CN106834175A - A kind of lactic acid bacteria protective agent for adding pholiota nameko polysaccharide - Google Patents
A kind of lactic acid bacteria protective agent for adding pholiota nameko polysaccharide Download PDFInfo
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- CN106834175A CN106834175A CN201710063489.0A CN201710063489A CN106834175A CN 106834175 A CN106834175 A CN 106834175A CN 201710063489 A CN201710063489 A CN 201710063489A CN 106834175 A CN106834175 A CN 106834175A
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- lactic acid
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Abstract
The invention discloses a kind of lactic acid bacteria protective agent for adding pholiota nameko polysaccharide.The present invention is comprised the following steps：Pholiota nameko polysaccharide is prepared first；Then the bacterial sediment thing addition pholiota nameko polysaccharide after being centrifuged to lactic acid bacteria, Boratex, sodium glutamate, glutathione, vitamin C, its adding proportion are respectively 0.001-0.04%, 0.01-0.03%, 0.01-0.03%, 0.02-0.08% and 0.01-1.0% of the bacterial sediment thing weight after lactic acid bacteria centrifugation.The pholiota nameko polysaccharide of protection lactic acid bacteria is mainly contained in formula.Wherein lactic acid bacteria includes Lactobacillus casei, streptococcus thermophilus, lactobacillus bulgaricus, Lactobacillus plantarum, lactobacillus acidophilus, Lactobacillus rhamnosus and Bifidobacterium.Using the survival rate of lactic acid bacteria bacterium powder obtained in the protective agent all more than 60%, viable count is all 1010More than.
Description
Technical field
The preparation of spray-dried lactic bacterium bacterium powder and the addition of vacuum freeze-drying method cunning mushroom are applied to the present invention relates to a kind of
The lactic acid bacteria protective agent of polysaccharide.
Background technology
Lactic acid bacteria (Lactic acid bacteria, LAB) is that a class can utilize carbohydrate to produce the leather of lactic acid blue
Family name's positive bacteria is generally called.Mainly include lactobacillus, lactococcus, more than 20 category such as streptococcus and Bifidobacterium.Mesh
Preceding much researchs confirm that many lactic acid bacterias are all normal host bacterium in the stomach and intestine and colon of humans and animals.Some of them lactic acid bacteria is
The probiotics of the intestines and stomach of human body, can produce many prebiotic functions.Probiotic lactobacillus have to be improved intestines and stomach, alleviates lactose not
Resistance to disease, improve body nutrition condition, regulation is immune, improve antioxidation activity, hypotensive, reduce cholesterol and nitrite and
Suppress pathogen to many functions such as the field planting of host.Lactic acid bacteria is widely used as important probiotics (Probiotic)
In food, biological medicine, the industry such as agricultural and feed.
Vacuum Freezing ＆ Drying Technology and spray drying technology are frequently utilized for the production of probiotic lactobacillus.In the process
Low temperature, high temperature and the dry pressure that lactic acid bacteria experiences can cause cellular damage.It is dry in order to reduce vacuum freeze drying and spraying
The dry damage to lactic acid bacteria, need to add certain protective agent before lyophilized, to improve cell survival rate, and can improve lactic acid
The storage-stable of bacterium.Temperature in process influences maximum to microbial survival, particularly high temperature.High temperature can destroy lactic acid
Bacterium maintains the enzyme system of vital movement, its activity is reduced or is lost.The important biomolecule macromolecule such as protein, nucleic acid and enzyme system
Also can hydrogen bond destroyed, cause bacterium protein solidify denaturation, nucleic acid occur degraded deactivation, so as to destroy cell
Composition；High temperature also results in lipoids composition on cell membrane and forms minimum hole, leaks cellular content, also to cell membrane,
Cell membrane, cytoplasm and nucleic acid cause damage, so as to cause death.Low temperature can also produce ice crystal to cause damage cell, its damage
Wound mostlys come from the change and the change of related protein structure of cell membrane fat acid physical behavior.Vacuum freeze drying is operated
With spray-drying operation all along with the drying of lactic acid bacteria, to dry also cause lactic acid bacteria and damage, lactic acid mycoderm can be caused
Fusion and the phase in version of lipid.These basic function and structures are protected for cell survival and keeps feature most important.
