CN111820364A - Biodegradation method for aflatoxin in corn steep liquor - Google Patents
Biodegradation method for aflatoxin in corn steep liquor Download PDFInfo
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- CN111820364A CN111820364A CN201910310426.XA CN201910310426A CN111820364A CN 111820364 A CN111820364 A CN 111820364A CN 201910310426 A CN201910310426 A CN 201910310426A CN 111820364 A CN111820364 A CN 111820364A
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- steep liquor
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- aflatoxin
- corn steep
- corn kernels
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- XWIYFDMXXLINPU-UHFFFAOYSA-N Aflatoxin G Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1C1C=COC1O2 XWIYFDMXXLINPU-UHFFFAOYSA-N 0.000 title claims abstract description 49
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 16
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 12
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 12
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
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- 235000015278 beef Nutrition 0.000 claims description 4
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- 229940041514 candida albicans extract Drugs 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 4
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 4
- 239000012153 distilled water Substances 0.000 claims description 4
- 239000000284 extract Substances 0.000 claims description 4
- 238000004108 freeze drying Methods 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- 239000001963 growth medium Substances 0.000 claims description 4
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 4
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 4
- 229940099596 manganese sulfate Drugs 0.000 claims description 4
- 239000011702 manganese sulphate Substances 0.000 claims description 4
- 235000007079 manganese sulphate Nutrition 0.000 claims description 4
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 4
- 230000001376 precipitating effect Effects 0.000 claims description 4
- 238000001556 precipitation Methods 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
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- 238000001228 spectrum Methods 0.000 description 3
- 229930132918 Aflatoxin B2 Natural products 0.000 description 2
- 229930063498 Aflatoxin G1 Natural products 0.000 description 2
- XWIYFDMXXLINPU-WNWIJWBNSA-N Aflatoxin G1 Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1[C@@H]1C=CO[C@@H]1O2 XWIYFDMXXLINPU-WNWIJWBNSA-N 0.000 description 2
- 229930166256 Aflatoxin G2 Natural products 0.000 description 2
- WPCVRWVBBXIRMA-WNWIJWBNSA-N Aflatoxin G2 Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1[C@@H]1CCO[C@@H]1O2 WPCVRWVBBXIRMA-WNWIJWBNSA-N 0.000 description 2
- 239000002115 aflatoxin B1 Substances 0.000 description 2
- OQIQSTLJSLGHID-WNWIJWBNSA-N aflatoxin B1 Chemical compound C=1([C@@H]2C=CO[C@@H]2OC=1C=C(C1=2)OC)C=2OC(=O)C2=C1CCC2=O OQIQSTLJSLGHID-WNWIJWBNSA-N 0.000 description 2
- 239000002097 aflatoxin B2 Substances 0.000 description 2
- WWSYXEZEXMQWHT-WNWIJWBNSA-N aflatoxin B2 Chemical compound C=1([C@@H]2CCO[C@@H]2OC=1C=C(C1=2)OC)C=2OC(=O)C2=C1CCC2=O WWSYXEZEXMQWHT-WNWIJWBNSA-N 0.000 description 2
- 239000002098 aflatoxin G1 Substances 0.000 description 2
- 239000002100 aflatoxin G2 Substances 0.000 description 2
- 229930020125 aflatoxin-B1 Natural products 0.000 description 2
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- 238000007781 pre-processing Methods 0.000 description 2
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- 241000228197 Aspergillus flavus Species 0.000 description 1
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- 210000001835 viscera Anatomy 0.000 description 1
Classifications
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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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B9/00—Preservation of edible seeds, e.g. cereals
- A23B9/08—Drying; Subsequent reconstitution
-
- 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
-
- 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
- A23L5/30—Physical treatment, e.g. electrical or magnetic means, wave energy or irradiation
- A23L5/36—Physical treatment, e.g. electrical or magnetic means, wave energy or irradiation using irradiation with frequencies of more than 10 MHz
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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
- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
- A23L7/10—Cereal-derived products
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention relates to the technical field of biodegradation, and discloses a biodegradation method for aflatoxin in corn steep liquor, which comprises the following steps: harvesting corns in sunny weather, selecting fresh corns without pollution and mildew as raw materials, and separating corn kernels and stalks by using a thresher; and irradiating the corn kernels by using an ultraviolet lamp and a small solar lamp tube, removing moisture in the corn kernels, carrying out ultraviolet radiation illumination on the corn kernels for 80-300W/m cultivation, and carrying out 4000-7500W/m cultivation on the light wave power emitted by the small solar lamp tube. According to the method, aflatoxin on the surfaces of corn kernels is degraded through ultraviolet rays, the degradation effect of the ultraviolet rays on the aflatoxin is enhanced through light waves emitted by a small solar lamp tube, the aflatoxin is enzymatically degraded by bacillus subtilis, the aflatoxin in the corn steep liquor is degraded by a biological method, the method has the characteristics of high efficiency, mildness, no toxicity and safety, and the aflatoxin in the corn steep liquor is degraded and removed from the outside to the inside, so that the aflatoxin in the corn steep liquor is completely removed.
