CN101919879B - Method for preparing probiotics preparation by taking attapulgite as carrier - Google Patents
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- CN101919879B CN101919879B CN2010102488365A CN201010248836A CN101919879B CN 101919879 B CN101919879 B CN 101919879B CN 2010102488365 A CN2010102488365 A CN 2010102488365A CN 201010248836 A CN201010248836 A CN 201010248836A CN 101919879 B CN101919879 B CN 101919879B
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Abstract
The invention discloses a method for preparing a probiotics preparation by taking attapulgite as a carrier, comprising the following steps of: firstly, purifying the attapulgite; secondly, adsorbing a probiotics suspension to the surface of the attapulgite under a certain condition; and finally, preparing the probiotics preparation by vacuum freeze-drying. By using the attapulgite to adsorb the probiotics to prepare the probiotics preparation, the preparation method is simple; and the body of the probiotics preparation has high survival rate and long survival time in the freeze-drying process and in the intestinal canal, and can be well absorbed by human body.
Description
Technical field
The present invention relates to the preparation method of probiotics preparation, being specifically related to a kind of is the preparation method of the probiotics preparation of carrier with recessed soil.
Background technology
Probiotic bacterium is called probiotics, probiotic, live bacteria agent or probiotics again.Be meant the active bacteria formulation and the meta-bolites thereof that can promote colony balance in the enteron aisle, the host played beneficial effect; Clinically; Probiotics preparation is mainly used in prevention and treats various gastrointestinal tract disease, also can be used for the assisting therapy of hepatopathy, is hepatocellular regeneration and the good interior environment of reparation creation.
Probiotic bacterium comprises bifidus bacillus, thermophilus streptococcus, Lactobacterium acidophilum, lactobicillus bulgaricus, lactobacillus rhamnosus, subtilis, streptococcus faecium, faecalis, leukonid, sheet coccus, Bacillus cereus and bacillus licheniformis etc.
Research of probiotic bacterium and utilization, more than ten country's researchs are comparatively specialized and systemic in Japan, France, the U.S., Germany, Russia etc.At present in the world the research of probiotic bacteriums such as milk-acid bacteria is seemed very active; The product consumption achievement is surprising; At a rough estimate, the year product total output value in the whole world is about about 30,000,000,000 dollars, and the milk-product gross output value of wherein using probiotic bacterium production is above 5,000,000,000 dollars; And growth indices is the rising developing state rising year by year.Milk-product as Japan utilizes probiotic bacterium to make have successfully been squeezed into international milk products beverage market, have built 18 tame source mill in 13 countries, and whole world people's every days at least 2,500 ten thousand is drinking this product at present.
Recessed soil is claimed attapulgite clay, attapulgite etc. again; Be to be a kind of clay mineral of staple with attapulgite (Attapulgite); Staple is polygorskite (Palygorskite) or sloping thin white silk used in ancient China stone, is a kind of rich magnesium silicate, belongs to 2: 1 type layer chain crystalline structure; Because microtexture and character that itself is unique; Give recessed soil many premium propertiess,, thereby be widely used in fields such as chemical industry, building materials, medicine, agricultural and environment protection like adsorptivity, colloidality, cohesiveness, catalytic etc.Do not utilize the report of recessed soil but see so far as the probiotic bacterium carrier.
Summary of the invention
The objective of the invention is to: providing a kind of is the preparation method of the probiotics preparation of carrier with recessed soil, utilizes recessed soil absorption probiotic bacterium to prepare active bacteria formulation, improves the survival rate that probiotic bacterium arrives enteron aisle, and prolongs the survival time of viable bacteria greatly.
Technical solution of the present invention is: at first be the recessed soil of purifying, then the probiotics bacterial suspension be adsorbed to recessed soil surface under certain conditions, last vacuum lyophilization is prepared into probiotics preparation.