Lactic acid bacteria protective agent is to improve a kind of most important measure of lactic bacteria activity, by the suitable protection agent prescription of research, Ke Yi great
The big survival rate for improving lactic acid bacteria lactic acid bacteria in spray-drying process and vacuum freeze drying process.
The content of the invention
The technical problems to be solved by the invention are to overcome above-mentioned the deficiencies in the prior art, there is provided one kind addition
The lactic acid bacteria protective agent of pholiota nameko polysaccharide.
A kind of lactic acid bacteria protective agent for adding pholiota nameko polysaccharide of the present invention, it is comprised the following steps：
1. the preparation of pholiota nameko polysaccharide
(1) selecting for mushroom is slided
Fresh sliding mushroom is chosen, surface is not rotted, shape is complete, organize scar without obvious；Mushroom handle tubbiness, mushroom lid circle and light
Sliding, diameter is thick in 3cm or so, capping, edge meat, and do not turn sour smelly taste；The homogeneous and clean excellent sliding mushroom of size is standby；
(2) weighing for mushroom is slided
The fresh sliding mushrooms of 1000g are weighed for using with electronic balance；
(3) crushing and beating of sliding mushroom
Add distilled water in sliding mushroom is weighed, sliding mushroom is 2 with the mass ratio of water:3；Sliding mushroom together pours into high-speed set with water
Knit in bruisher, interval crushing and beating is carried out to sliding mushroom, 10000 revs/min of mashing at a high speed in 30 seconds stops 2 minutes, totally 4 times；Must be sliding
Mushroom serum of tissue；
(4) digest
Above-mentioned sliding mushroom serum of tissue, is subsequently adding cellulase 10U/mg, and addition is the 0.08% of grind slurries quality；55
3 hours are incubated at DEG C；
(5) water-bath extraction
The previous step sliding mushroom serum of tissue of gained is put into water-bath heating, then the beaker of sliding mushroom serum of tissue will be filled and be placed in perseverance
In warm water bath, heating water bath 4h at 85 DEG C；
(6) filter
With 100 mesh stainless steel sieves to water-bath after sliding mushroom serum of tissue filter, filtrate mixing retain；
(7) washery slag
Previous step gained filter residue adds 1 times of water in water-bath extraction 1h at 85 DEG C, is filtered with 100 mesh stainless steel sieves, retains
Filtrate, and mix with previous step filtrate；
(8) alcohol analysis
Gained mixing filtrate presses 1:2 ratios add absolute ethyl alcohol alcohol to analyse, 4 DEG C of standing 24h；Then 20min is centrifuged in 4000rpm
After remove liquid, obtain the solid of pholiota nameko polysaccharide；
(9) dry
Pholiota nameko polysaccharide solid after the analysis of above-mentioned alcohol vacuumizes drying through 80 DEG C, obtain pholiota nameko polysaccharide in following steps for
Make protective agent；
2. the composition of the protection agent prescription of pholiota nameko polysaccharide is included
Bacterial sediment thing addition pholiota nameko polysaccharide after being centrifuged to lactic acid bacteria, Boratex, sodium glutamate, glutathione, dimension life
Plain C, its adding proportion be respectively the 0.001-0.04% of bacterial sediment thing weight after lactic acid bacteria centrifugation, 0.01-0.03%,
0.01-0.03%, 0.02-0.08% and 0.01-1.0% (w/w).
The present invention has the advantages that：
Lactic acid bacteria of the invention includes Lactobacillus casei, streptococcus thermophilus, lactobacillus bulgaricus, Lactobacillus plantarum, thermophilic
Lactobacillus lactis, Lactobacillus rhamnosus and Bifidobacterium.Lactic acid bacteria protective agent of the invention contains pholiota nameko polysaccharide or its mixture,
Can be used for spray-dried lactic bacterium bacterium powder to prepare and the preparation of vacuum freeze-drying method lactic acid bacteria bacterium powder, the method can produce viable bacteria
Number leavening high, simple to operate, low cost can large-scale production.
