CN113995082A - Aflatoxin B1Preparation of the degradation agent - Google Patents

Aflatoxin B1Preparation of the degradation agent Download PDF

Info

Publication number
CN113995082A
CN113995082A CN202111206577.4A CN202111206577A CN113995082A CN 113995082 A CN113995082 A CN 113995082A CN 202111206577 A CN202111206577 A CN 202111206577A CN 113995082 A CN113995082 A CN 113995082A
Authority
CN
China
Prior art keywords
juice
afb
mass
tween
aloe
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202111206577.4A
Other languages
Chinese (zh)
Inventor
王乐
邬有胜
焦冰玉
刘建光
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Henan University of Technology
Original Assignee
Henan University of Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Henan University of Technology filed Critical Henan University of Technology
Priority to CN202111206577.4A priority Critical patent/CN113995082A/en
Publication of CN113995082A publication Critical patent/CN113995082A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • 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
    • A23L5/25—Removal of unwanted matter, e.g. deodorisation or detoxification using enzymes
    • 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
    • A23L5/27—Removal of unwanted matter, e.g. deodorisation or detoxification by chemical treatment, by adsorption or by absorption
    • 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

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Microbiology (AREA)
  • Immobilizing And Processing Of Enzymes And Microorganisms (AREA)

Abstract

The invention discloses an aflatoxin B1（AFB1) Preparation of degradation agent belonging to AFB1The technical field of degradation. Preparation of AFB by the invention1The degrading agent is prepared by squeezing and filtering fresh aloe and 42-67% of fresh lemon, wherein the fresh aloe and the fresh lemon are 32-57% of the degrading agent by mass, respectively adding inorganic salt and 1-2% of glucose, which are 0.1-0.3% of the degrading agent by mass, into juice, respectively inoculating bacillus amyloliquefaciens and saccharomyces cerevisiae, and finally mixing fermentation liquor, and uniformly mixing the fermentation liquor with cinnamaldehyde, 0.04-0.4% of immobilized laccase and 0.04-0.1% of tween, which are 0.2-0.7% of the degrading agent by mass. The aloe polysaccharide in the aloe has an antioxidation effect, and the cinnamaldehyde has reducibility and can act on AFB1The immobilized laccase can destroy AFB1The chemical bond, Tween and lemon juice capable of providing acidic environment can promote AFB1High efficiency degradation. AFB of the invention1The degradation agent can be applied to grain and oil processing, and the degradation process has the beneficial effects of greenness, safety, high efficiency, mild reaction and the like.

Description

Aflatoxin B1Preparation of the degradation agent
Technical Field
The invention belongs to aflatoxins B1(AFB1) The technical field of degradation, in particular to an efficient degradation AFB1The method of (1).
Background
Studies of Chenkang et al show that polyhydroxy compounds such as Tween and the like can change the conformation of the immobilized laccase by increasing hydrogen bonds among molecules of the immobilized laccase, and prevent the adverse change of the secondary structure of the immobilized laccase caused by the temperature rise. Therefore, the low-concentration Tween can be used as an additive for improving the thermal stability of the immobilized laccase. As shown in the research results of Petr Baldrian et al, the activity of immobilized laccase produced by oyster mushroom is improved to different degrees by adding copper and cadmium in an oyster mushroom culture medium. Therefore, the copper and cadmium contained in the inorganic salt are beneficial to enhancing the activity of the immobilized laccase.
The saccharomyces cerevisiae is rich in nutrients such as protein, nucleic acid, vitamins, polysaccharide and the like, and the inactive form and cell components of the saccharomyces cerevisiae can effectively adsorb mycotoxins (such as aflatoxin and zearalenone) through hydrogen bonds, ionic bonds, hydrophobic acting force and the like due to the special spatial structure of the cell wall of the saccharomyces cerevisiae, and can be used as a mycotoxin adsorbent. The bacillus amyloliquefaciens can generate various antibacterial substances such as antibacterial protein substances and lipopeptide antibiotics in the growth process of the bacillus amyloliquefaciens as a probiotic, and provides possibility for wide application of the bacillus amyloliquefaciens in animal and plant production. Therefore, in this patent, after mixing with the fermented ingredients of Saccharomyces cerevisiae, if the fermented ingredients contain a small fraction of Saccharomyces cerevisiae, AFB will also be treated1The degradation agent produces a beneficial effect. The aloe mucilage contains polysaccharides such as arboran A Baloe mannanase as core component, has antioxidant and antiaging effects, and contains multiple hydroxyl groups in aloe gel1Lactone ring in the structure, thereby achieving the purpose of degrading AFB1The effect of (1).