Preparing method of the present invention comprises following concrete steps:
The first step, the purifying of recessed soil: the deionized water that adopts specific conductivity to be lower than 5 μ s/cm in recessed soil soaks 20～28h, and the mass ratio of recessed soil and deionized water is 10～20: 1; Whisking appliance is with 1000～1500rpm rotating speed restir, 20～28h, and standing demix is got the recessed soil suspension in middle layer; 5～20% amount with recessed soil suspension quality continues to add above-mentioned deionized water, stirs, layering operates repetition 2～3 times equally, in 105～120 ℃ of oven dry; Grind, sieve, it is subsequent use to get the recessed native fine powder of 200 order to 140 orders;
Second step; The preparation of probiotics bacterial suspension: bifidus bacillus, Lactobacterium acidophilum, thermophilus streptococcus, lactobicillus bulgaricus, lactobacillus rhamnosus, streptococcus faecium or bacillus licheniformis are cultured to the logarithmic phase later stage by conventional means; Collect thalline; Adopt the SPSS washing to remove substratum, and cell concentration is adjusted to 10 with SPSS
9～10
10CFU/ml is subsequent use;
The 3rd step; Recessed soil absorption probio: 10L probiotics bacterial suspension adds the recessed native fine powder of 100～1000g and constitutes absorption system; Regulate 20～35 ℃ of absorption system pH3～6, temperature; Adsorbed 20～50 minutes; Adopt the concussion method to increase contacting of recessed soil and probio during absorption, concussion speed is 90～180rpm;
In the 4th step, vacuum lyophilization: the absorption system of step 3 moves into freeze drier lyophilize 20～28 hours rapidly again in-40～-75 ℃ of pre-freezes 2～3 hours, probiotics preparation, preparation water cut≤2% detects that viable count is 10 in the live bacteria agent
9～10
11CFU/g.
Wherein, bifidus bacillus culture condition:
Substratum: peptone 15g, glucose 15g, yeast extract paste 4g, Carnis Bovis seu Bubali cream 4g, NaCl 5g, Na
2HPO
42.5g, zero(ppm) water 1000ml, pH7; 35 ℃ leave standstill cultivation 22 hours.
Wherein, Lactobacterium acidophilum culture condition:
Substratum: peptone 10g, Carnis Bovis seu Bubali cream 10g, yeast extract paste 5g, diammonium hydrogen citrate 2g, glucose 20g, tween-80 1ml, CH
3COONa3H
2O 5g, K
2HPO
43H
2O 2g, MgSO
47H
2O 0.58g, MnSO
4H
2O 0.25g, zero(ppm) water 1000ml transfers pH6.2-6.4; 37 ℃ leave standstill cultivation 20 hours.
Wherein, thermophilus streptococcus culture condition: the same Lactobacterium acidophilum of substratum; 40 ℃ leave standstill cultivation 20 hours.
Wherein, lactobicillus bulgaricus culture condition: the same Lactobacterium acidophilum of substratum; 37 ℃ leave standstill cultivation 20 hours.
Wherein, lactobacillus rhamnosus culture condition: the same Lactobacterium acidophilum of substratum.37 ℃ leave standstill cultivation 20 hours.
Wherein, streptococcus faecium culture condition:
Substratum: peptone 2g, Carnis Bovis seu Bubali cream 2g, lactose 2g, zero(ppm) water 1000ml, pH6.8; 37 ℃ of shaking culture 6h, shaking speed is 100rpm.
Wherein, bacillus licheniformis culture condition:
Substratum: glucose 5g, peptone 5g, K
2HPO
42g, MgSO
42g, zero(ppm) water 1000ml, pH 7.0; 37 ℃ of shaking culture 40h, shaking speed is 180rpm.
Preparing method of the present invention is simple, and production cost is low, and probiotic bacterium is high in the survival rate of enteron aisle, and the survival time of viable bacteria is long, and absorption of human body is respond well.
Description of drawings
Fig. 1 is to be the recessed native probiotics preparation of example with the Lactobacterium acidophilum and to be that viable count is relatively in manual simulation's intestines environment for protectant probiotics preparation with skimmed milk powder.