Protective agent of the present invention can be used for spray drying and vacuum freeze drying produces lactic acid bacteria bacterium powder.Mainly contain in formula
There is the pholiota nameko polysaccharide of protection lactic acid bacteria.Wherein lactic acid bacteria includes Lactobacillus casei, streptococcus thermophilus, lactobacillus bulgaricus, plant
Thing lactobacillus, lactobacillus acidophilus, Lactobacillus rhamnosus and Bifidobacterium.Using the survival of lactic acid bacteria bacterium powder obtained in the protective agent
All more than 60%, viable count is all 10 for rate10More than.
Specific embodiment
A kind of protectant preparation method of lactic acid bacteria for adding pholiota nameko polysaccharide of the present invention, it is comprised the following steps：
1. the preparation of pholiota nameko polysaccharide
(1) selecting for mushroom is slided
Fresh sliding mushroom is chosen, surface is not rotted, shape is complete, organize scar without obvious；Mushroom handle tubbiness, mushroom lid circle and light
Sliding, diameter is thick in 3cm or so, capping, edge meat, and do not turn sour smelly taste；The homogeneous and clean excellent sliding mushroom of size is standby；
(2) weighing for mushroom is slided
The fresh sliding mushrooms of 1000g are weighed for using with electronic balance；
(3) crushing and beating of sliding mushroom
Add distilled water in sliding mushroom is weighed, sliding mushroom is 2 with the mass ratio of water:3；Sliding mushroom together pours into high-speed set with water
Knit in bruisher, interval crushing and beating is carried out to sliding mushroom, 10000 revs/min of mashing at a high speed in 30 seconds stops 2 minutes, totally 4 times；Must be sliding
Mushroom serum of tissue；
(5) digest
Above-mentioned sliding mushroom serum of tissue, is subsequently adding cellulase 10U/mg, and addition is the 0.08% of grind slurries quality；55
3 hours are incubated at DEG C；
(6) water-bath extraction
The previous step sliding mushroom serum of tissue of gained is put into water-bath heating, then the beaker of sliding mushroom serum of tissue will be filled and be placed in perseverance
In warm water bath, heating water bath 4h at 85 DEG C；
(7) filter
With 100 mesh stainless steel sieves to water-bath after sliding mushroom serum of tissue filter, filtrate mixing retain；
(8) washery slag
Previous step gained filter residue adds 1 times of water in water-bath extraction 1h at 85 DEG C, is filtered with 100 mesh stainless steel sieves, retains
Filtrate, and mix with previous step filtrate；
(9) alcohol analysis
Gained mixing filtrate presses 1:2 ratios add absolute ethyl alcohol alcohol to analyse, 4 DEG C of standing 24h；Then 20min is centrifuged in 4000rpm
After remove liquid, obtain the solid of pholiota nameko polysaccharide；
(10) dry
Pholiota nameko polysaccharide solid after the analysis of above-mentioned alcohol vacuumizes drying through 80 DEG C, obtain pholiota nameko polysaccharide in following steps for
Make protective agent；
2. the composition of the protection agent prescription of pholiota nameko polysaccharide is included
Bacterial sediment thing addition pholiota nameko polysaccharide after being centrifuged to lactic acid bacteria, Boratex, sodium glutamate, glutathione, dimension life
Plain C, its adding proportion be respectively the 0.001-0.04% of bacterial sediment thing weight after lactic acid bacteria centrifugation, 0.01-0.03%,
0.01-0.03%, 0.02-0.08% and 0.01-1.0% (w/w).
