CN110583964A - Biological removal method for efficiently removing four aflatoxins in peanut meal - Google Patents
Biological removal method for efficiently removing four aflatoxins in peanut meal Download PDFInfo
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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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- 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
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Abstract
The invention discloses a biological removal method for efficiently removing four aflatoxins in peanut meal, belonging to the technical field of biological detoxification. According to the invention, 10 with corresponding proportion is inoculated in the peanut meal sample6cfu/mL Aspergillus niger spore suspension is added with sterile distilled water in a corresponding proportion, and solid state fermentation is carried out for a certain time at a proper temperature, so that four aflatoxins in peanut meal are effectively removed, wherein AFTB1The removal rate reaches 95.01 percent, and the total Aflatoxin (AFTB)1、AFTB2、AFTG1、AFTG2) The removal rate reaches 72.90 percent. AFTB in peanut meal treated by the method1The residual quantity not only meets the domestic limit standard, but also meets the European Union established standard (<2 mug/kg), the method has high detoxification efficiency, is environment-friendly and safe, and can be used for food and feedHas application prospect in the aspects of material and biological detoxification, and can provide basis for the formulation of the limit standard of the total quantity of aflatoxin.
Description
Technical Field
The invention relates to a biological removal method for efficiently removing four aflatoxins in peanut meal, belonging to the technical field of biological detoxification.
Background
Aflatoxins (Aflatoxin) are secondary metabolites produced by fungi of the genus aspergillus, which grow naturally in food products and produce a wide range of toxic effects on animals, including humans. Aflatoxins can contaminate agricultural crops and produce in a variety of ways, such as forage cereals, dried fruit products, cakes and the like. In parts of developing countries and regions such as Africa and southeast Asia, the aflatoxin pollution rate of crops and feeds is higher, the aflatoxin pollution of feeds and agricultural products in China is in a rising trend at present, particularly in the Yangtze river and the regions in the south of the Yangtze river, and in the crops, peanuts, corns and corresponding products of the peanuts, the corns and the aflatoxin B are particularly used1The pollution rate is highest, and the standard exceeding rate is also highest. The united nations food and agriculture organization reports show that 25% of grains are polluted by mycotoxins worldwide, about 50 hundred million people are at risk of aflatoxin chronic exposure all over the world, and continuous aflatoxin accumulation can cause cirrhosis and liver cancer. The extremely strong carcinogenicity of the mycotoxin brings huge hidden dangers to the development of agriculture and animal husbandry, limits the development of import and export trade of agriculture and animal husbandry products to a certain extent, and can cause huge economic losses to grain crops, aquatic products, livestock raising, breeding industry and the like.
Peanuts are the main oil crops in China, and the yield is the first of various oil crops. The peanut meal is used as a byproduct of peanut oil pressing, has rich protein content up to 48 percent, can be used as a protein functional food, is easy to infect aspergillus and generate aflatoxin, and greatly limits the wide application of the peanut meal. The aflatoxin is mainly metabolized by aspergillus flavus and aspergillus parasiticus to generate a compound capable of polluting various crops, and the aflatoxin is mainly divided into four types which are aflatoxin B1、B2、G1、G2A compound of this typeThe chemical structures of the materials are similar, the toxic groups are approximately the same, and the research shows that the aflatoxin is arranged into AFTB according to the sequence of toxicity from large to small1>AFTG1>AFTB2>AFTG2Edible quilt AFTB for people1Contaminated food can cause the effects of immune function suppression, malformation, gene mutagenesis, carcinogenesis, etc. of the human body. At present, the drug is only applied to AFTB1The limit is definitely specified, peanuts and products thereof do not exceed 20 mug/kg, the total amount of aflatoxin is a focus of attention at home and abroad at present, but the limit standard of the total amount of aflatoxin is lacked at present at home.
Aflatoxin control methods are broadly divided into three categories: physical, chemical and biological methods. Compared with a physical and chemical method for eliminating the defects and limitations of aflatoxin, the biological removal method has the advantages of safety, high efficiency, environmental protection and the like, so that the method becomes a more potential detoxification method.
Therefore, the biological removal method for removing the four aflatoxins in the peanut meal can remove the four aflatoxins and utilize the peanut meal, and has important significance for food, feed, biological detoxification and the establishment of limit standards of total amount of aflatoxins.
