CN101948775A - High-density cultivation of functional bifidobacterium and preparation method of freeze-drying products thereof - Google Patents
High-density cultivation of functional bifidobacterium and preparation method of freeze-drying products thereof Download PDFInfo
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Abstract
The invention relates to a high-density cultivation of functional bifidobacterium and the preparation method of freeze-drying products thereof, which relates to the cultivation of bifidobacterium and the preparation method of the freeze-drying products thereof and solves the problems of less viable bacterium content, large usage, poor fermentation effect, high production and use cost of the freeze-drying preparations. The cultivation comprises the following steps of (1) inoculating the functional bifidobacterium after oxygen resistance acclimatization to an enrichment medium to prepare fermentation liquor; (2) eluting and flocculating; and (3) film concentration processing. The preparation method comprises the following steps of (1) mixing and prefreezing film concentration products and a cryoprotectant to prepare the mixture; (2) prefreezing the mixture, carrying out primary drying and analysis drying, and then crushing, packaging and storing The invention can ensure that the viable bacterium count of the bifidobacterium is 5.4*10<10>CFU/g; the method has the advantages of usage reduction, good fermentation effect, low production cost of the freeze-drying products, simple technology and high activity; and the purpose for directly using can be realized.
Description
Technical field
The present invention relates to the preparation method of bifidus bacillus cultivation and freeze-drying prods thereof.
Background technology
Bifidus bacillus is a flora the most useful in the intestines, can produce lactic acid and acetic acid after the fermentation in the human body intestines, can improve the utilization ratio of calcium, phosphorus, iron, promotes the absorption of iron and vitamins D; The bifidobacterium fermentation lactose produces semi-lactosi, is to constitute cerebronic composition in the cerebral nervous system, with the ramp of baby due hindbrain substantial connection is arranged; Bifidus bacillus can produce the nutritive substance of needed by human such as VITMAIN B1, B2, B6, B12 and L-Ala, Xie Ansuan, aspartic acid and Threonine, has the important trophism that can not be ignored for human body; And have anti-ageing, antianaphylaxis, antitumor, adjust intestinal function and improve effect of nutrition etc.
At present, bifidus bacillus produces, and is mostly to adopt centrifugal spissated mode, has that treatment capacity is less, whizzer is got the shortcoming that the material program is numerous and diverse, the thalline mortality ratio is higher and yield rate is lower, causes that viable bacteria content is little in the product, consumption big, ferment effect is poor; And its product is mainly freeze-dried preparation, and manufacturing cost and use cost is all higher.
Summary of the invention
The objective of the invention is to produce and have that viable bacteria content is little in the product, consumption big, ferment effect is poor in order to solve existing bifidus bacillus, and the equal problem of higher of the manufacturing cost and use cost of its freeze-dried preparation, and provide the preparation method of functional bifid bacterium high-density culture and freeze-drying prods thereof.
The functional bifid bacterium high-density culture is carried out according to the following steps: one, will be inoculated in the enrichment medium of fermentor tank through the functional bifid bacterium of oxytolerant domestication, in temperature is that 35～37 ℃, pH value are to cultivate 15～17h under 6.5～6.8 the condition, fermented liquid; Two, fermented liquid is in the time of 0～20 ℃, add mass concentration with weight such as fermented liquid and be 10% sodium citrate solution or mass concentration and be 10% sodium radio-phosphate,P-32 solution fermented liquid is carried out proteic wash-out, add mass concentration again and be 0.2～2% chitosan and carry out the flocculation of thalline; Three, to use the aperture be that 100～200nm, area are 0.5m to the fermented liquid after the flocculation
2Ceramic membrane, be that 110KPa carries out membrane concentration and handles the cycles of concentration of the fermented liquid to the flocculation and reach 6 at transmembrane pressure, obtain containing the membrane concentration product of functional bifid bacterium, promptly finish the functional bifid bacterium high-density culture; Wherein inoculum size is 2% in the step 1; Enrichment medium is by the skimmed milk powder of 80～120g in the step 1, the trypsin hydrolyzing whey powder of 40～60g, the VITAMIN B4 of the yeast extract paste of the tomato juice of 30～60g, the wort of 30～60g, 10～30g lactose, 0.50～0.70g or yeast powder, 6.8～7.2mg and the L-cysteine hydrochloride of 50～54mg add distilled water and are settled to 1000mL and form; The mode that membrane concentration is handled in the step 3 is two ceramic-film tube series connection.
