CN105602875A - Preparation method of lactic acid bacteria freeze-dried powder suitable for storage at room temperature - Google Patents
Preparation method of lactic acid bacteria freeze-dried powder suitable for storage at room temperature Download PDFInfo
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Abstract
The invention discloses a preparation method of lactic acid bacteria freeze-dried powder suitable for storage at room temperature, comprising the following steps: carrying out fermentation culture on lactic acid bacteria strain liquid to obtain fermentation liquid, carrying out concentrating treatment on the fermentation liquid to obtain mycelium concentrated liquid and fermentation waste liquid, adding a freeze-drying protecting agent into the mycelium concentrated liquid, uniformly mixing, carrying out freeze drying, grinding under a vacuum and nitrogen charged condition and packaging. According to the preparation method disclosed by the invention, lactobacillus casei strain liquid, lactobacillus acidophilus strain liquid or a mixed strain liquid of lactobacillus casei and lactobacillus acidophilus in proportion of 1:1 is used as a strain, the freeze-drying protecting agent containing special contents of polydextrose, maltose, trehalose and the like which are compounded is adopted, water remaining in the freeze-dried powder is effectively restrained, and simultaneously a vacuum and nitrogen charged grinding and crushing technology is combined and can effectively prevent the freeze-dried powder from absorbing environmental water in the crushing process and the water activity of the freeze-dried powder from exceeding 0.15, thereby preparing the lactic acid bacteria freeze-dried powder suitable for storage at room temperature, of which the viable count is above 2*1011 CFU/g, the water content is below 5% and the water activity is below 0.15.
Description
Technical field
The present invention relates to a kind of preparation method of lactic acid bacteria freeze drying powder, particularly a kind of preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits.
Background technology
Lactic acid bacteria is a class fermentable carbohydrate, mainly generates the bacterium general name of lactic acid. Along with lactic acid bacteria deepening continuously and the enhancing of people's health care consciousness to the research of health beneficial effect mechanism, lactic acid bacteria is more and more paid close attention to and is researched and developed, and has been widely used at present in the industries such as nutrition and health care, food, medicine, feed addictive.
Lactic acid bacteria is mainly with the form supply market of freeze-dried powder at present; but lactic acid bacteria is easy to the inactivation that fails while directly carrying out freeze drying; viable bacteria stability is low; and freeze drying protectant can reduce the damage that freezing dry process causes for microbial cell; the biologically active of Cell protection, thus viable bacteria rate effectively improved. The lactic acid bacteria freeze drying protective agent of domestic extensive employing at present has skimmed milk powder, trehalose, lactose, maltodextrin, inulin, tween etc.; The bacterial classification of selecting has Bifidobacterium, moral formula lactobacillus etc. And in fact, freeze drying protectant role can show certain otherness with the composition of the variation of bacterial classification and freeze drying protectant and proportioning.
In addition, lactic acid bacteria freeze drying powder is higher to requirement for environmental conditions, if freeze-dried powder has absorbed the moisture in environment in crushing process, cause the water activity rising of freeze-dried powder to exceed 0.15, this freeze-dried powder is unsuitable for normal temperature and deposits, and has seriously limited its application in the industries such as nutrition and health care, food, medicine, feed addictive.
Therefore, exploitation one can be controlled water activity lower than 0.15, and the preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits needs the problem of solution at present badly.
Summary of the invention
In order to overcome the shortcoming and deficiency of prior art, the object of this invention is to provide a kind of viable count higher than 2 × 1011CFU/g, moisture lower than 5%, water activity is lower than 0.15 the preparation method who is suitable for the lactic acid bacteria freeze drying powder that room temperature deposits, the viable count that the lactic acid bacteria freeze drying powder preparing is deposited 6 months under normal temperature (24 ~ 26 DEG C) does not reduce substantially, and the viable count of depositing under normal temperature (24 ~ 26 DEG C) 1 year approximately declines 25 ~ 50%(viable count from higher than 2 × 1011CFU/g is down to 1 ~ 1.5 × 1011CFU/g）。
Above-mentioned purpose of the present invention is achieved by following technical solution:
A preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits, comprises the steps:
Lactic acid bacteria culturers liquid is carried out to fermented and cultured; obtain zymotic fluid; described zymotic fluid is carried out to concentration; obtain mycelium concentrate and fermented waste fluid, in described mycelium concentrate, add freeze drying protectant, mix; freeze-drying; vacuum-fill nitrogen to grind, packaging, must be suitable for the lactic acid bacteria freeze drying powder that normal temperature is deposited.
