CN103160490A - Preparation method of lactobacillus johnsonii microcapsule, microcapsule and application of additive - Google Patents
Preparation method of lactobacillus johnsonii microcapsule, microcapsule and application of additive Download PDFInfo
- Publication number
- CN103160490A CN103160490A CN2011104154791A CN201110415479A CN103160490A CN 103160490 A CN103160490 A CN 103160490A CN 2011104154791 A CN2011104154791 A CN 2011104154791A CN 201110415479 A CN201110415479 A CN 201110415479A CN 103160490 A CN103160490 A CN 103160490A
- Authority
- CN
- China
- Prior art keywords
- microcapsule
- lactobacillus johnsonii
- preparation
- kiln
- drying
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention relates to a preparation method of lactobacillus johnsonii microcapsule, the microcapsule, and application of the microcapsule in preparation of an additive; and belongs to technical field of lactobacillus high density fermentation and microcapsule coating. The preparation method helps to achieve drying of lactobacillus johnsonii at room temperature, the preparation process is easy to control, the bacteria survival rate is high, the obtained product is long in storage period, and the storage time can be significantly extended. An ultrasonic spray nozzle is used in the present invention for spraying the material to make capsules and to obtain microcapsules of a uniform size, and the microcapsules are dried under vacuum at room temperature in the atmosphere of a protective gas. The effects of low temperature of low temperature freeze drying and high temperature of high temperature spray drying on lactobacillus johnsonii in the microcapsules are prevented, the damage of temperature for the material in high temperature steam drying process is prevented, product quality is improve, energy consumption and environmental pollution are reduced, and the problem that continuous drying cannot be conducted in the vacuum freeze drying process is also solved.
Description
Technical field
The present invention relates to a kind of preparation method of Lactobacillus johnsonii microcapsule and microcapsule thereof and its application in the preparation fodder additives, belong to milk-acid bacteria high density fermentation and microcapsule coating technique field.
Background technology
Milk-acid bacteria is the resident probiotics in animal body, and it can the antagonism pathogenic micro-organism, regulates animal gastrointestinal microflora balance; Stimulating immune system, strengthening immunity; In addition, milk-acid bacteria can also prevent livestock and poultry diarrhea, improve the livestock and poultry weightening finish, is bringing into play important physiological function in enteron aisle.Along with domestic and international people deepen continuously to understanding and the research of milk-acid bacteria, increasing lactobacillus product nutrition, health care and medical aspect be widely applied.
But, be mainly live bacteria agent take milk-acid bacteria as main probiotics, and most lactic acid bacteria do not produce gemma, mostly belong to obligatory anaerobic bacteria, very easily be subject to the impact of the external environment factors such as temperature, oxygen, moisture and soda acid, the bacterium activity keeping is more difficult.Milk-acid bacteria can be subject to the rugged environments such as hydrochloric acid in gastric juice, bile after entering the digestive tube of animal, to such an extent as to most of viable lactic acid bacteria does not arrive just death of target site.In order to make milk-acid bacteria can play corresponding effect, people screen thalline and directed domestication on the one hand, make it possess good tolerance and resistance, adopt on the other hand some materials and methods that thalline is coated with and processes, can pass through smoothly stomach, small intestine, arrive smoothly large intestine.And the coated milk-acid bacteria of microcapsule is the most effective and most potential a kind of mode.What existing microcapsule preparation method mainly adopted is interfacial polymerization, phase separation method, extrusion process, dual emulsion method and spray-drying process.(1) the existing technique of interfacial polymerization exists operation more complicated, technique to be difficult to the problem of controlling, being difficult to realize scale operation and can't obtaining dry lactobacillus micro-capsule powder-product.(2) extrusion process is the most traditional prevailing method, use at most as the immobilization of material with sodium alginate, but shortcoming is also to be difficult to obtain dry lactobacillus micro-capsule powder-product.(3) existing spray-drying process generally adopts high temperature spray-drying, with single operation, the suspension of thalline and protective material, carrier is carried out spraying drying, obtains the Powdered microcapsule product of drying.But this method exists milk-acid bacteria disperses the high problem of milk-acid bacteria Yin Gaowen mortality ratio in inhomogeneous and preparation process.(4) interfacial polymerization and dual emulsion method all will adopt the linking agent with certain toxicity can the activity of milk-acid bacteria be damaged, and are difficult to be approved widely.
Lactobacillus johnsonii is one of edible lactobacillus bacterial classification of the relevant departments such as China Ministry of Health approval, the related products of Lactobacillus johnsonii has been arranged on market, but be mainly for medical aspect, and feeding aspect is fewer, and this bacterium is not also the fodder additives kind of Ministry of Agriculture's approval so far.
Summary of the invention
The preparation method who the purpose of this invention is to provide a kind of Lactobacillus johnsonii microcapsule, this technique have realized that Lactobacillus johnsonii carries out drying at normal temperatures, and preparation technology is convenient to control, the bacterium survival rate is high, product storage period of making is long.
Another object of the present invention has been to provide microcapsule product and the application thereof that the preparation method of these Lactobacillus johnsonii microcapsule makes, Lactobacillus johnsonii CGMCC No.4926 in this microcapsule product has good benefit and gives birth to effect, compare with other Lactobacillus johnsonii, the characteristic of anti-hydrochloric acid in gastric juice, anti-cholate is preferably arranged.
