CN108925983A - A kind of preparation method of enzyme microb - Google Patents
A kind of preparation method of enzyme microb Download PDFInfo
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- CN108925983A CN108925983A CN201810781956.8A CN201810781956A CN108925983A CN 108925983 A CN108925983 A CN 108925983A CN 201810781956 A CN201810781956 A CN 201810781956A CN 108925983 A CN108925983 A CN 108925983A
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- enzyme microb
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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Abstract
The present invention provides a kind of preparation method of enzyme microb, and the preparation method of the enzyme microb is the following steps are included: prepare bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium and saccharomycete fluid nutrient medium；Fluid nutrient medium is sterilized；It is cultivated for use after accessing bacillus subtilis, lactic acid bacteria, saccharomycete；Raw material powder of edible fungus, DEXTROSE ANHYDROUS, potassium dihydrogen phosphate and water are placed in container and are sterilized；Access bacillus subtilis fluid nutrient medium is simultaneously cultivated；It accesses the saccharomycete cultivated, lactic acid bacteria and cultivates；It is filtered by the enzyme microb fermented absorbent cotton net filtration, then with suction funnel, further removes filter residue and obtain enzyme microb；It will be placed in container, sterilize 121 DEG C after sealing, 20~40min is placed at 4 DEG C and saves to enzyme microb.The preparation method step of enzyme microb of the present invention is simple, easy, and yield is high, affected by environment small, can be with large-scale production.
Description
Technical field
The present invention relates to fermentation technique more particularly to a kind of preparation methods of enzyme microb.
Background technique
Edible mushroom is a kind of important living resources, provenance very abundant.China is that domestic fungus resource is the most in the world
Country abundant has research shows that edible mushroom contains the nutritional ingredient there are many needed by human body and improves the immunity of the human body, is anti-swollen
The multiple efficacies such as tumor, enhancing liver function, anti-oxidant.A large amount of mycelium can be obtained in a short time using edible fungus liquid fermentation
And its tunning has wide since this Period Process is short, yield is high, cost is relatively low in Edible Fungi
Application prospect.
Mushroom is known as the title of the king of mountain delicacy, is the nutritional health food of high protein, low fat.The edible part of dried thin mushroom accounts for
72%, every 100g eat aqueous 13g, fat 1.8g, carbohydrate 54g, crude fibre 7.8g, ash content 4.9g, calcium in part
124mg, phosphorus 415mg, iron 25.3mg, vitamin B1 0.07mg, vitamin B2 1.13mg, niacin 18.9mg.Lentinan
The vigor of helper T lymphocyte can be improved and enhance human body fluid immune function.Largely it was verified that the range of the anti-curing cancers of mushroom
Extensively, clinical treatment is had been used for.Mushroom also contains multivitamin, minerals, and to human metabolism is promoted, it is suitable to improve body
Stress has great role.Mushroom also plays therapeutic effect to diabetes, pulmonary tuberculosis, catarrhal jaundice, neuritis etc., and can be used for disappearing
Change bad, constipation, weight-reducing etc..Mushroom " QI invigorating is not hungry, controls wind blood-breaking and beneficial stomach helps food " is recorded in Chinese many ancient books.Civil use
The induction of variola, morbilli is helped, controls headache, dizziness.Modern study proves that lentinan, which is adjusted in human body, immune function
T cell activity, can reduce the ability of methyl cholanthrene induced tumor.Mushroom has strong inhibiting effect to cancer cell, to small white mouse
The inhibiting rate of sarcoma 180 is 97.5%, and the inhibiting rate to ehrlich carcinoma is 80%.Mushroom also contains double stranded RNA, can induce
Interferon is generated, there is anti-virus ability.Just take bait mushroom in chinese tradition medicine Shennong's Herbal and " wisdom can be increased ",
The record of " intelligence development is happy ".Modern medicine believes that, the increasing intelligence effect of mushroom is arginine and lysine rich in, often eat
Can be healthy and strong intelligence development.