Wherein, described lactic acid bacteria culturers liquid is Lactobacillus casei strain liquid, lactobacillus acidophilus strain liquid or strain liquid that both mix according to 1:1.
Wherein, the percent by volume that the lactic acid bacteria culturers liquid inoculum concentration of described fermented and cultured accounts for culture medium is 2% ~ 5%, fermentation temperature is 36 ~ 40 DEG C, fermentation time 20 ~ 25 hours, in fermented and cultured process, automatic powder adding adds ammoniacal liquor, the pH value that maintains fermented and cultured process is controlled at 5.6 ~ 5.8, and the zymotic fluid viable count of fermentation termination is 5 × 1010~1×1011CFU/mL；
Wherein, the preparation of the culture medium of fermented and cultured:
The composition of A, culture medium: enzymolysis skimmed milk powder 4.5wt%, enzymatic hydrolysis of soybean albumen 1.8wt%, yeast extract 1.2wt%, glucose 3.2wt%, sodium chloride 0.2wt%, ascorbic acid 0.1wt%, epsom salt 0.04wt%, manganese sulfate monohydrate 0.01wt%, natrium citricum 0.04wt%, Tween-80 0.1wt%, thiamine 0.01wt%, water 88.8wt%;
The sterilization of B, culture medium: above-mentioned various batchings are dissolved and are scattered in water, and stir 20 ~ 30 minutes under 58 ~ 62 DEG C of conditions of temperature, and the pH value of culture medium is adjusted to 5.6 ~ 5.8, be then warming up to 120 ~ 122 DEG C of insulation sterilizations 28 ~ 32 minutes, be cooled to subsequently 36 ~ 38 DEG C, obtain culture medium.
Wherein, described concentration is that the hollow-fibre membrane that is 0.1 ~ 0.2 μ m by aperture carries out, and the volume cycles of concentration of the mycelium concentrate of concentration gained is 20 ~ 30 times.
Wherein, described freeze drying protectant, by weight percentage, comprises following component:
Skimmed milk powder 2 ~ 4wt%;
Sodium glutamate 2 ~ 4wt%;
Trehalose 5 ~ 8wt%;
Maltitol 2 ~ 5wt%;
Polydextrose 12 ~ 18wt%;
Zymotic fluid carries out the fermented waste fluid 68 ~ 72wt% of concentration generation;
The preparation method of described freeze drying protectant; comprise the steps: said components to dissolve and be scattered in fermented waste fluid according to proportioning, and stir 12 ~ 18 minutes under 58 ~ 62 DEG C of conditions of temperature, be warming up to 120 ~ 122 DEG C of insulation sterilizations 28 ~ 32 minutes; then be cooled to 3 ~ 5 DEG C, obtain freeze drying protectant.
Wherein, in described mycelium concentrate, add the process conditions of freeze drying protectant to be: pH value is adjusted into 6.4 ~ 6.6, weight ratio 1:1 ~ 2 of mycelium concentrate and freeze drying protectant.
Wherein, described freeze-drying refers to that being placed in freeze drier carries out frozen dried, and the exhausted degree vacuum of freeze-drying is 5 ~ 100Pa, and freeze temperature is-45 ~-70 DEG C, freeze-drying time is 42 ~ 48h, the moisture in the freeze-dried material that freeze-drying obtains lower than 5%, water activity is lower than 0.15.