Description
Technical Field
The invention relates to the technical field of biodegradation, in particular to a biodegradation method for aflatoxin in corn steep liquor.
Background
Aflatoxins are a class of secondary metabolites produced primarily by aspergillus flavus and aspergillus parasiticus fungi and are the most toxic fungal toxins found to date. It can contaminate food and feed by several means, and can be directly or indirectly fed into human food chain, and can threaten human health and life safety, and can seriously damage internal organs of human body and animal. The united states centers for disease control and prevention estimate that 45 million people in developing countries have chronic exposure to aflatoxins by diet. According to one analysis, chronic exposure to aflatoxin results in 25200 to 155000 cases of liver cancer each year, especially in asia and sub-saharan africa. It is roughly estimated that on average at least about 2% of grains worldwide per year are inedible for food and feed because of mold contamination, and 25% are contaminated with mycotoxins. The strong toxicity, carcinogenicity and mutagenicity of aflatoxin seriously threaten the production performance of animals and the health of human beings, and cause huge economic loss to the food industry, the feed industry and the animal husbandry every year.
China is one of countries with a lot of corns planted in the world, the China is affected by aflatoxin pollution to different degrees every year, the aflatoxin in the corn steep liquor can affect the health of people when the corns are prepared into the corn steep liquor for eating, at present, reports about adopting ultraviolet rays to control the aflatoxin are reported, the aflatoxin is decomposed and removed by ultraviolet ray irradiation, but the sterilization range of the ultraviolet rays is only limited to the part which can be irradiated by the ultraviolet rays, the interior of the corns can not be sterilized, and the ultraviolet radiation intensity of the ultraviolet rays of an ultraviolet lamp is gradually weakened, so the aflatoxin in the corns can not be completely removed by adopting the ultraviolet ray irradiation.
Disclosure of Invention
Technical problem to be solved
Aiming at the defects of the prior art, the invention provides a biodegradation method for aflatoxin in corn steep liquor, which has the characteristics of high efficiency, mildness, no toxicity and safety, and adopts the combined action of ultraviolet rays, light waves and bacillus subtilis to treat so as to completely remove the aflatoxin in the corn steep liquor.
(II) technical scheme
In order to achieve the purposes of convenient installation and good waterproofness, the invention provides the following technical scheme: a biodegradation method for aflatoxin in corn steep liquor comprises the following steps:
the method comprises the following steps: degradation by ultraviolet light
1) Harvesting corns in sunny weather, selecting fresh corns without pollution and mildew as raw materials,
separating corn kernels and stalks by using a thresher;
2) and irradiating the corn kernels by using an ultraviolet lamp and a small solar lamp tube, removing moisture in the corn kernels, carrying out ultraviolet radiation illumination on the corn kernels for 80-300W/m cultivation, and carrying out 4000-7500W/m cultivation on the light wave power emitted by the small solar lamp tube.
Step two: corn steep liquor preparation
1) Cleaning the corn kernels with clear water, and then soaking the corn kernels in a basin for 5-10 hours;
2) taking out the soaked corn kernels from the water basin, adding the corn kernels into a stirrer, adding clear water into the stirrer, and beating the corn kernels into corn steep liquor by the stirrer;
3) and filtering the beaten corn steep liquor by using a filter screen.
