CN108456648A - A kind of method of aflatoxin B1 in degradation peanut meal - Google Patents
A kind of method of aflatoxin B1 in degradation peanut meal Download PDFInfo
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- CN108456648A CN108456648A CN201810177061.3A CN201810177061A CN108456648A CN 108456648 A CN108456648 A CN 108456648A CN 201810177061 A CN201810177061 A CN 201810177061A CN 108456648 A CN108456648 A CN 108456648A
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- aflatoxin
- peanut meal
- bacillus subtilis
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/28—Removal of unwanted matter, e.g. deodorisation or detoxification using microorganisms
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Abstract
The invention be related to it is a kind of degradation peanut meal in aflatoxin B1 method, it includes that the bacillus subtilis of preservation is trained bacillus subtilis bacterium solution：Prepare LB liquid medium：（Every liter）For 10 g of peptone, 10 g of sodium chloride, 5 g of yeast extract, pH value is adjusted to 7.2, and 50 mL culture mediums are dispensed into 250 ml triangular flasks, adds 4 layers of newspaper to seal using 16 layers of gauze, then sterilize 20min in high-pressure sterilizing pot, the bacillus subtilis of preservation is linked into sterilized culture medium, 37 DEG C, it is spare after 200r cultures for 24 hours；Therefore, the present invention have the advantages that have a wide range of application, the aflatoxin in peanut meal of effectively degrading and easy to operate.
Description
Technical field
The invention belongs to the aflatoxin technical fields in peanut meal of degrading, and in particular to a kind of microbial solid fermentation
The method of aflatoxin B1 in degradation peanut meal.
Background technology
The peanut protein of peanut meal almost includes eight kinds of amino acid needed by human, and glutamic acid, aspartic acid are higher, and dynamic
Object albumen is close, and is free of cholesterol, and digestibility is high, so there is abundant nutritive value.Therefore, peanut meal is raised as animal
Material is widely used；Peanut meal is the good carrier that aspergillus flavus grows, if peanut meal moisture is higher or preserves ring
Border humidity is excessively high, proper temperature（25~40 DEG C）, it is easy to aspergillus flavus is grown, causes aflatoxin exceeded, therefore, is used
Aflatoxin B1 in the safe and effective degradation peanut meal of microbial solid fermentation, improves the utility value and warp of peanut meal
Ji benefit, has very big application prospect.
Invention content
The purpose of the invention is to overcome the deficiencies in the prior art, and provide one kind and have a wide range of application, flower of effectively degrading
The method of aflatoxin in the raw dregs of rice and the aflatoxin B1 in easy to operate microbial solid fermentation degradation peanut meal.
The object of the present invention is achieved like this：A kind of method of aflatoxin B1 in degradation peanut meal, it includes
The bacillus subtilis of preservation is trained bacillus subtilis bacterium solution：Prepare LB liquid medium：（Every liter）For peptone 10
G, 10 g of sodium chloride, 5 g of yeast extract, pH value are adjusted to 7.2, and 50 mL culture mediums are dispensed into 250 ml triangular flasks, use
16 layers of gauze add 4 layers of newspaper sealing, and then sterilize in high-pressure sterilizing pot 20min, and the bacillus subtilis of preservation is linked into
It is 37 DEG C, spare after 200r cultures for 24 hours in sterilized culture medium.
It is described that bacillus subtilis bacterium solution is trained bacillus subtilis fermentation liquor：Fermentation medium components used：Pancreas
Peptone 10g, yeast extract 2g, glucose 2g, beef extract 3g, sodium chloride 4g, disodium hydrogen phosphate 3g, epsom salt 1.0g,
1000 ml of distilled water, pH value 7.2 dispense 50 mL culture mediums into 250 ml triangular flasks, add 4 layers of report using 16 layers of gauze
Paper seals, and then sterilize in high-pressure sterilizing pot 20min, takes the bacillus subtilis bacteria culture fluid 1ml inoculations after being cultivated in step 1
In 50ml fermentation mediums, 37 DEG C, 200r cultivate 2h after it is spare.
In proportion by peanut meal and water（Moisture 90%）（g/ml）After mixing, 121 DEG C of high pressure sterilization 10-
30min, ph to 7-6 is adjusted after cooling, then accesses the bacillus subtilis fermentation liquor in step 2, inoculum concentration 10%, stirring
After uniformly, 37 DEG C of cultures in incubator are put in, and primary every 12h stirrings, it is primary every microscopy for 24 hours, after cultivating 72h, take out
Dry, drying temperature is not higher than 60 DEG C, and moisture is not higher than 12% after drying, detects aflatoxin B1 content therein.
The content of the aflatoxin B1 in the peanut meal after unfermentable peanut meal and fermentation, detection method are detected respectively
Using affine in immunity column purification-high performance liquid chromatography detection, and degradation rate is calculated, degradation rate can reach 55%.