The literature proves that the aloe polysaccharide has good antioxidant activity. In vivo test, Wangli et al used 50mg/kg aloe polysaccharide to irrigate stomach for 30 days continuously to find that aloe polysaccharide has good oxidation resistance. The antioxidant ability can effectively prevent AFB1The oxidability of a lactone ring in the structure is realized, thereby achieving the purpose of degrading AFB1The effect of (1).
The study of Sunphen shows that cinnamaldehyde can play a role in AFB1Has antioxidant effect, and lactone ring is used as the active site to degrade AFB1The effect of (1).
The method of immobilizing an enzyme by adsorbing the enzyme or enzyme-containing bacterial cells on the surface of an adsorbent with an adsorbent is an adsorption-immobilized enzyme technique, and preferable adsorbents include activated carbon, diatomaceous earth and the like. The adsorption method is adopted to fix the enzyme, the operation is simple and convenient, the condition is mild, the enzyme denaturation or inactivation cannot be caused, the carrier is cheap and easy to obtain, and the enzyme can be repeatedly used.
In this patent, laccase is immobilized using activated carbon, chitosan, etc. to increase the stability of laccase, which keeps it active for a longer time. Immobilized laccases include the following classes: the laccase is copper-rich oxidase and can be used in AFB1Degradation of (2). Studies of Zhang Chu et al show that laccase is beneficial to improving enzyme activity under acidic conditions, and the acidic pH range provided by lemon in the patent can be beneficial to better exerting the immobilized laccase to degrade AFB1The function of (1). Weanxinxin research shows that the immobilized laccase can degrade AFB alone1And the copper ions in the immobilized laccase can catalyze the aloe polysaccharide and the cinnamaldehyde to react with AFB1Degradation of (2).
The research result of Arijit Das shows that the addition of certain surfactant such as Tween can improve the AFB of the oyster mushroom strain producing immobilized laccase1Enzymatic degradation of (1). Similarly, the results of the Wangjiaqiao et al study showed that the addition of Tween can promote AFB under the enzymatic reaction1Degradation of (2).
The following are the patents that have been published of primary relevance, but differ significantly from the present document, as in table 1:
TABLE 1 differentiation of the partially published patent from the application
Figure BDA0003307145210000021
Figure BDA0003307145210000031
Figure BDA0003307145210000041
Disclosure of Invention
To address the problems in the prior artThe invention aims to prepare safe and efficient aflatoxin B1(AFB1) A degradation agent.
In order to achieve the purpose, the technical scheme of the invention is as follows:
preparation of an AFB1A degradation agent is prepared by squeezing fresh aloe and fresh lemon together, filtering, pulverizing juice residue, adding inorganic salt solution, adding glucose into juice, inoculating Bacillus amyloliquefaciens and Saccharomyces cerevisiae into sterilized and cooled juice residue and juice, filtering fermented juice residue to remove residue, adding filtrate of Bacillus amyloliquefaciens fermented juice residue into juice fermented by Saccharomyces cerevisiae, adding cinnamaldehyde, immobilized laccase, and tween to obtain AFB1And the degrading agent comprises cinnamaldehyde and tween which are respectively used as an oil phase and a surfactant, and deionized water which is used as a water phase, wherein the cinnamaldehyde and the tween which are respectively used as the oil phase and the surfactant are mixed in a test tube, subjected to ultrasonic treatment for 30-60 min, and then kept stand in a water bath at the temperature of 25-35 ℃ for 24-36 h.