The objective of the invention is to be achieved through the following technical solutions:
The invention provides a kind of preparation method of Lactobacillus johnsonii microcapsule, the method key step is as follows:
(1) actication of culture: the Lactobacillus johnsonii (Lactobacillus johnsonii) of freezing preservation is inoculated in the MRS liquid nutrient medium, go down to posterity at 30 ℃-40 ℃ of temperature and cultivate 18-24h, so go down to posterity and cultivate 1-2 time, obtain described activation Lactobacillus johnsonii bacterial classification;
(2) preparation of strain fermentating liquid: preparation fermention medium, take fermention medium, take the bacterial classification that step (1) obtains, the weight of bacterial classification is the 1-3% of fermention medium weight, the bacterial classification that weighs up is inoculated in the fermention medium that takes, cultivate 8-10h 30 ℃-40 ℃ of temperature, obtain the Lactobacillus johnsonii strain fermentating liquid;
(3) preparation of microcapsule: preparation microcapsule wall material solution, with the Lactobacillus johnsonii strain fermentating liquid of step (2) preparation with wall material solution in 1: 1-1: 2 ratio is mixed, and gets mixing suspension; Mixing suspension by ultrasonic atomization drying plant spraying drying, is namely got the Lactobacillus johnsonii microcapsule.
Wherein, the compound method of described fermention medium is as follows: take respectively following raw material: 90-120 weight part brown sugar, 8-12 weight part yeast soak powder, 30-50 weight part soy peptone, 3.0-3.5 weight part dipotassium hydrogen phosphate, 13-15 weight part sodium acetate, 0.8-1.0 weight part MgSO
47H
2O, 0.04-0.08 weight part MnSO
45H
2O； Above-mentioned raw materials is added in 1000 weight parts waters, stir, use neutralizing agent that its pH value is adjusted to 6.4-6.6, sterilization is 15 minutes under temperature 115-121 ℃, obtains described fermention medium; Described neutralizing agent is a kind of, two or more the mixing solutions in sodium hydroxide, potassium hydroxide, sodium carbonate, salt of wormwood, and the percent weight in volume concentration of solution is 30%-40%.
Wherein, described preparation microcapsule wall material solution methods is as follows: take the wall material, the percent weight in volume concentration of wall material in solution is 30-40%, after distilled water is heated to 93-95 ℃, the wall material is fully stirred in water or with mulser emulsification 60-75min, mixes, the pH nature, to be cooled to room temperature, obtain described microcapsule wall material solution; Described wall material is any two or two or more the mixing in trehalose, lactose, maltodextrin, W-Gum, Resistant starch, glucose dry powder, skimmed milk powder.
Ultrasonic atomization drying plant of the present invention obtains for applicant invention, and this equipment has solved the problem of the inhomogeneous and drying operation infringement material of existing spray drying unit spraying; And the further problem that solves dry discontinuous work.This equipment, microcapsule capsulating material and technique of the present invention have realized that Lactobacillus johnsonii carries out drying at normal temperatures, and this spray drying device is composed as follows: comprise kiln, ultrasonic nozzle and vacuum pump; The below of described kiln is provided with fluidized-bed, and described fluidized-bed is connected with the bottom of kiln by the material collection pipeline; Described ultrasonic nozzle is arranged on the top of kiln, and is connected with outside material pipe; Described vacuum pump is connected with kiln by pipeline, and described vacuum pump comprises the first vacuum pump and the second vacuum pump, and the first vacuum pump is connected to the top of kiln by the first vacuum pipe, and the second vacuum pump is connected to the bottom of kiln by the second vacuum pipe.
Further, described ultrasonic atomization drying plant forms and also comprises aircooling cabinet, and described aircooling cabinet is used for the ducted aqueous vapor that the condensate vacuum pump is connected with kiln between vacuum pump and kiln; The bottom of described kiln is provided with the backgauge dividing plate, and described backgauge dividing plate is positioned at the pipeline below that is connected with the bottom of kiln; Described kiln also comprises the material collection chamber, and described material collection chamber is positioned at the below of backgauge dividing plate, and the bottom in described material recycle chamber is provided with baffle plate; In the middle part of the outer wall of described kiln, well heater is installed, the outer wall at middle part is provided with the described ultrasonic nozzle of viewing window and is connected with ultrasonic wave controller, and described ultrasonic wave controller is regulated hyperacoustic frequency to control the discharge rate of ultrasonic nozzle.
Excusing from death ripple drying plant of the present invention can be also 201120503311.1 Chinese patent referring to applicant's application number.
In the preparation method of Lactobacillus johnsonii microcapsule of the present invention, that mixing suspension is as follows by ultrasonic atomization drying plant spray-drying process:
(1) open the ultrasonic atomization drying plant, after indoor vacuum pressure to be dried maintains 2.3-3.7kpa, start product pump, utilize distilled water to try spraying, and to adjust flow velocity be 10-25ml/min, the temperature in kiln is 20-30 ℃ of left and right; Fill shielding gas in drying fluidized-bed, temperature maintains 10-15 ℃;
(2) after device parameter is stable, suspension is carried out feeding by product pump, material enters vacuum drying chamber after atomizing by ultrasonic nozzle, namely begins one-level vacuum-drying; Enter the vibrations fluidized-bed, namely carry out secondary vibrations fluidised bed drying, through 20-60min, collect at discharge port, namely obtain the Lactobacillus johnsonii microcapsule.
Wherein, the frequency of described ultrasonic nozzle is 20kHz-30kHz; Described shielding gas is N
2, CO
2In one or both mix.
The preparation method of Lactobacillus johnsonii microcapsule of the present invention, the preferred 35-37 of the culture temperature in the preparation of actication of culture and strain fermentating liquid ℃.
