation liquid;
(5) preparation of active microbiological preparations: above-mentioned probiotic microcapsule 20g is mixed with oligoisomaltose 50g, FOS 29.75g, VB12mg, VB22mg, VB62mg, VC200mg, folic acid 44mg and obtains active microbiological preparations.
Embodiment 2
A kind of mixed type enteral microecological formulation, containing probiotic microcapsule 5g, oligoisomaltose 60g, FOS 34.75g, VB12mg, VB22mg, VB62mg, VC200mg, folic acid 44mg in every 100g preparation.
Its preparation method, comprises the following steps:
(1) Miyarisan 5 parts, lactobacillus acidophilus 5 parts, bacillus coagulans 5 parts, bacillus licheniformis 5 parts, as seed bacterial strain after mixing, add in culture medium and carry out three grade fermemtation, every grade of fermentation time is 48h, and the bacterial classification amount of fermentation at different levels is 3% of culture volume at different levels; After cultivation terminates, centrifugal and wash 2 times with sterilized water, obtain active bacteria mud; Culture medium percentage at different levels consists of: glucose 2%, tryptone 2%, yeast extract 1.5%, beef extract 0.5%, sodium chloride 0.2%, ammonium sulfate 0.1%, dipotassium hydrogen phosphate 0.4%, ferrous sulfate 0.1%, magnesium sulfate 0.05%, sodium acid carbonate 0.2%, calcium carbonate 0.1%, Tween.800.02%, all the other are water;
(2) freeze drying protectant freeze-drying is added: the weight ratio of active bacteria mud and freeze drying protectant being pressed 100:35 mixes, and at vacuum 0.5Pa, at-40 DEG C, freeze-drying obtains freeze-dried powder, and freeze-drying time is 8h; Freeze drying protectant is made by the raw material of following weight portion: skimmed milk power 30 parts, sodium glutamate 5 parts, lactose 15 parts, starch 50 parts, vitamine C sodium 0.5 part, add water and be settled to 100mL, adding thermal agitation makes it dissolve, and then 105 DEG C of sterilizing 15min, are cooled to 10 ~ 35 DEG C;
(3) embedding of embedding liquid is added: to be mixed by the weight ratio of 100:35 with embedding liquid by freeze-dried powder and make embedding mixed liquor; Embedding liquid is made by the raw material of following weight portion: sodium alginate 50 parts, 20 parts, gelatin, 30 parts, aliphatic acid, add water and be settled to 100mL, stirs 10min, and leave standstill 2 ~ 6h, then 105 DEG C of sterilizing 15min, are cooled to 10 ~ 35 DEG C;
(4) preparation of microcapsules: embedding mixed liquor is poured in embedding injector, embedding consolidation liquid is injected under pressure 0.1MPa condition, constantly stir simultaneously, after the embedding solidification process of 5h, after filtration, after sterilized water washs 2 times, at vacuum 0.5Pa, at cryogenic temperature-40 DEG C, freeze-drying obtains probiotic microcapsule, and freeze-drying time is 8h; Embedding consolidation liquid is prepared by the following method: take calcium chloride 0.5g, potassium chloride 0.5g, adding water is settled to 100mL, thoroughly dissolve, then 105 DEG C of sterilizing 15min, are cooled to 10 ~ 35 DEG C, and embedding mixed liquor is 1:1 with the volume proportion of embedding consolidation liquid;
(5) preparation of active microbiological preparations: above-mentioned probiotic microcapsule 5g is mixed with oligoisomaltose 60g, FOS 34.75g, VB12mg, VB22mg, VB62mg, VC200mg, folic acid 44mg and obtains active microbiological preparations.
Embodiment 3
A kind of mixed type enteral microecological formulation, containing probiotic microcapsule 60g, oligoisomaltose 30g, FOS 9.9g, VB13mg, VB25mg, VB62mg, VC60mg, folic acid 30mg in every 100g preparation.
