CN101486986B - Preparation of freeze-dried Lactobacillus acidophilus powder - Google Patents
Preparation of freeze-dried Lactobacillus acidophilus powder Download PDFInfo
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
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Abstract
The present invention discloses a preparation method of freeze dried microbial powder of Lactobacillus acidophilus, which comprises the technical processes of fermented cultivation of Lactobacillus acidophilus, separation of a fermented fluid, emulsification of microbial soil and vacuum freeze drying of an emulsified solution. The preparation method is characterized in that the strain fermented cultivation comprises the steps as follows: a lactobacillus strain is inoculated into a fluid culture medium in a triangular flask, cultivated at the temperature of 30 to 40 DEG C for 10 to 30 hours, inoculated into a large fermentation flask and fermentation cultivated at the temperature of 35 to 40 DEG C for 10 to 30 hours, and sterilized at the temperature of 115 DEG C for 15 minutes; and fermentation and augmentation cultivation is carried out for 10 to 30 hours, the pH value of the fermented fluid is 5.5 to 6.5, and OD600 is over 2.0; the vacuum freeze drying of the emulsified solution comprises the steps: the emulsified solution is pre-frozen at the temperature of minus 60 to minus 40 DEG C for 1 to 5 hours, and then vacuum frozen and dried at the temperature of minus 60 DEG C and with the vacuum degree of 1 to 8 Pa for 10 to 30 hours. The freeze dried microbial powder of Lactobacillus acidophilus with the total plate count of 1.0 multiplied by 10<11> cfu/g can be preserved for 18 months at the temperature of minus 18 DEG C with unaffected liveness and stability.
Description
Technical field
The present invention discloses a kind of preparation method of freeze-dried Lactobacillus acidophilus powder, be applied to biological technical field, be a kind of efficiently, freeze-dried Lactobacillus acidophilus powder manufacturing technology cheaply, for Lactobacterium acidophilum Products Development and the application at aspects such as milk-product, food, medicine thereof provide necessary technical support.
Background technology
Lactobacterium acidophilum (Lactobacillus acidophilus) is lactobacillus (Lactobacillus), Gram-positive, not producing bud embraces, do not move, the elongated bacillus of atrichia, pod membrane, gemma, do not move, with single, become the form of two or short chain to exist, fermentable sugars produces the anaerobism or the amphimicrobian microorganism of lactic acid, ethanol, hydrogen peroxide etc.G+C content is 36.0%～37.4%, optimal pH 5.5～6.0, and 35～38 ℃ of optimum growth temperatures are not grown below 20 ℃ substantially, and poor heat resistance can utilize glucose, fructose, lactose, sucrose to carry out homofermentation, produces DL type lactic acid.Lactobacterium acidophilum is one of major microorganisms kind in the humans and animals enteron aisle, be from baby's ight soil, to separate at first, being a kind of probiotic bacterium very important in the humans and animals intestinal microflora, also is the probiotic bacterium of very paying attention to research and development in the present milk-acid bacteria family.Lactobacterium acidophilum is fermented in enteron aisle, produces lactic acid, acetic acid and antibiotic, has whole intestines effect, and intestinal tract diseases such as control and treatment chronic constipation, diarrhoea are had certain curative effect.In recent years, find that also Lactobacterium acidophilum has cholesterol level and the effect that suppresses some growth of cancer cells in the reduction blood, makes the research of Lactobacterium acidophilum and uses more noticeable.
In countries such as Japan, France, America and Europes, century-old history so that the milk-product of lactobacillus-fermented are existing, and occupy sizable ratio at dairy market.Be mainly used in functional foodstuff and health care food production at present, the application on US and European market is seen.According to Britain investigation company investigation, EU member country is to the consumption of lactobacillary milk goods, annually all increases with about 17% ratio, and wherein the product that contains Lactobacterium acidophilum and bifidus bacillus in France has increased by 300%, accounts for 4% of cow's milk total sales volume.The active lactobacillus fermented sour milk in North America accounts for 30% of milk-product.And in China Taiwan, the consumption of active lactobacillus fermented sour milk also surpasses 70%.In addition, the research report, many kinds of food can be as the carrier of Lactobacterium acidophilum, as candy, jelly etc.