The preparation method of functional bifid bacterium freeze-drying prods carries out according to the following steps: one, mix with cryoprotectant by weight 1: 1～2 membrane concentration products that will contain functional bifid bacterium, under the condition of 0～10 ℃ of temperature, carry out precooling treatment before 10～12h freezing, mixture; Two, mixture is placed the refrigerator tray of-70 ℃ of refrigerators to carry out pre-freeze, temperature until mixture reaches-45 ℃, keep 30min, speed with 5 ℃/30min is warming up to-28 ℃ then, and keep this temperature to vacuum tightness and reach 0.05MPa and finish the dry stage of trunk, be warming up to 10 ℃ and keep this temperature to vacuum tightness and reach 0.03MPa and finish the parsing-desiccation stage again, taking out the back pulverizes with granulator, with aseptic aluminium foil carton package and place-20 ℃ environment to preserve, promptly finish the preparation of functional bifid bacterium freeze-drying prods again; Wherein the trehalose of the sucrose of the L-glutamic acid of the vitamins C of the glycerine of the skimmed milk powder of cryoprotectant 10～20g, 1～3g, 1～2g, 0.5～1g or glycine, 5g and 3～15g adds distilled water and is settled to the 1000mL composition in the step 1; Mixture places the size of the refrigerator tray of-70 ℃ of refrigerators to be long 40cm * wide 50cm * high 20mm in the step 2.
The viable count that functional bifid bacterium high-density culture among the present invention, gained contain bifidus bacillus in the membrane concentration product of functional bifid bacterium is 5.4 * 10
10CFU/g, improve more than 10 times than existing viable count, the Bacterium lacticum consumption reduces, only getting 0.005%～0.010% is used with ferment agent for sour milk and can satisfies the requirement that probiotics fermention breast is produced, can be used as fine probiotic bacterium throw type leaven, and ferment effect is good, and used VITAMIN B4 in the enrichment medium has simultaneously increased the survival rate of functional bifid bacterium in the preparation process for follow-up freeze-drying prods.
The preparation method of functional bifid bacterium freeze-drying prods among the present invention, the membrane concentration product of functional bifid bacterium high-density culture gained is processed, reduce the load of freezing treatment like this, simultaneously, cryoprotectant after the optimization also can improve the survival rate of functional bifid bacterium, compares with freeze-dried products to have the low and active high advantage of production cost, reach direct application target, and technology is simple; In addition, functional beneficial natural disposition breast bar freeze-drying prods can also add in the milk powder as auxiliary material among the present invention, and the viable bacteria number average of the bifidus bacillus of probiotic bacterium milk powder is greater than 1.0 * 10 in 12 months quality guaranteed period
6CFU/g.
Embodiment
Technical solution of the present invention is not limited to following cited embodiment, also comprises the arbitrary combination between each embodiment.
Embodiment one: present embodiment functional bifid bacterium high-density culture is carried out according to the following steps: one, will be inoculated in the enrichment medium of fermentor tank through the functional bifid bacterium of oxytolerant domestication, in temperature is that 35～37 ℃, pH value are to cultivate 15～17h under 6.5～6.8 the condition, fermented liquid; Two, fermented liquid is in the time of 0～20 ℃, add mass concentration with weight such as fermented liquid and be 10% sodium citrate solution or mass concentration and be 10% sodium radio-phosphate,P-32 solution fermented liquid is carried out proteic wash-out, add mass concentration again and be 0.2～2% chitosan and carry out the flocculation of thalline; Three, to use the aperture be that 100～200nm, area are 0.5m to the fermented liquid after the flocculation
2Ceramic membrane, be that 110KPa carries out membrane concentration and handles the cycles of concentration of the fermented liquid to the flocculation and reach 6 at transmembrane pressure, obtain containing the membrane concentration product of functional bifid bacterium, promptly finish the functional bifid bacterium high-density culture; Wherein inoculum size is 2% in the step 1; Enrichment medium is by the skimmed milk powder of 80～120g in the step 1, the trypsin hydrolyzing whey powder of 40～60g, the VITAMIN B4 of the yeast extract paste of the tomato juice of 30～60g, the wort of 30～60g, 10～30g lactose, 0.50～0.70g or yeast powder, 6.8～7.2mg and the L-cysteine hydrochloride of 50～54mg add distilled water and are settled to 1000mL and form; The mode that membrane concentration is handled in the step 3 is two ceramic-film tube series connection.