Lactic acid bacteria protective agent of the invention contains pholiota nameko polysaccharide or its mixture, can be used for spray-dried lactic bacterium bacterium
Prepared by powder and prepared by vacuum freeze-drying method lactic acid bacteria bacterium powder, and specific embodiment is as follows：
Embodiment 1：Spray drying process produces Lactobacillus casei bacterium powder
1. High Density Cultivation
Cheese breast bar lactic acid bacteria thalline culture after activation is connected to MRS cultures 24 by 1% inoculum concentration of volume fraction
Hour, the MRS is the MRS culture mediums that Lactobacillus casei is cultivated after optimum experimental, can make maximum culture viable count up to 1012。
2. agent prescription and adding method are protected
A kind of protection agent prescription containing pholiota nameko polysaccharide, adding proportion is respectively the hundred of the thalline centrifugal sediment weight collected
Divide ratio：Pholiota nameko polysaccharide 0.01%, Boratex 0.02%, glutathione 0.08% and vitamin C 0.02% (w/w).Protective agent system
Standby and adding method：The protective agent that bacterial sediment addition after Lactobacillus casei centrifugation is prepared.
Collect the lactic acid bacteria thalline centrifugal sediment of Lactobacillus casei.After protective agent is weighed by above-mentioned formula, add its heavy
The water dissolves of 5 times of amount, 8000r/min homogeneous 5 minutes, 121 DEG C sterilize 15 minutes, are cooled to 40 DEG C.Add 0.1%Tween 80
Second homogenate, processing condition 5000r/min, 10 minutes, after bacterial sediment thing and protective agent are mixed, are gently mixed uniformly afterwards,
Then next step spray drying is carried out.
3. the technological parameter being spray-dried：1. feed rate 15ml/min (using 10L small-sized spray drier)；Charging
Concentration 30%；2. feeding temperature is 28 DEG C；3. 130 DEG C of EAT；4. nebulizer pressure 150kPa.
Using the survival rate of Lactobacillus casei bacterium powder obtained in the method all more than 60%, viable count is all 1010More than.
Embodiment 2：Vacuum freeze-drying method produces lactobacillus acidophilus bacterium powder
1. High Density Cultivation
Cheese breast bar lactic acid bacteria thalline culture after activation is connected to MRS cultures 24 by 1% inoculum concentration of volume fraction
Hour, the MRS is the MRS culture mediums that Lactobacillus casei is cultivated after optimum experimental, can make maximum culture viable count up to 1012。
2. agent prescription and adding method are protected
A kind of protection agent prescription containing pholiota nameko polysaccharide, adding proportion is respectively the hundred of the thalline centrifugal sediment weight collected
Divide ratio：Pholiota nameko polysaccharide 0.008%, Boratex 0.01%, sodium glutamate 0.02% and vitamin C 0.04% (w/w).Protective agent
Prepare and adding method：The protective agent that bacterial sediment thing addition after Lactobacillus casei centrifugation is prepared.
Collect the lactic acid bacteria thalline centrifugal sediment of Lactobacillus casei.After protective agent is weighed by above-mentioned formula, add its heavy
The water dissolves of 5 times of amount, 8000r/min homogeneous 5 minutes, 121 DEG C sterilize 15 minutes, are cooled to 40 DEG C.Add 0.1%Tween 80
Second homogenate, processing condition 5000r/min, 10 minutes, after bacterial sediment thing and protective agent are mixed, are gently mixed uniformly afterwards,
Then next step vacuum freeze drying is carried out.
3. the technological parameter of vacuum freeze drying：1. -30 DEG C of precooling temperature；2. 0.6 DEG C/min of pre-freeze speed；3. hothouse
Pressure 62Pa；4. 40 DEG C of temperature of heating plate；5. water content is below 5%.
Using the survival rate of Lactobacillus casei bacterium powder obtained in the method all more than 65%, viable count is all 1010More than.
Survival rate after storage half a year is all more than 50%.