Contain Lactobacterium acidophilum probiotics and the medicine various places of also being popular all over the world just gradually, calendar year 2001, in the probiotic bacterium list that can be used for protective foods that China Ministry of Health announces, Lactobacterium acidophilum is listed.Live lactobacillus acidophilus preparation has been widely used in clinical treatment.Pro-Bionate and microbiotic are taken simultaneously, can obviously alleviate gastrointestinal side effect and dependency diarrhoea that oral Broad spectrum antibiotics causes, also can be used for treating trichomonal vaginitis.The mechanism that Lactobacterium acidophilum suppresses the pathogenic bacteria growth has: secretion antibacterial substance, occupancy protection, nutrient competition and promotion host immune function etc.In recent years, more abroad studies show that: Lactobacterium acidophilum energy secreting bacteria element, the growth of leopards such as strongly inhibited dysentery bacterium, gonococcus, Salmonella enteritidis, vagina flagellate; The culture supernatant of people Lactobacterium acidophilum LB bacterial strain can obviously reduce the survival rate of helicobacter pylori (Helicobacterpylori) and to the ability of sticking of people's mucous membrane cell.
Abroad, carry out early for the research work of probiotic bacteriums such as milk-acid bacteria, bifidus bacillus, the product that contains probiotic bacterium is very general on Japan and European market, simultaneously, physiological function and the mechanism of action of probiotic bacterium is also compared comprehensive and deep research.The research of not restraining oneself aspects such as disease, inhibition diarrhoea, enhance immunity function, reducing cholesterol level and the generation of inhibition tumour at the alleviation lactose of probiotic bacterium has all obtained certain progress.In nearest decades, the health-promoting of probiotic bacteriums such as relevant milk-acid bacteria acts on China and more and more is subject to people's attention.Though research of probiotic bacterium and duration of service are not long, its make fast progress and effect also clearly.
In recent years, the special physiological activity of probiotic bacterium is studied widely by various ways such as cultured milk prod and probioticses, and its research field progressively widened, and this series products also increases just year by year.Though China is to the research comparative heat of probiotic bacterium, the research of Lactobacterium acidophilum also had certain distance compared with developed countries.Therefore, the research and development probiotic products has the bigger market space, will have very important significance to revitalizing industries such as China's food, feed, medicine and healthcare products.
At present, the produce market of probiotic bacterium that contains Lactobacterium acidophilum is very active, and kind is more and more.In food service industry, comprise milk-product, meat product and the fruit and vegetable product etc. of probiotics fermention; Medicine and health products trade all contain different types of probiotic bacterium as whole intestines life, beautiful pearl Chang Le, ANGLI No.1 etc., and throw in various ways such as capsule, tablet, pulvis and oral liquids.Yet can the stability of probiotic bacterium be directly connected to product and bring into play its health-promoting effect in the product, also is one of key index of examining the quality of the production.Studies show that in order to reach the health-promoting effect of serious hope, in the product based on probiotic bacteriums such as Lactobacterium acidophilums, number of viable should remain on 10
6More than the cfu/mL.Therefore, in the processing of probiotic bacterium based article, storage and transport process, should take measures as far as possible the to reduce inactivation of probiotic bacterium.Simultaneously, the pH value of human body hydrochloric acid in gastric juice changes between 1.5～5.0 usually, and the content of biliary salts is sometimes up to 4% in the enteron aisle, and many microorganisms can't be stood the poor environment in the human intestinal, causes arriving the interior probiotics viable bacteria number of also surviving of human body seldom.Studies show that, the ability that Lactobacterium acidophilum is stood low pH and high biliary salts environment changes with bacterial strain is different, therefore, the probiotic bacteriums such as Lactobacterium acidophilum that selection can be stood high hydrochloric acid in gastric juice and high biliary salts environment and have a potential health effect make it arrive host's enteron aisle and field planting be got off with higher viable count, be to guarantee that they give full play to a major criterion of health-promoting effect, this has become one of new direction of this field development.
The research and development of Lactobacterium acidophilum more and more get more and more people's extensive concerning.
Because Lactobacterium acidophilum is with health role, the research about this respect in recent years gets more and more, and has now become the research focus of active bacteria formulation.Active bacteria formulation should be kept higher viable bacteria level before consumption, and current many products all can not prevent the decay of viable count effectively.How to improve the viable bacteria amount and prolong the viable bacteria preservation term, become the key problem in technology of active bacteria formulation product research exploitation.Utilizing Vacuum Freezing ﹠ Drying Technology to produce active bacteria formulation is that comparatively ideal is a kind of in the multiple method for preserving; but freezing and drying process can cause the passivation of damage, death and some zymoprotein molecule of part microorganism cells; if the protective material selection is improper in the freeze-dry process, then fermentative activity descends thereupon.