Its preparation method, comprises the following steps:
(1) Miyarisan 25 parts, lactobacillus acidophilus 25 parts, bacillus coagulans 25 parts, bacillus licheniformis 25 parts, as seed bacterial strain after mixing, add in culture medium and carry out three grade fermemtation, every grade of fermentation time is 24h, and the bacterial classification amount of fermentation at different levels is 8% of culture volume at different levels; After cultivation terminates, centrifugal and wash 2 times with sterilized water, obtain active bacteria mud; Culture medium percentage at different levels consists of: glucose 5%, tryptone 5%, yeast extract 5%, beef extract 0.5%, sodium chloride 0.3%, ammonium sulfate 0.2%, dipotassium hydrogen phosphate 0.5%, ferrous sulfate 0.2%, magnesium sulfate 0.1%, sodium acid carbonate 0.5%, calcium carbonate 0.05%, Tween.800.05%, all the other are water;
(2) freeze drying protectant freeze-drying is added: the weight ratio of active bacteria mud and freeze drying protectant being pressed 100:60 mixes, and at vacuum 0.5Pa, at-40 DEG C, freeze-drying obtains freeze-dried powder, and freeze-drying time is 8h; Freeze drying protectant is made by the raw material of following weight portion: skimmed milk power 55 parts, sodium glutamate 25 parts, lactose 5 parts, starch 20 parts, vitamine C sodium 0.5 part, add water and be settled to 100mL, adding thermal agitation makes it dissolve, and then 105 DEG C of sterilizing 15min, are cooled to 10-35 DEG C;
(3) embedding of embedding liquid is added: to be mixed by the weight ratio of 100:60 with embedding liquid by freeze-dried powder and make embedding mixed liquor; Embedding liquid is made by the raw material of following weight portion: sodium alginate 10 parts, 40 parts, gelatin, 50 parts, aliphatic acid, add water and be settled to 100mL, stirs 10min, and leave standstill 2 ~ 6h, then 105 DEG C of sterilizing 15min, are cooled to 10 ~ 35 DEG C;
(4) preparation of microcapsules: embedding mixed liquor is poured in embedding injector, embedding consolidation liquid is injected under pressure 0.4MPa condition, constantly stir simultaneously, after the embedding solidification process of 8h, after filtration, after sterilized water washs 2 times, at vacuum 0.5Pa, at cryogenic temperature-40 DEG C, freeze-drying obtains probiotic microcapsule, and freeze-drying time is 8h; Embedding consolidation liquid is prepared by the following method: take calcium chloride 20g, potassium chloride 15g, adding water is settled to 100mL, thoroughly dissolve, then 105 DEG C of sterilizing 15min, are cooled to 10-35 DEG C, and embedding mixed liquor is 1:6 with the volume proportion of embedding consolidation liquid;
(5) preparation of active microbiological preparations: above-mentioned probiotic microcapsule 60g is mixed with oligoisomaltose 30g, FOS 9.9g, VB13mg, VB25mg, VB62mg, VC60mg, folic acid 30mg and obtains active microbiological preparations.
Performance measurement
1, microencapsulated forms, size
The microcapsules of the middle preparation of the step (4) of Example 1-3 do Gram's staining, microscopy (16 × 40 times) respectively, and in the visual field, microcapsules are high-visible, diameter 50 ~ 200 μm, and each microcapsules comprise 20 ~ 50 bacteriums.Illustrate that microcapsule size prepared by the present invention is suitable, thalline bag is by evenly.
2, high temperature resistant, sour, alkalescence inspection
The microcapsules getting preparation in the step (4) of a certain amount of embodiment 1-3 respectively process under following three kinds of conditions:
Condition is 85 DEG C of 10min 1.; Condition 2. 37 DEG C, pH=2, HCL solution 30min; Condition 3. 37 DEG C, pH=13, NaOH solution 30min, centrifuging and taking sediment dyeing microscopic examination after process, its sporoderm-broken rate situation is shown in Table 1.
High temperature resistant, sour, the alkaline assay of table 1 embodiment 1-3
。
As shown in Table 1, probiotics prepared by the present invention is high temperature resistant, acidproof, alkali resistance good, well can resist the impact of extraneous undesirable element.