Wherein, the process conditions that described vacuum-fill nitrogen grinds are: the freeze-dried material that freeze-drying is obtained is placed in ball mill cavity, under cavity airtight, vacuumize, vacuum is between-0.09 ~-0.095MPa, then spray into liquid nitrogen, the mass volume ratio example of freeze-dried material and liquid nitrogen is 1:1 ~ 1.5, starts ball mill and grinds, and freeze-dried material is ground to form to 60 ~ 100 object freeze-dried powders; The viable count of obtained freeze-drying powder is higher than 2 × 1011CFU/g。
Wherein, described packaging be by the freeze-dried powder of gained relative humidity lower than 35% environment under, with aluminium film packaging seal.
The present invention compared with prior art, has following beneficial effect:
The preparation method who is suitable for the lactic acid bacteria freeze drying powder that room temperature deposits of the present invention, by adopting Lactobacillus casei strain liquid, lactobacillus acidophilus strain liquid or strain liquid that both mix according to 1:1 as bacterial classification, and adopt the composite freeze drying protectant such as polydextrose, maltose and trehalose that contains certain content, because polydextrose, maltose and trehalose belong to polyhydroxy substance, even if the moisture of freeze-dried powder is higher than 5%, all can effectively fetter the moisture remaining in freeze-dried powder, thereby the water activity of controlling lactic acid bacteria freeze drying powder is lower than 0.15; In addition,, in conjunction with the vacuum after freeze-drying-fill nitrogen to grind technology, can effectively avoid freeze-dried powder moisture of absorbing environmental in crushing process to cause the water activity of freeze-dried powder to rise and exceed 0.15, thereby prepare viable count higher than 2 × 1011CFU/g, moisture lower than 5%, water activity is lower than 0.15 the lactic acid bacteria freeze drying powder that room temperature is deposited that is suitable for, the viable count that the lactic acid bacteria freeze drying powder that this prepares is deposited 6 months under normal temperature (24 ~ 26 DEG C) does not reduce substantially, and the viable count of depositing under normal temperature (24 ~ 26 DEG C) 1 year approximately declines 25 ~ 50%(viable count from higher than 2 × 1011CFU/g is down to 1 ~ 1.5 × 1011CFU/g）。
Detailed description of the invention
Further illustrate the present invention below by detailed description of the invention, following examples are preferably embodiment of the present invention, but embodiments of the present invention are not subject to the restriction of following embodiment.
Embodiment 1:
A preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits, comprises the steps:
(1) preparation of the culture medium of fermented and cultured:
The component of A, culture medium comprises: enzymolysis skimmed milk powder 4.5wt%, enzymatic hydrolysis of soybean albumen 1.8wt%, yeast extract 1.2wt%, glucose 3.2wt%, sodium chloride 0.2wt%, ascorbic acid 0.1wt%, epsom salt 0.04wt%, manganese sulfate monohydrate 0.01wt%, natrium citricum 0.04wt%, Tween-80 0.1wt%, thiamine 0.01wt%, water 88.8wt%;
The sterilization of B, culture medium: said components is dissolved and is scattered in water according to proportioning, and under temperature 60 C condition, stir 25 minutes, and the pH value of culture medium is adjusted to 5.7, be then warming up to 121 DEG C of insulation sterilizations 30 minutes, be cooled to subsequently 37 DEG C, obtain culture medium.
(2) fermented and cultured: the strain liquid mixing according to 1:1 with lactobacillus acidophilus strain liquid through the Lactobacillus casei strain liquid of activating pretreatment is under agitation added and carries out fermented and cultured in culture medium, the percent by volume that strain liquid inoculum concentration accounts for culture medium is 3%, be 37 DEG C at fermentation temperature and carry out high density fermentation cultivation 20 hours, in fermented and cultured process, automatic powder adding adds ammoniacal liquor, the pH value that maintains fermented and cultured process is controlled at 5.7, and the zymotic fluid viable count of fermentation termination is 1 × 1011CFU/mL；
(3) zymotic fluid is concentrated: the hollow-fibre membrane that is 0.1 μ m by aperture by gained zymotic fluid carries out concentration, obtains volume cycles of concentration and be mycelium concentrate and the fermented waste fluid of 25 times.