There are low yields for current edible fungus fermented liquid (enzyme microb) preparation method, affected by environment big, it is difficult to scale
Active constituent composition and content have the shortcomings that larger difference between metaplasia production and different batches.
In view of this, the present invention is specifically proposed.
Summary of the invention
It is an object of the present invention to it is directed to current enzyme microb low yield, it is affected by environment big, it is difficult to large-scale production
The problem of, proposing a kind of preparation method of enzyme microb, this method step is simple, easy, and yield is high, affected by environment small,
It can be with large-scale production.
To achieve the above object, the technical solution adopted by the present invention is that: a kind of preparation method of enzyme microb, including with
Lower step:
Step 1 prepares bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium and saccharomycete fluid nutrient medium；
Step 2 will be filled with bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium and saccharomycete liquid respectively
The container closure of culture medium, 121 DEG C of 20~40min of sterilizing, pot subject to sterilization take when cooling to 65~80 DEG C in high-pressure sterilizing pot
It is put into 55~65 DEG C of drying in baking oven out；
After step 3,20~40min that ultraviolet super-clean bench sterilizes, bacillus subtilis, lactic acid are accessed with oese respectively
Bacterium, saccharomycete；
Step 4 connects bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium is placed in 37 DEG C of shaking tables after bacterium,
180r/min takes out after cultivating 24~48h respectively；Saccharomycete fluid nutrient medium is placed in 30 DEG C of shaking tables, 180r/min cultivates 48h
After take out, cryo-conservation is stand-by；
Raw material powder of edible fungus, DEXTROSE ANHYDROUS, potassium dihydrogen phosphate and water are placed in container, the edible mushroom by step 5
Powder, DEXTROSE ANHYDROUS, potassium dihydrogen phosphate mass ratio be 3~4:1~2:0.2~0.3, the amount ratio of the powder of edible fungus and water
It (g/ml) is 3~4:100~120；
Container described in step 5 is put into 121 DEG C of 20~40min of sterilizing of high pressure sterilization by step 6, and pot subject to sterilization cools to 65
It is taken out at~80 DEG C and is put into 55~65 DEG C of drying in baking oven；
After step 7,20~40min that ultraviolet super-clean bench sterilizes, bacillus subtilis fluid nutrient medium is accessed, it is described edible
The amount ratio (g/ml) of bacterium powder and bacillus subtilis fluid nutrient medium is 3.5~3.7:1.5~2.5；
Fermentation liquid is sealed after connecing bacterium and is put into 37 DEG C of shaking tables by step 8, shakes 5~7d under the conditions of 160r/min；
Step 9, sterilize ultraviolet super-clean bench 30min, accesses lactic acid bacteria fluid nutrient medium and the training of saccharomycete liquid of culture
The amount ratio (g/ml) of feeding base, the powder of edible fungus and saccharomycete is 3.5~3.7:1.5~2.5；The powder of edible fungus and lactic acid
The amount ratio (g/ml) of bacterium is 3.5~3.7:1.5~2.5, and container is sealed and is placed in 37 DEG C of shaking tables, in 160r/min condition
Under shake 6~7d after take out the enzyme microb fermented；
Enzyme microb is obtained by filtration in the enzyme microb fermented by step 10, and described be filtered into is filtered with centrifuge
Or with absorbent cotton net filtration, then filtered with suction funnel, further removes filter residue；It will be placed in container to enzyme microb, it is close
It is honored as a queen and sterilizes 121 DEG C, 20~40min is placed at 4 DEG C and saves.
Further, the edible mushroom is mushroom, Pleurotus eryngii, needle mushroom, hedgehog hydnum, rainbow conk, coprinus comatus, agaricus bisporus, tea
Set mushroom, antrodia, agaric, Cordyceps Militaris, parasol mushroom, long root mushroom, straw mushroom, oyster mushroom, phoenix-tail mushroom, flower mushroom, grey mushroom (abalonelike), red mushroom,
One of small mushroom and matsutake are a variety of.
Further, the mushroom is the good mushroom of gold.