3, wrap and contrasted by survival rate after the drying of bacterium liquid with not wrapping
(1) method of inspection: colony counting method
(2) preparation of the external shell-broken liquid of gastrointestinal environment is simulated:
No. 1 liquid: it is appropriate that dilute hydrochloric acid solution pH is adjusted to 2.0+ acid protease (purchased from certain biotech firm);
No. 2 liquid: get No. 1 liquid 750mL and add 250mL0.2mol/L metabisulfite solution, adjust pH to 6.8.
(3) process:
Microcapsules broken wall:
The microcapsules drying sample of the middle preparation of the step (4) of a certain amount of embodiment 1 of accurate weighing is placed in No. 1 liquid of certain volume respectively, and 37 DEG C of water bath heat preservation 30min, adjust pH to 6.8, add isopyknic No. 2 liquid, 37 DEG C of insulation 30min.Centrifugal, taking precipitate, microscopy sporoderm-broken rate.
Mixed bacterium freeze-dried powder without microcapsules bag quilt: process with reference to microcapsules broken wall.
(4) sediment in (3) is done to plate count and compares viable count with the mixed bacterium freeze-dried powder without microcapsules bag quilt.
Result shows, compared with mixed bacterium freeze-dried powder, wrap by after drying sample be respectively 98% with the mixed bacterium freeze-dried powder beneficial bacterium average viability not wrapping quilt, 50%, this illustrates the resistivity adopting coating technique of the present invention significantly can strengthen the poor environment factors such as gastrointestinal environment, extends the survival rate of active thalline.
To sum up, mixed type enteral microecological formulation of the present invention, the microcapsule size of preparation is suitable, thalline bag is effective, effectively enhance the resistivity of microorganism to poor environment factors such as high temperature, acidity, alkalescence, well can resist the impact of extraneous undesirable element, improve the survival rate of probio and the stability of product, be convenient to transport, storage and use.
The shelf-life of stability test different disposal probiotics compares
Laboratory sample: three kinds of probioticses obtained by embodiment 1-3;
Comparative sample: mixed bacterium freeze-dried powder 1, by the mixed bacterium freeze-dried powder directly obtained in the step (1) of embodiment 1 and step (2); Mixed bacterium freeze-dried powder 2, preparation method's difference from Example 1 is, does not comprise step (3) and step (4).
By laboratory sample and comparative sample, constant temperature (25 DEG C) under air-proof condition is preserved, and periodic detection viable count, the results are shown in Table 2.
In table 2 three kinds of probioticses and mixed bacterium freeze-dried powder storage life, viable count compares
。
Table 2 data show, in two years, the probiotics viable count storage rate of embodiment 1 reaches 95.68%, the probiotics viable count storage rate of embodiment 2 is 90.91%, the probiotics viable count storage rate of embodiment 3 is 98.47%, and is 37.21% without mixed bacterium freeze-dried powder 1 and 2 storage rate of bag quilt.Result shows, probiotics good stability of the present invention, and the coating technique of employing can play a protective role to the survival of probio Miyarisan and lactobacillus acidophilus, bacillus coagulans, bacillus licheniformis, can its storage life of significant prolongation.
Pharmacodynamic experiment: mixed type enteral microecological formulation of the present invention is on the impact of nonspecific immunity of mice
The healthy mice of 20-25g is about as animal model using body weight, using chicken red blood cell as phagocyte, animal is divided at random experimental group, control group 1, control group 2, control group 3 and blank group (often organizing 10), conventional raising is after 3 days, according to the medicining condition in table 3 to experimental group and the medication respectively of each control animals, dose be every day every gavage 0.5g(in effective content), come into operation continuously 15 days; Blank treated animal then raises the physiological saline with same volume.After 15 days, disconnected neck process animal also collects abdominal cavity phagocytic, using the washed chicken red blood cell of physiological saline as target cell, to count under high-power microscope and the red blood cell recording experimental group and each control animals peritoneal macrophage engulfs number, and by the percentage phagocytic rate (%) of calculating macrophage and phagocytic index, shown in the following list 4 of result.