The present invention also provides the preparation method of these Lactobacillus johnsonii microcapsule prepared Lactobacillus johnsonii microcapsule, described Lactobacillus johnsonii (Lactobacillus johnsonii) can be other Lactobacillus johnsonii, but the preferred deposit number of the present invention is the Lactobacillus johnsonii of CGMCC No.4926.
Lactobacillus johnsonii microcapsule of the present invention can be prepared into fodder additives to be added in feed.
Lactobacillus johnsonii of the present invention (Lactobacillus johnsonii) CGMCC No.4926 is that the applicant screens in the kind Radix Polygalae Crotalarioidis of Beijing, the domestication of process hydrochloric acid in gastric juice, biliary salts and seed selection obtain, this strain Lactobacillus johnsonii resistance is more intense, has good acid resistance, artificial gastrointestinal fluid tolerance and cholate tolerance.This bacterial strain is in the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms, preservation date is on June 8th, 2011, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, be called for short CGMCC, preserving number is CGMCC No.4926.
Beneficial effect:
1, technique of the present invention has realized that Lactobacillus johnsonii carries out drying at normal temperatures, and preparation technology is convenient to control, the bacterium survival rate is high, product storage period of making is long, can significantly extend the storage time of product, preserves after 90 days, and survival rate is still more than 40%.2, ultrasonic atomization drying installation used in the present invention adopts ultrasonic nozzle to carry out material spray, has realized that material spray is even, and ultrasonic nozzle can be sent the suspension homogeneous forth, obtains 20-40 μ m small droplets of uniform size; Make the encapsulated complete of the coated or medicine of microcapsule; And utilize vacuum and protective gas to carry out at normal temperatures drying, avoided the high temperature of the low temperature of low-temperature freeze drying and high temperature spray-drying on the impact of Lactobacillus johnsonii activity; Avoid the damage of temperature to material in the high-temperature steam drying process, improved the quality of product, reduced energy consumption and environmental pollution.Further, adopt fluidized-bed to carry out material collection, solved the problem that can not carry out continuously drying in the vacuum lyophilization process.3, Lactobacillus johnsonii CGMCC No.4926 of the present invention has the living effect of good benefit, the characteristic of anti-hydrochloric acid in gastric juice, anti-cholate is preferably arranged; After fermentation, in fermented liquid, viable count can reach 5.0-8.0 * 10
9/ mL, recycling trehalose, maltodextrin, sodium alginate and/or skimmed milk powder adopt that ultrasonic equipment of the present invention atomizes etc. as the wall material, have realized that Lactobacillus johnsonii carries out drying at normal temperatures, and the bacterium survival rate of product can reach more than 90%; And trehalose and maltodextrin are all natural polysaecharides, directly feeding to animal without any toxic side effect.
Description of drawings
Fig. 1 is excusing from death ripple drying equipment structure schematic diagram of the present invention.
Wherein, 1, kiln; 2, ultrasonic nozzle; 3, material pipe; 4, product pump; 5, fluidized-bed; 6, material collection pipeline; 7a, the first vacuum pump; 7b, the second vacuum pump; 8a, the first vacuum pipe; 8b, the second vacuum pipe; 9, aircooling cabinet; 10, backgauge dividing plate; 11, material collection chamber; 12, viewing window.
Embodiment
Following examples only are used for explanation the present invention, but are not to limit by any way protection scope of the present invention.
The present invention's instrument model used is as follows:
Recirculated water machine: Beijing long stream scientific instrument DX-208 of company type;
Air compressor: Hospitality Franchise, Beijing mechanical ﹠ electrical technology Application and Development HF-6100C of company limited type;
Mulser: the SAII-2 of Shanghai Suowei Electromechanical Equipment Co., Ltd. type;
Microscope: OLYMPUS CX31 type;
Trehalose, maltodextrin are asked chemical industry limited liability company available from Beijing essence, and other reagent are all available from Beijing extensive and profound in meaning star biotechnology responsibility company limited.
Excusing from death ripple spray drying device: ultrasonic atomization drying equipment structure of the present invention as shown in Figure 1.This ultrasonic atomization drying plant comprises kiln 1, ultrasonic nozzle 2 and vacuum pump.Ultrasonic nozzle 2 is arranged on the top of kiln 1, and is connected with outside material pipe 3, and material pipe 3 is provided with product pump 4 and connects.Vacuum pump is connected with kiln 1 by pipeline.Adopt ultrasonic nozzle 3 to carry out material spray and realized that material spray is even, make the encapsulated complete of the coated or medicine of microcapsule; And the kiln 1 that adopts vacuum pump to vacuumize before dry materials has been avoided the damage of temperature to material in the high-temperature steam drying process, has improved the quality of product, has reduced energy consumption and environmental pollution.
Wherein, ultrasonic nozzle 2 is connected with ultrasonic wave controller, and ultrasonic wave controller is used for regulating hyperacoustic frequency to control the discharge rate of ultrasonic nozzle 2, to regulate the size of material according to needs of production.In the middle part of the outer wall of kiln 1, well heater is installed, this well heater maintains in the temperature range that is suitable for dry materials the temperature of kiln 1, is generally the room temperature of 20-30 ℃, to guarantee the characteristic of material.The outer wall at kiln 1 middle part is provided with viewing window 12, so that the working condition in the chamber 1 that sees drying.