Disclosure of Invention
Aiming at the defects of the existing method, the invention provides an optimized biological removal method capable of removing four aflatoxins in peanut meal with high efficiency aiming at the aspergillus niger inoculation amount, the material-water ratio, the solid-state fermentation temperature and the solid-state fermentation time.
The technical scheme of the invention is as follows:
a method for removing four aflatoxins in peanut meal comprises inoculating 1-20% of 1 × 10 aflatoxin in peanut meal6cfu/mL Aspergillus niger spore suspension, adding sterile water to a certain feed-water ratio (the ratio of the mass of the peanut meal to the volume of the sterile water, g/m L), and performing solid-state fermentation; the Aspergillus niger is preserved in China center for type culture Collection with the preservation number of CCTCC M 2013703。
In one embodiment of the present invention, the solid state fermentation temperature is 20 to 40 ℃.
In one embodiment of the present invention, the solid state fermentation time is 24-72 hours.
In one embodiment of the invention, the Aspergillus niger spore suspension is prepared by eluting spores with 0.9% normal saline containing 0.05% Tween-80 and diluting.
In one embodiment of the invention, the method comprises: culturing Aspergillus niger; solid-state fermenting peanut meal, pulverizing peanut meal containing aflatoxin, sieving with 40 mesh sieve, sterilizing at high temperature and high pressure, transferring to a fermentation vessel, adding 10 of inoculating agent at corresponding ratio6cfu/mL Aspergillus niger spore suspension, adding sterile distilled water in a corresponding proportion, uniformly stirring, and performing solid state fermentation at a proper fermentation temperature for a certain time.
Solid State Fermentation (SSF) refers to a process of Fermentation of one or more microorganisms performed in a State where the medium is in a Solid State, although rich in water, but has no or little free flowing water. Solid state fermentation is one of the longest techniques in the human history for producing products using microorganisms.
In one embodiment of the present invention, the method for culturing aspergillus niger comprises: inoculating Aspergillus niger in a glycerin pipe taken out at the temperature of minus 80 ℃ into a PDA solid culture medium for activation for 3-7 d.
In one embodiment of the invention, the peanut meal is crushed and then sieved by a 40-mesh sieve.
In one embodiment of the invention, the peanut meal is sterilized by a high-temperature high-pressure sterilizer at 121 ℃ for 20 min.
In one embodiment of the present invention, the 1 × 106cfu/mL Aspergillus niger spore suspension is prepared by eluting spore with 0.9% normal saline containing 0.05% Tween-80 and diluting.
In one embodiment of the invention, the inoculation ratio of the aspergillus niger spore suspension is 1% -20%, wherein the inoculation ratio refers to the ratio of the volume of the bacterial liquid to the mass of the peanut meal, and is mL/g.
In one embodiment of the invention, the sterile water is obtained by sterilizing at 121 ℃ for 20min in a high-temperature high-pressure sterilizing pot.
In one embodiment of the invention, the ratio of the material to the water added with the sterile water is 1: 0-1: 2, wherein the ratio of the material to the liquid refers to the ratio of the mass of the peanut meal to the volume of the sterile water, g/m L.
The second purpose of the invention is to provide a peanut meal product by using the method.
The invention has the following beneficial technical effects:
the invention applies the aspergillus niger (CCTCC M2013703) with the effect of removing the aflatoxin, which is separated from the soy sauce mash, to the removal of the four aflatoxins in the peanut meal for the first time, and can ensure that the AFTB can be caused by matching with the specific fermentation conditions1The removal rate of the product reaches 95.01 percent, and the total Aflatoxin (AFTB)1、AFTB2、AFTG1And AFTG2) The removal rate reaches 72.90 percent, and the AFTB in the treated peanut meal1The residual quantity not only meets the domestic limit standard, but also meets the European Union established standard (<2. mu.g/kg); meanwhile, the aspergillus niger can remove aflatoxin in the solid-state fermentation process, and also generates special aroma in the fermentation process.