The functional bifid bacterium of oxytolerant domestication in the present embodiment step 1, be to isolate functional bifid bacterium from spontaneous fermentation food or human intestine, then it carried out ultraviolet mutagenesis, mutagenic condition is: the 15-25W UV-lamp, bacterial strain suspension shines 2mic apart from ultraviolet source 30cm.
Used ceramic membrane can be regenerated in the present embodiment step 3, reclaiming process is: 50 ℃ hot water cleans 5min, mass concentration is 0.5% sodium hydroxide solution cleaning 10min, 50 ℃ hot water cleans 5min, mass concentration is 0.5% nitric acid cleaning 10min, 50 ℃ hot water cleans 5min, promptly finishes ceramic membrane regeneration.
Present embodiment adopts the stream addition to replenish enrichment medium.
Embodiment two: what present embodiment and embodiment one were different is to adopt neutralizing agent to regulate the pH value of enrichment medium in the step 1, and neutralizing agent is that mass concentration is that 10～15% sodium hydroxide, mass concentration are that 10～15% ammoniacal liquor or mass concentration are 20% Na
2CO
3Cultivate in the step 1 in the process of 15～17h and need add lactose, the lactose additional amount is 3～5 times of the neutralizing agent neutral and alkali material molar weight that consumed, adopts the sodium salt that produces in the D301 type resin absorption culturing process simultaneously.Other step and parameter are identical with embodiment one.
Embodiment three: present embodiment is different with embodiment one or two is to be that 35 ℃, pH value are to cultivate 17h under 6.5 the condition in temperature in the step 1, fermented liquid.Other step and parameter are identical with embodiment one or two.
Embodiment four: present embodiment is different with embodiment one or two is to be that 37 ℃, pH value are to cultivate 15h under 6.8 the condition in temperature in the step 1, fermented liquid.Other step and parameter are identical with embodiment one or two.
Embodiment five: present embodiment is different with embodiment one or two is to be that 36 ℃, pH value are to cultivate 16h under 6.6 the condition in temperature in the step 1, fermented liquid.Other step and parameter are identical with embodiment one or two.
Embodiment six: present embodiment is different with one of embodiment one to five be in the step 1 enrichment medium by the skimmed milk powder of 80g, the trypsin hydrolyzing whey powder of 40g, the VITAMIN B4 of the yeast extract paste of the tomato juice of 30g, the wort of 30g, 10g lactose, 0.50g or yeast powder, 6.8mg and the L-cysteine hydrochloride of 50mg add distilled water and are settled to 1000mL and form.Other step and parameter are identical with one of embodiment one to five.
Embodiment seven: present embodiment is different with one of embodiment one to five be in the step 1 enrichment medium by the skimmed milk powder of 120g, the trypsin hydrolyzing whey powder of 60g, the VITAMIN B4 of the yeast extract paste of the tomato juice of 60g, the wort of 60g, 30g lactose, 0.70g or yeast powder, 7.2mg and the L-cysteine hydrochloride of 54mg add distilled water and are settled to 1000mL and form.Other step and parameter are identical with one of embodiment one to five.