This equipment is provided with fluidized-bed 5 and carries out material collection in order to realize carrying out continuously drying and material is carried out redrying in the vacuum lyophilization process below kiln 1, fluidized-bed 5 is connected with the bottom of kiln 1 by material collection pipeline 6.Wherein, the bottom of kiln 1 also is provided with backgauge dividing plate 10, and this backgauge dividing plate 10 is positioned at the pipeline below that is connected with the bottom of kiln 1, and this backgauge dividing plate 10 is set to be easy to open and closed structure.Kiln 1 also comprises material collection chamber 11, and material collection chamber 11 is positioned at the below of backgauge dividing plate 10, and the bottom of material collection chamber 11 is provided with and is easy to open and closed baffle plate.When this ultrasonic atomization drying plant carries out spray-dired work, the baffle plate of material collection chamber 11 is closed, backgauge dividing plate 10 in opened condition, material falls into material collection chamber 11, after spraying drying work is completed, close baffle plate that backgauge dividing plate 10 opens material collection chamber 11 to discharge material, close backgauge dividing plate 10 so that kiln 1 keeps vacuum state to enter the dry work of next batch material spray always at this.When connecting fluidized-bed 5, fluidized-bed 5 closely is connected with bottom with kiln 1 by material collection pipeline 6, and the baffle plate of backgauge dividing plate 10 and material collection chamber 11 is in open mode always and carries out continuously drying and discharge for material.
In order to make material be in a kind of suspended state of slow whereabouts, be able to fully drying and cooling to extend time of drying, with the damage of temperature to material in further minimizing high-temperature steam drying process, the vacuum pump of this equipment comprises the first vacuum pump 7a and the second vacuum pump 7b, the first vacuum pump 7a is connected to the top of kiln 1 by the first vacuum pipe 8a, the second vacuum pump 7b is connected to the bottom of kiln 1 by the second vacuum pipe 8b.Wherein, can regulate according to needs of production the power of the first vacuum pump 7a and the second vacuum pump 7b, to regulate the falling velocity of material in kiln 1.
Ultrasonic atomization drying plant of the present invention also comprises aircooling cabinet 9, aircooling cabinet 9 is between vacuum pump and kiln 1, be used for the aqueous vapor that pipeline that the condensate vacuum pump is connected with kiln 1 is extracted out from kiln 1, with the speed of cooling of quickening kiln 1 interior material.Wherein, the first vacuum pump 7a is disc distribution by the first vacuum pipe 8a in aircooling cabinet 9, with fully cooling.
When excusing from death ripple drying plant of the present invention carries out the spraying drying operation, after carrying out emulsification wait spraying with dry material, the first vacuum pump 7a and the second vacuum pump 7b are opened, give vacuumizing of kiln 1, open simultaneously the well heater of kiln 1 outer wall, make temperature maintain 20-30 ℃, during vacuum pressure 2.27-3.35kpa, in frequency modulation 20-30khz scope with ultrasonic nozzle 2, utilize water to spray in advance, and after the power of regulating the first vacuum pump 7a and the second vacuum pump 7b makes air-flow velocity in kiln 1 stable.Treat the equipment operational excellence, squeezing into material sprays, material is introduced into the upper subregion of kiln 1 after ultrasonic nozzle 2 ejections, moisture evaporates rapidly, power difference by the first vacuum pump 7a and the second vacuum pump 7b, make material be in a kind of suspended state of slow whereabouts, extend time of drying with abundant drying, material enters kiln after spraying by ultrasonic nozzle 2, dehydration rapidly in kiln forms microcapsule and concentrates at the lower region of kiln 1; The moisture that evaporates in this process reclaims after being extracted out by the first vacuum pump 7a and the second vacuum pump 7b.After one batch of end, the baffle plate of opening material collection chamber 11 directly enters fluidized-bed 5 with material collection or material and carries out redrying, obtains at last dried microcapsule product.
Wherein, after being evacuated in kiln 1, material only needs coutroi velocity just can enter ultrasonic nozzle 2; 2 pairs of materials of ultrasonic nozzle are sprayed, and the size of the material particulate that not only obtains is even, but also can regulate by regulating ultrasonic frequency the size of particulate; Adopt vacuum-drying, the moisture in material can be reclaimed, reduced the waste at water source; Whole drying process is carried out under normal temperature (20-30 ℃) simultaneously, has protected the characteristic of material, has also reduced energy consumption simultaneously.
A, actication of culture:
The Lactobacillus johnsonii of freezing preservation is inoculated in the MRS liquid nutrient medium, cultivates 18h, the Lactobacillus johnsonii bacterial classification that so goes down to posterity and obtain activating for 1 time at 37 ℃ of temperature.
The preparation of b, substratum:
At first, take respectively following raw material: 90 weight part brown sugar, 8 weight part yeast soak powder, 30 weight part soy peptones, 3 weight part dipotassium hydrogen phosphates, 13 weight part sodium acetates, 0.8 weight part MgSO
47H
2O, 0.04 weight part MnSO
45H
2O； Above-mentioned raw materials is added in 1000 weight parts waters, stir, use 30% sodium hydroxide that its pH value is adjusted to 6.4, then sterilized 15 minutes at 115 ℃ of temperature, obtain described fermention medium;
The preparation of c, strain fermentating liquid:
To use described substratum weighing scale, the activation Lactobacillus johnsonii bacterial classification that ratio according to 1% obtains step a is inoculated in the described substratum that step b obtains, cultivate 12h 37 ℃ of temperature and obtain the Lactobacillus johnsonii strain fermentating liquid, in fermented liquid, viable count is 6.3 * 10
9Cfu/mL.
The preparation of d, microcapsule wall material solution:
The composition of wall material: select trehalose: maltodextrin is 1: 10;
Prepare by weight 30% wall material solution, after distilled water is heated to 93 ℃, with the wall material fully mix in water, pH nature, to be cooled to room temperature, obtain described microcapsule wall material solution.