Embodiment
Further specify technical solution of the present invention below in conjunction with specific embodiment, these embodiment can not be interpreted as it is the restriction to technical scheme.
Embodiment 1: prepare probiotics preparation according to following concrete steps:
The first step, the purifying of recessed soil: the deionized water that adopts specific conductivity to be lower than 5 μ s/cm in recessed soil soaks 28h with mass ratio at 10: 1, and whisking appliance is with 1000rpm rotating speed restir 28h; Standing demix is got the recessed soil suspension in middle layer, continues to add above-mentioned deionized water with 5% amount of recessed soil suspension quality; Stirring, layering are operated repetition 2 times equally, in 105 ℃ of oven dry, grind; Sieve, it is subsequent use to get the recessed native fine powder of 200 order to 140 orders;
Second step, the preparation of probiotics bacterial suspension: bifidus bacillus is cultured to the logarithmic phase later stage by conventional means, collects thalline, adopt the SPSS washing to remove substratum, and cell concentration is adjusted to 10 with SPSS
9～10
10CFU/ml is subsequent use; Wherein, bifidus bacillus culture condition: substratum: peptone 15g, glucose 15g, yeast extract paste 4g, Carnis Bovis seu Bubali cream 4g, NaCl 5g, Na
2HPO
42.5g, zero(ppm) water 1000ml, pH7; 35 ℃ leave standstill cultivation 22 hours;
The 3rd step; Recessed soil absorption probiotic bacterium: 10L probiotics bacterial suspension adds the recessed native fine powder of 100g and constitutes absorption system, regulates 35 ℃ of absorption system pH3, temperature, adsorbs 20 minutes; Adopt the concussion method to increase contacting of recessed soil and probiotic bacterium during absorption, concussion speed is 180rpm;
In the 4th step, vacuum lyophilization: the absorption system of step 3 moves into freeze drier lyophilize 20 hours rapidly again in-75 ℃ of pre-freezes 2 hours, probiotics preparation, preparation water cut≤2% detects that viable count is 10 in the live bacteria agent
9CFU/g.
Embodiment 2: prepare probiotics preparation according to following concrete steps:
The first step, the purifying of recessed soil: the deionized water that adopts specific conductivity to be lower than 5 μ s/cm in recessed soil soaks 24h with mass ratio at 15: 1, and whisking appliance is with 1250rpm rotating speed restir 24h; Standing demix is got the recessed soil suspension in middle layer, continues to add above-mentioned deionized water with 12.5% amount of recessed soil suspension quality; Stirring, layering are operated repetition 3 times equally, in 112 ℃ of oven dry, grind; Sieve, it is subsequent use to get the recessed native fine powder of 200 order to 140 orders;
Second step, the preparation of probiotics bacterial suspension: Lactobacterium acidophilum is cultured to the logarithmic phase later stage by conventional means, collects thalline, adopt the SPSS washing to remove substratum, and cell concentration is adjusted to 10 with SPSS
9～10
10CFU/ml is subsequent use; Wherein, Lactobacterium acidophilum culture condition: substratum: peptone 10g, Carnis Bovis seu Bubali cream 10g, yeast extract paste 5g, diammonium hydrogen citrate 2g, glucose 20g, tween-80 1ml, CH
3COONa3H
2O 5g, K
2HPO
43H
2O2g, MgSO
47H
2O 0.58g, MnSO
4H
2O 0.25g, zero(ppm) water 1000ml transfers pH6.2-6.4; 37 ℃ leave standstill cultivation 20 hours;
The 3rd step; Recessed soil absorption probiotic bacterium: 10L probiotics bacterial suspension adds the recessed native fine powder of 550g and constitutes absorption system, regulates 27.5 ℃ of absorption system pH4.5, temperature, and adsorption time is 35 minutes; Adopt the concussion method to increase contacting of recessed soil and probiotic bacterium during absorption, concussion speed is 135rpm;
In the 4th step, vacuum lyophilization: the absorption system of step 3 moved into the freeze drier lyophilize rapidly 24 hours in-58 ℃ of pre-freezes 2.5 hours, probiotics preparation, preparation water cut≤2% detects that viable count is 10 in the live bacteria agent
10CFU/g.