Further, the preparation method of the enzyme microb, comprising the following steps:
Step 1 prepares bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium and saccharomycete fluid nutrient medium；
Step 2 will be filled with bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium and saccharomycete liquid respectively
The container closure of culture medium, 121 DEG C of sterilizing 30min in high-pressure sterilizing pot take out when pot subject to sterilization cools to 70 DEG C and are put into baking
60 DEG C of drying in case；
After step 3,30min that ultraviolet super-clean bench sterilizes, bacillus subtilis, lactic acid bacteria, ferment are accessed with oese respectively
Female bacterium；
Step 4 connects bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium is placed in 37 DEG C of shaking tables after bacterium,
180r/min takes out after cultivating 24~48h respectively, saccharomycete fluid nutrient medium is placed in 30 DEG C of shaking tables, 180r/min cultivates 48h
After take out, cryo-conservation is stand-by；
Raw material powder of edible fungus, DEXTROSE ANHYDROUS, potassium dihydrogen phosphate and water are placed in container, the edible mushroom by step 5
Powder, DEXTROSE ANHYDROUS, potassium dihydrogen phosphate mass ratio be 3.6:1.188:0.2376, the amount ratio of the powder of edible fungus and water
It (g/ml) is 3.6:110.
Container described in step 5 is put into 121 DEG C of sterilizing 30min of high pressure sterilization by step 6, when pot subject to sterilization cools to 70 DEG C
Taking-up is put into 60 DEG C of drying in baking oven；
After step 7,30min that ultraviolet super-clean bench sterilizes, bacillus subtilis fluid nutrient medium, the powder of edible fungus are accessed
Amount ratio (g/ml) with bacillus subtilis fluid nutrient medium is 3.6:2；
Fermentation liquid is sealed after connecing bacterium and is put into 37 DEG C of shaking tables by step 8, shakes 6d under the conditions of 160r/min；
Step 9, sterilize ultraviolet super-clean bench 30min, accesses lactic acid bacteria fluid nutrient medium and the training of saccharomycete liquid of culture
The amount ratio (g/ml) of feeding base, the powder of edible fungus and saccharomycete is 3.6:2；The amount ratio of the powder of edible fungus and lactic acid bacteria
(g/ml) it is 3.6:2, container is sealed in and is placed in 37 DEG C of shaking tables, shakes to take out after 6~7d under the conditions of 160r/min and ferment
Enzyme microb；
Enzyme microb is obtained by filtration in the enzyme microb fermented by step 10, and described be filtered into is filtered with centrifuge
Or with absorbent cotton net filtration, then filtered with suction funnel, further removes filter residue；It will be placed in container to enzyme microb, it is close
It is honored as a queen and sterilizes 121 DEG C, 30min is placed at 4 DEG C and saves.
Further, the bacillus subtilis fluid nutrient medium includes the following each component of weight percent: yeast powder
0.2~4%, beef extract 0.2~4%, peptone 0.9~1.1%, NaC10~11%, glucose 0.8~1.1%, remaining is
Deionized water.
Further, the bacillus subtilis fluid nutrient medium includes the following each component of weight percent: yeast powder
0.3%, beef extract 0.3%, peptone 1%, NaCl 0.5%, glucose 1%, remaining is deionized water.
Further, the lactic acid bacteria is MRS culture medium.
Further, the saccharomycete is YPD fluid nutrient medium.
Further, raw material described in step 5 is powder of edible fungus, DEXTROSE ANHYDROUS, potassium dihydrogen phosphate and water, the edible mushroom
Powder, DEXTROSE ANHYDROUS, potassium dihydrogen phosphate mass ratio be 3.6:1.188:0.2376, the amount ratio of the powder of edible fungus and water
It (g/ml) is 3.6:110.
Another object of the present invention also discloses a kind of enzyme microb, is prepared using the above method.
A kind of enzyme microb formula of the present invention is scientific, reasonable, and preparation method is simple, easy, compared with prior art
Relatively have the advantage that
1) enzyme microb preparation method of the present invention is simple, easy, and yield is high, affected by environment small, can be with scale metaplasia
It produces.