E, microcapsule preparation:
Method 1 (the inventive method): with above-mentioned fermented liquid c with after wall material solution d mixed by 1: 1, this mixing suspension is carried out uniform atomizing by ultrasonic nozzle, by one-level vacuum-drying and secondary vibrations fluidised bed drying, the step that concrete ultrasonic vacuum spray is done is as follows:
(1) start the ultrasonic atomization drying plant, after indoor vacuum pressure to be dried maintains 2.5kpa, first try spray with distilled water, the adjustment flow velocity is 15ml/min, and the temperature in kiln is 22 ℃ of left and right.Fill shielding gas N in the secondary drying fluidized-bed
2, temperature maintains 12 ℃.
(2) mixed suspension in e after ultrasonic nozzle atomizes, is entered vacuum drying chamber.
(3) to enter vacuum-drying by spraying gun indoor as mixed solution e, just can enter the secondary drying fluidized-bed through the 3s left and right, and this moment, most of moisture was volatilized.After entering the vibrations fluidized-bed, through 30min, just can collect product at the collection discharge port, obtain the Lactobacillus johnsonii microcapsule product.
(4) detect collecting product: after taking the microcapsule product of receiving quantitatively, insert 105 ℃, after 3h, then take weight, obtain water content.
The total viable count of described Lactobacillus johnsonii is the measuring method that adopts flat board that those skilled in the art know to pour into, for example with MRS substratum pour plate counting.
Method 2 (contrast: adopt the high temperature spray-drying method this mixed solution to be sprayed and dry) with above-mentioned fermented liquid c with after wall material solution d mixed by 1: 1, adopt the high temperature spray-drying method this mixed solution to be sprayed and drying, its spray-dired technique is: 160 ℃ of air intakes, 70 ℃ of air-out, flow velocity 0.2L/min, its concrete operation step is as follows:
(1) start spray drying device, with fan starting, regulate the speed of air intake and air-out, treat that inlet temperature is stabilized in 160 ℃, after 70 ℃ of air outlet temperatures, first try spray with distilled water, the adjustment flow velocity is 200ml/min.
(2) treat temperature-stable, after operating steadily, mixed solution by peristaltic pump, is squeezed in drying tower, then flow velocity 180mL/min collects dried material at tower body rewinding mouth.
(3) detect collecting product: after taking the microcapsule product of receiving quantitatively, insert 105 ℃, after 3h, then take weight, obtain water content.
The total viable count of described Lactobacillus johnsonii is the measuring method that adopts flat board that those skilled in the art know to pour into, for example with MRS substratum pour plate counting.
Result: the inventive method: (1) is detected collecting product, and the water content of microcapsule is 8%, bacterium survival rate 90%; (2) under drying, normal temperature environment, preserve after 90 days, the Lactobacillus johnsonii survival rate is 42%.And the survival rate of the survival rate of the high temperature spray-drying method bacterial classification of contrast and the active microcapsule that all obtain than the present invention and active low, its result: (1) is detected collecting product, and the water content of microcapsule is 10%, bacterium survival rate 23%; (2) under drying, normal temperature environment, preserve after 90 days, the Lactobacillus johnsonii survival rate is 10%, has reduced an order of magnitude.
Embodiment 2
A, actication of culture:
Lactobacillus johnsonii (Lactobacillus johnsonii) the CGMCC No.4926 of freezing preservation is inoculated in the MRS liquid nutrient medium, cultivates 20h at 37 ℃ of temperature, so going down to posterity obtains described activation Lactobacillus johnsonii bacterial classification for 2 times.
The preparation of b, substratum:
At first, take respectively following raw material: 115 weight part brown sugar, 10 weight part yeast soak powder, 40 weight part soy peptones, 3.2 weight part dipotassium hydrogen phosphates, 14 weight part sodium acetates, 0.9 weight part MgSO
47H
2O, 0.06 weight part MnSO
45H
2O； In above-mentioned raw materials and 1000 weight parts waters, stir, use 35%NaOH that its pH value is adjusted to 6.6, then sterilized 15 minutes at 121 ℃ of temperature, obtain described fermention medium;
The preparation of c, strain fermentating liquid:
To use described substratum weighing scale, according to weight ratio 2%, the activation Lactobacillus johnsonii bacterial classification that step a) obtains is inoculated at step b) in the described substratum that obtains, cultivate 10h 37 ℃ of temperature and obtain the Lactobacillus johnsonii strain fermentating liquid, in fermented liquid, viable count is 8.0 * 10
9Cfu/mL.
The preparation of d, microcapsule wall material solution:
Select trehalose: W-Gum is 1: 10;
Account for 36% preparation of solution weight by the wall material: after distilled water is heated to 93 ℃, with the wall material fully mix in water, pH nature, to be cooled to room temperature, obtain described microcapsule wall material solution.
E, microcapsule preparation:
Above-mentioned fermented liquid c with after wall material solution d mixed by 1: 1, is carried out uniform atomizing with this mixing suspension by ultrasonic nozzle, and by one-level vacuum-drying and secondary vibrations fluidised bed drying, the step that concrete ultrasonic vacuum spray is done is as follows:
(1) start the ultrasonic atomization drying plant, after indoor vacuum pressure to be dried maintains 2.3kpa, first try spray with distilled water, the adjustment flow velocity is 10ml/min, and the temperature in kiln is 20 ℃ of left and right.Fill N in the secondary drying fluidized-bed
2, temperature maintains 15 ℃.