Table 3 experimental group and each control group medicining condition
。
Table 4 enteral microecological formulation of the present invention is on the impact of Turnover of Mouse Peritoneal Macrophages phagocytic activity
。
The ability of macrophage phagocytic foreign matter represents one of mammal non-specific immune function, and as can be seen from data shown in table 4, compared with the single culture freeze-dried powder that directly comes into operation, the Mixed Microbes that comes into operation performance has better non-specific immune function enhanced activity; With directly adopt compared with mixed bacterium freeze-dried powder, probiotic microcapsule performance prepared by the present invention has better non-specific immune function enhanced activity, therefore can be used in Immune-enhancing effect clinically.
Claims (9)
1. a mixed type enteral microecological formulation, is characterized in that, containing probiotic microcapsule 5 ~ 60g, oligoisomaltose 30 ~ 60g, FOS 9.9 ~ 34.75g, vitamin 0.1 ~ 0.5g in every 100g preparation; Described probiotic microcapsule is made up of the raw material of following weight portion: Miyarisan 1 ~ 25 part, lactobacillus acidophilus 1 ~ 25 part, bacillus coagulans 1 ~ 25 part, bacillus licheniformis 1 ~ 25 part.
2. a kind of mixed type enteral microecological formulation as claimed in claim 1, is characterized in that, described vitamin is one or more the mixing in vitamin B1, vitamin B2, vitamin B6, vitamin C and folic acid.
3. a preparation method for mixed type enteral microecological formulation, is characterized in that, comprises the following steps:
(1) preparation of active bacteria mud: take each raw material by following parts by weight: Miyarisan 1 ~ 25 part, lactobacillus acidophilus 1 ~ 25 part, bacillus coagulans 1 ~ 25 part, bacillus licheniformis 1 ~ 25 part, as seed bacterial strain after mixing, add in culture medium and carry out three grade fermemtation, every grade of fermentation time is 24 ~ 48h, and the bacterial classification amount that fermentation at different levels is added is 3 ~ 8% of culture volume at different levels; After cultivation terminates, centrifugal and with sterilized water washing 2 ~ 3 times, obtain active bacteria mud;
(2) freeze drying protectant freeze-drying is added: after the weight ratio of active bacteria mud and freeze drying protectant being pressed 100:35 ~ 60 mixes, freeze-drying obtains freeze-dried powder;
(3) embedding of embedding liquid is added: to be mixed by the weight ratio of 100:35 ~ 60 with embedding liquid by freeze-dried powder and make embedding mixed liquor;
(4) preparation of microcapsules: embedding mixed liquor is poured in embedding injector, high pressure injection embedding consolidation liquid, constantly stirs simultaneously, after the embedding solidification process of 2 ~ 10h, filtration, sterilized water washing 2 ~ 3 times, freeze-drying obtains probiotic microcapsule; Described embedding mixed liquor is 1:1 ~ 1:6 with the volume ratio of embedding consolidation liquid;
(5) preparation of active microbiological preparations: mixed with oligoisomaltose, FOS, vitamin by above-mentioned probiotic microcapsule and obtain active microbiological preparations, containing probiotic microcapsule 5 ~ 60g, oligoisomaltose 30 ~ 60g, FOS 10 ~ 34.5g, vitamin 0.1 ~ 0.5g in every 100g preparation.
4. the preparation method of mixed type enteral microecological formulation as claimed in claim 3, it is characterized in that, the percentage by weight of described culture medium at different levels consists of: weight hundred score of glucose, tryptone, yeast extract is 1 ~ 5%, beef extract, sodium chloride, ammonium sulfate, dipotassium hydrogen phosphate, ferrous sulfate, magnesium sulfate, sodium acid carbonate, calcium carbonate, Tween.80 weight hundred score are 0.01 ~ 0.5%, and all the other are water.