Embodiment eight: present embodiment is different with one of embodiment one to five be in the step 1 enrichment medium by the skimmed milk powder of 90～110g, the trypsin hydrolyzing whey powder of 45～55g, the VITAMIN B4 of the yeast extract paste of the tomato juice of 40～50g, the wort of 40～50g, 15～25g lactose, 0.55～0.65g or yeast powder, 6.9～7.1mg and the L-cysteine hydrochloride of 51～53mg add distilled water and are settled to 1000mL and form.Other step and parameter are identical with one of embodiment one to five.
Embodiment nine: present embodiment is different with one of embodiment one to five be in the step 1 enrichment medium by the skimmed milk powder of 100g, the trypsin hydrolyzing whey powder of 50g, the VITAMIN B4 of the yeast extract paste of the tomato juice of 45g, the wort of 45g, 20g lactose, 0.60g or yeast powder, 7mg and the L-cysteine hydrochloride of 52mg add distilled water and are settled to 1000mL and form.Other step and parameter are identical with one of embodiment one to five.
Embodiment ten: present embodiment is different with one of embodiment one to nine is that to add mass concentration in the step 2 be that 0.2% chitosan carries out the flocculation of thalline.Other step and parameter are identical with one of embodiment one to nine.
Embodiment 11: present embodiment is different with one of embodiment one to nine is that to add mass concentration in the step 2 be that 1% chitosan carries out the flocculation of thalline.Other step and parameter are identical with one of embodiment one to nine.
Embodiment 12: present embodiment is different with one of embodiment one to nine is that to add mass concentration in the step 2 be that 0.5～1.5% chitosan carries out the flocculation of thalline.Other step and parameter are identical with one of embodiment one to nine.
Embodiment 13: present embodiment is different with one of embodiment one to nine is that to add mass concentration in the step 2 be that 2% chitosan carries out the flocculation of thalline.Other step and parameter are identical with one of embodiment one to nine.
Embodiment 14: the preparation method of present embodiment functional bifid bacterium freeze-drying prods carries out according to the following steps: one, mix with cryoprotectant by weight 1: 1～2 membrane concentration products that will contain functional bifid bacterium, under the condition of 0～10 ℃ of temperature, carry out precooling treatment before 10～12h freezing, mixture; Two, mixture is placed the refrigerator tray of-70 ℃ of refrigerators to carry out pre-freeze, temperature until mixture reaches-45 ℃, keep 30min, speed with 5 ℃/30min is warming up to-28 ℃ then, and keep this temperature to vacuum tightness and reach 0.05MPa and finish the dry stage of trunk, be warming up to 10 ℃ and keep this temperature to vacuum tightness and reach 0.03MPa and finish the parsing-desiccation stage again, taking out the back pulverizes with granulator, with aseptic aluminium foil carton package and place-20 ℃ environment to preserve, promptly finish the preparation of functional bifid bacterium freeze-drying prods again; Wherein the trehalose of the sucrose of the L-glutamic acid of the vitamins C of the glycerine of the skimmed milk powder of cryoprotectant 10～20g, 1～3g, 1～2g, 0.5～1g or glycine, 5g and 3～15g adds distilled water and is settled to the 1000mL composition in the step 1; Mixture places the size of the refrigerator tray of-70 ℃ of refrigerators to be long 40cm * wide 50cm * high 20mm in the step 2.
Embodiment 15: what present embodiment and embodiment 14 were different is to mix with cryoprotectant by weight the membrane concentration product that will contain functional bifid bacterium at 1: 1 in the step 1; under the condition of 0 ℃ of temperature, carry out precooling treatment before 12h freezing, mixture.Other step and parameter are identical with embodiment 14.
Embodiment 16: what present embodiment and embodiment 14 were different is to mix with cryoprotectant by weight the membrane concentration product that will contain functional bifid bacterium at 1: 2 in the step 1; under the condition of 10 ℃ of temperature, carry out precooling treatment before 10h freezing, mixture.Other step and parameter are identical with embodiment 14.
Embodiment 17: what present embodiment and embodiment 14 were different is to mix with cryoprotectant by weight the membrane concentration product that will contain functional bifid bacterium at 1: 1.5 in the step 1; under the condition of 5 ℃ of temperature, carry out precooling treatment before 11h freezing, mixture.Other step and parameter are identical with embodiment 14.