Therefore, seek the new breakthrough mouth technically, screening has the lactobacillus acidophilus strains of premium properties, improve the vigor of Lactobacterium acidophilum, increase the stability of Lactobacterium acidophilum, reduce of the influence of various processing conditionss, strengthen research, provide support for the Lactobacterium acidophilum Products Development at aspects such as basic theory and production technologies to the Lactobacterium acidophilum product forms to Lactobacterium acidophilum growth and survival.
Summary of the invention
The objective of the invention is to disclose a kind of preparation method of freeze-dried Lactobacillus acidophilus powder, make the fermented liquid cell concentration reach 1.0 * 109cfu/g by 30～40 ℃ of 10～30 hours high-density culture, make bacterium mud concentration reach 1.0 * 1010cfu/g by efficient fermented liquid isolation technique, by obtaining 1.0 * 10 after the optimization technologies such as high reactivity emulsification and vacuum lyophilization
11The method of the freeze-dried Lactobacillus acidophilus powder of cfu/g, the highly active freeze-dried Lactobacillus acidophilus powder of preparation high-density high stability.
The present invention is on the basis to the lactobacillus acidophilus strains characteristic research; to its high density and activity fermentation culture; technology and technologies such as the separation of high stability high reactivity, emulsification, freeze-drying; the composition and the concentration of freezeproof protectant are optimized; in the hope of fermentation culture and the activity stability defencive function that obtains higher density to greatest extent; reduce the death of thalline in preparation process, improve the survival rate of cell, for the preparation of efficient freeze-dried vaccine powder provides technology and technology.
The present invention is achieved in that a kind of preparation method of freeze-dried Lactobacillus acidophilus powder, its technical process comprises the Lactobacterium acidophilum fermentation culture, fermented liquid separates, emulsification of bacterium mud and emulsion vacuum lyophilization, it is characterized in that: described strain fermentation technology is the Lactobacterium acidophilum bacterial classification to be inserted in the triangular flask liquid nutrient medium cultivate, the bacterial classification weight percentage that inserts in the substratum is 0.1%～0.01%, cultivated 10～30 hours for 30～40 ℃, the weight percent of described triangular flask liquid nutrient medium consists of: peptone 0.5～1.5%, glucose 0.2～0.6%, yeast extract paste 0.5～1.5%, extractum carnis 0.4～0.8%, sal epsom 0.02%, manganous sulfate 0.02%, sodium-chlor 0.003%, ferrous sulfate 0.001%, sodium-acetate 0.4%, citric acid diamines 0.2%, potassium primary phosphate 0.2%, tween-80 0.001%, all the other are water, sterilize 20 minutes for 121 ℃.Insert big fermentor tank again and carry out fermentation culture, inoculation weight percentage in the substratum is 1%～5%, 30～40 ℃ to be cultivated 10～30 hours, and the weight percent of described big ferment tank substratum consists of: peptone 0.5～1.5%, glucose 0.2～0.6%, yeast extractive substance 0.5～1.5%, sodium acetate 0.067%, sal epsom 0.001%, manganous sulfate 0.001%, ferrous sulfate 0.001%, sodium-chlor 0.001%, all the other are water; 115 ℃ of sterilizations 15 minutes, and to add sodium hydroxide solution control to cultivate acidity be pH4.0～6.5, making fermented liquid pH value is 4.0～5.0, OD600 is more than 2.0, microscopy Lactobacterium acidophilum form is normal, does not have assorted bacterium, stops fermenting.
Described fermented liquid separation method, with pore size at 0.01～1 μ m, 0.05MPa～0.5MPa hydraulic condition is down by the ceramic membrane filter separating thallus, or to carry out under 4000～10000rpm condition with tubular type or disc centrifuge separating thallus, and, obtain bacterium mud with sterilized water washing 1～2 time.
Described bacterium mud emulsifying process; according to viable bacteria bacterium mud: the ratio of bacterium mud protective material=0.1: 1～1: 0.1 is with bacterium mud and bacterium mud protective material mixing and emulsifying; bacterium mud protective material prescription each component weight percent is Sodium Glutamate 0.1～5.0%, lactose 0.1～8.0%, trehalose 1～15%, antioxidant 0.1～1%, skim-milk 1～20%, glycerine or tween-80 0.1～5.0%; all the other are water, and sterilize under 105 ℃～115 ℃ conditions 15 minutes postcooling to 5～20 of bacterium mud protective material are ℃ standby.Described antioxidant is selected a kind of in isoascorbic acid, L-glutamic acid and glycine.