(4) preparation of freeze drying protectant:
The component of A, freeze drying protectant comprises: the fermented waste fluid 69.5wt% that skimmed milk powder 3wt%, sodium glutamate 2.5wt%, trehalose 6.5wt%, maltitol 3.5wt%, polydextrose 15wt%, step (3) Fermented Condensed produce;
B, said components is dissolved and is scattered in fermented waste fluid according to proportioning, and stir 15 minutes under temperature 60 C condition, be warming up to 121 DEG C of insulation sterilizations 30 minutes, be then cooled to 4 DEG C, obtain freeze drying protectant;
(5) freeze drying (freeze-drying): the freeze drying protectant that mycelium concentrate prepared step (3) is prepared with step (4) mixes according to the ratio of weight ratio 1:1, be placed in freeze drier and carry out freeze drying processing, the exhausted degree vacuum of freeze-drying is 5 ~ 100Pa, freeze temperature is-45 ~-70 DEG C, freeze-drying time is 42 ~ 48h, the moisture in the freeze-dried material that freeze-drying obtains lower than 5%, water activity is lower than 0.15;
(6) vacuum-fill nitrogen to grind: the freeze-dried material that freeze-drying is obtained is placed in ball mill cavity, under cavity airtight, vacuumize, vacuum is-0.09MPa, then spray into liquid nitrogen, the mass volume ratio example of freeze-dried material and liquid nitrogen is 1:3, start ball mill and grind, freeze-dried material is ground to form to 80 object freeze-dried powders;
(7) packaging: by the freeze-dried powder of gained relative humidity lower than 35% environment under, with aluminium film packaging seal, must be suitable for the lactic acid bacteria freeze drying powder that normal temperature is deposited, the viable count of the lactic acid bacteria freeze drying powder of gained is higher than 2 × 1011CFU/g, moisture lower than 5%, water activity is lower than 0.15; The viable count of depositing 6 months under normal temperature (24 ~ 26 DEG C) does not reduce substantially, and the viable count of depositing 1 year under normal temperature (24 ~ 26 DEG C) approximately declines 25%(viable count from higher than 2 × 1011CFU/g is down to 1.5 × 1011CFU/g）。
Embodiment 2:
A preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits, comprises the steps:
(1) preparation of the culture medium of fermented and cultured:
The component of A, culture medium comprises: enzymolysis skimmed milk powder 4.5wt%, enzymatic hydrolysis of soybean albumen 1.8wt%, yeast extract 1.2wt%, glucose 3.2wt%, sodium chloride 0.2wt%, ascorbic acid 0.1wt%, epsom salt 0.04wt%, manganese sulfate monohydrate 0.01wt%, natrium citricum 0.04wt%, Tween-80 0.1wt%, thiamine 0.01wt%, water 88.8wt%;
The sterilization of B, culture medium: said components is dissolved and is scattered in water according to proportioning, and under temperature 60 C condition, stir 25 minutes, and the pH value of culture medium is adjusted to 5.7, be then warming up to 121 DEG C of insulation sterilizations 30 minutes, be cooled to subsequently 37 DEG C, obtain culture medium.
(2) fermented and cultured: carry out fermented and cultured in culture medium under agitation adding through the Lactobacillus casei strain liquid of activating pretreatment, the percent by volume that strain liquid inoculum concentration accounts for culture medium is 5%, be 37 DEG C at fermentation temperature and carry out high density fermentation cultivation 20 hours, in fermented and cultured process, automatic powder adding adds ammoniacal liquor, the pH value that maintains fermented and cultured process is controlled at 5.7, and the zymotic fluid viable count of fermentation termination is 1 × 1011CFU/mL；
(3) zymotic fluid is concentrated: the hollow-fibre membrane that is 0.1 μ m by aperture by gained zymotic fluid carries out concentration, obtains volume cycles of concentration and be mycelium concentrate and the fermented waste fluid of 20 times.