Step three: method for culturing bacillus subtilis
1) Carrying out fermentation culture on bacillus subtilis; the culture medium consists of the following components: 10g of tryptone, 2g of yeast extract, 20g of glucose, 3g of beef extract, 4g of sodium chloride, 3g of disodium hydrogen phosphate, 1.0g of magnesium sulfate heptahydrate, 1.0g of manganese sulfate, 1000mL of distilled water and pH value of 7.2; the conditions for shake flask fermentation were: the fermentation temperature is 30-40 ℃, the fermentation time is 20-30h, the pH value is 7.0-7.4, and the rotation speed is 180-220 r/min.
2) After fermentation is finished, centrifuging to remove thallus, extracting active protein in the supernatant by using an extracting agent, precipitating for 1-24 hours at 0 ℃, and then centrifuging for 10min at 5000r/min to obtain protein precipitate, wherein the extracting agent is 95% methanol, 95% ethanol, 40% ammonium sulfate solution, 60% ammonium sulfate solution or 80% ammonium sulfate solution; preferably, ethanol is used for precipitation at 0 ℃ for 1 hour;
3) dissolving the obtained protein precipitate in PBS buffer solution, freeze-drying and concentrating to obtain the activity.
Step four: biodegradation process
Taking fermentation liquor of bacillus subtilis or PBS solution of active protein (the concentration of the PBS solution is 0.01 mol/L), dissolving the active protein in the corn steep liquor solution to obtain the PBS solution of the active protein with the concentration of 0.01 g/ml) 600-.
(III) advantageous effects
Compared with the prior art, the invention provides a biodegradation method for aflatoxin in corn steep liquor, which has the following beneficial effects:
according to the method for biologically degrading the aflatoxin in the corn steep liquor, the corn steep liquor is irradiated by the ultraviolet lamp and the small solar lamp tube, so that the evaporation of water in the corn steep liquor is accelerated, the corn steep liquor is quickly dried, the mildew of the corn steep liquor is prevented, the aflatoxin on the surface of the corn steep liquor is degraded by the ultraviolet ray, the degradation effect of the ultraviolet ray on the aflatoxin is enhanced by the light wave emitted by the small solar lamp tube, in the process of heating the corn steep liquor, the added bacillus subtilis is used for carrying out enzymatic degradation on the aflatoxin, the aflatoxin in the corn steep liquor is degraded by a biological method, the method has the characteristics of high efficiency, mildness, no toxicity and safety, the aflatoxin in the corn steep liquor is degraded and removed from the outside to the inside, and the aflatoxin in the corn steep liquor is.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example one
A biodegradation method for aflatoxin in corn steep liquor comprises the following steps:
the method comprises the following steps: degradation by ultraviolet light
1) Harvesting corns in sunny weather, selecting fresh corns without pollution and mildew as raw materials,
separating corn kernels and stalks by using a thresher;
2) and irradiating the corn kernels by using an ultraviolet lamp and a small solar lamp tube, removing moisture in the corn kernels, carrying out ultraviolet radiation illumination on the corn kernels for 80-300W/m cultivation, and carrying out 4000-7500W/m cultivation on the light wave power emitted by the small solar lamp tube.
Step two: corn steep liquor preparation
1) Cleaning the corn kernels with clear water, and then soaking the corn kernels in a basin for 5-10 hours;
2) taking out the soaked corn kernels from the water basin, adding the corn kernels into a stirrer, adding clear water into the stirrer, and beating the corn kernels into corn steep liquor by the stirrer;
3) and filtering the beaten corn steep liquor by using a filter screen.