2) active material in ethyl acetate and extracting n-butyl alcohol mushroom (lentinus edodes) fermentation liquid is used, into
Row antibacterial tests.Using filter paper enzyme measurement extract to Escherichia coli, tobacco Ralstonia solanacearum, Candida albicans antibacterial activity,
As a result, it has been found that Lentinus edodes fermented liquid ethyl acetate phase and n-butanol are with respect to Escherichia coli, the growth of tobacco Ralstonia solanacearum and Candida albicans
There is good inhibiting effect, ethyl acetate extraction phase (0.1g/mL) is respectively to the average inhibition zone of these three bacterium
15.52mm, 16.75mm and 16.38mm: n-butanol phase (0.2g/mL) is respectively to the average inhibition zone of these three bacterium
16.72mm, 19.01mm and 17.89mm.
3) acute toxicity testing, cumulative toxicity and mutagenic test, subchronic toxicity testing have been carried out to enzyme microb
Deng three stage toxicological experiments, the results showed that, mycelium and supernatant are non-toxic, and long-term consumption is safely and reliably.Mushroom
A variety of nutrition, health-care components possessed by fermentation liquid and safe and reliable toxicological evaluation produce to develop Lentinus edodes fermented liquid downstream
Product provide reliable basis.
Specific embodiment
The present invention is further described with reference to embodiments:
Embodiment 1
Present embodiment discloses a kind of preparation methods of Lentinus edodes fermented liquid, comprising the following steps:
1, with bacillus subtilis, lactic acid bacteria, saccharomycete fluid nutrient medium.
Note: bacillus subtilis fluid nutrient medium: yeast powder 0.3%, beef extract 0.3%, peptone 1%, NaCl
0.5%, glucose 1%, deionized water 50ml.
Lactic acid bacteria: MRS culture medium.
Saccharomycete: YPD fluid nutrient medium.
2, it is sealed after the completion of configuration, 121 DEG C of sterilizing 30min, pot subject to sterilization cool to 70 DEG C or so in high-pressure sterilizing pot
When take out and be put into baking oven 60 DEG C of drying.
3, after ultraviolet super-clean bench sterilizing 30min, bacillus subtilis, lactic acid bacteria, saccharomycete are accessed with oese respectively.
4, it connects and bacillus subtilis, lactic acid bacteria fluid nutrient medium is placed in 37 DEG C of shaking tables after bacterium, 180r/min is cultivated respectively
24, it is taken out after 48h, saccharomycete fluid nutrient medium is placed in 30 DEG C of shaking tables, takes out after 180r/min culture 48h, low temperature is protected when not used
It deposits.
5, raw material respectively weighs 8 parts and is placed in 250ml conical flask: 3.6g mushroom powder, 1.188g DEXTROSE ANHYDROUS, 0.2376g
Potassium dihydrogen phosphate adds 110ml deionized water.
6, the conical flask of step 5 is put into 121 DEG C of sterilizing 30min of high-pressure sterilizing pot.Pot subject to sterilization cools to 70 DEG C or so
When take out and be put into baking oven 60 DEG C of drying.
7, after ultraviolet super-clean bench sterilizing 30min, the fluid nutrient medium of bacillus subtilis is cultivated in access, and each conical flask is each
2ml。
8, it connects fermentation liquid to be sealed after bacterium and be put into 37 DEG C of shaking tables, shake 6d under the conditions of 160r/min.
9, after 6d, after ultraviolet super-clean bench sterilizes 30min, access shifts to an earlier date cultured lactic acid bacteria fluid nutrient medium and yeast
Bacteria liquid culture medium, is separately added into 2ml, after be sealed in place 37 DEG C of shaking tables in, taken out after 6~7d is shaken under the conditions of 160r/min
The fermentation liquid fermented.
10, it by the fermentation liquid fermented absorbent cotton net filtration, then is filtered with suction funnel, further removes filter residue.