(4) preparation of freeze drying protectant:
The component of A, freeze drying protectant comprises: the fermented waste fluid 71wt% that skimmed milk powder 2wt%, sodium glutamate 2wt%, trehalose 8wt%, maltitol 5wt%, polydextrose 12wt%, step (3) Fermented Condensed produce;
B, said components is dissolved and is scattered in fermented waste fluid according to proportioning, and stir 15 minutes under temperature 60 C condition, be warming up to 121 DEG C of insulation sterilizations 30 minutes, be then cooled to 4 DEG C, obtain freeze drying protectant;
(5) freeze drying (freeze-drying): the freeze drying protectant that mycelium concentrate prepared step (3) is prepared with step (4) mixes according to the ratio of weight ratio 1:1.5, be placed in freeze drier and carry out freeze drying processing, the exhausted degree vacuum of freeze-drying is 5 ~ 100Pa, freeze temperature is-45 ~-70 DEG C, freeze-drying time is 42 ~ 48h, the moisture in the freeze-dried material that freeze-drying obtains lower than 5%, water activity is lower than 0.15;
(6) vacuum-fill nitrogen to grind: the freeze-dried material that freeze-drying is obtained is placed in ball mill cavity, under cavity airtight, vacuumize, vacuum is-0.09MPa, then spray into liquid nitrogen, the mass volume ratio example of freeze-dried material and liquid nitrogen is 1:1, start ball mill and grind, freeze-dried material is ground to form to 60 object freeze-dried powders;
(7) packaging: by the freeze-dried powder of gained relative humidity lower than 35% environment under, with aluminium film packaging seal, must be suitable for the lactic acid bacteria freeze drying powder that normal temperature is deposited, the viable count of the lactic acid bacteria freeze drying powder of gained is higher than 2 × 1011CFU/g, moisture lower than 5%, water activity is lower than 0.15; The viable count of depositing 6 months under normal temperature (24 ~ 26 DEG C) does not reduce substantially, and the viable count of depositing 1 year under normal temperature (24 ~ 26 DEG C) approximately declines 35%(viable count from higher than 2 × 1011CFU/g is down to 1.3 × 1011CFU/g）。
Embodiment 3:
A preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits, comprises the steps:
(1) preparation of the culture medium of fermented and cultured:
The component of A, culture medium comprises: enzymolysis skimmed milk powder 4.5wt%, enzymatic hydrolysis of soybean albumen 1.8wt%, yeast extract 1.2wt%, glucose 3.2wt%, sodium chloride 0.2wt%, ascorbic acid 0.1wt%, epsom salt 0.04wt%, manganese sulfate monohydrate 0.01wt%, natrium citricum 0.04wt%, Tween-80 0.1wt%, thiamine 0.01wt%, water 88.8wt%;
The sterilization of B, culture medium: said components is dissolved and is scattered in water according to proportioning, and under temperature 60 C condition, stir 25 minutes, and the pH value of culture medium is adjusted to 5.7, be then warming up to 121 DEG C of insulation sterilizations 30 minutes, be cooled to subsequently 37 DEG C, obtain culture medium.
(2) fermented and cultured: carry out fermented and cultured in culture medium under agitation adding through the lactobacillus acidophilus strain liquid of activating pretreatment, the percent by volume that strain liquid inoculum concentration accounts for culture medium is 2%, be 37 DEG C at fermentation temperature and carry out high density fermentation cultivation 20 hours, in fermented and cultured process, automatic powder adding adds ammoniacal liquor, the pH value that maintains fermented and cultured process is controlled at 5.7, and the zymotic fluid viable count of fermentation termination is 1 × 1011CFU/mL；
(3) zymotic fluid is concentrated: the hollow-fibre membrane that is 0.1 μ m by aperture by gained zymotic fluid carries out concentration, obtains volume cycles of concentration and be mycelium concentrate and the fermented waste fluid of 30 times.