Described emulsion vacuum freeze-drying technique is characterized in that drying conditions is: in the final vacuum lyophilize in 1～5 hour of-60～-40 ℃ of pre-freezes, the lyophilize temperature is-60 ℃, and vacuum tightness is 1～8pa, vacuum lyophilization 10～30 hours; Obtain viable count after the drying greater than 1.0 * 10
11The freeze-dried Lactobacillus acidophilus powder of cfu/g.
Described strain fermentation, fermented liquid separation and bacterium mud emulsive operating environment conditional request are for carrying out in the clean environment that is lower than 25 degree, humidity RH＜35% in temperature.
Bacterium lacticum such as described Lactobacterium acidophilum, its screening strain excellent all passes through following test: 1. to the resistibility of acid, bile and osmotic pressure, 2. sticking the human intestinal cell, 3. digest the security of taking the photograph the people, 4. to the inhibition of pathogenic bacterium such as streptococcus aureus, pathogenic colon bacillus, Candida albicans and Gardnerella, 5. in the of short duration field planting of colon, 6. confirm the clinical study of health effect etc.
The use of freeze-dried Lactobacillus acidophilus powder of the present invention:
1, in dairy production, does the application of starter and probiotic bacterium;
2, aspect food, feed adding component, do the application of whole intestines effect;
3, do the application of microecosystem regulation environment effect at aspects such as medicine of health products.
The invention has the beneficial effects as follows: the freeze-dried Lactobacillus acidophilus powder that the inventive method prepares greater than 1.0 * 1011cfu/g; it is active and stability is unaffected to obtain storing 18 months at-18 degree by relevant resist technology, for Lactobacterium acidophilum Products Development and the application at aspects such as milk-product, food, medicine thereof provide necessary technical support.
Embodiment
Below in conjunction with specific embodiment, further illustrate the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in the restriction the scope of protection of the invention that the scope of protection of the invention is said being as the criterion of setting forth with claims of the present patent application.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, or the condition of advising according to manufacturer.Ratio and per-cent are based on weight unit, unless stated otherwise.
Embodiment 1:
The Lactobacterium acidophilum bacterial classification that filters out is inserted the triangular flask liquid nutrient medium earlier, the back is inserted in the fermentor tank and is cultivated 10～30 hours fermentation culture that increase step by step down at 30～40 ℃ respectively, the weight percent of triangular flask liquid nutrient medium consists of: peptone 0.5%, glucose 0.2%, yeast extract paste 0.5%, extractum carnis 0.4%, sal epsom 0.02%, manganous sulfate 0.02%, sodium-chlor 0.003%, ferrous sulfate 0.001%, sodium-acetate 0.4%, citric acid diamines 0.2%, potassium primary phosphate 0.2%, tween-80 0.001%, all the other are water, and the sodium hydroxide adjust pH is 6.0,121 ℃ of sterilizations 20 minutes.The weight percent of big ferment tank substratum consists of: peptone 0.5%, glucose 0.2%, yeast extractive substance 0.5%, sodium acetate 0.067%, sal epsom 0.001%, manganous sulfate 0.001%, ferrous sulfate 0.001%, sodium-chlor 0.001%, all the other are water.And to add NaOH solution control to cultivate acidity pH value be 4.0～6.5, making fermented liquid pH value is 4.0～5.0, OD600 is more than 2.0, microscopy Lactobacterium acidophilum form is normal, does not have assorted bacterium, termination is fermented.At 0.01～1 μ m, 0.05MPa～0.5MPa hydraulic condition by the ceramic membrane filter separating thallus, obtains bacterium mud down with pore size.
Bacterium mud protective material prescription each component weight percent is Sodium Glutamate 0.1%, lactose 0.1%, trehalose 1%, isoascorbic acid 0.1%, skim-milk 1%, tween-80 0.1%; all the other are water, and sterilize under 105 ℃～115 ℃ conditions 15 minutes postcooling to 5～20 of bacterium mud protective material are ℃ standby.According to viable bacteria bacterium mud: the ratio of bacterium mud protective material=1: 10 is with bacterium mud and bacterium mud protective material mixing and emulsifying.In the final vacuum lyophilize in 1～5 hour of-60～-40 ℃ of pre-freezes, the lyophilize temperature is-60 ℃, and vacuum tightness is 1～8pa, obtains the high reactivity freeze-dried Lactobacillus acidophilus powder after the drying under 10～30 hours conditions of vacuum lyophilization.
