CN103834598B - A kind of method of screening special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian - Google Patents
A kind of method of screening special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian Download PDFInfo
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Abstract
Screen a method for special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian, relate to vinegar.Take wine of rice fermented with red yeast as the special acetic bacteria of carbon source enrichment Fujian Monas cuspurpureus Went vinegar; The special acetic acid bacteria strain of primary dcreening operation Fujian Monas cuspurpureus Went vinegar; Sieve the special acetic acid bacteria strain of Fujian Monas cuspurpureus Went vinegar again, the acid producing ability of more different primary dcreening operation bacterial strain, the bacterial strain that can produce acid in the substratum higher than 4% ethanol is fast special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian.Described bacterial strain is Y5052 and Y5072.Through qualification, Y5052 and Y5072 is wood sugar glyconic acid acetobacter.Adopt two gordian techniquies.One is adopt to add aimed strain in wine of rice fermented with red yeast enrichment mother of vinegar, and two is adopt high concentration ethanol culture medium flat plate, makes acetic bacteria bacterial strain can form hydrolysis transparent circle on flat board, increases the success ratio of screening.The enriching method adding wine of rice fermented with red yeast in mother of vinegar adopted and add the screening method of high concentration ethanol in flat board, can simplify screening process, improve screening efficiency.
Description
Technical field
The present invention relates to vinegar, especially relate to a kind of method of screening special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian.
Background technology
Vinegar is the traditional brewing seasonings of China, can improve a poor appetite, suppresses germ, help digest, indispensable in people's dietetic life.Vinegar is raw material with starchiness such as grains (Cereals class, potato class etc.), completes the stage brews such as saccharification, zymamsis, acetic fermentation form through different types of microorganisms.The main component of vinegar is acetic acid (3% ~ 5%), also containing nutritive ingredient and flavor compound such as each seed amino acid, organic acid, carbohydrate, VITAMIN, alcohol and esters, has unique color.
The production technique of vinegar can be divided into solid state fermentation, the large class of liquid state fermentation two.Folk tradition vinegar adopts solid-state fermentation process, and also there is employing liquid-state fermentation technology in medium and small enterprise, and these techniques generally can only produce medium-sized vinegar.Along with socio-economic development and people's living standard improve, the high-grade vinegar seasonings of market demand.Fujian Monas cuspurpureus Went vinegar is one of China four great tradition name vinegar, has unique local flavor and health-care effect.The high-grade vinegar of traditional characteristics generally all has the features such as technique uniqueness, output are lower, cannot realize industrial-scale production, can not meet the demand in market.Need to adopt modern biotechnology transformation traditional technology, improve product quality and yield, meet social needs.
Many bacterial strains acetic bacteria that conventional solid-state zymotechnique utilizes nature to inoculate produces vinegar, and modern liquid-state fermentation technology adopts single pure tungus inoculation to ferment, but local flavor is poor.Fujian Monas cuspurpureus Went vinegar also for liquid state fermentation, but adopts for many years that mother of vinegar is as Inoculant, and product special flavour is unique, in this mother of vinegar, obviously there are some specific acetic bacteria bacterial strains.It is existing that much about the research of separating acetic acid bacterium, general extensive sampling, comprises the soil sample of mother of vinegar, vinegar sample, vinegar production environment, etc.In physical environment, microorganism is of a great variety, and each microbe species proportion is very low.For being separated certain specified strain, investigator needs first to take enrichment measure, to increase the ratio of this bacterial classification in sample microbial total quantity, before separation and purification, therefore adopts suitable beneficiation technologies will to be the key successfully obtaining object bacterial classification.In the work of separating acetic acid bacterium bacterial classification, mostly investigator is to take interpolation glucose or ethanol with enrichment acetic bacteria.Brewageing of Fujian Monas cuspurpureus Went vinegar have employed unique preservation mother of vinegar for many years, this mother of vinegar acid content reaches 6% ~ 8%, be far longer than the content of acetic acid 3% ~ 5% in general vinegar, original dominant acetic bacteria itself is also suppressed in this environment, and quantity significantly declines; If take this mother of vinegar as sample, dilution-plate method separating acetic acid bacterium is directly adopted to be difficult to succeed.