(4) preparation of freeze drying protectant:
The component of A, freeze drying protectant comprises: the fermented waste fluid 68wt% that skimmed milk powder 4wt%, sodium glutamate 4wt%, trehalose 5wt%, maltitol 2wt%, polydextrose 17wt%, step (3) Fermented Condensed produce;
B, said components is dissolved and is scattered in fermented waste fluid according to proportioning, and stir 15 minutes under temperature 60 C condition, be warming up to 121 DEG C of insulation sterilizations 30 minutes, be then cooled to 4 DEG C, obtain freeze drying protectant;
(5) freeze drying (freeze-drying): the freeze drying protectant that mycelium concentrate prepared step (3) is prepared with step (4) mixes according to the ratio of weight ratio 1:2, be placed in freeze drier and carry out freeze drying processing, the exhausted degree vacuum of freeze-drying is 5 ~ 100Pa, freeze temperature is-45 ~-70 DEG C, freeze-drying time is 42 ~ 48h, the moisture in the freeze-dried material that freeze-drying obtains lower than 5%, water activity is lower than 0.15;
(6) vacuum-fill nitrogen to grind: the freeze-dried material that freeze-drying is obtained is placed in ball mill cavity, under cavity airtight, vacuumize, vacuum is-0.095MPa, then spray into liquid nitrogen, the mass volume ratio example of freeze-dried material and liquid nitrogen is 1:5, start ball mill and grind, freeze-dried material is ground to form to 100 object freeze-dried powders;
(7) packaging: by the freeze-dried powder of gained relative humidity lower than 35% environment under, with aluminium film packaging seal, must be suitable for the lactic acid bacteria freeze drying powder that normal temperature is deposited, the viable count of the lactic acid bacteria freeze drying powder of gained is higher than 2 × 1011CFU/g, moisture lower than 5%, water activity is lower than 0.15; The viable count of depositing 6 months under normal temperature (24 ~ 26 DEG C) does not reduce substantially, and the viable count of depositing 1 year under normal temperature (24 ~ 26 DEG C) approximately declines 50%(viable count from higher than 2 × 1011CFU/g is down to 1 × 1011CFU/g）。
Comparative example 1:
The component of freeze drying protectant comprises: the fermented waste fluid 88wt% that skimmed milk powder 3wt%, sodium glutamate 2.5wt%, trehalose 6.5wt%, step (3) Fermented Condensed produce; Other are with embodiment 1, and the viable count of the lactic acid bacteria freeze drying powder preparing is higher than 2 × 1011CFU/g, moisture is lower than 5%; But owing to there is no polydextrose and maltitol in freeze drying protectant, the water activity of the lactic acid bacteria freeze drying powder preparing is generally higher, is everlasting between 0.3 ~ 0.4, be difficult to accomplish that water activity is lower than 0.15, even be also difficult to realize lower than 0.2; The viable count that this lactic acid bacteria freeze drying powder is deposited 6 months under normal temperature (24 ~ 26 DEG C) does not reduce substantially, but significant decline appears in the viable count of depositing 1 year under normal temperature (24 ~ 26 DEG C), and the 10%(viable count of being down to original viable count is from higher than 2 × 1011CFU/g is down to 2.5 × 1010CFU/g）。
Comparative example 2:
Without the vacuum of step (6)-fill nitrogen to grind; Other are with embodiment 1, and the viable count of the lactic acid bacteria freeze drying powder preparing is higher than 2 × 1011CFU/g, moisture is lower than 5%, but owing to not adopting vacuum-fill nitrogen to grind, in the crushing process after freeze-drying, the water activity of lactic acid bacteria freeze drying powder can be higher, water activity is everlasting more than 0.3, and the height of water activity is the key factor that affects the viable count of storage at normal temperature process, thereby before pulverizing, the water activity of freeze-dried material may be below 0.15, once freeze-dried material is ground into the lactic acid bacteria freeze drying powder of powdery, water activity will directly rise to more than 0.3; The viable count that this lactic acid bacteria freeze drying powder is deposited 3 months under normal temperature (24 ~ 26 DEG C) does not reduce substantially, and the viable count of depositing 6 months under normal temperature (24 ~ 26 DEG C) will there will be slight decline, and the 25%(viable count of being down to original viable count is from higher than 2 × 1011CFU/g is down to 5.0 × 1010CFU/g), but there is significant decline in the viable count of depositing 1 year under normal temperature (24 ~ 26 DEG C), and the 5%(viable count of being down to original viable count is from higher than 2 × 1011CFU/g is down to 1.0 × 1010CFU/g）。
Claims (9)
1. a preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits, is characterized in that, comprises the steps:
Lactic acid bacteria culturers liquid is carried out to fermented and cultured; obtain zymotic fluid; described zymotic fluid is carried out to concentration; obtain mycelium concentrate and fermented waste fluid, in described mycelium concentrate, add freeze drying protectant, mix; freeze-drying; vacuum-fill nitrogen to grind, packaging, must be suitable for the lactic acid bacteria freeze drying powder that normal temperature is deposited.
