CN103937679B - A kind of aspergillosis strain of vinegar processed - Google Patents
A kind of aspergillosis strain of vinegar processed Download PDFInfo
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Abstract
A kind of aspergillosis strain, this strain is in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC) preservation, and preserving number is CGMCC NO.8847.The strain of the present invention is a kind of composite bacteria, only need to add this strain the most permissible in solid-state vinegar production process, it is not necessary to adding yeast wine, acetic acid bacteria strain, production operation process is the most fairly simple.Saccharifying, alcohol fermentation, acetic fermentation need not individually designed technique, their biochemical reaction is carried out at the same time.Strain solid state process of the present invention produces vinegar, need not fall pond and turn over unstrained spirits, thus reduce labor intensity, improve labor efficiency.
Description
Technical field
The present invention relates to strain and the application of a kind of vinegar processed.
Background technology
At present, what China's Solid-state fermentation vinegar technique commonly used is aspergillus niger, and this kind of strain enzyme system is more single, only
Containing amylase and saccharifying enzyme, saccharifying power is stronger, but also needs to add yeast wine, acetic acid bacteria strain during Vinegar Fermentation.Behaviour
Making process more complicated, labor intensity is relatively big, and labor efficiency is low, and quality control there is also bigger difficulty, and yield rate is relatively low.
Summary of the invention
An object of the present invention, is to provide one to produce admittedly for the situation that current solid-state vinegar production process is more complicated
The compound strain of state vinegar；The two of the purpose of the present invention, are to provide the preparation method of aforementioned strain.
The mesh of the present invention is achieved in that
A kind of aspergillosis strain, this strain is at China Committee for Culture Collection of Microorganisms's common micro-organisms center
(CGMCC) preservation, preserving number is CGMCC NO. 8847.
The preparation method of this strain is as follows:
A, the separation of strain:
(1) taking 5 test tubes equipped with physiological saline solution, labelling 1,2,3,4,5, goes bail for and deposits this strain test tube respectively
Bacterium colony 2 ring, puts into No. 1 test tube filling physiological saline solution, and firmly shaking, makes microorganism suspend in water uniformly;
(2) with 1ml aseptic straw by aseptic manipulation, from No. 1 test tube, draw 1ml suspension inject in No. 2 pipes, and will
Manage quick shake well, make uniform suspension for No. 2.Equally by No. 2 pipes being drawn in 1ml suspension No. 3 pipes of injection, and by No. 3
Manage rapid shake well, make uniform suspension.By that analogy until No. 5 pipes;
(3) from No. 4 and No. 5 pipes, respectively take 1ml suspension respectively with two aseptic straws, and be injected separately into two aseptic trainings
Support in ware, add culture medium 15ml being cooled to 45 DEG C, slight rocking-turn rapidly, note fully shaking of suspension and culture medium
Even, it is frozen into flat board after standing, then culture dish paper using is fixed, and be inverted in cultivation in 30 DEG C of calorstats, after 4 days therefrom
Select single bacterium colony, and transplant cultivation in test tube slant culture medium, if a kind of bacteria growing, obtain pure culture;
B, the preparation of aspergillosis strain
(1) slant medium is prepared
1. culture medium prescription
Fermented bean drink (3 ° of B é concentration) 100ml
Potassium dihydrogen phosphate (KH2PO4) 0.1g
Sodium chloride (NaCl) 0.1g
Magnesium sulfate (MgSO4) 0.2g
Citric acid 0.1g
Sucrose 5.0g
Agar 2.5g
PH value 5.5～6.0
2. prepared by culture medium
Accurately weigh medicine, fermented bean drink add medicine and boils, making medicine fully dissolve, the culture medium will boiled while hot
It is sub-packed in test tube, will be equipped with the test tube of culture medium and fix and put into autoclave sterilizer, put inclined-plane after taking-up while hot, treat
After culture medium solidifying strain inclined plane culture medium;
(2) prepared by aspergillosis strain
1. inoculate
By the operating process operation of inoculation, the spore access using Inoculating needle picking growing way good in an aseptic environment has solidified
In slant medium, it is respectively connected to 10 culture medium.