Embodiment 3: prepare probiotics preparation according to following concrete steps:
The first step, the purifying of recessed soil: the deionized water that adopts specific conductivity to be lower than 5 μ s/cm in recessed soil soaks 28h with mass ratio at 20: 1, and whisking appliance is with 1000rpm rotating speed restir 28h; Standing demix is got the recessed soil suspension in middle layer, continues to add above-mentioned deionized water with 20% amount of recessed soil suspension quality; Stirring, layering are operated repetition 3 times equally, in 120 ℃ of oven dry, grind; Sieve, it is subsequent use to get the recessed native fine powder of 200 order to 140 orders;
Second step, the preparation of probiotics bacterial suspension: thermophilus streptococcus is cultured to the logarithmic phase later stage by conventional means, collects thalline, adopt the SPSS washing to remove substratum, and cell concentration is adjusted to 10 with SPSS
9～10
10CFU/ml is subsequent use; Wherein, thermophilus streptococcus culture condition: the same Lactobacterium acidophilum of substratum; 40 ℃ leave standstill cultivation 20 hours.
The 3rd step; Recessed soil absorption probiotic bacterium: 10L probiotics bacterial suspension adds the recessed native fine powder of 1000g and constitutes absorption system, and adjusting absorption system pH6, temperature are 20 ℃, adsorb 50 minutes; Adopt the concussion method to increase contacting of recessed soil and probiotic bacterium during absorption, concussion speed is 90rpm;
In the 4th step, vacuum lyophilization: the absorption system of step 3 moves into freeze drier lyophilize 28 hours rapidly again in-40 ℃ of pre-freezes 3 hours, probiotics preparation, preparation water cut≤2% detects that viable count is 10 in the live bacteria agent
11CFU/g.
Embodiment 4:
The preparation process of probiotics preparation is with embodiment 2, and wherein, probiotic bacterium is a lactobicillus bulgaricus, the lactobicillus bulgaricus culture condition: the same Lactobacterium acidophilum of substratum; 37 ℃ leave standstill cultivation 20 hours.
Embodiment 5:
The preparation process of probiotics preparation is with embodiment 2, and wherein, probiotic bacterium is a lactobacillus rhamnosus, the lactobacillus rhamnosus culture condition: the same Lactobacterium acidophilum of substratum, 37 ℃ leave standstill cultivation 20 hours.
Embodiment 6:
The preparation process of probiotics preparation is with embodiment 1, and wherein, probiotic bacterium is a streptococcus faecium, streptococcus faecium culture condition: substratum: peptone 2g, Carnis Bovis seu Bubali cream 2g, lactose 2g, zero(ppm) water 1000ml, pH6.8; 37 ℃ of shaking culture 6h, shaking speed is 100rpm.
Embodiment 7:
The preparation process of probiotics preparation is with embodiment 3, and wherein, probiotic bacterium is a bacillus licheniformis, bacillus licheniformis culture condition: substratum: glucose 5g, peptone 5g, K
2HPO
42g, MgSO
42g, zero(ppm) water 1000ml, pH 7.0; 37 ℃ of shaking culture 40h, shaking speed is 180rpm.
Experimental result shows; Add probiotics preparation that recessed soil makes than the active bacteria formulation that makes with conventional skimmed milk powder in the vacuum freezing process and the survival rate in manual simulation's gastric environment much higher, in artificial intestines environment, can keep high viable bacteria level for a long time.
Part experimental data such as table 1, table 2 and shown in Figure 1.