Embodiment 18: present embodiment is different with one of embodiment 14 to 17 is that the trehalose of the sucrose of the L-glutamic acid of vitamins C, 0.5g of glycerine, the 1g of skimmed milk powder, the 1g of cryoprotectant 10g in the step 1 or glycine, 5g and 3g adds distilled water and is settled to 1000mL and forms.Other step and parameter are identical with one of embodiment 14 to 17.
Embodiment 19: present embodiment is different with one of embodiment 14 to 17 is that the trehalose of the sucrose of the L-glutamic acid of vitamins C, 1g of glycerine, the 2g of skimmed milk powder, the 3g of cryoprotectant 20g in the step 1 or glycine, 5g and 15g adds distilled water and is settled to 1000mL and forms.Other step and parameter are identical with one of embodiment 14 to 17.
Embodiment 20: present embodiment is different with one of embodiment 14 to 17 is that the trehalose of the sucrose of the L-glutamic acid of vitamins C, 0.6～0.9g of glycerine, the 1.2～1.8g of skimmed milk powder, the 1.5～2.5g of cryoprotectant 12～18g in the step 1 or glycine, 5g and 5～10g adds distilled water and is settled to 1000mL and forms.Other step and parameter are identical with one of embodiment 14 to 17.
Embodiment 21: present embodiment is different with one of embodiment 14 to 17 is that the trehalose of the sucrose of the L-glutamic acid of vitamins C, 0.8g of glycerine, the 1.5g of skimmed milk powder, the 2g of cryoprotectant 15g in the step 1 or glycine, 5g and 8g adds distilled water and is settled to 1000mL and forms.Other step and parameter are identical with one of embodiment 14 to 17.
Embodiment 22: present embodiment functional bifid bacterium high-density culture is carried out according to the following steps: one, will be inoculated in the enrichment medium of fermentor tank through the functional bifid bacterium of oxytolerant domestication, in temperature is that 36 ℃, pH value are to cultivate 16h under 6.6 the condition, fermented liquid; Two, fermented liquid is in the time of 10 ℃, add mass concentration with weight such as fermented liquid and be 10% sodium citrate solution or mass concentration and be 10% sodium radio-phosphate,P-32 solution fermented liquid is carried out proteic wash-out, add mass concentration again and be 1% chitosan and carry out the flocculation of thalline; Three, the fermented liquid after the flocculation uses the aperture to be 0.5m as 150nm, area
2Ceramic membrane, be that 110KPa carries out the fermented liquid cycles of concentration that membrane concentration handles to the flocculation and reaches 6 at transmembrane pressure, obtain containing the membrane concentration product of functional bifid bacterium, promptly finish the functional bifid bacterium high-density culture; Wherein inoculum size is 2% in the step 1; Enrichment medium is by the skimmed milk powder of 100g in the step 1, the trypsin hydrolyzing whey powder of 50g, the VITAMIN B4 of the yeast extract paste of the tomato juice of 45g, the wort of 45g, 20g lactose, 0.60g or yeast powder, 6.8mg and the L-cysteine hydrochloride of 52mg add distilled water and are settled to 1000mL and form; The mode that membrane concentration is handled in the step 3 is two ceramic-film tube series connection.
The preparation method of functional bifid bacterium freeze-drying prods carries out according to the following steps: one, mix with cryoprotectant by weight the membrane concentration product that will contain functional bifid bacterium at 1: 1.5, under the condition of 6 ℃ of temperature, carry out precooling treatment before 12h freezing, mixture; Two, mixture is placed the refrigerator tray of-70 ℃ of refrigerators to carry out pre-freeze, temperature until mixture reaches-45 ℃, keep 30min, speed with 5 ℃/30mi n is warming up to-28 ℃ then, and keep this temperature to vacuum tightness and reach 0.05MPa and finish the dry stage of trunk, be warming up to 10 ℃ and keep this temperature to vacuum tightness and reach 0.03MPa and finish the parsing-desiccation stage again, taking out the back pulverizes with granulator, with aseptic aluminium foil carton package and place-20 ℃ environment to preserve, promptly finish the preparation of functional bifid bacterium freeze-drying prods again; Wherein the trehalose of the sucrose of the L-glutamic acid of the vitamins C of the glycerine of the skimmed milk powder of cryoprotectant 15g, 2g, 1.5g, 0.8g or glycine, 5g and 10g adds distilled water and is settled to the 1000mL composition in the step 1; Mixture places the size of the refrigerator tray of-70 ℃ of refrigerators to be long 40cm * wide 50cm * high 20mm in the step 2.