Drawings
FIG. 1: the fermentation condition is the influence of different aspergillus niger inoculum sizes on the removal rates of the four aflatoxins;
FIG. 2: the fermentation conditions are the influence of different fermentation temperatures on the removal rate of the four aflatoxins;
FIG. 3: the fermentation conditions are the influence of different fermentation time on the removal rate of the four aflatoxins;
FIG. 4: the fermentation condition is the influence of different feed water ratios on the removal rates of the four aflatoxins;
FIG. 5: interaction of factors on AFB1Response surface diagram of removing rate influence; wherein, a is an interaction response surface diagram of the inoculum size and the fermentation temperature, b is an interaction response surface diagram of the inoculum size and the fermentation time, c is an interaction response surface diagram of the inoculum size and the feed-water ratio, and d is an interaction response surface diagram of the fermentation temperature and the fermentation timeThe graph is shown in the specification, wherein e is an interaction response surface graph of fermentation temperature and a feed-water ratio, and f is an interaction response surface graph of the feed-water ratio and fermentation time;
FIG. 6: a response surface diagram of the influence of interaction of all factors on the total AFT removal rate; wherein a is an interaction response surface map of fermentation temperature and inoculation amount, b is an interaction response surface map of fermentation time and inoculation amount, c is an interaction response surface map of inoculation amount and material-water ratio, d is an interaction response surface map of fermentation time and fermentation temperature, e is an interaction response surface map of material-water ratio and fermentation temperature, and f is an interaction response surface map of material-water ratio and fermentation time.
Detailed Description
For a better understanding of the present invention, the following examples are included to further illustrate the present invention, but the present invention is not limited to the following examples.
The method for screening the aspergillus niger in the soy sauce mash comprises the following steps: see chinese patent CN 103937681B. The strain is preserved in China center for type culture Collection in 2013, 12 and 24 months, with the preservation number of CCTCC M2013703 and the preservation address of Wuhan university in China.
Preparation of aspergillus niger CCTCC M2013703 spore suspension:
(1) culturing Aspergillus niger CCTCC M2013703: inoculating Aspergillus niger CCTCC M2013703 from a preserved glycerin pipe into a PDA solid culture medium to activate for 3-7d until the state is good;
(2) potato dextrose agar medium (PDA medium): 300g of potato, 20g of glucose, 20g of agar, 0.1g of chloramphenicol and 1L of distilled water, and autoclaving at 121 ℃ for 20 min;
(3) then eluting with 0.9% normal saline containing 0.05% Tween-80, diluting, and activating to obtain 106cfu/mL Aspergillus niger CCTCC M2013703 spore suspension.
Example 1
Crushing a peanut meal sample containing aflatoxin, sieving with a 40-mesh sieve, uniformly mixing, sampling by adopting a GB/T14699.1-2005 method, weighing 5.00g of the sample, placing the sample in a glass container, sealing a four-layer gauze newspaper, and sterilizing at 121 ℃ for 20min in a high-temperature high-pressure sterilization pot.Transferring sterilized peanut meal to a fermentation dish, and adding 10% of the sterilized peanut meal into the treated sample at a ratio of 20%6cfu/mL Aspergillus niger CCTCC M2013703 spore suspension (eluted and diluted by 0.9% physiological saline containing 0.05% Tween-80), sterile distilled water with the material-water ratio of 1:1.5 is added, the mixture is stirred and mixed uniformly, constant-temperature solid state fermentation is carried out at the fermentation temperature of 32.5 ℃, and a sample after four aflatoxins are removed after the solid state fermentation is carried out for 36 hours is obtained.
Example 2
Crushing a peanut meal sample containing aflatoxin, sieving with a 40-mesh sieve, uniformly mixing, sampling by adopting a GB/T14699.1-2005 method, weighing 5.00g of the sample, placing the sample in a glass container, sealing a four-layer gauze newspaper, and sterilizing at 121 ℃ for 20min in a high-temperature high-pressure sterilization pot. Transferring sterilized peanut meal to a fermentation dish, and adding 10 with the proportion of 12.5% into the treated sample6cfu/mL Aspergillus niger CCTCC M2013703 spore suspension (eluted and diluted by 0.9% physiological saline containing 0.05% Tween-80), sterile distilled water with the material-water ratio of 1:1.5 is added, the mixture is stirred and mixed uniformly, constant-temperature solid state fermentation is carried out at the fermentation temperature of 25 ℃, and a sample after four aflatoxins are removed after the solid state fermentation is carried out for 36 hours is obtained.