2. cultivate
Putting into constant incubator to cultivate finished 10, cultivation temperature is set to 30 DEG C, and incubation time is set to 72 hours.
After colony growth is good, 5 conducts preserve strain, and 5 flags are for producing strain.
Three, the preservation of aspergillosis strain
Including following sequence and step:
Using Storage in refrigerator method, put in refrigerator by 5 preservation strains and preserve, temperature controls between 4～5 DEG C, the time
Must not exceed 6 months.
D, the application of aspergillosis strain:
(1) amplification culture of strain
1. culture medium is prepared
Take rice, be soaked in water 20 hours, then rinse well with water, clean swimming to drench, put into steam-boiler by its steaming and decocting
Become rice, by rice spreading for cooling after taking the dish out of the pot, after temperature cools down 30 DEG C, in being loaded in triangular flask, last sterilizing.
2. amplification culture
In an aseptic environment the strain in test tube is accessed in triangular flask, be then placed in constant incubator carrying out piling up training
Supporting, temperature is 30 DEG C, and incubation time is about 18 hours.The long mycelia that turns white of internal material at the bottom for the treatment of bottle carries out shake-flask culture for the first time,
Bottle intramatrical mycelium is shaken up and shakeouts, carry out second time shaking flask again after material internal at the bottom of the 4～5 hours bottles long mycelia that turns white, treat mycelia
Growing up to Testa oryzae color is spore shape, and after 96 hours cultivate, bottle outlet is standby.
(2) preparation of saccharifying koji
1. raw material: with Sorghum vulgare Pers. and wheat bran as raw material.
2. technique: using aerated koji making technique to make saccharifying koji, access strain after material cooking cooling, temperature controls
37-39 DEG C, the yeast production time is 66 hours.
(3) solid state process produces vinegar
The most former, auxiliary material: with Sorghum vulgare Pers. as raw material, with wheat bran as adjuvant, with rice husk as inserts.
2. technical process: steaming and decocting → cooling → access strain → fermentation → pouring vinegar.
E, the preservation of aspergillosis strain
Using Storage in refrigerator method, put in refrigerator by 5 preservation strains and preserve, temperature controls between 4-5 DEG C, and the time is not
Must be more than 6 months.
Beneficial effect:
1, the strain of the present invention is a kind of composite bacteria, only need to add this strain the most permissible in solid-state vinegar production process,
Without adding yeast wine, acetic acid bacteria strain, production operation process is the most fairly simple.Saccharifying, alcohol fermentation, acetic fermentation need not individually set
Meter technique, their biochemical reaction is carried out at the same time, the main body of the most different its biochemical reactions of fermentation period different and
?.
2, strain solid state process of the present invention produces vinegar, need not fall pond and turn over unstrained spirits, thus reduce labor intensity, improve work
Efficiency.
3, the present invention uses fermented bean drink to manufacture slant medium, has the advantage that strain growing way is good.
4, using the vinegar product that strain of the present invention is produced, product is limpid, nothing precipitation, and sour and sweet palatability, product quality compares
Stable.
5, strain vinegar-producing rate of the present invention is higher, and yield rate reaches 10kg/kg(major ingredient), and other yield rate is at 8.5-
9kg/kg(major ingredient).
Detailed description of the invention:
The present invention is described in further detail to combine example in detail below.
Bacterium classification is named: aspergillosis
Latin literary fame: Aspergillus sp
Depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC).
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: February 24 in 2014
Deposit number: CGMCC NO. 8847
One, the separation of aspergillosis strain, including following sequence and step:
1,5 test tubes equipped with physiological saline solution (every 9ml) are taken, respectively labelling 1,2,3,4,5.Go bail for and deposit strain
Bacterium colony 2 ring of test tube, puts into No. 1 test tube filling physiological saline solution, and firmly shaking, makes microorganism be suspended in water uniformly
In.