Summary of the invention
The object of the present invention is to provide a kind of method of screening special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian.
Another object of the present invention is to provide special acetic bacteria in the Monas cuspurpureus Went vinegar of a kind of Fujian.
The method of special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of described screening Fujian, comprises the following steps:
1) take wine of rice fermented with red yeast as the special acetic bacteria of carbon source enrichment Fujian Monas cuspurpureus Went vinegar:
Glutinous rice is added first time water soaking, eluriate, drain, steam heating, load in altar after spreading for cooling, add red colouring agent for food, also used as a Chinese medicine and second time water, stir evenly rear Zha Tankou, control temperature, lower than 34 DEG C, maintains 40 ~ 45 days, intermediate agitation 4 ~ 5 times, i.e. obtained wine of rice fermented with red yeast fermented glutinous rice; Again after filtration, after heating, i.e. obtained wine of rice fermented with red yeast; In wine of rice fermented with red yeast, add Monas cuspurpureus Went vinegar mother of vinegar, keep temperature lower than 34 DEG C, 5 ~ 35 days time, be i.e. the enrichment culture thing of the obtained special acetic acid bacteria strain of enrichment Fujian Monas cuspurpureus Went vinegar;
2) the special acetic acid bacteria strain of primary dcreening operation Fujian Monas cuspurpureus Went vinegar:
First prepare acetic bacteria selective medium, then it is dull and stereotyped to prepare acetic bacteria selective medium; Dilution step 1) the enrichment culture thing of the obtained special acetic acid bacteria strain of enrichment Fujian Monas cuspurpureus Went vinegar, the concentration of dilution is followed successively by: 10
-1, 10
-2... to 10
-8getting each concentration dilution liquid 100 μ L is respectively coated on acetic bacteria selective medium flat board, be inverted and cultivate, select the bacterium colony that acetic bacteria selective medium flat board crossed existing hydrolysis last week, at the flat lining out purifying of acetic bacteria selective medium, then be inoculated on acetic bacteria selective medium inclined-plane, namely obtain the special acetic acid bacteria strain of Fujian Monas cuspurpureus Went vinegar of primary dcreening operation;
3) the special acetic acid bacteria strain of Fujian Monas cuspurpureus Went vinegar is sieved again:
Sour substratum is produced in preparation, by step 2) the Fujian Monas cuspurpureus Went vinegar special acetic acid bacteria strain of primary dcreening operation is inoculated in and fills 100mL and produce in the triangular flask of sour substratum, first time shaking table cultivate, obtain primary dcreening operation bacterial strain seed liquor; Get 100mL again and produce sour substratum in triangular flask, access 1mL primary dcreening operation bacterial strain seed liquor, second time shaking table is cultivated, and obtains primary dcreening operation bacterial strain bacterium liquid, then measures the acid producing ability of primary dcreening operation bacterial strain bacterium liquid, concrete grammar is: draw primary dcreening operation bacterial strain bacterium liquid 200 μ L, add 1.8mL sterilized water and dilute 10 times, drip the alcohol phenolphthalein solution of 1 ~ 2 0.1%, be titrated to micro mist look with 0.1mol/LNaOH solution, record the volume of the 0.1mol/LNaOH solution of consumption, primary dcreening operation bacterial strain produces acid amount and is:
X（g/L)=V
NaOH×C
NaOH×0.060×100/V
Wherein, V
naOH---measurement sample consumes NaOH reference liquid volume, mL;
C
naOH---the concentration of NaOH standardized solution, mol/L;
0.060---the quality of acetic acid suitable for 1.0mLNaOH standardized solution [C (NaOH)=1.000mol/L], g;
V---volume of sample, mL;
Produce acid amount X---total acid content (with acetometer) in sample, g/L;
The acid producing ability of more different primary dcreening operation bacterial strain, the bacterial strain that can produce acid in the substratum higher than 4% ethanol is fast special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian.