11, the liquid for taking coarse filtration to cross is put into blue lid bottle, sterilizes 121 DEG C after being sealed, 30min is placed at 4 DEG C and saves.
Contain 18 kinds of amino acid in Lentinus edodes fermented liquid of the present invention, test shows that mycelial amino acid content is 9800mg/
100g, the amino acid content of supernatant are 460mg/100ml.
The present embodiment has also carried out acute toxicity testing, cumulative toxicity and mutagenic test, sub- chronic to Lentinus edodes fermented liquid
Three stage such as toxicity test toxicological experiment, the results showed that, mycelium and supernatant are non-toxic, and long-term consumption is safe and reliable
Ground.A variety of nutrition, health-care components possessed by Lentinus edodes fermented liquid and safe and reliable toxicological evaluation, to develop Lentinus edodes fermented liquid
Downstream product provides reliable basis.
Embodiment 2
Present embodiment discloses a kind of preparation methods of Pleurotus eryngii fermentation liquid, comprising the following steps:
1, with bacillus subtilis, lactic acid bacteria, saccharomycete fluid nutrient medium.
Note: bacillus subtilis fluid nutrient medium: yeast powder 0.3%, beef extract 0.3%, peptone 1%, NaCl
0.5%, glucose 1%, deionized water 50ml.
Lactic acid bacteria: MRS culture medium.
Saccharomycete: YPD fluid nutrient medium.
2, it is sealed after the completion of configuration, 121 DEG C of sterilizing 40min, pot subject to sterilization cool to 70 DEG C or so in high-pressure sterilizing pot
When take out and be put into baking oven 60 DEG C of drying.
3, after ultraviolet super-clean bench sterilizing 30min, bacillus subtilis, lactic acid bacteria, saccharomycete are accessed with oese respectively.
4, it connects and bacillus subtilis, lactic acid bacteria fluid nutrient medium is placed in 37 DEG C of shaking tables after bacterium, 180r/min is cultivated respectively
24, it is taken out after 48h, saccharomycete fluid nutrient medium is placed in 30 DEG C of shaking tables, takes out after 180r/min culture 48h, low temperature is protected when not used
It deposits.
5, raw material respectively weighs 8 parts and is placed in 250ml conical flask: 3.7g pleurotus eryngii powder, 1.2g DEXTROSE ANHYDROUS, 0.22g phosphorus
Acid dihydride potassium adds 110ml deionized water.
6, the conical flask of step 5 is put into 121 DEG C of sterilizing 40min of high-pressure sterilizing pot.Pot subject to sterilization cools to 70 DEG C or so
When take out and be put into baking oven 60 DEG C of drying.
7, after ultraviolet super-clean bench sterilizing 40min, the fluid nutrient medium of bacillus subtilis is cultivated in access, and each conical flask is each
2ml。
8, it connects fermentation liquid to be sealed after bacterium and be put into 37 DEG C of shaking tables, shake 6d under the conditions of 160r/min.
9, after 6d, after ultraviolet super-clean bench sterilizes 40min, access shifts to an earlier date cultured lactic acid bacteria fluid nutrient medium and yeast
Bacteria liquid culture medium, is separately added into 2ml, after be sealed in place 37 DEG C of shaking tables in, taken out after 6~7d is shaken under the conditions of 160r/min
The fermentation liquid fermented.
10, it by the fermentation liquid fermented absorbent cotton net filtration, then is filtered with suction funnel, further removes filter residue.
11, the liquid for taking coarse filtration to cross is put into blue lid bottle, sterilizes 121 DEG C after being sealed, 40min is placed at 4 DEG C and saves.
The present embodiment has also carried out acute toxicity testing, cumulative toxicity and mutagenic test, Asia slowly to Pleurotus eryngii fermentation liquid
Property the three stage toxicological experiments such as toxicity test, the results showed that, mycelium and supernatant are non-toxic, and long-term consumption is that safety can
By ground.A variety of nutrition, health-care components possessed by Pleurotus eryngii fermentation liquid and safe and reliable toxicological evaluation, to develop Pleurotus eryngii
Fermentation liquid downstream product provides reliable basis.