Obtained freeze-drying product in the present embodiment, wherein viable count is 5.4 * 10 after tested
10CFU/g, the amount that is used with 0.005%-0.010% with ferment agent for sour milk is inoculated in sterilization skimming milk and the fresh milk, condition bottom fermentation 3.5h at 41～43 ℃, solid and the 70 ° of T of product acid of curdling, structural state, excellent flavor, the viable bacteria number average of probiotic bacterium is greater than 1.0 * 10 in the quality guaranteed period of 21 days sour milks
6CFU/g can be used as fine probiotic bacterium throw type leaven; In addition, can also add in the milk powder as auxiliary material, the viable bacteria number average of the bifidus bacillus of probiotic bacterium milk powder is greater than 1.0 * 10 in 12 months quality guaranteed period
6CFU/g.
Claims (10)
1. functional bifid bacterium high-density culture, it is characterized in that the functional bifid bacterium high-density culture carries out according to the following steps: one, will be inoculated in the enrichment medium of fermentor tank through the functional bifid bacterium of oxytolerant domestication, in temperature is that 35～37 ℃, pH value are to cultivate 15～17h under 6.5～6.8 the condition, fermented liquid; Two, fermented liquid is in the time of 0～20 ℃, add mass concentration with weight such as fermented liquid and be 10% sodium citrate solution or mass concentration and be 10% sodium radio-phosphate,P-32 solution fermented liquid is carried out proteic wash-out, add mass concentration again and be 0.2～2% chitosan and carry out the flocculation of thalline; Three, to use the aperture be that 100～200nm, area are 0.5m to the fermented liquid after the flocculation
2Ceramic membrane, be that 110KPa carries out membrane concentration and handles the cycles of concentration of the fermented liquid to the flocculation and reach 6 at transmembrane pressure, obtain containing the membrane concentration product of functional bifid bacterium, promptly finish the functional bifid bacterium high-density culture; Wherein inoculum size is 2% in the step 1; Enrichment medium is by the skimmed milk powder of 80～120g in the step 1, the trypsin hydrolyzing whey powder of 40～60g, the VITAMIN B4 of the yeast extract paste of the tomato juice of 30～60g, the wort of 30～60g, 10～30g lactose, 0.50～0.70g or yeast powder, 6.8～7.2mg and the L-cysteine hydrochloride of 50～54mg add distilled water and are settled to 1000mL and form; The mode that membrane concentration is handled in the step 3 is two ceramic-film tube series connection.
2. functional bifid bacterium high-density culture according to claim 1, it is characterized in that in the step 1 adopting neutralizing agent to regulate the pH value of enrichment medium, neutralizing agent is that mass concentration is that 10～15% sodium hydroxide, mass concentration are that 10～15% ammoniacal liquor or mass concentration are 20% Na
2CO
3Cultivate in the step 1 in the process of 15～17h and need add lactose, the lactose additional amount is 3～5 times of the neutralizing agent neutral and alkali material molar weight that consumed, adopts the sodium salt that produces in the D301 type resin absorption culturing process simultaneously.
3. functional bifid bacterium high-density culture according to claim 1 and 2 is characterized in that in the step 1 in temperature being that 36 ℃, pH value are to cultivate 16h under 6.6 the condition, fermented liquid.