The recessed native probiotics preparation of table 1 and relatively with skimmed milk powder probiotics preparation survival rate
The recessed native probiotics preparation of table 2 and be protectant probiotics preparation with skimmed milk powder
In manual simulation's gastric environment (3h) survival rate relatively
Claims (8)
1. being the preparation method of the probiotics preparation of carrier with recessed soil, at first is the recessed soil of purifying, then the probiotics bacterial suspension is adsorbed to recessed soil surface under certain conditions, and last vacuum lyophilization is prepared into probiotics preparation; It is characterized in that this preparation method comprises following concrete steps:
The first step, the purifying of recessed soil: the deionized water that adopts specific conductivity to be lower than 5 μ s/cm in recessed soil soaks 20～28h, and the mass ratio of recessed soil and deionized water is 10～20: 1; Whisking appliance is with 1000～1500rpm rotating speed restir, 20～28h, and standing demix is got the recessed soil suspension in middle layer; 5～20% amount with recessed soil suspension quality continues to add above-mentioned deionized water, stirs, layering operates repetition 2～3 times equally, in 105～120 ℃ of oven dry; Grind, sieve, it is subsequent use to get the recessed native fine powder of 200 order to 140 orders;
Second step; The preparation of probiotics bacterial suspension: bifidus bacillus, Lactobacterium acidophilum, thermophilus streptococcus, lactobicillus bulgaricus, lactobacillus rhamnosus, streptococcus faecium or bacillus licheniformis are cultured to the logarithmic phase later stage by conventional means; Collect thalline; Adopt the SPSS washing to remove substratum, and cell concentration is adjusted to 10 with SPSS
9～10
10CFU/ml is subsequent use;
The 3rd step; Recessed soil absorption probio: 10L probiotics bacterial suspension adds the recessed native fine powder of 100～1000g and constitutes absorption system; Regulate 20～35 ℃ of absorption system pH3～6, temperature; Adsorbed 20～50 minutes; Adopt the concussion method to increase contacting of recessed soil and probio during absorption, concussion speed is 90～180rpm;
In the 4th step, vacuum lyophilization: the absorption system of step 3 moves into freeze drier lyophilize 20～28 hours rapidly again in-40～-75 ℃ of pre-freezes 2～3 hours, probiotics preparation, preparation water cut≤2% detects that viable count is 10 in the live bacteria agent
9～10
11CFU/g.
2. according to claim 1 is the preparation method of the probiotics preparation of carrier with recessed soil, it is characterized in that: wherein, and bifidus bacillus culture condition: substratum: peptone 15g, glucose 15g, yeast extract paste 4g, Carnis Bovis seu Bubali cream 4g, NaCl5g, Na
2HPO
42.5g, zero(ppm) water 1000ml, pH7; 35 ℃ leave standstill cultivation 22 hours.
3. according to claim 1 is the preparation method of the probiotics preparation of carrier with recessed soil, it is characterized in that: wherein, and Lactobacterium acidophilum culture condition: substratum: peptone 10g; Carnis Bovis seu Bubali cream 10g, yeast extract paste 5g, diammonium hydrogen citrate 2g; Glucose 20g, tween-80 1ml, CH
3COONa3H
2O 5g, K
2HPO
43H
2O 2g, MgSO
47H
2O 0.58g, MnSO
4H
2O 0.25g, zero(ppm) water 1000ml transfers pH6.2-6.4; 37 ℃ leave standstill cultivation 20 hours.
According to claim 1 or 3 described be the preparation method of the probiotics preparation of carrier with recessed soil, it is characterized in that: wherein, thermophilus streptococcus culture condition: the same Lactobacterium acidophilum of substratum; 40 ℃ leave standstill cultivation 20 hours.
According to claim 1 or 3 described be the preparation method of the probiotics preparation of carrier with recessed soil, it is characterized in that: wherein, lactobicillus bulgaricus culture condition: the same Lactobacterium acidophilum of substratum; 37 ℃ leave standstill cultivation 20 hours.