In step 1), the mass ratio of described glutinous rice, for the first time water, red colouring agent for food, also used as a Chinese medicine, second time water, Monas cuspurpureus Went vinegar mother of vinegar and wine of rice fermented with red yeast can be 20: 40: (1 ~ 2): (30 ~ 40): (20 ~ 40): (10 ~ 20); The time of described immersion can be 15 ~ 18h; Described elutriation can adopt clear water to eluriate 3 ~ 4 times; The described steam-heated time can be 15 ~ 25min; Described spreading for cooling can spreading for cooling to 40 ~ 45 DEG C; Described red colouring agent for food, also used as a Chinese medicine can adopt commercially available red colouring agent for food, also used as a Chinese medicine; Described Zha Tankou can adopt newspaper Zha Tankou; Described filtration can adopt cloth bag squeeze and filter; The condition of described heating can heat 5 ~ 20min at 80 ~ 95 DEG C.
In step 2) in, the composition of described acetic bacteria selective medium can be by mass percentage: glucose 5% ~ 10%, peptone 0.5%, yeast powder 1%, CaCO
31%, agar 1.5% ~ 2%, dehydrated alcohol 1.5% ~ 8%, surplus is water; The preparation method of described acetic bacteria selective medium can be: glucose, peptone, yeast powder, CaCO
3, agar and water mixing, 121 DEG C of sterilizing 20min, are cooled to 45 ~ 55 DEG C, then add dehydrated alcohol after sterilizing, obtain acetic bacteria selective medium; Described dilution can adopt decimal dilution method; The described condition of cultivating of being inverted can be inverted cultivation 3 ~ 7 days at 25 ~ 32 DEG C.
In step 3), the sour substratum of described product composition by mass percentage can be: glucose 1% ~ 4%, yeast powder 1%, dehydrated alcohol 1% ~ 8%, and surplus is water, pH5.5 ~ 7.5; Described first time the shaking table condition of cultivating can be 25 ~ 32 DEG C, cultivate 24h in the shaking table of 100 ~ 180r/min; The condition that described second time shaking table is cultivated can be 25 ~ 32 DEG C, cultivate 48 ~ 120h in the shaking table of 100 ~ 180r/min.
In step 3), the preparation method of described 0.1mol/LNaOH solution can be: take 0.4gNaOH solid, with volumetric flask is settled to 100mL after dissolving with pure water; The preparation method of the alcohol phenolphthalein solution of described 0.1% can be: take 0.025g phenolphthalein powder and be dissolved in dehydrated alcohol, be settled to 25mL with volumetric flask.
In the Monas cuspurpureus Went vinegar of described Fujian, special acetic bacteria bacterial strain is:
1.(Gluconacetobacterswingsii) Y5052, described (Gluconacetobacterswingsii) Y5052 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 21st, 2013, address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101, registers on the books and is numbered CGMCCNo.8494 in preservation center.
2.(Gluconacetobacterswingsii) Y5072, described (Gluconacetobacterswingsii) Y5072 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 21st, 2013, address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101, registers on the books and is numbered CGMCCNo.8495 in preservation center.
Through qualification, (Gluconacetobacterswingsii) Y5052 and (Gluconacetobacterswingsii) Y5072 is wood sugar glyconic acid acetobacter.Physiology and biochemistry Epidemiological Analysis finds, the fermentation and acid speed of (Gluconacetobacterswingsii) Y5052 is comparatively slow, the fermentation and acid speed of (Gluconacetobacterswingsii) Y5072.