Shine the kernel of corn through ultraviolet lamp and little solar lamp for the evaporation of the inside moisture of kernel of corn for the quick drying of kernel of corn prevents that the kernel of corn from taking place to milden and rot, degrades the aflatoxin on kernel of corn surface through the ultraviolet ray simultaneously, and the degradation effect to aflatoxin of the ultraviolet ray of light wave reinforcing that sends through little solar lamp is to aflatoxin
Example two
A biodegradation method for aflatoxin in corn steep liquor comprises the following steps:
the method comprises the following steps: corn steep liquor preparation
1) Harvesting corns in sunny weather, selecting fresh corns without pollution and mildew as raw materials,
separating corn kernels and stalks by using a thresher;
2) cleaning the corn kernels with clear water, and then soaking the corn kernels in a basin for 5-10 hours;
3) taking out the soaked corn kernels from the water basin, adding the corn kernels into a stirrer, adding clear water into the stirrer, and beating the corn kernels into corn steep liquor by the stirrer;
4) and filtering the beaten corn steep liquor by using a filter screen.
Step two: method for culturing bacillus subtilis
1) Carrying out fermentation culture on bacillus subtilis; the culture medium consists of the following components: 10g of tryptone, 2g of yeast extract, 20g of glucose, 3g of beef extract, 4g of sodium chloride, 3g of disodium hydrogen phosphate, 1.0g of magnesium sulfate heptahydrate, 1.0g of manganese sulfate, 1000mL of distilled water and pH value of 7.2; the conditions for shake flask fermentation were: the fermentation temperature is 30-40 ℃, the fermentation time is 20-30h, the pH value is 7.0-7.4, and the rotation speed is 180-220 r/min.
2) After fermentation is finished, centrifuging to remove thallus, extracting active protein in the supernatant by using an extracting agent, precipitating for 1-24 hours at 0 ℃, and then centrifuging for 10min at 5000r/min to obtain protein precipitate, wherein the extracting agent is 95% methanol, 95% ethanol, 40% ammonium sulfate solution, 60% ammonium sulfate solution or 80% ammonium sulfate solution; preferably, ethanol is used for precipitation at 0 ℃ for 1 hour;
3) dissolving the obtained protein precipitate in PBS buffer solution, freeze-drying and concentrating to obtain the activity.
Step three: biodegradation process
Taking fermentation liquor of bacillus subtilis or PBS solution of active protein (the concentration of the PBS solution is 0.01 mol/L), dissolving the active protein in the corn steep liquor solution to obtain the PBS solution of the active protein with the concentration of 0.01 g/ml) 600-.
The bacillus subtilis has high efficiency of degrading aflatoxin, strong specificity, higher degradation rate of aflatoxin and mild effect, and does not destroy nutrient components in corn steep liquor.
EXAMPLE III
A biodegradation method for aflatoxin in corn steep liquor is characterized by comprising the following steps:
the method comprises the following steps: degradation by ultraviolet light
1) Harvesting corns in sunny weather, selecting fresh corns without pollution and mildew as raw materials,
separating corn kernels and stalks by using a thresher;
2) and irradiating the corn kernels by using an ultraviolet lamp and a small solar lamp tube, removing moisture in the corn kernels, carrying out ultraviolet radiation illumination on the corn kernels for 80-300W/m cultivation, and carrying out 4000-7500W/m cultivation on the light wave power emitted by the small solar lamp tube.
Step two: corn steep liquor preparation
1) Cleaning the corn kernels with clear water, and then soaking the corn kernels in a basin for 5-10 hours;
2) taking out the soaked corn kernels from the water basin, adding the corn kernels into a stirrer, adding clear water into the stirrer, and beating the corn kernels into corn steep liquor by the stirrer;
3) and filtering the beaten corn steep liquor by using a filter screen.
Step three: method for culturing bacillus subtilis
1) Carrying out fermentation culture on bacillus subtilis; the culture medium consists of the following components: 10g of tryptone, 2g of yeast extract, 20g of glucose, 3g of beef extract, 4g of sodium chloride, 3g of disodium hydrogen phosphate, 1.0g of magnesium sulfate heptahydrate, 1.0g of manganese sulfate, 1000mL of distilled water and pH value of 7.2; the conditions for shake flask fermentation were: the fermentation temperature is 30-40 ℃, the fermentation time is 20-30h, the pH value is 7.0-7.4, and the rotation speed is 180-220 r/min.
