CN101649302B - Method for producing high-activity lactic acid bacteria agent by two-step drying method - Google Patents
Method for producing high-activity lactic acid bacteria agent by two-step drying method Download PDFInfo
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- CN101649302B CN101649302B CN2009101158897A CN200910115889A CN101649302B CN 101649302 B CN101649302 B CN 101649302B CN 2009101158897 A CN2009101158897 A CN 2009101158897A CN 200910115889 A CN200910115889 A CN 200910115889A CN 101649302 B CN101649302 B CN 101649302B
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Abstract
The invention relates to a method for producing a high-activity lactic acid bacteria agent by a two-step drying method, belonging to the technical field of microorganisms of preservation or activity maintenance. The method comprises steps of lactobacillus mire preparation and vacuum freeze drying. The method is characterized in that: before the step of vacuum freeze drying, the prepared lactobacillus mire is put in a vacuum drying box to carry out vacuum drying for 4-20 hours under a vacuum degree below 500Pa and a vacuum drying temperature of 25-45 DEG C. The survival rate of thalli in the method is higher than that of thalli of a method which directly carries out freeze drying, and the load and the operation time of the vacuum freeze drying can be reduced. The invention has small equipment loss and low energy consumption and can effectively save cost and improve the production efficiency.
Description
Technical field
The invention belongs to the microorganism preparing technical field, particularly the preparation of agent of lactic acid bacteria (production).
Technical background
Vacuum Freezing ﹠ Drying Technology originated from for 19 beginnings of the century, but its substantial development is at 20 middle of century, it is used for biological medicine industries such as biological active materials such as kept dry human plasma, vaccine, microbiotic and medicine the earliest, and has obtained development rapidly.It is slightly late that freeze drying equipment is used for foodstuffs industry, and main at present food or some flavoured foodstuff with dry some special purpose are as instant convenient food, aerospace food, high value protective foods, seasonings etc.
The advantage of Vacuum Freezing ﹠ Drying Technology is that it can keep the nutritive ingredient and the color of fresh provisions to greatest extent; Constant substantially, the instant of the volume shape of food, its quality is identical substantially with bright product; Dehydration is thorough, and water content is low, and is in light weight, and storing is convenient; Can preserve for a long time, can keep never degenerating in 5 years; Can avoid surface hardening and nutritive loss phenomenon in the general drying means; During the microorganism vacuum lyophilization, dry before fast with the freezing temperature below eutectic point of sample, the cold crystalline substance of formation is tiny evenly, and is lighter to the microbial cell damage, the contained water soluble component change in concentration that is evenly distributed is little.The drying process sample is in frozen state all the time, is moisturely deviate from by solid-state direct distillation being higher than under the 133Pa vacuum it.
But vacuum lyophilization also has its weak point: 1. freeze-dried vaccine powder surviving rate is lower after the vacuum lyophilization, microorganism water content height causes the phenomenon of ice crystal expansion, aqtocytolysis in the cell to take place easily when precooling, has reduced the survival rate of freeze-dried vaccine powder in the freezing dry process significantly; 2. freeze dried microorganism needs to handle through water activation before use, and concrete activation method is decided according to the specificity of microorganism; 3. the vacuum lyophilization microorganism is consuming time longer, is generally 6~18h, and expense is higher.
Other desiccating methods as boulton process, warm air drying and spraying drying, not too be fit to dehydrating of microorganism, or dehydration are not thorough, or the microorganism surviving rate are reduced greatly.
Summary of the invention
The objective of the invention is to propose a kind of two step of agent of lactic acid bacteria desiccating method, make and produce the moisture of as far as possible sloughing in the agent of lactic acid bacteria process wherein, and prevent that in its process ice crystal expands in the microorganism cells, the phenomenon of aqtocytolysis takes place, keep the high reactivity of milk-acid bacteria.
The present invention is achieved by the following technical solutions.
1, prepares milk-acid bacteria bacterium mud with ordinary method;
2, the vacuum-drying of milk-acid bacteria bacterium mud.To carry out vacuum-drying in the milk-acid bacteria bacterium mud placement vacuum drying oven that prepare, vacuum tightness remains on below the 500Pa, 25-45 ℃ of vacuum-drying temperature, vacuum-drying 4-20 hour.