Further, the immobilized laccase includes the following species: the laccase is immobilized by activated carbon or chitosan, wherein the laccase immobilization method comprises the following steps: adding 0.2-0.8 g of activated carbon or chitosan into a laccase phosphate buffer solution (0.1-0.5 mg/mL and 2-5 mL) with the pH value of 3.0-6.0, oscillating for 6-12 h at 20-35 ℃, filtering, washing with a phosphate buffer solution (0.02-0.08 mol/L and pH of 5.0-7.0) until no laccase can be detected in the supernatant, and thus obtaining the immobilized laccase.
Further, the fresh aloe comprises the following species: aloe vera, aloe arborescens, aloe vera, aloe saponaria, aloe vera, aloe barbadensis, added in an amount of 32-57 mass% based on the mass of the degradant, wherein the aloe polysaccharide content is 0.60-1.15 mass% based on the mass of the degradant.
Further, the fresh lemons include the following categories: the lemon juice is prepared from lemon, perfume lemon, Taiwan lime and Beijing lime, wherein the addition amount of the lemon is 42-67 percent (mass fraction) of the mass of the degradation agent, and fresh aloe and fresh lemon are squeezed together and filtered to obtain juice residue and juice: crushing the juice residues to 40-80 meshes, adding 0.1-0.3% of inorganic salt solution, and uniformly stirring; adding glucose accounting for 1-2% of the juice by mass, and uniformly stirring; and (4) respectively sterilizing the juice residues and the juice by low-temperature steam at 80-100 ℃ for 15-20 min, and cooling to room temperature.
Further, the formula of the inorganic salt solution is as follows (mass fraction of the degradation agent): (NH)4)2SO4 0.03～0.09％、KH2PO4 0.03～0.09％、MgSO4 0.01～0.03％、NaCl 0.01～0.03％、CuSO4 0.01～0.03％、CdCl2 0.001～0.01％。
Further, the juice residue after sterilization and cooling is added with a bacillus amyloliquefaciens (preservation number is CGMCC NO: 1.8719) bacterial liquid with the initial concentration of 109～1010CFU/mL, culturing at 30-37 deg.C for 36-48 h to make the bacterial liquid concentration reach 1010～1011CFU/mL。
Adding Saccharomyces cerevisiae (preservation number of CGMCC NO: 2.3889) bacterial liquid into the sterilized and cooled juice, wherein the initial concentration is 107～108CFU/mL, 28-33 ℃, 150-180 rpm, fermenting for 48-60 hours to make the concentration of the saccharomyces cerevisiae bacterial liquid reach 108～109CFU/mL。
Further, filtering fermented juice residues to remove residues, adding filtrate into the fermented juice, and then adding cinnamaldehyde accounting for 0.2-0.7% (mass fraction) of the mass of the degradation agent, immobilized laccase accounting for 0.04-0.4% (mass fraction), and tween accounting for 0.04-0.1% (mass fraction), wherein the tween comprises the following types: tween-20, tween-40, tween-60, tween-80 and tween-85. After being mixed evenly, AFB is prepared1The degrading agent comprises immobilized laccase with enzyme activity of 48-56U/mL, Tween with purity of more than 99%, and all three are commercial reagents and can be obtained from reagent companies.
Unless otherwise indicated, all reagents and materials used above are commercially available.
The invention has the advantages of
AFB of the invention1Degradation agent to aflatoxin B1The degradation is safe and efficient, the reaction is mild, and the degradation is resistant to AFB1The degradation rate can reach 80-87%, so that the content of the material in the grain reaches the present levelThe national standard is: AFB1The limit standard GB 2761-.
The aloe of the invention contains a plurality of effective active ingredients and can effectively degrade AFB1(ii) a The fermentation of amylolytic bacillus and Saccharomyces cerevisiae can fully release the effective components of Aloe, and their metabolites and various enzyme active components also act on AFB1Has the degradation function; laccase itself has the function of degrading AFB1The stability of the laccase is enhanced after immobilization, the cinnamaldehyde has reducibility, the copper and cadmium in the inorganic salt are beneficial to enhancing the activity of the immobilized laccase, and the acidic environment and the Tween provided by the lemon juice can promote the immobilized laccase to degrade the AFB1Let the AFB1The degradation agent obtains better degradation effect compared with the degradation agent with single component.