According to claim 1 or 3 described be the preparation method of the probiotics preparation of carrier with recessed soil, it is characterized in that: wherein, the lactobacillus rhamnosus culture condition: the same Lactobacterium acidophilum of substratum, 37 ℃ leave standstill and cultivated 20 hours.
7. according to claim 1 is the preparation method of the probiotics preparation of carrier with recessed soil, it is characterized in that: wherein, and streptococcus faecium culture condition: substratum: peptone 2g, Carnis Bovis seu Bubali cream 2g, lactose 2g, zero(ppm) water 1000ml, pH6.8; 37 ℃ of shaking culture 6h, shaking speed is 100rpm.
8. according to claim 1 is the preparation method of the probiotics preparation of carrier with recessed soil, it is characterized in that: wherein, and bacillus licheniformis culture condition: substratum: glucose 5g, peptone 5g, K
2HPO
42g, MgSO
42g, zero(ppm) water 1000ml, pH 7.0; 37 ℃ of shaking culture 40h, shaking speed is 180rpm.
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|---|---|---|---|
| CN2010102488365A CN101919879B (en) | 2010-08-06 | 2010-08-06 | Method for preparing probiotics preparation by taking attapulgite as carrier |
Applications Claiming Priority (1)
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|---|---|---|---|
| CN2010102488365A CN101919879B (en) | 2010-08-06 | 2010-08-06 | Method for preparing probiotics preparation by taking attapulgite as carrier |
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| Publication Number | Publication Date |
|---|---|
| CN101919879A CN101919879A (en) | 2010-12-22 |
| CN101919879B true CN101919879B (en) | 2012-09-05 |
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| CN102266308B (en) * | 2011-07-27 | 2012-10-24 | 淮阴工学院 | Method for preparing odor masking sustained-release garlicin composite micro-capsules |
| CN102515452A (en) * | 2011-12-07 | 2012-06-27 | 江苏金梓环境科技有限公司 | Attapulgite-carried sediment repairing agent and preparation method thereof |
| CN103098649B (en) * | 2013-01-31 | 2014-02-19 | 上海市农业科学院 | Method capable of rapidly evaluating extent of low temperature injury of straw mushroom |
| CN104857519A (en) * | 2015-04-28 | 2015-08-26 | 淮阴工学院 | Method for preparing probiotic preparations with porous starch composite attapulgite serving as protection agents |
| CN104911171A (en) * | 2015-05-25 | 2015-09-16 | 淮阴工学院 | Method for preparing probiotics microcapsule by compounding sodium alginate, gelatin and attapulgite |
| CN105063007B (en) * | 2015-07-16 | 2018-11-23 | 江苏天晟药业股份有限公司 | A kind of recessed native adsorption method to Lactococcus |
| CN105661546A (en) * | 2016-01-18 | 2016-06-15 | 西安木本贞生物科技有限公司 | Oral preparation of probiotics as well as preparation method thereof |
| CN109293000A (en) * | 2018-10-18 | 2019-02-01 | 无锡康立斯科技发展有限公司 | A kind of suspend mode storage application method of suspended biological active filler |
| CN110804569A (en) * | 2019-11-27 | 2020-02-18 | 江苏财经职业技术学院 | Quadruple bacteria direct vat type pickle starter culture and preparation method and application thereof |
| CN112742861B (en) * | 2020-12-18 | 2022-03-01 | 兰州大学 | Remediation method for heavy metal contaminated soil |
| CN114601169B (en) * | 2022-04-11 | 2023-04-21 | 上海明现生物科技有限公司 | Preparation method of ready-to-eat high-activity probiotic freeze-dried powder |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1096416A (en) * | 1993-12-02 | 1994-12-21 | 王厚德 | Soil fodder and special production technology thereof |
-
2010
- 2010-08-06 CN CN2010102488365A patent/CN101919879B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1096416A (en) * | 1993-12-02 | 1994-12-21 | 王厚德 | Soil fodder and special production technology thereof |
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