Beneficial effects of the present invention：The present invention is using the aflatoxin in microbial solid fermentation method degradation peanut meal
B1 does not influence the nutritive value of peanut meal, and detoxicating method is safe efficient, has compared to other physics, chemical detoxicating method apparent excellent
Gesture；The bacterial strain used in fermentation process of the present invention is bacillus subtilis, through screening obtain can be with efficient degradation aspergillus flavus poison
The bacterial strain of plain B1, and there is prebiotic effect, a variety of harmful bacterias can be inhibited；Microbial solid fermentation method and process is simple, cost
It is relatively low, and than other poison-removing method temperature；Therefore, the present invention have have a wide range of application, effectively degrade peanut meal in aspergillus flavus
Toxin and easy to operate advantage.
Specific implementation mode
Embodiment 1
A kind of method of aflatoxin B1 in degradation peanut meal, it includes that the bacillus subtilis of preservation is trained withered grass
Bacillus bacterium solution：Prepare LB liquid medium：（Every liter）For 10 g of peptone, 10 g of sodium chloride, 5 g of yeast extract, pH value
7.2 are adjusted to, 50 mL culture mediums are dispensed into 250 ml triangular flasks, adds 4 layers of newspaper to seal using 16 layers of gauze, then exists
Sterilize 20min in high-pressure sterilizing pot, the bacillus subtilis of preservation is linked into sterilized culture medium, 37 DEG C, 200r trainings
It is spare after supporting for 24 hours.
The present invention does not influence peanut meal using the aflatoxin B1 in microbial solid fermentation method degradation peanut meal
Nutritive value, detoxicating method is safe efficient, has a clear superiority compared to other physics, chemical detoxicating method；Fermentation process of the present invention
The middle bacterial strain used is bacillus subtilis, through screening obtain can be with the bacterial strain of efficient degradation aflatoxin B1, and have
Prebiotic effect can inhibit a variety of harmful bacterias；Microbial solid fermentation method and process is simple, and cost is relatively low, and than other detoxifications
Method temperature；Bacillus subtilis bacterium solution is trained bacillus subtilis fermentation liquor by specific step to be described：Fermentation used
Medium component：Tryptone 10g, yeast extract 2g, glucose 2g, beef extract 3g, sodium chloride 4g, disodium hydrogen phosphate 3g, seven
50 mL culture mediums are dispensed into 250 ml triangular flasks, use 16 by water magnesium sulfate 1.0g, 1000 ml of distilled water, pH value 7.2
Layer gauze adds 4 layers of newspaper sealing, and then sterilize in high-pressure sterilizing pot 20min, takes the bacillus subtilis after being cultivated in step 1
Culture solution 1ml is inoculated in 50ml fermentation mediums, 37 DEG C, spare after 200r cultures 2h；In proportion by peanut meal and water（Moisture
Content 90%）（g/ml）After mixing, 121 DEG C of high pressure sterilization 10-30min adjust ph to 7-6 after cooling, then access step
Bacillus subtilis fermentation liquor in 2, inoculum concentration 10% after stirring evenly, are put in 37 DEG C of cultures in incubator, and every 12h
Stirring is primary, primary every microscopy for 24 hours, after cultivating 72h, takes out drying, drying temperature is not higher than 60 DEG C, moisture after drying
Not higher than 12%, aflatoxin B1 content therein is detected；Therefore, the present invention have have a wide range of application, peanut meal of effectively degrading
In aflatoxin and easy to operate advantage.
Embodiment 2
A kind of method of aflatoxin B1 in degradation peanut meal, it includes that the bacillus subtilis of preservation is trained withered grass
Bacillus bacterium solution：Prepare LB liquid medium：（Every liter）For 10 g of peptone, 10 g of sodium chloride, 5 g of yeast extract, pH value
7.2 are adjusted to, 50 mL culture mediums are dispensed into 250 ml triangular flasks, adds 4 layers of newspaper to seal using 16 layers of gauze, then exists
Sterilize 20min in high-pressure sterilizing pot, the bacillus subtilis of preservation is linked into sterilized culture medium, 37 DEG C, 200r trainings
It is spare after supporting for 24 hours；It is described that bacillus subtilis bacterium solution is trained bacillus subtilis fermentation liquor：Fermentation medium used at
Point：Tryptone 10g, yeast extract 2g, glucose 2g, beef extract 3g, sodium chloride 4g, disodium hydrogen phosphate 3g, epsom salt
1.0g, 1000 ml of distilled water, pH value 7.2 dispense 50 mL culture mediums into 250 ml triangular flasks, are added using 16 layers of gauze
4 layers of newspaper sealing, then sterilize in high-pressure sterilizing pot 20min, takes the bacillus subtilis bacteria culture fluid after being cultivated in step 1
1ml is inoculated in 50ml fermentation mediums, 37 DEG C, spare after 200r cultures 2h；In proportion by peanut meal and water（Moisture
90%）（g/ml）After mixing, 121 DEG C of high pressure sterilization 10-30min adjust ph to 7-6 after cooling, then access in step 2
Bacillus subtilis fermentation liquor, inoculum concentration 10% after stirring evenly, is put in incubator 37 DEG C of cultures, and stir every 12h
Mix it is primary, it is primary every microscopy for 24 hours, after cultivating 72h, take out drying, drying temperature is not higher than 60 DEG C, and moisture is not after drying
Higher than 12%, aflatoxin B1 content therein is detected；It detects respectively in the peanut meal after unfermentable peanut meal and fermentation
The content of aflatoxin B1, detection method calculate degradation using affine in immunity column purification-high performance liquid chromatography detection
Rate, degradation rate can reach 55%.