Drawings
FIG. 1 shows different aflatoxins B1(AFB1) Addition amount of degradation agent to AFB1Degradation of (2).
FIG. 2 shows aflatoxin B1(AFB1) Degradation agent for AFB in mildewed peanuts1Degradation of (2).
Detailed Description
Specific embodiments of the present invention will be described in further detail with reference to examples. The following examples are intended to further illustrate the invention without limiting its scope. The related matters and modifications of the invention are within the scope of the invention.
Example 1
Aflatoxin B1(AFB1) A degradation agent is prepared by squeezing fresh aloe and fresh lemon together, filtering, pulverizing juice residue, adding inorganic salt solution, adding glucose into juice, inoculating Bacillus amyloliquefaciens and Saccharomyces cerevisiae into sterilized and cooled juice residue and juice, filtering fermented juice residue to remove residue, adding filtrate of Bacillus amyloliquefaciens fermented juice residue into juice fermented by Saccharomyces cerevisiae, adding cinnamaldehyde, immobilized laccase, and tween to obtain AFB1Degradation agents of cinnamic aldehyde and vomitThe temperature is respectively used as an oil phase and a surfactant, deionized water is used as a water phase, the three are mixed in a test tube, ultrasonic treatment is carried out for 30-60 min, and then standing is carried out for 24-36 h in a water bath at 25-35 ℃.
The immobilized laccase includes the following species: the laccase is immobilized by activated carbon or chitosan, wherein the laccase immobilization method comprises the following steps: adding 0.2-0.8 g of activated carbon or chitosan into a laccase phosphate buffer solution (0.1-0.5 mg/mL and 2-5 mL) with the pH value of 3.0-6.0, oscillating for 6-12 h at 20-35 ℃, filtering, and washing with a phosphate buffer solution (0.02-0.08 mol/L and pH value of 5.0-7.0) to obtain the immobilized laccase.
Fresh aloe includes the following species: aloe vera, aloe arborescens, aloe vera, aloe saponaria, aloe vera, aloe barbadensis, added in an amount of 32-57 mass% based on the mass of the degradant, wherein the aloe polysaccharide content is 0.60-1.15 mass% based on the mass of the degradant.
The fresh lemons include the following categories: the lemon juice is prepared from lemon, perfume lemon, Taiwan lime and Beijing lime, wherein the addition amount of the lemon is 42-67 percent (mass fraction) of the mass of the degradation agent, and fresh aloe and fresh lemon are squeezed together and filtered to obtain juice residue and juice: crushing the juice residues to 40-80 meshes, adding 0.1-0.3% of inorganic salt solution, and uniformly stirring; adding glucose accounting for 1-2% of the juice by mass, and uniformly stirring; and (4) respectively sterilizing the juice residues and the juice by low-temperature steam at 80-100 ℃ for 15-20 min, and cooling to room temperature.
The formula of the inorganic salt solution is as follows (mass fraction of the degradation agent): (NH)4)2SO4 0.03～0.09％、KH2PO40.03～0.09％、MgSO4 0.01～0.03％、NaCl 0.01～0.03％、CuSO4 0.01～0.03％、CdCl20.001～0.01％。
Adding Bacillus amyloliquefaciens (preservation number CGMCC NO: 1.8719) bacterial liquid into the sterilized and cooled juice residue, wherein the initial concentration is 109～1010CFU/mL, culturing at 30-37 deg.C for 36-48 h to make the bacterial liquid concentration reach 1010～1011CFU/mL。
Adding Saccharomyces cerevisiae (preservation number of CGMCC NO: 2.3889) bacterial liquid into the sterilized and cooled juice, wherein the initial concentration is 107～108CFU/mL, 28-33 ℃, 150-180 rpm, fermenting for 48-60 hours to make the concentration of the saccharomyces cerevisiae bacterial liquid reach 108～109CFU/mL。
Filtering fermented juice residues to remove residues, adding filtrate obtained after bacillus amyloliquefaciens fermented juice residues into juice fermented by saccharomyces cerevisiae, and then adding cinnamaldehyde accounting for 0.2-0.7% (mass fraction) of the mass of the degradation agent, immobilized laccase accounting for 0.04-0.4% (mass fraction), and tween accounting for 0.04-0.1% (mass fraction), wherein the tween comprises the following types: mixing Tween-20, Tween-40, Tween-60, Tween-80, and Tween-85, and making into AFB1And the enzymatic activity of the immobilized laccase is 48-56U/mL, and the purity of the Tween is more than 99%.