Example 3
Crushing a peanut meal sample containing aflatoxin, sieving with a 40-mesh sieve, uniformly mixing, sampling by adopting a GB/T14699.1-2005 method, weighing 5.00g of the sample, placing the sample in a glass container, sealing a four-layer gauze newspaper, and sterilizing at 121 ℃ for 20min in a high-temperature high-pressure sterilization pot. Transferring sterilized peanut meal to a fermentation dish, and adding 10% of the sterilized peanut meal into the treated sample at a ratio of 20%6cfu/mL Aspergillus niger CCTCC M2013703 spore suspension (eluted and diluted by 0.9% physiological saline containing 0.05% Tween-80), sterile distilled water with the material-water ratio of 1:1 is added, the mixture is stirred and mixed uniformly, constant-temperature solid state fermentation is carried out at the fermentation temperature of 32.5 ℃, and a sample after four aflatoxins are removed after solid state fermentation is carried out for 54 hours.
Example 4
Crushing a peanut meal sample containing aflatoxin, sieving with a 40-mesh sieve, uniformly mixing, sampling by a GB/T14699.1-2005 method, andweighing 5.00g of the mixture, placing the mixture in a glass container, sealing the four layers of gauze newspaper, and sterilizing the mixture at the temperature of 121 ℃ for 20min in a high-temperature high-pressure sterilization pot. Transferring sterilized peanut meal to a fermentation dish, and adding 10% of 5% of the treated peanut meal into the treated peanut meal6cfu/mL Aspergillus niger CCTCC M2013703 spore suspension (eluted and diluted by 0.9% physiological saline containing 0.05% Tween-80), sterile distilled water with the material-water ratio of 1:1.5 is added, the mixture is stirred and mixed uniformly, constant-temperature solid state fermentation is carried out at the fermentation temperature of 40 ℃, and a sample after four aflatoxins are removed after solid state fermentation is carried out for 54 hours.
Example 5
Crushing a peanut meal sample containing aflatoxin, sieving with a 40-mesh sieve, uniformly mixing, sampling by adopting a GB/T14699.1-2005 method, weighing 5.00g of the sample, placing the sample in a glass container, sealing a four-layer gauze newspaper, and sterilizing at 121 ℃ for 20min in a high-temperature high-pressure sterilization pot. Transferring sterilized peanut meal to a fermentation dish, and adding 10% of the treated peanut meal into the fermentation dish6cfu/mL Aspergillus niger CCTCC M2013703 spore suspension (eluted and diluted by 0.9% physiological saline containing 0.05% Tween-80), sterile distilled water with the material-water ratio of 1:1.56 is added, the mixture is stirred and mixed uniformly, constant-temperature solid state fermentation is carried out at the fermentation temperature of 31.8 ℃, and a sample after four aflatoxins are removed after solid state fermentation is carried out for 63.3 hours.
Example 6
Crushing a peanut meal sample containing aflatoxin, sieving with a 40-mesh sieve, uniformly mixing, sampling by adopting a GB/T14699.1-2005 method, weighing 5.00g of the sample, placing the sample in a glass container, sealing a four-layer gauze newspaper, and sterilizing at 121 ℃ for 20min in a high-temperature high-pressure sterilization pot. Transferring sterilized peanut meal to a fermentation dish, and adding 10% of the sterilized peanut meal into the treated sample6cfu/mL Aspergillus niger CCTCC M2013703 spore suspension (eluted and diluted by 0.9% physiological saline containing 0.05% Tween-80), sterile distilled water with the material-water ratio of 1:1 is added, the mixture is stirred and mixed uniformly, constant-temperature solid state fermentation is carried out at the fermentation temperature of 30 ℃, and a sample after four aflatoxins are removed after solid state fermentation is obtained after 48 hours of fermentation.
Effect of removing Aflatoxin by the methods of examples 1 to 6And (3) detection: the aflatoxin B1 and the total aflatoxin removal rate (AFTB)1+AFTB2+AFTG1+AFTG2) Is a representative detection index.
Collecting fermented samples, drying, respectively introducing each part of peanut meal into a centrifugal tube, adding acetonitrile-water solution (70+30) for extraction, washing a fermentation vessel, and introducing into the centrifugal tube together, so as to avoid the influence of uneven distribution of the toxin in the meal on the experimental result; mixing, ultrasonic treating for 10min, oscillating for 30min, centrifuging at 8000r/min for 10min, collecting the filtered supernatant, purifying with solid phase purification column, collecting the liquid, freeze concentrating, re-dissolving with methanol-water solution (50+50), filtering, and detecting with LC-MS/MS. Chromatographic conditions are as follows: a chromatographic column: BEH C18Column (2.1 mm. times.100 mm, 1.7 μm) mobile phase A was acetonitrile, B was ammonium formate solution, column temperature: 45 ℃, flow rate: 0.3mL/min, sample size: 5 μ L. The elution conditions are shown in Table 1.