Embodiment 3
Present embodiment discloses a kind of preparation methods of oyster mushroom fermentation liquid, comprising the following steps:
1, with bacillus subtilis, lactic acid bacteria, saccharomycete fluid nutrient medium.
Note: bacillus subtilis fluid nutrient medium: yeast powder 0.3%, beef extract 0.3%, peptone 1%, NaCl
0.5%, glucose 1%, deionized water 50ml.
Lactic acid bacteria: MRS culture medium.
Saccharomycete: YPD fluid nutrient medium.
2, it is sealed after the completion of configuration, 121 DEG C of sterilizing 35min, pot subject to sterilization cool to 70 DEG C or so in high-pressure sterilizing pot
When take out and be put into baking oven 60 DEG C of drying.
3, after ultraviolet super-clean bench sterilizing 30min, bacillus subtilis, lactic acid bacteria, saccharomycete are accessed with oese respectively.
4, it connects and bacillus subtilis, lactic acid bacteria fluid nutrient medium is placed in 37 DEG C of shaking tables after bacterium, 180r/min is cultivated respectively
24, it is taken out after 48h, saccharomycete fluid nutrient medium is placed in 30 DEG C of shaking tables, takes out after 180r/min culture 48h, low temperature is protected when not used
It deposits.
5, raw material respectively weighs 8 parts and is placed in 250ml conical flask: 3.5g oyster mushroom powder, 1.5g DEXTROSE ANHYDROUS, 0.3g di(2-ethylhexyl)phosphate
Hydrogen potassium adds 110ml deionized water.
6, the conical flask of step 5 is put into 121 DEG C of sterilizing 35min of high-pressure sterilizing pot.Pot subject to sterilization cools to 70 DEG C or so
When take out and be put into baking oven 60 DEG C of drying.
7, after ultraviolet super-clean bench sterilizing 35min, the fluid nutrient medium of bacillus subtilis is cultivated in access, and each conical flask is each
2ml。
8, it connects fermentation liquid to be sealed after bacterium and be put into 37 DEG C of shaking tables, shake 6d under the conditions of 160r/min.
9, after 6d, after ultraviolet super-clean bench sterilizes 35min, access shifts to an earlier date cultured lactic acid bacteria fluid nutrient medium and yeast
Bacteria liquid culture medium, is separately added into 2ml, after be sealed in place 37 DEG C of shaking tables in, taken out after 6~7d is shaken under the conditions of 160r/min
The fermentation liquid fermented.
10, the fermentation liquid fermented is filtered with centrifuge, further removes filter residue.
11, the liquid for taking coarse filtration to cross is put into blue lid bottle, sterilizes 121 DEG C after being sealed, 35min is placed at 4 DEG C and saves.
The present embodiment has also carried out acute toxicity testing, cumulative toxicity and mutagenic test, sub- chronic to oyster mushroom fermentation liquid
Three stage such as toxicity test toxicological experiment, the results showed that, mycelium and supernatant are non-toxic, and long-term consumption is safe and reliable
Ground.A variety of nutrition, health-care components possessed by oyster mushroom fermentation liquid and safe and reliable toxicological evaluation, to develop oyster mushroom fermentation liquid
Downstream product provides reliable basis.
Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Pipe present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that: its according to
So be possible to modify the technical solutions described in the foregoing embodiments, or to some or all of the technical features into
Row equivalent replacement；And these are modified or replaceed, various embodiments of the present invention technology that it does not separate the essence of the corresponding technical solution
The range of scheme.