2) After fermentation is finished, centrifuging to remove thallus, extracting active protein in the supernatant by using an extracting agent, precipitating for 1-24 hours at 0 ℃, and then centrifuging for 10min at 5000r/min to obtain protein precipitate, wherein the extracting agent is 95% methanol, 95% ethanol, 40% ammonium sulfate solution, 60% ammonium sulfate solution or 80% ammonium sulfate solution; preferably, ethanol is used for precipitation at 0 ℃ for 1 hour;
3) dissolving the obtained protein precipitate in PBS buffer solution, freeze-drying and concentrating to obtain the activity.
Step four: biodegradation process
Taking fermentation liquor of bacillus subtilis or PBS solution of active protein (the concentration of the PBS solution is 0.01 mol/L), dissolving the active protein in the corn steep liquor solution to obtain the PBS solution of the active protein with the concentration of 0.01 g/ml) 600-.
According to the method for biologically degrading the aflatoxin in the corn steep liquor, the corn steep liquor is irradiated by the ultraviolet lamp and the small solar lamp tube, so that the evaporation of water in the corn steep liquor is accelerated, the corn steep liquor is quickly dried, the mildew of the corn steep liquor is prevented, the aflatoxin on the surface of the corn steep liquor is degraded by the ultraviolet ray, the degradation effect of the ultraviolet ray on the aflatoxin is enhanced by the light wave emitted by the small solar lamp tube, in the process of heating the corn steep liquor, the added bacillus subtilis is used for carrying out enzymatic degradation on the aflatoxin, the aflatoxin in the corn steep liquor is degraded by a biological method, the method has the characteristics of high efficiency, mildness, no toxicity and safety, the aflatoxin in the corn steep liquor is degraded and removed from the outside to the inside, and the aflatoxin in the corn steep liquor is.
Example three: a biodegradation method for aflatoxin in corn steep liquor comprises the following steps:
the method comprises the following steps: degradation by ultraviolet light
1) Harvesting corns in sunny weather, selecting fresh corns without pollution and mildew as raw materials,
separating corn kernels and stalks by using a thresher;
2) and irradiating the corn kernels by using an ultraviolet lamp and a small solar lamp tube, removing moisture in the corn kernels, carrying out ultraviolet radiation illumination on the corn kernels for 80-300W/m cultivation, and carrying out 4000-7500W/m cultivation on the light wave power emitted by the small solar lamp tube.
Step two: corn steep liquor preparation
1) Cleaning the corn kernels with clear water, and then soaking the corn kernels in a basin for 5-10 hours;
2) taking out the soaked corn kernels from the water basin, adding the corn kernels into a stirrer, adding clear water into the stirrer, and beating the corn kernels into corn steep liquor by the stirrer;
3) and filtering the beaten corn steep liquor by using a filter screen.
Step three: biodegradation
1) Pouring the corn steep liquor into a pot, and boiling for 5-10 minutes with strong fire;
2) in the process of cooking the corn steep liquor, adding bacillus subtilis into a pot, and then uniformly stirring the bacillus subtilis, wherein the addition amount of the bacillus subtilis is 0.01-10% of the mass of the corn steep liquor;
3) and (5) turning off the fire to obtain the corn steep liquor after the aflatoxin degradation.
Step four: the method for detecting the aflatoxin content in the corn steep liquor comprises the following specific steps:
1) performing spectrum detection, namely scanning the spectrum information of the corn steep liquor sample by using a Fourier transform near-infrared spectrometer, taking part of the screened sample, and determining the levels of aflatoxins B1, B2, G1 and G2 and the total amount of aflatoxins in the corn steep liquor sample by using a multifunctional column purification-high performance liquid chromatography-fluorescence method;
2) data preprocessing, namely preprocessing the original spectrum information of the sample obtained in the previous step to eliminate interference;
3) quantitative prediction analysis, namely establishing a correlation relation model between the true content level and the prediction level of aflatoxin in the corn steep liquor sample based on a partial least squares regression analysis method (PLSR) according to the corresponding relation between the levels of aflatoxin B1, B2, G1 and G2 in the corn steep liquor sample and the corresponding spectral absorption values of the aflatoxin B1, B2, G1 and G2;
4) and (3) rapidly determining, namely outputting aflatoxins B1, B2, G1 and G2 and total amount levels thereof based on the spectral information of the corn steep liquor to be determined by utilizing the established model.