3, carry out vacuum lyophilization with the milk-acid bacteria bacterium mud of ordinary method after to vacuum-drying.
Ordinary method of the present invention prepares milk-acid bacteria bacterium mud; can be following condition: milk-acid bacteria be through after the high-density culture; 3000-5000 rev/min of tubular-bowl centrifuge of fermented liquid; centrifugal 5-15 minute; topple over supernatant liquor and obtain bacterium mud; take out bacterium mud to container, and add protective material to bacterium mud according to processing requirement, mixing.
Conventional vacuum lyophilization of the present invention can be following condition: the milk-acid bacteria bacterium mud after the vacuum-drying is carried out precooling in cryogenic refrigerator, freezing temp is-60 to-80 ℃, and freezing time is 30-90 minute.Through the bacterium mud that precooling is handled, put into the vacuum chamber of the vacuum freeze drier of precooling rapidly, vacuum pump is opened in sealing.Vacuum tightness remains on below the 133Pa, and condenser temperature-40 is to-55 ℃, vacuum lyophilization 6-22 hour.
Advantage of the present invention and beneficial effect:
If bacterium mud is directly carried out vacuum lyophilization, because the moisture that contains in the bacterium mud is more, will form bigger ice crystal to a certain extent, the big more pair cell destructive of ice crystal degree is big more, make the mortality ratio of the thalline after the freeze-drying improve, thereby the reduction viable count does not reach the needed target of expection.On the contrary, before freeze-drying, carry out vacuum lyophilization, the moisture content of reduction bacterium mud that can be to a certain degree, the thalline surface that can be placed on after the freeze-drying forms big ice crystal, can significantly improve the thalline survival rate.
By vacuum-drying the water content of cell is reduced to 15-30% at normal temperatures, be reduced to below 5% by the water content of vacuum lyophilization at low temperatures at last cell, can prevent effectively that ice crystal expands in the cell, the phenomenon of aqtocytolysis takes place, increased substantially the survival rate of freeze-drying microbial inoculum in the freezing dry process, the viable lactic acid bacteria number can reach 10
12Cfu/g surpasses all commercially available lactic fermentation microbial inoculum product viable counts at present.
Because thalline is first through freezing before the distillation dehydration, form stable solid skeletal, so after the water sublimed, solid skeletal remains unchanged substantially, thalline can keep original form and structure, and the goods after the freeze-drying have very ideal instantly-soluble and fast rehydrating.
Because of bacterium mud is in frozen state, temperature is very low, and is less demanding to the heat source temperature of heat supply, adopts normal temperature or the high slightly termite of temperature to meet the demands.Whole kiln does not often need thermal insulation, does not have a lot of thermosteresis, heat energy utilization economy yet.
Through after the vacuum-drying, the active directly vacuum lyophilization of microbial inoculum wants high, and the survival rate of thalline is than direct lyophilize height, and can reduce the load and the operating time of vacuum lyophilization, and equipment loss is little, and it is low to consume energy, can effectively save cost, enhance productivity.
Embodiment
The present invention will be described further by following examples.
Embodiment 1.
1, the preparation of milk-acid bacteria bacterium mud.After the milk-acid bacteria process high-density culture, 3500 rev/mins of tubular-bowl centrifuges of fermented liquid, centrifugal 15 minutes, topple over supernatant liquor and obtain bacterium mud, take out bacterium mud to container, and add protective material to bacterium mud according to processing requirement, mixing.
2, the vacuum-drying of milk-acid bacteria bacterium mud.To carry out vacuum-drying in the milk-acid bacteria bacterium mud placement vacuum drying oven that prepare, vacuum tightness remains on below the 500Pa, 40 ℃ of vacuum-drying temperature, vacuum-drying 6 hours.
3, the vacuum lyophilization of milk-acid bacteria bacterium mud.Milk-acid bacteria bacterium mud after the vacuum-drying is carried out precooling in Ultralow Temperature Freezer, freezing temp is-70 ℃, and freezing time is 40 minutes.Through the bacterium mud that precooling is handled, put into the vacuum chamber of the vacuum freeze drier of precooling rapidly, vacuum pump is opened in sealing.Vacuum tightness remains on below the 133Pa, condenser temperature-55 ℃, vacuum lyophilization 12 hours.