2, with 1ml aseptic straw by aseptic manipulation, from No. 1 test tube, draw 1ml suspension inject in No. 2 pipes, and by 2
Number manage quick shake well, make uniform suspension.Equally by No. 2 pipes being drawn in 1ml suspension No. 3 pipes of injection, and by No. 3 pipes
Shake well, makes uniform suspension rapidly.By that analogy until No. 5 pipes.
3, from No. 4 and No. 5 pipes, respectively take 1ml suspension respectively with two aseptic straws, and be injected separately into two aseptic trainings
Support in ware, add culture medium 15ml being cooled to 45 DEG C, slight rocking-turn rapidly, note fully shaking of suspension and culture medium
Even, it is frozen into flat board after standing, then culture dish paper using is fixed, and be inverted in cultivation in 30 DEG C of calorstats, after 4 days therefrom
Select single bacterium colony, and transplant cultivation in test tube slant culture medium, if a kind of bacteria growing, obtain pure culture.
Two, the preparation method of aspergillosis strain, including following sequence and step:
1, the preparation of slant medium
(1) formula of culture medium
Fermented bean drink (3 ° of B é) 100ml
Potassium dihydrogen phosphate (KH2PO4) 0.1g
Sodium chloride (NaCl) 0.1g
Magnesium sulfate (MgSO4) 0.2g
Citric acid 0.1g
Sucrose 5.0g
Agar 2.5g
PH value 5.5～6.0
(2) preparation of culture medium
Accurately weigh medicine, fermented bean drink add medicine and boils, making medicine fully dissolve.The culture medium will boiled while hot
It is sub-packed in test tube, every test tube about 15ml.Will be equipped with the test tube of culture medium to fix and put into autoclave sterilizer, steam
Pressure 0.1Mpa, sterilising temp 121 DEG C, sterilization time 30 minutes, put inclined-plane after taking-up while hot, be cost after culture medium solidifying
Invention strain inclined plane culture medium.
2, the preparation of aspergillosis strain
(1) inoculation
By the operating process operation of inoculation, the spore access using Inoculating needle picking growing way good in an aseptic environment has solidified
In slant medium, it is respectively connected to 10 culture medium.
(2) cultivate
Putting into constant incubator to cultivate finished 10, cultivation temperature is set to 30 DEG C, and incubation time is set to 72 hours.
After colony growth is good, 5 as preserve strain, 5 as produce strain.
Three, the preservation of aspergillosis strain, including following sequence and step:
Using Storage in refrigerator method, put in refrigerator by 5 preservation strains and preserve, temperature controls between 4-5 DEG C, and the time is not
Must be more than 6 months.
Four, the application of aspergillosis strain:
1, the amplification culture of strain
(1) preparation of culture medium
Produce 1000 kilograms of vinegars, need to use the secondary strain of 1 gram.Amplification culture strain 500 grams every time.
Weighing 500 grams of rice, be soaked in water 20 hours, then rinse well with water, clean swimming to drench, steaming and decocting becomes rice,
By rice spreading for cooling after taking the dish out of the pot, after temperature is cooled to 30 DEG C, in being loaded in triangular flask.Last sterilizing, steam pressure is
0.1MPa, sterilization time is 30 minutes.
(2) amplification culture
By the operating process of inoculation, in an aseptic environment the strain in test tube is accessed in triangular flask, be then placed in constant temperature
Carrying out trick layer culture in incubator, temperature is 30 DEG C, and incubation time is about 18 hours.The long mycelia that turns white enters internal material at the bottom for the treatment of bottle
Row for the first time shake-flask culture, shakes up bottle intramatrical mycelium and shakeouts, and carries out after material internal at the bottom of the 45 hours bottles long mycelia that turns white again
Shaking flask for the second time, treating that mycelia grows up to Testa oryzae color is spore shape, and after 96 hours cultivate, bottle outlet is standby.