The present invention adopts two gordian techniquies.One is adopt to add aimed strain in wine of rice fermented with red yeast enrichment mother of vinegar, and two is adopt high concentration ethanol culture medium flat plate, makes acetic bacteria bacterial strain can form hydrolysis transparent circle on flat board, increases the success ratio of screening.The enriching method adding wine of rice fermented with red yeast in mother of vinegar that the present invention adopts and add the screening method of high concentration ethanol in flat board, can simplify screening process, improve screening efficiency.
Embodiment
The present invention is further illustrated for following examples.
Embodiment 1:
The screening of bacterial strain, concrete operation step is as follows:
1. be Monas cuspurpureus Went vinegar special acetic bacteria in carbon source enrichment Fujian with wine of rice fermented with red yeast
1.1. wine of rice fermented with red yeast fermented glutinous rice is prepared
Get glutinous rice 20kg, clear water is eluriated, and the 40kg that adds water soaks 18h; Clear water eluriate 4 times, drain; By glutinous rice steam heating 20min in steamer, spreading for cooling to 42 DEG C; Move in altar by the glutinous rice after boiling, add commercially available red colouring agent for food, also used as a Chinese medicine 1.5kg, water 30kg, stirs evenly; Use newspaper Zha Tankou, control temperature, lower than 32 DEG C, maintains 45 days, intermediate agitation 4 ~ 5 times, i.e. obtained wine of rice fermented with red yeast fermented glutinous rice.
1.2. wine of rice fermented with red yeast is prepared
By aforementioned wine of rice fermented with red yeast fermented glutinous rice through cloth bag squeeze and filter, 85 DEG C of heating 10min, i.e. obtained wine of rice fermented with red yeast.
1.3. the special acetic bacteria of enrichment Fujian Monas cuspurpureus Went vinegar
Get traditional Monas cuspurpureus Went vinegar mother of vinegar 30kg, the wine of rice fermented with red yeast 15kg obtained by interpolation, keep temperature lower than 30 DEG C, 15 days time, the enrichment culture thing of the obtained special acetic acid bacteria strain of enrichment Fujian Monas cuspurpureus Went vinegar.
2. screen the special acetic acid bacteria strain of Fujian Monas cuspurpureus Went vinegar
2.1. acetic bacteria selective medium is prepared
Selective medium is filled a prescription: glucose 7%, peptone 0.5%, yeast powder 1%, CaCO
31%, agar 1.5%7,121 DEG C of sterilizing 20min, are cooled to 50 DEG C after sterilizing, add 6% dehydrated alcohol, and obtained acetic bacteria selective medium, prepares acetic bacteria selective medium flat board for subsequent use.
2.2. primary dcreening operation process
Adopt decimal dilution method, dilute the enrichment culture thing of the special acetic bacteria of above-mentioned enrichment Fujian Monas cuspurpureus Went vinegar, the concentration of dilution is followed successively by: 10
-1, 10
-2... to 10
-8, get each concentration dilution liquid 100 μ L respectively and be coated on selective medium flat board, be inverted cultivation 7 days for 32 DEG C; Select the bacterium colony that selective medium flat board crossed existing hydrolysis last week, at the flat lining out purifying of selective medium, be then inoculated on selective medium inclined-plane, obtain 12 strain Fujian Monas cuspurpureus Went vinegar special acetic bacteria primary dcreening operation bacterial strain.
3. sieve the special acetic bacteria strain excellent of Fujian Monas cuspurpureus Went vinegar again
3.1. sour substratum is produced
Produce sour culture medium prescription as follows: glucose 1% ~ 4%, yeast powder 1%, dehydrated alcohol 6%, pH6.0.
3.2. seed liquor is prepared
From the selective medium inclined-plane of primary dcreening operation bacterial strain, a small amount of test strains of picking is inoculated in and fills the 150mL triangular flask that 100mL produces sour substratum; 32 DEG C, cultivate 24h, i.e. obtained primary dcreening operation bacterial strain seed liquor in the shaking table of 150r/min.
3.3. bacterium liquid is prepared
Get 100mL and produce sour substratum to 150mL triangular flask, access 1mL primary dcreening operation bacterial strain seed liquor, 30 DEG C, cultivate 72h, i.e. obtained primary dcreening operation bacterial strain bacterium liquid in the shaking table of 150r/min.