Claims (10)
1. a kind of preparation method of enzyme microb, which comprises the following steps:
Step 1 prepares bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium and saccharomycete fluid nutrient medium；
Step 2 will be filled with bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium and saccharomycete Liquid Culture respectively
The container closure of base, 121 DEG C of 20~40min of sterilizing in high-pressure sterilizing pot take out when pot subject to sterilization cools to 65~80 DEG C and put
Enter 55~65 DEG C of drying in baking oven；
After step 3,20~40min that ultraviolet super-clean bench sterilizes, bacillus subtilis, lactic acid bacteria, ferment are accessed with oese respectively
Female bacterium；
Step 4 connects bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium is placed in 37 DEG C of shaking tables, 180r/ after bacterium
Min takes out after cultivating 24~48h respectively；Saccharomycete fluid nutrient medium is placed in 30 DEG C of shaking tables, is taken after 180r/min culture 48h
Out, cryo-conservation is stand-by；
Raw material powder of edible fungus, DEXTROSE ANHYDROUS, potassium dihydrogen phosphate and water are placed in container, the powder of edible fungus, nothing by step 5
Water glucose, potassium dihydrogen phosphate mass ratio be 3~4:1~2:0.2~0.3, the amount ratio of the powder of edible fungus and water is 3~
4:100~120；
Container described in step 5 is put into 121 DEG C of 20~40min of sterilizing of high pressure sterilization by step 6, and pot subject to sterilization cools to 65~80
DEG C when take out and be put into baking oven 55~65 DEG C of drying；
After step 7,20~40min that ultraviolet super-clean bench sterilizes, bacillus subtilis fluid nutrient medium, the powder of edible fungus are accessed
Amount ratio with bacillus subtilis fluid nutrient medium is 3.5~3.7:1.5~2.5；
Fermentation liquid is sealed after connecing bacterium and is put into 37 DEG C of shaking tables by step 8, shakes 5~7d under the conditions of 160r/min；
Step 9, sterilize ultraviolet super-clean bench 30min, accesses the lactic acid bacteria fluid nutrient medium and saccharomycete fluid nutrient medium of culture,
The amount ratio of the powder of edible fungus and saccharomycete is 3.5~3.7:1.5~2.5；The amount ratio of the powder of edible fungus and lactic acid bacteria
For 3.5~3.7:1.5~2.5, container is sealed and is placed in 37 DEG C of shaking tables, is taken out after 6~7d is shaken under the conditions of 160r/min
The enzyme microb fermented；
Step 10 is filtered by the enzyme microb fermented absorbent cotton net filtration, then with suction funnel, further removes filter residue
Obtain enzyme microb；It will be placed in container, sterilize 121 DEG C after sealing, 20~40min is placed at 4 DEG C and protects to enzyme microb
It deposits.
2. the preparation method of enzyme microb according to claim 1, which is characterized in that the edible mushroom is mushroom, apricot Bao
Mushroom, needle mushroom, hedgehog hydnum, rainbow conk, coprinus comatus, agaricus bisporus, agrocybe, antrodia, agaric, Cordyceps Militaris, parasol mushroom, long root
One of mushroom, straw mushroom, oyster mushroom, phoenix-tail mushroom, flower mushroom, grey mushroom (abalonelike), red mushroom, small mushroom and matsutake are a variety of.
3. the preparation method of enzyme microb according to claim 2, which is characterized in that the mushroom is the good mushroom of gold.