2, the preparation of saccharifying koji
(1) raw material: with Sorghum vulgare Pers. and wheat bran as raw material.
(2) technique: using aerated koji making technique to make saccharifying koji, access strain after material cooking cooling, temperature controls
37-39 DEG C, the yeast production time is 66 hours.
3, solid fermentation legal system vinegar
With Sorghum vulgare Pers. as raw material, with wheat bran as adjuvant, with rice husk as inserts, technical process: steaming and decocting → cool down → connect
Enter strain → fermentation → pouring vinegar.
Claims (2)
1. a vinegar processed aspergillosis strain (Aspergillus sp), belong to aspergillosis strain, it is characterised in that this strain is micro-in China
Biological inoculum preservation administration committee's common micro-organisms center preservation, preserving number is CGMCC NO.8847.
2. the preparation method of the aspergillosis strain of a vinegar processed as claimed in claim 1, it is characterised in that method is as follows:
A, the separation of strain
(1) taking 5 test tubes equipped with physiological saline solution, labelling 1,2,3,4,5, goes bail for and deposits the bacterium colony of this strain test tube respectively
2 rings, put into No. 1 test tube filling physiological saline solution, and firmly shaking, makes microorganism suspend in water uniformly；
(2) with 1ml aseptic straw by aseptic manipulation, from No. 1 test tube, draw 1ml suspension inject in No. 2 pipes, and by No. 2
Manage quick shake well, make uniform suspension, equally by No. 2 pipes being drawn in 1ml suspension No. 3 pipes of injection, and by fast for No. 3 pipes
Speed shake well, makes uniform suspension, by that analogy until No. 5 pipes；
(3) from No. 4 and No. 5 pipes, respectively take 1ml suspension respectively with two aseptic straws, and be injected separately into two sterile petri dish
In, add culture medium 15ml being cooled to 45 DEG C, quickly slight rocking-turn, note fully shaking up of suspension and culture medium,
It is frozen into flat board after standing, then culture dish paper using is fixed, and be inverted in cultivation in 30 DEG C of calorstats, therefrom choose after 4 days
Menu bacterium colony, and transplant cultivation in test tube slant culture medium, if a kind of bacteria growing, obtain pure culture；
B, the preparation of aspergillosis strain
(1) slant medium is prepared
1. culture medium prescription
2. prepared by culture medium
Accurately weigh medicine, fermented bean drink add medicine and boils, making medicine fully dissolve, the culture medium subpackage that will boil while hot
In test tube, will be equipped with the test tube of culture medium and fix and put into autoclave sterilizer, put inclined-plane after taking-up while hot, wait to cultivate
Base solidification after strain inclined plane culture medium；
2, prepared by aspergillosis strain
1. inoculate
By the operating process operation of inoculation, access, with the spore that Inoculating needle picking growing way is good, the inclined-plane solidified in an aseptic environment
In culture medium, it is respectively connected to 10 culture medium；
2. cultivate
Putting into constant incubator to cultivate finished 10, cultivation temperature is set to 30 DEG C, and incubation time is set to 72 hours, bacterium colony
After growth is good, 5 conducts preserve strain, and 5 flags are for producing strain；
C, the preservation of aspergillosis strain
Using Storage in refrigerator method, put in refrigerator by 5 preservation strains and preserve, temperature controls between 4～5 DEG C, and the time must not
More than 6 months.
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| CN201410109595.4A CN103937679B (en) | 2014-03-21 | 2014-03-21 | A kind of aspergillosis strain of vinegar processed |
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| CN1632098B (en) * | 2004-11-25 | 2010-09-08 | 吉林大学 | Vinegar containing organic selenium from aspergillus oryzae and method for preparing same |
| CN101979526A (en) * | 2010-11-05 | 2011-02-23 | 山西三盟实业发展有限公司 | Method for preparing complex enzyme preparation for vinegar |
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