Specific implementation mode is unrestricted to further explanation of the invention, is existed for those of ordinary skills
Further transformation is done to structure in the case of not departing from substantive content of the present invention, and all these transformation should all belong to institute of the present invention
Attached scope of the claims.
Claims (4)
1. it is a kind of degradation peanut meal in aflatoxin B1 method, it include the bacillus subtilis of preservation is trained it is withered
Careless bacillus bacterium solution：Prepare LB liquid medium：（Every liter）For 10 g of peptone, 10 g of sodium chloride, yeast extract 5 g, pH
Value is adjusted to 7.2, and 50 mL culture mediums are dispensed into 250 ml triangular flasks, adds 4 layers of newspaper to seal using 16 layers of gauze, then
Sterilize 20min in high-pressure sterilizing pot, the bacillus subtilis of preservation is linked into sterilized culture medium, 37 DEG C, 200r
It is spare after culture for 24 hours.
2. the method for the aflatoxin B1 in a kind of degradation peanut meal according to claim 1, it is characterised in that：It is described
Bacillus subtilis bacterium solution is trained bacillus subtilis fermentation liquor：Fermentation medium components used：Tryptone 10g, ferment
Female medicinal extract 2g, glucose 2g, beef extract 3g, sodium chloride 4g, disodium hydrogen phosphate 3g, epsom salt 1.0g, distilled water 1000
Ml, pH value 7.2 dispense 50 mL culture mediums into 250 ml triangular flasks, add 4 layers of newspaper to seal using 16 layers of gauze, then
Sterilize 20min in high-pressure sterilizing pot, and the bacillus subtilis bacteria culture fluid 1ml after being cultivated in step 1 is taken to be inoculated in 50ml fermentations
In culture medium, 37 DEG C, 200r cultivate 2h after it is spare.
3. the method for the aflatoxin B1 in a kind of ferment degradation peanut meal according to claim 1, it is characterised in that：It will
Peanut meal and water are in proportion（Moisture 90%）（g/ml）After mixing, 121 DEG C of high pressure sterilization 10-30min are adjusted after cooling
Ph to 7-6 is saved, the bacillus subtilis fermentation liquor in step 2 is then accessed, inoculum concentration 10% after stirring evenly, is put in culture
37 DEG C of cultures in case, and it is primary every 12h stirrings, it is primary every microscopy for 24 hours, after cultivating 72h, drying is taken out, drying temperature is not
Higher than 60 DEG C, moisture is not higher than 12% after drying, detects aflatoxin B1 content therein.
4. the method for the aflatoxin B1 in a kind of degradation peanut meal according to claim 1, it is characterised in that：Respectively
The content of the aflatoxin B1 in the peanut meal after unfermentable peanut meal and fermentation is detected, detection method uses affine in immunity
Column purification-high performance liquid chromatography detection, and degradation rate is calculated, degradation rate can reach 55%.
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| CN201810177061.3A CN108456648A (en) | 2018-03-04 | 2018-03-04 | A kind of method of aflatoxin B1 in degradation peanut meal |
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| CN201810177061.3A CN108456648A (en) | 2018-03-04 | 2018-03-04 | A kind of method of aflatoxin B1 in degradation peanut meal |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109331788A (en) * | 2018-10-29 | 2019-02-15 | 山东省农业科学院农业质量标准与检测技术研究所 | The adsorbent and preparation method thereof of aflatoxin in a kind of removal peanut oil |
| CN110872569A (en) * | 2018-08-29 | 2020-03-10 | 重庆市畜牧科学院 | Culture medium for culturing recombinant bacillus subtilis with melittin protein fragment displayed on surface |
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| CN102067957A (en) * | 2010-10-28 | 2011-05-25 | 江南大学 | Application of bacillus megaterium in degradation of peanut meal aflatoxin B1 |
| CN102031234A (en) * | 2010-11-09 | 2011-04-27 | 中国农业大学 | Bacillussubtilis for decomposing aflatoxin and active protein secreted by same |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110872569A (en) * | 2018-08-29 | 2020-03-10 | 重庆市畜牧科学院 | Culture medium for culturing recombinant bacillus subtilis with melittin protein fragment displayed on surface |
| CN109331788A (en) * | 2018-10-29 | 2019-02-15 | 山东省农业科学院农业质量标准与检测技术研究所 | The adsorbent and preparation method thereof of aflatoxin in a kind of removal peanut oil |
| CN109331788B (en) * | 2018-10-29 | 2021-09-28 | 山东省农业科学院农业质量标准与检测技术研究所 | Adsorbent for removing aflatoxin in peanut oil and preparation method thereof |
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Application publication date: 20180828 |