Strain fermentation, fermented liquid separation and bacterium mud emulsive operating environment conditional request are for carrying out in the clean environment that is lower than 25 degree, humidity RH＜35% in temperature.
Viable count is greater than 1.0 * 10 in the product for preparing
11Cfu/g, with two aluminium film phonograph seal vacuum packagings, every bag of 50g or pack according to customer requirement.
Embodiment 2:
The Lactobacterium acidophilum bacterial classification that filters out is inserted the triangular flask liquid nutrient medium earlier, the back is inserted in the fermentor tank and is cultivated 10～30 hours fermentation culture that increase step by step down at 30～40 ℃ respectively, the weight percent of triangular flask liquid nutrient medium consists of: peptone 1.0%, glucose 0.4%, yeast extract paste 1.0%, extractum carnis 0.6%, sal epsom 0.02%, manganous sulfate 0.02%, sodium-chlor 0.003%, ferrous sulfate 0.001%, sodium-acetate 0.4%, citric acid diamines 0.2%, potassium primary phosphate 0.2%, tween-80 0.001%, all the other are water, and the sodium hydroxide adjust pH is 6.0,121 ℃ of sterilizations 20 minutes.The weight percent of big ferment tank substratum consists of: peptone 1.5%, glucose 0.6%, yeast extractive substance 1.5%, sodium acetate 0.067%, sal epsom 0.001%, manganous sulfate 0.001%, ferrous sulfate 0.001%, sodium-chlor 0.001%, all the other are water, sterilize 15 minutes for 115 ℃.And to add NaOH solution control to cultivate acidity be pH4.0～6.5.Making fermented liquid pH value is 4.0～5.0, and OD600 is more than 2.0, and microscopy Lactobacterium acidophilum form is normal, does not have assorted bacterium, stops fermentation.Under the 8000rpm condition, carry out with tubular type or disc centrifuge separating thallus, and, obtain bacterium mud with sterilized water washing 1～2 time.
Bacterium mud protective material prescription each component weight percent is Sodium Glutamate 1.0%, lactose 2.0%, trehalose 5%, L-glutamic acid 1%, skim-milk 20%, glycerine 1.0%; all the other are water, and sterilize under 105 ℃～115 ℃ conditions 15 minutes postcooling to 5～20 of protective material are ℃ standby.According to viable bacteria bacterium mud: the ratio of bacterium mud protective material=1: 2 is with bacterium mud and bacterium mud protective material mixing and emulsifying.
Other are with embodiment 1.
Viable count is greater than 1.0 * 10 in the product for preparing
11Cfu/g, with two aluminium film phonograph seal vacuum packagings, every bag of 50g or pack according to customer requirement.
Embodiment 3:
The Lactobacterium acidophilum bacterial classification that filters out is inserted the triangular flask liquid nutrient medium earlier, the back is inserted in the fermentor tank and is cultivated 10～30 hours fermentation culture that increase step by step down at 30～40 ℃ respectively, the weight percent of triangular flask liquid nutrient medium consists of: peptone 1.5%, glucose 0.6%, yeast extract paste 1.0%, extractum carnis 0.4%, sal epsom 0.02%, manganous sulfate 0.02%, sodium-chlor 0.003%, ferrous sulfate 0.001%, sodium-acetate 0.4%, citric acid diamines 0.2%, potassium primary phosphate 0.2%, tween-80 0.001%, all the other are water, and the sodium hydroxide adjust pH is 6.0,121 ℃ of sterilizations 20 minutes.The weight percent of big ferment tank substratum consists of: peptone 1.5%, glucose 0.6%, yeast extractive substance 1.5%, sodium acetate 0.067%, sal epsom 0.001%, manganous sulfate 0.001%, ferrous sulfate 0.001%, sodium-chlor 0.001%, all the other are water, sterilize 15 minutes for 115 ℃.
Bacterium mud protective material prescription each component weight percent is Sodium Glutamate 5.0%, lactose 8.0%, trehalose 15%, L-glutamic acid 1%, skim-milk 20%, glycerine 5.0%; all the other are water, and sterilize under 105 ℃～115 ℃ conditions 15 minutes postcooling to 5～20 of protective material are ℃ standby.According to viable bacteria bacterium mud: the ratio of bacterium mud protective material=10: 1 is with bacterium mud and bacterium mud protective material mixing and emulsifying.
Other are with embodiment 1.