Embodiment 2.
1, the preparation of milk-acid bacteria bacterium mud.After the milk-acid bacteria process high-density culture, 4000 rev/mins of tubular-bowl centrifuges of fermented liquid, centrifugal 12 minutes, topple over supernatant liquor and obtain bacterium mud, take out bacterium mud to container, and add protective material to bacterium mud according to processing requirement, mixing.
2, the vacuum-drying of milk-acid bacteria bacterium mud.To carry out vacuum-drying in the milk-acid bacteria bacterium mud placement vacuum drying oven that prepare, vacuum tightness remains on below the 500Pa, 45 ℃ of vacuum-drying temperature, vacuum-drying 4 hours.
3, the vacuum lyophilization of milk-acid bacteria bacterium mud.Milk-acid bacteria bacterium mud after the vacuum-drying is carried out precooling in Ultralow Temperature Freezer, freezing temp is-60 ℃, and freezing time is 90 minutes.Through the bacterium mud that precooling is handled, put into the vacuum chamber of the vacuum freeze drier of precooling rapidly, vacuum pump is opened in sealing.Vacuum tightness remains on below the 133Pa, condenser temperature-55 ℃, vacuum lyophilization 10 hours.
Embodiment 3.
1, the preparation of milk-acid bacteria bacterium mud.After the milk-acid bacteria process high-density culture, 5000 rev/mins of tubular-bowl centrifuges of fermented liquid, centrifugal 5 minutes, topple over supernatant liquor and obtain bacterium mud, take out bacterium mud to container, and add protective material to bacterium mud according to processing requirement, mixing.
2, the vacuum-drying of milk-acid bacteria bacterium mud.To carry out vacuum-drying in the milk-acid bacteria bacterium mud placement vacuum drying oven that prepare, vacuum tightness remains on below the 500Pa, 25 ℃ of vacuum-drying temperature, vacuum-drying 20 hours.
3, the vacuum lyophilization of milk-acid bacteria bacterium mud.Milk-acid bacteria bacterium mud after the vacuum-drying is carried out precooling in Ultralow Temperature Freezer, freezing temp is-80 ℃, and freezing time is 30 minutes.Through the bacterium mud that precooling is handled, put into the vacuum chamber of the vacuum freeze drier of precooling rapidly, vacuum pump is opened in sealing.Vacuum tightness remains on below the 133Pa, condenser temperature-40 ℃, vacuum lyophilization 16 hours.
Below be the contrast of described three embodiment bacterial classification survival rates, viable count:
Annotate: two-step approach refers to adopt vacuum-drying and vacuum lyophilization combined techniques to carry out the method that milk-acid bacteria dehydrates generation bacterium powder (agent); Single stage method refers to only adopt in the prior art vacuum freeze-drying method to carry out the method that milk-acid bacteria dehydrates generation bacterium powder (agent).
Claims (1)
1. one kind two goes on foot the method that desiccating method is produced high-activity lactic acid bacteria agent, comprise that milk-acid bacteria (Lactobacillus) bacterium is clay fully, vacuum lyophilization, it is characterized in that before vacuum lyophilization, will carrying out vacuum-drying in the milk-acid bacteria bacterium mud placement vacuum drying oven that prepare, vacuum tightness remains on below the 500Pa, 25-45 ℃ of vacuum-drying temperature, vacuum-drying 4-20 hour.
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TWI486323B (en) * | 2013-07-31 | 2015-06-01 | Advanced Green Biotechnology Inc | Manufacturing method of a granular microbial fertilizer |
CN103968649B (en) * | 2014-05-13 | 2015-12-02 | 上海理工大学 | A kind of freeze-drying method and corollary equipment |
CN108179130A (en) * | 2018-03-21 | 2018-06-19 | 广州同心源生物科技有限公司 | A kind of preparation method of high activity Enterococcus faecalis microorganisms preparation dry powder |
CN114705000B (en) * | 2021-06-30 | 2023-06-02 | 海南大学 | Method and device for drying sea cucumber by combining solar energy, microwave and vacuum freezing |
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CN1648239A (en) * | 2004-12-13 | 2005-08-03 | 南京农业大学 | Method for producing fruit particle fixing lactic acid bacteria fermenting agent |
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