2. the preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits according to claim 1, is characterized in that, described lactic acid bacteria culturers liquid is Lactobacillus casei strain liquid, lactobacillus acidophilus strain liquid or strain liquid that both mix according to 1:1.
3. the preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits according to claim 1, it is characterized in that, the percent by volume that the lactic acid bacteria culturers liquid inoculum concentration of described fermented and cultured accounts for culture medium is 2% ~ 5%, fermentation temperature is 36 ~ 40 DEG C, fermentation time 20 ~ 25 hours, fermentation pH value is controlled at 5.6 ~ 5.8, and the zymotic fluid viable count of fermentation termination is 5 × 1010~1×1011CFU/mL; Wherein, the composition of culture medium: enzymolysis skimmed milk powder 4.5wt%, enzymatic hydrolysis of soybean albumen 1.8wt%, yeast extract 1.2wt%, glucose 3.2wt%, sodium chloride 0.2wt%, ascorbic acid 0.1wt%, epsom salt 0.04wt%, manganese sulfate monohydrate 0.01wt%, natrium citricum 0.04wt%, Tween-80 0.1wt%, thiamine 0.01wt%, water 88.8wt%.
4. the preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits according to claim 1, it is characterized in that, described concentration is that the hollow-fibre membrane that is 0.1 ~ 0.2 μ m by aperture carries out, and the volume cycles of concentration of the mycelium concentrate of concentration gained is 20 ~ 30 times.
5. the preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits according to claim 1, is characterized in that described freeze drying protectant by weight percentage, comprises following component:
Skimmed milk powder 2 ~ 4wt%;
Sodium glutamate 2 ~ 4wt%;
Trehalose 5 ~ 8wt%;
Maltitol 2 ~ 5wt%;
Polydextrose 12 ~ 18wt%;
Zymotic fluid carries out the fermented waste fluid 68 ~ 72wt% of concentration generation;
The preparation method of described freeze drying protectant; comprise the steps: said components to dissolve and be scattered in fermented waste fluid according to proportioning, and stir 12 ~ 18 minutes under 58 ~ 62 DEG C of conditions of temperature, be warming up to 120 ~ 122 DEG C of insulation sterilizations 28 ~ 32 minutes; then be cooled to 3 ~ 5 DEG C, obtain freeze drying protectant.
6. the preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits according to claim 1; it is characterized in that; describedly in described mycelium concentrate, add the process conditions of freeze drying protectant to be: pH value is adjusted into 6.4 ~ 6.6, weight ratio 1:1 ~ 2 of mycelium concentrate and freeze drying protectant.
7. the preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits according to claim 1, it is characterized in that, described freeze-drying refers to that being placed in freeze drier carries out frozen dried, and the exhausted degree vacuum of freeze-drying is 5 ~ 100Pa, freeze temperature is-45 ~-70 DEG C, and freeze-drying time is 42 ~ 48h.
8. the preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits according to claim 1, it is characterized in that, the process conditions that described vacuum-fill nitrogen grinds are: the freeze-dried material that freeze-drying is obtained is placed in ball mill cavity, under cavity airtight, vacuumize, vacuum, between-0.09 ~-0.095MPa, then sprays into liquid nitrogen, and the mass volume ratio example of freeze-dried material and liquid nitrogen is 1:1 ~ 1.5, start ball mill and grind, freeze-dried material is ground to form to 60 ~ 100 object freeze-dried powders.