TABLE 1 gradient elution Table
The method is adopted to detect the residual quantity of the four toxins in the samples after the aflatoxins are removed in the embodiments 1 to 6, and the corresponding removal rate of the four aflatoxins is calculated. See table 2 for results.
Table 2 examples 1-6 AFTB in samples after removal of aflatoxin1And total AFT removal rate
Example 7 fermentation Process optimization
According to a single-factor contrast test, carrying out optimization selection aiming at the aspergillus niger species, the inoculation amount, the material-water ratio, the solid-state fermentation temperature and the solid-state fermentation time:
FIG. 1 shows that the fermentation conditions are different inoculum sizes of 1%, 5%, 10%, 15% and 20%, and the feed-water ratio is controlled to be 1: spore suspension concentration of 106cfu/mL, culture temperature of 30 ℃, fermentation time of 48h, four yellowThe removal rate of the aflatoxin.
FIG. 2 shows that the fermentation conditions are different fermentation temperatures of 20 ℃, 25 ℃, 30 ℃, 35 ℃ and 40 ℃, and the ratio of material to water is controlled to be 1: spore suspension concentration of 106cfu/mL, the inoculation amount is 10%, and the removal rate of the four aflatoxins is 48 hours after fermentation.
FIG. 3 shows that the fermentation conditions are different fermentation times of 24h, 36h, 48h, 60h and 72h, and the ratio of feed to water is controlled to be 1: spore suspension concentration of 106cfu/mL, the inoculation amount is 10 percent, and the removal rate of the four aflatoxins is realized at the culture temperature of 30 ℃.
FIG. 4 shows the fermentation conditions of different feed-water ratios (ratio of mass of peanut meal to volume of sterile water, g/m L) 1: 0. 1: 0.5, 1:1. 1:1.5, 1:2, controlling the concentration of the spore suspension to be 106cfu/mL, the inoculation amount (the ratio of the volume of the bacterial liquid to the mass of the peanut meal, mL/g) is 10%, the culture temperature is 30 ℃, and the removal rate of the four aflatoxins is 48 hours when the fermentation time is 48 hours.
As can be seen from the condition optimization shown in fig. 1, 2, 3, and 4: by AFTB1、AFTB2、AFTG1、AFTG2Removing rate is used as index, and spore suspension concentration is controlled to be 106cfu/mL, wherein the inoculation amount is 1, 5, 10, 15 and 20%, the fermentation temperature is 20, 25, 30, 35 and 40 ℃, the fermentation time is 24, 36, 48, 60h and 72h, and the feed-water ratio (the ratio of the mass of the peanut meal to the volume of the sterile water, g/mL) is 1: 0. 1: 0.5, 1:1. 1:1.5, 1:2, performing an experiment, and selecting proper fermentation conditions to optimize the response surface according to the influence of various factors on the removal condition of the aflatoxin.
FIG. 5 is the interaction of factors on AFB1Response surface diagram of removing rate influence; wherein a is an interaction response surface diagram of the inoculation amount and the fermentation temperature, b is an interaction response surface diagram of the inoculation amount and the fermentation time, c is an interaction response surface diagram of the inoculation amount and the material-water ratio, d is an interaction response surface diagram of the fermentation temperature and the fermentation time, e is an interaction response surface diagram of the fermentation temperature and the material-water ratio, and f is an interaction response surface diagram of the material-water ratio and the fermentation time.
FIG. 6 is a response surface plot of the effect of the interaction of factors on the overall AFT removal rate; wherein a is an interaction response surface map of fermentation temperature and inoculation amount, b is an interaction response surface map of fermentation time and inoculation amount, c is an interaction response surface map of inoculation amount and material-water ratio, d is an interaction response surface map of fermentation time and fermentation temperature, e is an interaction response surface map of material-water ratio and fermentation temperature, and f is an interaction response surface map of material-water ratio and fermentation time.
Specific results discussion analysis was performed in conjunction with fig. 5 and 6: reflects four factors to AFTB1There was a pairwise interaction with the effect of total AFT removal, which is seen for AFTB1The method has the advantages of obvious removal effect, obvious interaction of inoculation amount and material-water ratio, obvious interaction of temperature, time and material-water ratio, obvious interaction of time, temperature and material-water ratio, and obvious interaction of material-water ratio and other three factors. For the total AFT removal effect, the central points of the contour lines are all in the condition range, and the total AFT removal rate can reach more than 70%.