5. the preparation method of mixed type enteral microecological formulation as claimed in claim 3; it is characterized in that; described freeze drying protectant is made by the raw material of following weight portion: skimmed milk power 30 ~ 55 parts, sodium glutamate 5 ~ 25 parts, lactose 5 ~ 35 parts, starch 20 ~ 50 parts, vitamine C sodium 0.5 ~ 5 part; add water and be settled to 100mL; adding thermal agitation makes it dissolve, sterilizing.
6. the preparation method of mixed type enteral microecological formulation as claimed in claim 3, it is characterized in that, described embedding liquid is made by the raw material of following weight portion: sodium alginate 10 ~ 50 parts, 20 ~ 40 parts, gelatin, 30 ~ 50 parts, aliphatic acid, add water and be settled to 100mL, stir 10min, leave standstill 2 ~ 6h, sterilizing.
7. the preparation method of mixed type enteral microecological formulation as claimed in claim 3, it is characterized in that, described embedding consolidation liquid is prepared by the following method: take calcium chloride 0.5 ~ 20.0g, potassium chloride 0.5 ~ 15.0g, add water and be settled to 100mL, sterilizing.
8. the preparation method of mixed type enteral microecological formulation as claimed in claim 3, is characterized in that, step (2) and the middle freeze-drying of step (4) are carried out under the following conditions: vacuum 0.5 ~ 1.3Pa, temperature-30 ~-50 DEG C, the time is 7 ~ 10h.
9. the preparation method of mixed type enteral microecological formulation as claimed in claim 3, is characterized in that, the pressure injecting embedding consolidation liquid in step (2) is 0.1 ~ 0.4MPa.
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| CN107129946A (en) * | 2017-04-27 | 2017-09-05 | 长春中俄科技园股份有限公司 | A kind of microorganism formulation of high field planting and its application |
| CN107299071A (en) * | 2017-08-16 | 2017-10-27 | 济南青瑞生物科技有限公司 | A kind of enteric probiotics preparation |
| US20180185397A1 (en) * | 2017-01-04 | 2018-07-05 | Jamieson Laboratories Ltd. | Isomalto-Oligosaccharide Prebiotic Formulations |
| CN111743875A (en) * | 2019-09-17 | 2020-10-09 | 河南金百合生物科技股份有限公司 | Preparation method of clostridium butyricum microencapsulated viable bacteria preparation |
| CN113521013A (en) * | 2020-04-20 | 2021-10-22 | 青岛海益康健康科技有限公司 | Embedded probiotic product with planting effect and preparation process thereof |
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| CN101138573A (en) * | 2007-01-29 | 2008-03-12 | 青岛东海药业有限公司 | Application of clostridium butyricum and condensate bacillus in the preparation of medicament for treating inflammatory bowel disease |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US20180185397A1 (en) * | 2017-01-04 | 2018-07-05 | Jamieson Laboratories Ltd. | Isomalto-Oligosaccharide Prebiotic Formulations |
| CN107129946A (en) * | 2017-04-27 | 2017-09-05 | 长春中俄科技园股份有限公司 | A kind of microorganism formulation of high field planting and its application |
| CN107129946B (en) * | 2017-04-27 | 2021-06-08 | 长春中俄科技园股份有限公司 | High-planting microbial preparation and application thereof |
| CN107299071A (en) * | 2017-08-16 | 2017-10-27 | 济南青瑞生物科技有限公司 | A kind of enteric probiotics preparation |
| CN111743875A (en) * | 2019-09-17 | 2020-10-09 | 河南金百合生物科技股份有限公司 | Preparation method of clostridium butyricum microencapsulated viable bacteria preparation |
| CN111743875B (en) * | 2019-09-17 | 2022-07-29 | 河南金百合生物科技股份有限公司 | Preparation method of clostridium butyricum microencapsulated viable bacteria preparation |
| CN113521013A (en) * | 2020-04-20 | 2021-10-22 | 青岛海益康健康科技有限公司 | Embedded probiotic product with planting effect and preparation process thereof |
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