(2) mixed suspension in e after ultrasonic nozzle atomizes, is entered vacuum drying chamber.
(3) to enter vacuum-drying by spraying gun indoor as mixed solution e, just can enter the secondary drying fluidized-bed through the 3.5s left and right, and this moment, most of moisture was volatilized.After entering the vibrations fluidized-bed, through 20min, just can collect product at the collection discharge port, obtain the Lactobacillus johnsonii microcapsule product.
(4) detect collecting product: after taking the microcapsule product of receiving quantitatively, insert 105 ℃, after 3h, then take weight, obtain water content.
Result: (1) is detected collecting product, and the water content of microcapsule is 6%, bacterium survival rate 92%; (2) under drying, normal temperature environment, preserve after 90 days, the Lactobacillus johnsonii survival rate is 43%.
Embodiment 3
A, actication of culture:
Lactobacillus johnsonii (Lactobacillus johnsonii) the CGMCC No.4926 of freezing preservation is inoculated in the MRS liquid nutrient medium, cultivates 20h at 37 ℃ of temperature, so going down to posterity obtains described activation Lactobacillus johnsonii bacterial classification for 2 times.
The preparation of b, substratum:
At first, take respectively following raw material: 120 weight part brown sugar, 12 weight part yeast soak powder, 50 weight part soy peptones, 3.5 weight part dipotassium hydrogen phosphates, 15 weight part sodium acetates, 1.0 weight part MgSO
47H
2O, 0.08 weight part MnSO
45H
2O； Above-mentioned raw materials is added in 1000 weight parts waters, stir, use 40%NaOH that its pH value is adjusted to 6.6, then sterilized 20 minutes at 115 ℃ of temperature, obtain described fermention medium;
The preparation of c, strain fermentating liquid:
To use described substratum weighing scale, the activation Lactobacillus johnsonii bacterial classification that step a is obtained according to the amount of weight ratio 3% is inoculated in the described substratum that step b obtains, cultivate 8h 37 ℃ of temperature and obtain the Lactobacillus johnsonii strain fermentating liquid, the viable count that contains in fermented liquid is 7.2 * 10
9/ mL.
The preparation of d, microcapsule wall material solution:
The wall material is selected lactose: maltodextrin is 1: 9.
Account for 40% preparation of liquor capacity by the wall material, after distilled water is heated to 93 ℃, with the wall material fully mix in water, pH nature, to be cooled to room temperature, obtain described microcapsule wall material solution.
E, microcapsule are made:
Above-mentioned fermented liquid c with after wall material solution d mixed by 1: 1.2, is carried out uniform atomizing with this mixing suspension by ultrasonic nozzle, and by one-level vacuum-drying and secondary vibrations fluidised bed drying, the step that concrete ultrasonic vacuum spray is done is as follows:
(1) start the ultrasonic atomization drying plant, after indoor vacuum pressure to be dried maintains 3.7kpa, first try spray with distilled water, the adjustment flow velocity is 25ml/min, and the temperature in kiln is 27 ℃ of left and right.Fill shielding gas in the secondary drying fluidized-bed, temperature maintains 15 ℃.
(2) mixed suspension in e after ultrasonic nozzle atomizes, is entered vacuum drying chamber.
(3), to enter vacuum-drying by spraying gun indoor as mixed solution e, just can enter the secondary drying fluidized-bed through 4s, this moment, most of moisture was volatilized.After entering the vibrations fluidized-bed, through 25min, just can collect product at the collection discharge port, obtain the Lactobacillus johnsonii microcapsule product.
(4) detect collecting product:
After taking the microcapsule product of receiving quantitatively, insert 105 ℃, after 3h, then take weight, obtain water content.
Result: (1) is detected collecting product, and the water content of microcapsule is 7%, bacterium survival rate 90%; (2) under drying, normal temperature environment, preserve after 90 days, the Lactobacillus johnsonii survival rate is 45%.
Claims (10)
1. the preparation method of Lactobacillus johnsonii microcapsule is characterized in that the method key step is as follows:
(1) actication of culture: the Lactobacillus johnsonii (Lactobacillus johnsonii) of freezing preservation is inoculated in the MRS liquid nutrient medium, go down to posterity at 30 ℃-40 ℃ of temperature and cultivate 18-24h, so go down to posterity and cultivate 1-2 time, obtain described activation Lactobacillus johnsonii bacterial classification;
(2) preparation of strain fermentating liquid: preparation fermention medium, take fermention medium, take the bacterial classification that step (1) obtains, the weight of bacterial classification is the 1-3% of fermention medium weight, the bacterial classification that weighs up is inoculated in the fermention medium that takes, cultivate 8-10h 30 ℃-40 ℃ of temperature, obtain the Lactobacillus johnsonii strain fermentating liquid;
(3) preparation of microcapsule: preparation microcapsule wall material solution, with the Lactobacillus johnsonii strain fermentating liquid of step (2) preparation with wall material solution in 1: 1-1: 2 ratio is mixed, and gets mixing suspension; Mixing suspension by ultrasonic atomization drying plant spraying drying, is namely got the Lactobacillus johnsonii microcapsule.