9. the preparation method who is suitable for the lactic acid bacteria freeze drying powder that normal temperature deposits according to claim 1, is characterized in that, described packaging be by the freeze-dried powder of gained relative humidity lower than 35% environment under, with aluminium film packaging seal.
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| CN201610191460.6A CN105602875B (en) | 2016-03-30 | 2016-03-30 | A kind of preparation method of lactic acid bacteria freeze drying powder suitable for room temperature storage |
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| CN201610191460.6A CN105602875B (en) | 2016-03-30 | 2016-03-30 | A kind of preparation method of lactic acid bacteria freeze drying powder suitable for room temperature storage |
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| CN106983134A (en) * | 2017-04-10 | 2017-07-28 | 北华大学 | The preparation method of northern Chinese caterpillar Fungus bacterium freeze-dried powder |
| CN108753656A (en) * | 2018-06-19 | 2018-11-06 | 苏州汉德瑞生物工程有限公司 | A kind of lactic acid bacteria and the compound fresh-keeping strain freeze-dried powder and application of bacterium acidi propionici |
| CN109541205A (en) * | 2018-12-07 | 2019-03-29 | 河南省商业科学研究所有限责任公司 | The detection method of lactic acid bacteria in a kind of probiotics |
| CN110373351A (en) * | 2019-07-09 | 2019-10-25 | 华中农业大学 | It a kind of freeze drying protectant and its is applied in preparing lactic acid bacteria freeze drying powder |
| CN110754594A (en) * | 2019-09-30 | 2020-02-07 | 中国热带农业科学院农产品加工研究所 | Active probiotic coconut powder stored at normal temperature and preparation method thereof |
| CN112553128A (en) * | 2020-12-30 | 2021-03-26 | 瞿瀚鹏 | Probiotic freeze-dried powder and preparation method and application thereof |
| CN112779195A (en) * | 2021-03-24 | 2021-05-11 | 湖南农业大学 | Lactobacillus freeze separation and dry powder preparation process |
| US11396559B2 (en) * | 2019-07-31 | 2022-07-26 | Jiangnan University | Amylopectin-based cyclic glucan and method for processing the same |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106983134A (en) * | 2017-04-10 | 2017-07-28 | 北华大学 | The preparation method of northern Chinese caterpillar Fungus bacterium freeze-dried powder |
| CN108753656A (en) * | 2018-06-19 | 2018-11-06 | 苏州汉德瑞生物工程有限公司 | A kind of lactic acid bacteria and the compound fresh-keeping strain freeze-dried powder and application of bacterium acidi propionici |
| CN109541205A (en) * | 2018-12-07 | 2019-03-29 | 河南省商业科学研究所有限责任公司 | The detection method of lactic acid bacteria in a kind of probiotics |
| CN109541205B (en) * | 2018-12-07 | 2022-01-28 | 河南省商业科学研究所有限责任公司 | Method for detecting lactic acid bacteria in probiotics |
| CN110373351A (en) * | 2019-07-09 | 2019-10-25 | 华中农业大学 | It a kind of freeze drying protectant and its is applied in preparing lactic acid bacteria freeze drying powder |
| CN110373351B (en) * | 2019-07-09 | 2021-02-02 | 华中农业大学 | Freeze-drying protective agent and application thereof in preparation of lactic acid bacteria freeze-dried powder |
| US11396559B2 (en) * | 2019-07-31 | 2022-07-26 | Jiangnan University | Amylopectin-based cyclic glucan and method for processing the same |
| CN110754594A (en) * | 2019-09-30 | 2020-02-07 | 中国热带农业科学院农产品加工研究所 | Active probiotic coconut powder stored at normal temperature and preparation method thereof |
| CN112553128A (en) * | 2020-12-30 | 2021-03-26 | 瞿瀚鹏 | Probiotic freeze-dried powder and preparation method and application thereof |
| CN112553128B (en) * | 2020-12-30 | 2023-08-18 | 瞿瀚鹏 | Probiotic freeze-dried powder and preparation method and application thereof |
| CN112779195A (en) * | 2021-03-24 | 2021-05-11 | 湖南农业大学 | Lactobacillus freeze separation and dry powder preparation process |
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