4. the preparation method of enzyme microb according to claim 1, which is characterized in that the preparation side of the enzyme microb
Method, comprising the following steps:
Step 1 prepares bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium and saccharomycete fluid nutrient medium；
Step 2 will be filled with bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium and saccharomycete Liquid Culture respectively
The container closure of base, 121 DEG C of sterilizing 30min in high-pressure sterilizing pot take out when pot subject to sterilization cools to 70 DEG C and are put into baking oven
60 DEG C of drying；
After step 3,30min that ultraviolet super-clean bench sterilizes, bacillus subtilis, lactic acid bacteria, saccharomycete are accessed with oese respectively；
Step 4 connects bacillus subtilis fluid nutrient medium, lactic acid bacteria fluid nutrient medium is placed in 37 DEG C of shaking tables, 180r/ after bacterium
Min takes out after cultivating 24~48h respectively, and saccharomycete fluid nutrient medium is placed in 30 DEG C of shaking tables, is taken after 180r/min culture 48h
Out, cryo-conservation is stand-by；
Raw material powder of edible fungus, DEXTROSE ANHYDROUS, potassium dihydrogen phosphate and water are placed in container, the powder of edible fungus, nothing by step 5
Water glucose, potassium dihydrogen phosphate mass ratio be 3.6:1.188:0.2376, the amount ratio of the powder of edible fungus and water is 3.6:
110。
Container described in step 5 is put into 121 DEG C of sterilizing 30min of high pressure sterilization, taking-up when pot subject to sterilization cools to 70 DEG C by step 6
It is put into 60 DEG C of drying in baking oven；
Step 7, will ultraviolet super-clean bench sterilize 30min after, access bacillus subtilis fluid nutrient medium, the powder of edible fungus with it is withered
The amount ratio of careless bacillus liquid culture medium is 3.6:2；
Fermentation liquid is sealed after connecing bacterium and is put into 37 DEG C of shaking tables by step 8, shakes 6d under the conditions of 160r/min；
Step 9, sterilize ultraviolet super-clean bench 30min, accesses the lactic acid bacteria fluid nutrient medium and saccharomycete fluid nutrient medium of culture,
The amount ratio of the powder of edible fungus and saccharomycete is 3.6:2；The powder of edible fungus and the amount ratio of lactic acid bacteria are 3.6:2, will be held
Device, which is sealed in, to be placed in 37 DEG C of shaking tables, takes out the enzyme microb fermented after 6~7d is shaken under the conditions of 160r/min；
Step 10 is filtered by the enzyme microb fermented absorbent cotton net filtration, then with suction funnel, further removes filter residue
Obtain enzyme microb；It will be placed in container, sterilize 121 DEG C after sealing, 30min is placed at 4 DEG C and saves to enzyme microb.
5. the preparation method of enzyme microb according to claim 1, which is characterized in that the bacillus subtilis bacteria liquid training
Feeding base includes the following each component of weight percent: yeast powder 0.2~4%, beef extract 0.2~4%, and peptone 0.9~
1.1%, NaC10~11%, glucose 0.8~1.1%, remaining is deionized water.
6. the preparation method of enzyme microb according to claim 1, which is characterized in that the bacillus subtilis bacteria liquid training
Feeding base includes the following each component of weight percent: yeast powder 0.3%, beef extract 0.3%, peptone 1%, NaCl 0.5%,
Glucose 1%, remaining is deionized water.
7. the preparation method of enzyme microb according to claim 1, which is characterized in that the lactic acid bacteria is MRS culture medium.
8. the preparation method of enzyme microb according to claim 1, which is characterized in that the saccharomycete is the training of YPD liquid
Support base.
9. the preparation method of enzyme microb according to claim 1, which is characterized in that raw material described in step 5 is edible mushroom
Powder, DEXTROSE ANHYDROUS, potassium dihydrogen phosphate and water, the powder of edible fungus, DEXTROSE ANHYDROUS, potassium dihydrogen phosphate mass ratio be 3.6:
1.188:0.2376 the amount ratio of the powder of edible fungus and water is 3.6:110.
10. a kind of enzyme microb, which is characterized in that be prepared using claim 1~9 any one the method.
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| CN111685310A (en) * | 2020-06-04 | 2020-09-22 | 包选平 | Composite edible fungus enzyme and preparation method thereof |
| CN112826082A (en) * | 2021-01-20 | 2021-05-25 | 武汉工程大学 | Preparation method of mushroom enzyme |
| CN113456563A (en) * | 2021-07-19 | 2021-10-01 | 河南城建学院 | Aloe lysozyme gel rich in pleurotus djamor fruiting body polypeptide and preparation method thereof |
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| CN113456563A (en) * | 2021-07-19 | 2021-10-01 | 河南城建学院 | Aloe lysozyme gel rich in pleurotus djamor fruiting body polypeptide and preparation method thereof |
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