3.4. the acid producing ability of primary dcreening operation bacterial strain is measured
The method of conventional mensuration total acid is adopted to measure the acid producing ability of primary dcreening operation bacterial strain.
Preparation 0.1mol/LNaOH solution: take 0.4gNaOH solid, with volumetric flask is settled to 100mL after dissolving with pure water.Prepare 0.1% alcohol phenolphthalein: take 0.025g phenolphthalein powder and be dissolved in dehydrated alcohol, be settled to 25mL with volumetric flask.
Draw primary dcreening operation bacterial strain bacterium liquid 0.2mL, add 1.8mL sterilized water and dilute 10 times, drip the alcohol phenolphthalein solution of 1 ~ 2 0.1%, be titrated to micro mist look with 0.1mol/LNaOH solution, record the volume of the 0.1mol/LNaOH solution of consumption.
Primary dcreening operation bacterial strain produces acid amount X(g/L)=V
naOH× C
naOH× 0.060 × 100/V
V
naOH---measure and consume NaOH reference liquid volume, mL
C
naOH---the concentration of NaOH standardized solution, mol/L
0.060---the quality of acetic acid suitable for 1.0mLNaOH standardized solution [C (NaOH)=1.000mol/L], g
V---primary dcreening operation bacterial strain bacteria liquid amasss, mL
Produce acid amount X---total acid content (with acetometer) in sample, g/L
Obtain the sour amount of product of 12 strain primary dcreening operation bacterial strains, compare the product acid amount of 12 strain primary dcreening operation bacterial strains, find that a wherein strain is numbered Y5052 strain culturing 6 days product acid amounts and is greater than 10g/L, Ethanol-torlerance reaches 6%, acidproof; Y5052 bacterial strain is the special acetic bacteria of Fujian Monas cuspurpureus Went vinegar.Through identifying that Y5052 bacterial strain is wood sugar glyconic acid acetobacter (Gluconacetobacterswingsii).
Embodiment 2:
Concrete operation step is as follows:
1. be Monas cuspurpureus Went vinegar special acetic bacteria in carbon source enrichment Fujian with wine of rice fermented with red yeast
1.1. wine of rice fermented with red yeast fermented glutinous rice is prepared
The same, slightly.
1.2. wine of rice fermented with red yeast is prepared
The same, slightly.
1.3. the special acetic bacteria of enrichment Fujian Monas cuspurpureus Went vinegar
The same, slightly, but 25 days terminate enrichment process.
2. screen the special acetic acid bacteria strain of Fujian Monas cuspurpureus Went vinegar
2.1 prepare selective medium
Fill a prescription the same, slightly; Add 5% ethanol, preparation is dull and stereotyped for subsequent use.
2.2. primary dcreening operation process
Fill a prescription the same, slightly.Obtain 14 strain acetic bacteria primary dcreening operation bacterial strains
3. multiple sieve
3.1. sour substratum is produced
Fill a prescription the same, but dehydrated alcohol 7%, pH5.5.
3.2. seed liquor is prepared
Process is the same, slightly.
3.3. bacterium liquid is prepared
Get 100mL and produce sour substratum to 150mL triangular flask, access 1mL primary dcreening operation bacterial strain seed liquor, 32 DEG C, cultivate 96h, i.e. obtained primary dcreening operation bacterial strain bacterium liquid in the shaking table of 180r/min.
3.4. the acid producing ability of primary dcreening operation bacterial strain is measured
Measure the product acid amount of 14 strain primary dcreening operation bacterial strains, find that wherein numbering Y5072 strain culturing produces acid amount for 6 days and be greater than 24g/L, Ethanol-torlerance is greater than 6%, acidproof, and Y5072 bacterial strain is the special acetic bacteria of Fujian Monas cuspurpureus Went vinegar.Through identifying that Y5072 bacterial strain is wood sugar glyconic acid acetobacter (Gluconacetobacterswingsii).