2. the preparation method of Lactobacillus johnsonii microcapsule claimed in claim 1, it is characterized in that: the compound method of described fermention medium is as follows: take respectively following raw material: 90-120 weight part brown sugar, 8-12 weight part yeast soak powder, 30-50 weight part soy peptone, 3.0-3.5 weight part dipotassium hydrogen phosphate, 13-15 weight part sodium acetate, 0.8-1.0 weight part MgSO
47H
2O, 0.04-0.08 weight part MnSO
45H
2O； Above-mentioned raw materials is added in 1000 weight parts waters, stir, use neutralizing agent that its pH value is adjusted to 6.4-6.6, sterilization is 15 minutes under temperature 115-121 ℃, obtains described fermention medium; Described neutralizing agent is a kind of, two or more the mixing solutions in sodium hydroxide, potassium hydroxide, sodium carbonate, salt of wormwood, and the percent weight in volume concentration of solution is 30%-40%.
3. the preparation method of Lactobacillus johnsonii microcapsule claimed in claim 1, it is characterized in that: preparation microcapsule wall material solution methods is as follows: take the wall material, the percent weight in volume concentration of wall material in solution is 30-40%, after distilled water is heated to 93-95 ℃, the wall material is fully stirred in water or with mulser emulsification 60-75min, mixes, the pH nature, to be cooled to room temperature, obtain described microcapsule wall material solution; Described wall material is any two or two or more the mixing in trehalose, lactose, maltodextrin, W-Gum, Resistant starch, glucose, skimmed milk powder.
4. the preparation method of Lactobacillus johnsonii microcapsule claimed in claim 1, it is characterized in that: described ultrasonic atomization drying plant is composed as follows: comprise kiln, ultrasonic nozzle and vacuum pump; The below of described kiln is provided with fluidized-bed, and described fluidized-bed is connected with the bottom of kiln by the material collection pipeline; Described ultrasonic nozzle is arranged on the top of kiln, and is connected with outside material pipe; Described vacuum pump is connected with kiln by pipeline, and described vacuum pump comprises the first vacuum pump and the second vacuum pump, and the first vacuum pump is connected to the top of kiln by the first vacuum pipe, and the second vacuum pump is connected to the bottom of kiln by the second vacuum pipe.
5. the preparation method of Lactobacillus johnsonii microcapsule as claimed in claim 4, it is characterized in that: described ultrasonic atomization drying plant forms and also comprises aircooling cabinet, described aircooling cabinet is used for the pipeline that the cooling vacuum pump is connected with kiln between vacuum pump and kiln; The bottom of described kiln is provided with the backgauge dividing plate, and described backgauge dividing plate is positioned at the pipeline below that is connected with the bottom of kiln; Described kiln also comprises the material collection chamber, and described material collection chamber is positioned at the below of backgauge dividing plate, and the bottom in described material recycle chamber is provided with baffle plate; In the middle part of the outer wall of described kiln, well heater is installed, the outer wall at middle part is provided with the described ultrasonic nozzle of viewing window and is connected with ultrasonic wave controller, and described ultrasonic wave controller is regulated hyperacoustic frequency to control the discharge rate of ultrasonic nozzle.
6. the preparation method of Lactobacillus johnsonii microcapsule as claimed in claim 1 is characterized in that: mixing suspension is as follows by ultrasonic atomization drying plant spray-drying process:
(1) open the ultrasonic atomization drying plant, after indoor vacuum pressure to be dried maintains 2.3-3.7kpa, first adopt distilled water to try spray, and to adjust flow velocity be 10-25ml/min, the temperature in kiln is 20-30 ℃ of left and right; Fill shielding gas in drying fluidized-bed, temperature maintains 10-15 ℃, after stable, advance material and begins to carry out spraying drying;
(2) with mixing suspension through ultrasonic nozzle after vacuum drying chamber atomizes, namely begin one-level vacuum-drying; Then enter the vibrations fluidized-bed, namely carry out secondary vibrations fluidised bed drying, through 20-60min, collect at discharge port, namely obtain the Lactobacillus johnsonii microcapsule.
7. the preparation method of Lactobacillus johnsonii microcapsule as claimed in claim 6, it is characterized in that: the frequency of described ultrasonic nozzle is 20kHz-30kHz; Described shielding gas is N
2, CO
2In one or both mix.
8. the preparation method of Lactobacillus johnsonii microcapsule as claimed in claim 1, is characterized in that: the preferred 35-37 of the culture temperature in the preparation of actication of culture and strain fermentating liquid ℃.
9. as the prepared Lactobacillus johnsonii microcapsule of preparation method of the described Lactobacillus johnsonii microcapsule of claim 1-8 any one, it is characterized in that, the deposit number of described Lactobacillus johnsonii (Lactobacillus johnsonii) is: CGMCC No.4926.