Viable count is greater than 1.0 * 10 in the product for preparing
11Cfu/g, with two aluminium film phonograph seal vacuum packagings, every bag of 50g or pack according to customer requirement.
The scope of protection of present invention is not subjected to the restriction of described specific embodiments; specific embodiment provided by the invention is only as further specifying example of the present invention; those skilled in the art can be easy to the present invention is made the replacement of multiple improvement and component with reference to the description of this case specification sheets, and these need not the improvement of creative work and replace also should fall within the protection domain of appended claims of the present invention.
Claims (5)
1. the preparation method of a freeze-dried Lactobacillus acidophilus powder, its technical process comprises the Lactobacterium acidophilum fermentation culture, fermented liquid separates, emulsification of bacterium mud and emulsion vacuum lyophilization, it is characterized in that: described strain fermentation technology is the Lactobacterium acidophilum bacterial classification to be inserted in the triangular flask liquid nutrient medium cultivate, cultivated 10～30 hours for 30～40 ℃, the weight percent of described triangular flask liquid nutrient medium consists of: peptone 0.5～1.5%, glucose 0.2～0.6%, yeast extract paste 0.5～1.5%, extractum carnis 0.4～0.8%, sal epsom 0.02%, manganous sulfate 0.02%, sodium-chlor 0.003%, ferrous sulfate 0.001%, sodium-acetate 0.4%, citric acid diamines 0.2%, potassium primary phosphate 0.2%, tween-80 0.001%, all the other are water, sterilize 20 minutes for 121 ℃; Insert big fermentor tank again and carry out fermentation culture, cultivated 10～30 hours for 30～40 ℃, the weight percent of described big ferment tank substratum consists of: peptone 0.5～1.5%, glucose 0.2～0.6%, yeast extractive substance 0.5～1.5%, sodium acetate 0.067%, sal epsom 0.001%, manganous sulfate 0.001%, ferrous sulfate 0.001%, sodium-chlor 0.001%, all the other are water; 115 ℃ of sterilizations 15 minutes, and to add sodium hydroxide solution control to cultivate acidity be pH4.0～6.5; Described bacterium mud emulsifying process, according to viable bacteria bacterium mud: the ratio of bacterium mud protective material=0.1: 1～1: 0.1 is with bacterium mud and bacterium mud protective material mixing and emulsifying, bacterium mud protective material prescription each component weight percent is Sodium Glutamate 0.1～5.0%, lactose 0.1～8.0%, trehalose 1～15%, antioxidant 0.1～1%, skim-milk 1～20%, glycerine or tween-80 0.1～5.0%, all the other are water, and sterilize under 105 ℃～115 ℃ conditions 15 minutes postcooling to 5～20 of bacterium mud protective material are ℃ standby; Described antioxidant is selected a kind of in isoascorbic acid, L-glutamic acid and glycine.
2. the preparation method of a kind of freeze-dried Lactobacillus acidophilus powder according to claim 1 is characterized in that: described big ferment tank is cultivated, and fermented liquid pH value is 4.0～5.0, OD600 is more than 2.0, microscopy Lactobacterium acidophilum form is normal, does not have assorted bacterium, stops fermentation.
3. the preparation method of a kind of freeze-dried Lactobacillus acidophilus powder according to claim 1, it is characterized in that: described fermented liquid separation method, with pore size at 0.01～1 μ m, 0.05MPa～0.5MPa hydraulic condition is down by the ceramic membrane filter separating thallus, or to carry out under 4000～10000rpm condition with tubular type or disc centrifuge separating thallus, and, obtain bacterium mud with sterilized water washing 1～2 time.
4. the preparation method of a kind of freeze-dried Lactobacillus acidophilus powder according to claim 1, it is characterized in that: described emulsion vacuum freeze-drying technique, it is characterized in that drying conditions is: in the final vacuum lyophilize in 1～5 hour of-60～-40 ℃ of pre-freezes, the lyophilize temperature is-60 ℃, vacuum tightness is 1～8pa, vacuum lyophilization 10～30 hours; Obtain viable count after the drying greater than 1.0 * 10
11The freeze-dried Lactobacillus acidophilus powder of cfu/g.
5. according to the preparation method of claim 1 or 3 described a kind of freeze-dried Lactobacillus acidophilus powders, it is characterized in that: described strain fermentation, fermented liquid separation and bacterium mud emulsive operating environment conditional request are for carrying out in the clean environment that is lower than 25 degree, humidity RH＜35% in temperature.
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