Claims (8)
1. screen a method for special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian, it is characterized in that comprising the following steps:
1) take wine of rice fermented with red yeast as the special acetic bacteria of carbon source enrichment Fujian Monas cuspurpureus Went vinegar:
Glutinous rice is added first time water soaking, eluriate, drain, steam heating, load in altar after spreading for cooling, add red colouring agent for food, also used as a Chinese medicine and second time water, stir evenly rear Zha Tankou, control temperature, lower than 34 DEG C, maintains 40 ~ 45 days, intermediate agitation 4 ~ 5 times, i.e. obtained wine of rice fermented with red yeast fermented glutinous rice; Again after filtration, after heating, i.e. obtained wine of rice fermented with red yeast; In wine of rice fermented with red yeast, add Monas cuspurpureus Went vinegar mother of vinegar, keep temperature lower than 34 DEG C, 5 ~ 35 days time, be i.e. the enrichment culture thing of the obtained special acetic acid bacteria strain of enrichment Fujian Monas cuspurpureus Went vinegar;
2) the special acetic acid bacteria strain of primary dcreening operation Fujian Monas cuspurpureus Went vinegar:
First prepare acetic bacteria selective medium, then it is dull and stereotyped to prepare acetic bacteria selective medium; Dilution step 1) the enrichment culture thing of the obtained special acetic acid bacteria strain of enrichment Fujian Monas cuspurpureus Went vinegar, the concentration of dilution is followed successively by: 10
-1, 10
-2... to 10
-8getting each concentration dilution liquid 100 μ L is respectively coated on acetic bacteria selective medium flat board, be inverted and cultivate, select the bacterium colony that acetic bacteria selective medium flat board crossed existing hydrolysis last week, at the flat lining out purifying of acetic bacteria selective medium, then be inoculated on acetic bacteria selective medium inclined-plane, namely obtain the special acetic acid bacteria strain of Fujian Monas cuspurpureus Went vinegar of primary dcreening operation; The composition of described acetic bacteria selective medium is by mass percentage: glucose 5% ~ 10%, peptone 0.5%, yeast powder 1%, CaCO
31%, agar 1.5% ~ 2%, dehydrated alcohol 1.5% ~ 8%, surplus is water;
3) the special acetic acid bacteria strain of Fujian Monas cuspurpureus Went vinegar is sieved again:
Sour substratum is produced in preparation, by step 2) the Fujian Monas cuspurpureus Went vinegar special acetic acid bacteria strain of primary dcreening operation is inoculated in and fills 100mL and produce in the triangular flask of sour substratum, first time shaking table cultivate, obtain primary dcreening operation bacterial strain seed liquor; Get 100mL again and produce sour substratum in triangular flask, access 1mL primary dcreening operation bacterial strain seed liquor, second time shaking table is cultivated, and obtains primary dcreening operation bacterial strain bacterium liquid, then measures the acid producing ability of primary dcreening operation bacterial strain bacterium liquid, concrete grammar is: draw primary dcreening operation bacterial strain bacterium liquid 200 μ L, add 1.8mL sterilized water and dilute 10 times, drip the alcohol phenolphthalein solution of 1 ~ 2 0.1%, be titrated to micro mist look with 0.1mol/LNaOH solution, record the volume of the 0.1mol/LNaOH solution of consumption, primary dcreening operation bacterial strain produces acid amount and is:
X(g/L)＝V
NaOH×C
NaOH×0.060×100/V
Wherein, V
naOH---measurement sample consumes NaOH reference liquid volume, mL;
C
naOH---the concentration of NaOH standardized solution, mol/L;
0.060---the quality of acetic acid suitable for 1.0mLNaOH standardized solution [C (NaOH)=1.000mol/L], g;
V---volume of sample, mL;
Produce acid amount X---total acid content in sample, with acetometer, g/L;
The acid producing ability of more different primary dcreening operation bacterial strain, the bacterial strain that can produce acid in the substratum higher than 4% ethanol is fast special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian;
The sour substratum of described product consisting of by mass percentage: glucose 1% ~ 4%, yeast powder 1%, dehydrated alcohol 1% ~ 8%, surplus is water, pH5.5 ~ 7.5; Described first time the shaking table condition of cultivating be 25 ~ 32 DEG C, cultivate 24h in the shaking table of 100 ~ 180r/min; The condition that described second time shaking table is cultivated be 25 ~ 32 DEG C, cultivate 48 ~ 120h in the shaking table of 100 ~ 180r/min.