10. the application of Lactobacillus johnsonii microcapsule as claimed in claim 9 in the preparation fodder additives.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110415479.1A CN103160490B (en) | 2011-12-13 | 2011-12-13 | The preparation method of Lactobacillus johnsonii microcapsule and the application of its microcapsule and additive |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110415479.1A CN103160490B (en) | 2011-12-13 | 2011-12-13 | The preparation method of Lactobacillus johnsonii microcapsule and the application of its microcapsule and additive |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103160490A true CN103160490A (en) | 2013-06-19 |
| CN103160490B CN103160490B (en) | 2016-02-10 |
Family
ID=48584025
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110415479.1A Active CN103160490B (en) | 2011-12-13 | 2011-12-13 | The preparation method of Lactobacillus johnsonii microcapsule and the application of its microcapsule and additive |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103160490B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104004670A (en) * | 2014-05-29 | 2014-08-27 | 广东省农业科学院动物科学研究所 | Feeding high temperature-resistant microcapsule yeast probiotics preparation as well as preparation method and application thereof |
| CN105533684A (en) * | 2015-12-09 | 2016-05-04 | 沈阳师范大学 | Intestinal tract probiotics microcapsule taking resistant starch as wall material and preparation method |
| CN111387291A (en) * | 2019-02-13 | 2020-07-10 | 南昌大学 | Method for producing grease microcapsule by low-temperature electrostatic spray drying |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101496555A (en) * | 2008-01-31 | 2009-08-05 | 北京大北农科技集团股份有限公司 | Lactobacillus micro-capsule as well as preparation method and use |
| CN101738063A (en) * | 2010-01-18 | 2010-06-16 | 山东天力干燥设备有限公司 | Intermittent drying system and method of multifunctional fluidized bed for freezing, spraying and pelleting |
| CN101914518A (en) * | 2010-08-02 | 2010-12-15 | 天津商业大学 | Method for preparing Lactobacillus casei microcapsule bacterial powder by spray drying |
-
2011
- 2011-12-13 CN CN201110415479.1A patent/CN103160490B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101496555A (en) * | 2008-01-31 | 2009-08-05 | 北京大北农科技集团股份有限公司 | Lactobacillus micro-capsule as well as preparation method and use |
| CN101738063A (en) * | 2010-01-18 | 2010-06-16 | 山东天力干燥设备有限公司 | Intermittent drying system and method of multifunctional fluidized bed for freezing, spraying and pelleting |
| CN101914518A (en) * | 2010-08-02 | 2010-12-15 | 天津商业大学 | Method for preparing Lactobacillus casei microcapsule bacterial powder by spray drying |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104004670A (en) * | 2014-05-29 | 2014-08-27 | 广东省农业科学院动物科学研究所 | Feeding high temperature-resistant microcapsule yeast probiotics preparation as well as preparation method and application thereof |
| CN104004670B (en) * | 2014-05-29 | 2016-03-09 | 广东省农业科学院动物科学研究所 | A kind of feeding thermostable microcapsule yeast probiotics preparation and preparation method thereof and application |
| CN105533684A (en) * | 2015-12-09 | 2016-05-04 | 沈阳师范大学 | Intestinal tract probiotics microcapsule taking resistant starch as wall material and preparation method |
| CN111387291A (en) * | 2019-02-13 | 2020-07-10 | 南昌大学 | Method for producing grease microcapsule by low-temperature electrostatic spray drying |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103160490B (en) | 2016-02-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103156063B (en) | The production technology of feeding lactobacillus microcapsules and microcapsules thereof and application and premix | |
| CN110151795B (en) | Clostridium butyricum viable bacteria preparation and production process thereof | |
| CN103704719B (en) | A kind of preparation method of the probiotic microcapsule with high viable count | |
| CN103602707B (en) | A kind of method and nano zine oxide composite feed additive utilizing lactic acid bacteria biological synthesis of nano-zinc oxide | |
| CN102286417B (en) | High-density Enterococcus faecalis and fermentation culture technology thereof | |
| CN109717481A (en) | A kind of preparation process of coating probiotics | |
| CN101857469B (en) | Preparation process for golf course lawn slow-release bio-organic fertilizer produced by three continuous fermentations | |
| CN102559560B (en) | Bacillus licheniformis for producing lactic acid at high yield after fermentation, and preparation and application of Bacillus licheniformis | |
| CN111534435B (en) | Freeze-drying protective agent capable of improving acid resistance of bifidobacteria and application thereof | |
| CN102409007A (en) | Bacillus microecological preparation and liquid-solid fermentation combining preparation process thereof | |
| CN102283334A (en) | Complex probiotic microcapsule capable of improving intestinal tract micro-ecology of small-sized dogs | |
| CN102102081A (en) | Pure-culture solid-state fermentation apparatus capable of gas-phase inoculation and feeding, and method thereof | |
| CN102559523A (en) | Selenium-rich yeast, selenium-rich yeast hydrolysate and preparation method of the hydrolysate | |
| CN109055276A (en) | A kind of mixed bacteria agent preparation method of liquid | |
| CN104904992A (en) | Method for producing protein feed raw materials by carrying out enzymolysis and fermentation on rapeseed dregs | |
| CN113549574A (en) | Bacillus coagulans and application thereof | |
| CN103160490B (en) | The preparation method of Lactobacillus johnsonii microcapsule and the application of its microcapsule and additive | |
| CN105941835A (en) | Method for preparing bacillus subtilis traditional Chinese medicine feed additive | |
| CN101851121B (en) | Compound bacterial agent for efficiently converting pig excrements and preparation method and application thereof | |
| CN104905013A (en) | Method for producing protein feed raw materials by carrying out enzymolysis and fermentation on white spirit vinasse | |
| CN107950753A (en) | A kind of method of aerobic-anaerobic coupled fermentation production biological feedstuff | |
| CN103555592B (en) | Preparation method of Aspergillus oryzae strain and spore powder | |
| CN104397324B (en) | A kind of Synbiotic feed additive and its application | |
| CN116751683B (en) | Application of plant source peptide in lactobacillus paracasei spray drying | |
| WO2023197743A1 (en) | Method for improving ability of bacillus species to remove red tide algae cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20170510 Address after: 100080 Beijing City, Haidian District Zhongguancun street, No. 27 Zhongguancun building 14 DaBeiNong group Co-patentee after: Yangjiang Chang Nong agriculture and animal husbandry technology Co., Ltd. Patentee after: Beijing Dabeinong Technology Group Co., Ltd. Address before: 100080 Beijing City, Haidian District Zhongguancun street, No. 27 Zhongguancun building 14 DaBeiNong group Patentee before: Beijing Dabeinong Technology Group Co., Ltd. |
|
| TR01 | Transfer of patent right |