By utilizing ATR-FTIR analysis technology, a rapid detection method system for the aflatoxin content level in the corn steep liquor is established, and the method has important significance for ensuring the quality safety of the corn steep liquor and the health of people.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (1)
1. A biodegradation method for aflatoxin in corn steep liquor is characterized by comprising the following steps:
the method comprises the following steps: degradation by ultraviolet light
1) Harvesting corns in sunny weather, selecting fresh corns without pollution and mildew as raw materials,
separating corn kernels and stalks by using a thresher;
2) irradiating the corn kernels by using an ultraviolet lamp and a small solar lamp tube to remove moisture in the corn kernels, carrying out high-speed cultivation under the condition that the irradiation illumination of the ultraviolet light is 80-300W/m, and carrying out high-speed cultivation under the condition that the light wave power emitted by the small solar lamp tube is 4000-7500W/m;
step two: corn steep liquor preparation
1) Cleaning the corn kernels with clear water, and then soaking the corn kernels in a basin for 5-10 hours;
2) taking out the soaked corn kernels from the water basin, adding the corn kernels into a stirrer, adding clear water into the stirrer, and beating the corn kernels into corn steep liquor by the stirrer;
3) filtering the milled corn steep liquor by using a filter screen;
step three: method for culturing bacillus subtilis
1) Carrying out fermentation culture on bacillus subtilis; the culture medium consists of the following components: 10g of tryptone, 2g of yeast extract, 20g of glucose, 3g of beef extract, 4g of sodium chloride, 3g of disodium hydrogen phosphate, 1.0g of magnesium sulfate heptahydrate, 1.0g of manganese sulfate, 1000mL of distilled water and pH value of 7.2; the conditions for shake flask fermentation were: the fermentation temperature is 30-40 ℃, the fermentation time is 20-30h, the pH value is 7.0-7.4, and the rotation speed is 180-;
2) after fermentation is finished, centrifuging to remove thallus, extracting active protein in the supernatant by using an extracting agent, precipitating for 1-24 hours at 0 ℃, and then centrifuging for 10min at 5000r/min to obtain protein precipitate, wherein the extracting agent is 95% methanol, 95% ethanol, 40% ammonium sulfate solution, 60% ammonium sulfate solution or 80% ammonium sulfate solution; preferably, ethanol is used for precipitation at 0 ℃ for 1 hour;
3) dissolving the obtained protein precipitate in PBS buffer solution, and freeze-drying and concentrating to obtain the activity;
step four: biodegradation process
Taking fermentation liquor of bacillus subtilis or PBS solution of active protein (the concentration of the PBS solution is 0.01 mol/L), dissolving the active protein in the corn steep liquor solution to obtain the PBS solution of the active protein with the concentration of 0.01 g/ml) 600-.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101238866A (en) * | 2008-03-05 | 2008-08-13 | 仲恺农业技术学院 | Degradation method of aflatoxin |
| CN102031234A (en) * | 2010-11-09 | 2011-04-27 | 中国农业大学 | Bacillussubtilis for decomposing aflatoxin and active protein secreted by same |
| CN109363058A (en) * | 2017-09-29 | 2019-02-22 | 海南泓缘生物科技股份有限公司 | The biodegradation method of vomitoxin in a kind of pair of corn pulp |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101238866A (en) * | 2008-03-05 | 2008-08-13 | 仲恺农业技术学院 | Degradation method of aflatoxin |
| CN102031234A (en) * | 2010-11-09 | 2011-04-27 | 中国农业大学 | Bacillussubtilis for decomposing aflatoxin and active protein secreted by same |
| CN109363058A (en) * | 2017-09-29 | 2019-02-22 | 海南泓缘生物科技股份有限公司 | The biodegradation method of vomitoxin in a kind of pair of corn pulp |
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