2. a kind of method of screening special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian as claimed in claim 1, it is characterized in that in step 1) in, the mass ratio of described glutinous rice, for the first time water, red colouring agent for food, also used as a Chinese medicine, second time water, Monas cuspurpureus Went vinegar mother of vinegar and wine of rice fermented with red yeast is 20: 40: (1 ~ 2): (30 ~ 40): (20 ~ 40): (10 ~ 20).
3. a kind of method of screening special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian as claimed in claim 1, is characterized in that in step 1) in, the time of described immersion is 15 ~ 18h; Described elutriation adopts clear water to eluriate 3 ~ 4 times; The described steam-heated time is 15 ~ 25min; Described spreading for cooling is spreading for cooling to 40 ~ 45 DEG C; Described red colouring agent for food, also used as a Chinese medicine adopts commercially available red colouring agent for food, also used as a Chinese medicine; Described Zha Tankou adopts newspaper Zha Tankou; Described filtration adopts cloth bag squeeze and filter; The condition of described heating heats 5 ~ 20min at 80 ~ 95 DEG C.
4. a kind of method of screening special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian as claimed in claim 1, is characterized in that in step 2) in, the preparation method of described acetic bacteria selective medium is: glucose, peptone, yeast powder, CaCO
3, agar and water mixing, 121 DEG C of sterilizing 20min, are cooled to 45 ~ 55 DEG C, then add dehydrated alcohol after sterilizing, obtain acetic bacteria selective medium.
5. a kind of method of screening special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian as claimed in claim 1, is characterized in that in step 2) in, described dilution adopts decimal dilution method; The described condition of cultivating of being inverted is inverted cultivation 3 ~ 7 days at 25 ~ 32 DEG C.
6. a kind of method of screening special acetic bacteria bacterial strain in the Monas cuspurpureus Went vinegar of Fujian as claimed in claim 1, it is characterized in that in step 3) in, the preparation method of described 0.1mol/LNaOH solution is: take 0.4gNaOH solid, with volumetric flask is settled to 100mL after dissolving with pure water; The preparation method of the alcohol phenolphthalein solution of described 0.1% is: take 0.025g phenolphthalein powder and be dissolved in dehydrated alcohol, be settled to 25mL with volumetric flask.
7. (Gluconacetobacterswingsii) Y5052, being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 21st, 2013, registers on the books and is numbered CGMCCNo.8494 in preservation center.
8. (Gluconacetobacterswingsii) Y5072, being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 21st, 2013, registers on the books and is numbered CGMCCNo.8495 in preservation center.
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| CN101475902A (en) * | 2008-11-27 | 2009-07-08 | 吴德辉 | Production method for edible vinegar |
| CN103421665A (en) * | 2013-07-17 | 2013-12-04 | 杭州清正生物科技有限公司 | Method for producing monascus fruit vinegar |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101475902A (en) * | 2008-11-27 | 2009-07-08 | 吴德辉 | Production method for edible vinegar |
| CN103421665A (en) * | 2013-07-17 | 2013-12-04 | 杭州清正生物科技有限公司 | Method for producing monascus fruit vinegar |
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| Title |
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| 高产红曲色素红曲霉菌株的筛选及抑菌试验的研究;郑虹;《中国酿造》;20131231;第32卷(第2期);70-72 * |
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