4. functional bifid bacterium high-density culture according to claim 3, it is characterized in that enrichment medium is by the skimmed milk powder of 90～110g in the step 1, the trypsin hydrolyzing whey powder of 45～55g, the VITAMIN B4 of the yeast extract paste of the tomato juice of 40～50g, the wort of 40～50g, 15～25g lactose, 0.55～0.65g or yeast powder, 6.9～7.1mg and the L-cysteine hydrochloride of 51～53mg add distilled water and are settled to 1000mL and form.
5. functional bifid bacterium high-density culture according to claim 3, it is characterized in that enrichment medium is by the skimmed milk powder of 100g in the step 1, the trypsin hydrolyzing whey powder of 50g, the VITAMIN B4 of the yeast extract paste of the tomato juice of 45g, the wort of 45g, 20g lactose, 0.60g or yeast powder, 7mg and the L-cysteine hydrochloride of 52mg add distilled water and are settled to 1000mL and form.
6. according to claim 4 or 5 described functional bifid bacterium high-density culture, it is characterized in that adding in the step 2 mass concentration and be 0.5～1.5% chitosan and carry out the flocculation of thalline.
7. according to claim 4 or 5 described functional bifid bacterium high-density culture, it is characterized in that adding in the step 2 mass concentration and be 2% chitosan and carry out the flocculation of thalline.
8. the method for preparing the gained functional bifid bacterium freeze-drying prods of functional bifid bacterium high-density culture as claimed in claim 1, the preparation method who it is characterized in that the functional bifid bacterium freeze-drying prods carries out according to the following steps: one, mix with cryoprotectant by weight 1: 1～2 membrane concentration products that will contain functional bifid bacterium, under the condition of 0～10 ℃ of temperature, carry out precooling treatment before 10～12h freezing, mixture; Two, mixture is placed the refrigerator tray of-70 ℃ of refrigerators to carry out pre-freeze, temperature until mixture reaches-45 ℃, keep 30min, speed with 5 ℃/30min is warming up to-28 ℃ then, and keep this temperature to vacuum tightness and reach 0.05MPa and finish the dry stage of trunk, be warming up to 10 ℃ and keep this temperature to vacuum tightness and reach 0.03MPa and finish the parsing-desiccation stage again, taking out the back pulverizes with granulator, with aseptic aluminium foil carton package and place-20 ℃ environment to preserve, promptly finish the preparation of functional bifid bacterium freeze-drying prods again; Wherein the trehalose of the sucrose of the L-glutamic acid of the vitamins C of the glycerine of the skimmed milk powder of cryoprotectant 10～20g, 1～3g, 1～2g, 0.5～1g or glycine, 5g and 3～15g adds distilled water and is settled to the 1000mL composition in the step 1; Mixture places the size of the refrigerator tray of-70 ℃ of refrigerators to be long 40cm * wide 50cm * high 20mm in the step 2.
9. the preparation method of functional bifid bacterium freeze-drying prods according to claim 8; it is characterized in that mixing with cryoprotectant by weight the membrane concentration product that will contain functional bifid bacterium at 1: 1.5 in the step 1; under the condition of 5 ℃ of temperature, carry out precooling treatment before 11h freezing, mixture.
10. according to Claim 8 or the preparation method of 9 described functional bifid bacterium freeze-drying prods, the trehalose that it is characterized in that the sucrose of the L-glutamic acid of vitamins C, 0.8g of glycerine, the 1.5g of skimmed milk powder, the 2g of cryoprotectant 15g in the step 1 or glycine, 5g and 8g adds distilled water and is settled to 1000mL and forms.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103898018A (en) * | 2014-03-28 | 2014-07-02 | 北京和美科盛生物技术有限公司 | High-density culture method of bifidobacterium lactis and preparation method of freeze-dried powder of bifidobacterium lactis |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103898018A (en) * | 2014-03-28 | 2014-07-02 | 北京和美科盛生物技术有限公司 | High-density culture method of bifidobacterium lactis and preparation method of freeze-dried powder of bifidobacterium lactis |
| CN103898018B (en) * | 2014-03-28 | 2016-05-25 | 北京科拓恒通生物技术开发有限公司 | A kind of high-density cultivation method of bifidobacterium lactis and the preparation of freeze-dried vaccine powder thereof |
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