Comparative example 1 Effect of different methods on the removal of four toxins
The method comprises the following steps: the Aspergillus niger CCTCC M2013703 solid fermentation removal method in example 5 is replaced by ozone treatment, and the specific ozone treatment process is the optimal method described in the publication of 2013 by the topic group (response surface optimizes the degradation effect of ozone on aflatoxin B1 in peanut meal, Journal of Food Science and Biotechnology.2013,32(5),494 500).
The method 2 comprises the following steps: the Aspergillus niger CCTCC M2013703 solid fermentation removal method in example 5 is replaced by weak base high-temperature treatment, and the specific weak base high-temperature treatment process is the optimal method recorded in the article published in 2007 by Caesalpinia japonica (the influence of the weak base high-temperature method on the detoxification and enzymolysis effects of peanut meal. Caesalpinia japonica, modern food science and technology, 2007,23(11), 4-6).
The method 3 comprises the following steps: the aspergillus niger CCTCC M2013703 solid fermentation removal method in example 5 is replaced by electron beam acceleration treatment, and the specific electron beam acceleration treatment process refers to the optimal method described in the article published in 2014 by royal gas (the influence of the electron beam acceleration aflatoxin removal B1 process on the quality of peanut meal, the royal gas, the chinese oil 2014,39(2), 33-36).
The method 4 comprises the following steps: referring to example 5, aspergillus niger CCTCC M2013703 was replaced with another Bacillus megaterium TRS-3, and the fermentation process was performed by the method described in the article published in 2013 of liu rui je (research on removing aflatoxin B1 in peanut meal by solid state fermentation of Bacillus megaterium in response, china oil, 38(3), 28-30).
The method 5 comprises the following steps: referring to example 5, aspergillus niger CCTCC M2013703 was replaced with another aspergillus niger CCTCC M2013553, and the peanut meal was subjected to fermentation removal treatment under the same conditions. The same test method as described above was used to determine the residual amounts of the four toxins in the treated samples, and the results are shown in Table 3.
TABLE 3 Effect of different methods for removing four toxins from peanut meal
Comparative examples 2 to 6
The concentration of the Aspergillus niger suspensions in examples 1-5 was replaced by 5X 10, respectively6cfu/mL, with other conditions unchanged, was used to remove toxins from the peanut meal, which was recorded as comparative examples 2-6. The results of removal were measured for the four toxins in the sample after removal of aflatoxin using the same method, as shown in table 4.
TABLE 4 Effect of different inoculum sizes on removal of toxins from peanut meal
In combination with the results of comparative examples 2 to 6 in Table 4, it can be seen that the inoculation amount is too large to be advantageous for the solid-state fermentation and the removal of aflatoxin, as compared with the corresponding examples 1 to 5.
Claims (10)
1. A method for removing four aflatoxins in peanut meal is characterized in that 1% -20% of 1 x 10 aflatoxins are inoculated into the peanut meal6cfu/mL Aspergillus niger spore suspension, adding sterile water for solid state fermentation; the Aspergillus niger is preserved in China center for type culture Collection with the preservation number of CCTCC M2013703.
2. The method according to claim 1, wherein the solid state fermentation temperature is 20-40 ℃.
3. The method according to claim 1, wherein the solid state fermentation time is 24-72 hours.
4. The method of any one of claims 1 to 3, wherein said 1 x 10 is selected from the group consisting of6The inoculation ratio of cfu/mL Aspergillus niger spore suspension is 1-20%.
5. The method according to any one of claims 1 to 4, wherein the sterile water is added at a ratio of 1:0 to 1: 2.
6. The method according to any one of claims 1 to 5, further comprising preliminarily pulverizing the peanut meal, sieving, and sterilizing.
7. The method according to any one of claims 1 to 6, wherein the Aspergillus niger spore suspension is prepared by culturing Aspergillus niger and activating followed by dilution to a suspension.
8. The method according to any one of claims 1 to 7, wherein said Aspergillus niger spore suspension is diluted by eluting the spores with 0.9% physiological saline containing 0.05% Tween-80.
9. The method according to claim 7, wherein Aspergillus niger is inoculated into PDA solid culture medium for activation culture for 3-7 d.
10. A peanut meal product produced by the method of any one of claims 1-9.
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