Claims (3)
1. it is a kind of add pholiota nameko polysaccharide lactic acid bacteria protective agent, it is characterized in that, follow the steps below：
A. the preparation of pholiota nameko polysaccharide
(1) selecting for mushroom is slided：Choose fresh sliding mushroom；
(2) crushing and beating of sliding mushroom：Add distilled water in sliding mushroom is weighed, sliding mushroom is 2 with the mass ratio of water:3；Sliding mushroom and water
Together pour into high-speed tissue mashing machine, interval crushing and beating is carried out to sliding mushroom, 10000 revs/min of mashing at a high speed in 30 seconds stops 2
Minute, totally 4 times；Mushroom serum of tissue must be slided；
(3) digest：Above-mentioned sliding mushroom serum of tissue, is subsequently adding cellulase 10U/mg, and addition is the 0.08% of grind slurries quality；
3 hours are incubated at 55 DEG C；
(4) water-bath extraction：The sliding mushroom serum of tissue of previous step gained is put into water-bath heating, then the burning that will fill sliding mushroom serum of tissue
Cup is placed in thermostat water bath, heating water bath 4h at 85 DEG C；
(5) filter：With 100 mesh stainless steel sieves to water-bath after sliding mushroom serum of tissue filter, filtrate mixing retain；
(6) washery slag：Previous step gained filter residue adds 1 times of water in water-bath extraction 1h at 85 DEG C, is filtered with 100 mesh stainless steel sieves,
Retain filtrate, and mix with previous step filtrate；
(7) alcohol analysis：Gained mixing filtrate presses 1:2 ratios add absolute ethyl alcohol alcohol to analyse, 4 DEG C of standing 24h；Then it is centrifuged in 4000rpm
Liquid is removed after 20min, the solid of pholiota nameko polysaccharide is obtained；
(8) dry：Pholiota nameko polysaccharide solid after above-mentioned alcohol analysis vacuumizes drying through 80 DEG C, and acquisition pholiota nameko polysaccharide is in following steps
For use as protective agent；
B. the composition of the protection agent prescription of pholiota nameko polysaccharide is included
Bacterial sediment thing addition pholiota nameko polysaccharide after being centrifuged to lactic acid bacteria, Boratex, sodium glutamate, glutathione, vitamin C,
Its adding proportion be respectively the 0.001-0.04% of bacterial sediment thing weight after lactic acid bacteria centrifugation, 0.01-0.03%,
0.01-0.03%, 0.02-0.08% and 0.01-1.0%.
2. it is according to claim 1 addition pholiota nameko polysaccharide lactic acid bacteria protective agent, it is characterized in that, be applied to spray drying breast
The preparation of sour bacterium bacterium powder, using the small-sized spray drier of 10L, spray-drying operation technological parameter：1. feed rate 15ml/
min；2. feeding temperature is 28 DEG C；3. 130 DEG C of EAT；4. nebulizer pressure 150kPa.
3. it is according to claim 1 addition pholiota nameko polysaccharide lactic acid bacteria protective agent, it is characterized in that, be applied to vacuum refrigeration do
The preparation of dry method lactic acid bacteria bacterium powder, wherein vacuum freeze drying Operating parameters：1. -30 DEG C of precooling temperature；2. pre-freeze speed
0.6℃/min；3. drying chamber pressure 55-75Pa；4. 40 DEG C of temperature of heating plate；5. water content is below 5%.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101863999A (en) * | 2010-07-22 | 2010-10-20 | 天津商业大学 | Preparation method of pholiota nameko polysaccharide extract |
| CN101914518A (en) * | 2010-08-02 | 2010-12-15 | 天津商业大学 | Method for preparing Lactobacillus casei microcapsule bacterial powder by spray drying |
| CN106070653A (en) * | 2016-06-08 | 2016-11-09 | 天津商业大学 | A kind of spray drying method produces the preparation method adding pholiota nameko polysaccharide fermented soybean milk powder |
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- 2017-02-03 CN CN201710063489.0A patent/CN106834175A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101863999A (en) * | 2010-07-22 | 2010-10-20 | 天津商业大学 | Preparation method of pholiota nameko polysaccharide extract |
| CN101914518A (en) * | 2010-08-02 | 2010-12-15 | 天津商业大学 | Method for preparing Lactobacillus casei microcapsule bacterial powder by spray drying |
| CN106070653A (en) * | 2016-06-08 | 2016-11-09 | 天津商业大学 | A kind of spray drying method produces the preparation method adding pholiota nameko polysaccharide fermented soybean milk powder |
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