Taking the above AFB10.1-0.5 mL of degradation agent is added into 2mL of AFB with initial concentration of 2 mu g/mL1In the standard solution, vortex, mix evenly, react for 20h at 30 ℃ under 180rpm in the dark, and AFB in the reaction system is measured by HPLC1The remaining amount. The results are shown in FIG. 1.
Example 2
AFB (active carbon boron)1A degradation agent is prepared by squeezing fresh aloe and fresh lemon together, filtering, pulverizing juice residue, adding inorganic salt solution, adding glucose into juice, inoculating Bacillus amyloliquefaciens and Saccharomyces cerevisiae into sterilized and cooled juice residue and juice, filtering fermented juice residue to remove residue, adding filtrate of Bacillus amyloliquefaciens fermented juice residue into juice fermented by Saccharomyces cerevisiae, adding cinnamaldehyde, immobilized laccase, and tween to obtain AFB1And the degrading agent comprises cinnamaldehyde and tween which are respectively used as an oil phase and a surfactant, and deionized water which is used as a water phase, wherein the cinnamaldehyde and the tween which are respectively used as the oil phase and the surfactant are mixed in a test tube, subjected to ultrasonic treatment for 30-60 min, and then kept stand in a water bath at the temperature of 25-35 ℃ for 24-36 h.
The immobilized laccase includes the following species: the laccase is immobilized by activated carbon or chitosan, wherein the laccase immobilization method comprises the following steps: adding 0.2-0.8 g of activated carbon or chitosan into a laccase phosphate buffer solution (0.1-0.5 mg/mL and 2-5 mL) with the pH value of 3.0-6.0, oscillating for 6-12 h at 20-35 ℃, filtering, and washing with a phosphate buffer solution (0.02-0.08 mol/L and pH value of 5.0-7.0) to obtain the immobilized laccase.
Fresh aloe includes the following species: aloe vera, aloe arborescens, aloe vera, aloe saponaria, aloe vera, aloe barbadensis, added in an amount of 32-57 mass% based on the mass of the degradant, wherein the aloe polysaccharide content is 0.60-1.15 mass% based on the mass of the degradant.
The fresh lemons include the following categories: the lemon juice is prepared from lemon, perfume lemon, Taiwan lime and Beijing lime, wherein the addition amount of the lemon is 42-67 percent (mass fraction) of the mass of the degradation agent, and fresh aloe and fresh lemon are squeezed together and filtered to obtain juice residue and juice: crushing the juice residues to 40-80 meshes, adding 0.1-0.3% of inorganic salt solution, and uniformly stirring; adding glucose accounting for 1-2% of the juice by mass, and uniformly stirring; and (4) respectively sterilizing the juice residues and the juice by low-temperature steam at 80-100 ℃ for 15-20 min, and cooling to room temperature.
The formula of the inorganic salt solution is as follows (mass fraction of the degradation agent): (NH)4)2SO4 0.03～0.09％、KH2PO40.03～0.09％、MgSO4 0.01～0.03％、NaCl 0.01～0.03％、CuSO4 0.01～0.03％、CdCl20.001～0.01％。
Adding Bacillus amyloliquefaciens (preservation number CGMCC NO: 1.8719) bacterial liquid into the sterilized and cooled juice residue, wherein the initial concentration is 109～1010CFU/mL, culturing at 30-37 deg.C for 36-48 h to make the bacterial liquid concentration reach 1010～1011CFU/mL。
Adding Saccharomyces cerevisiae (preservation number of CGMCC NO: 2.3889) bacterial liquid into the sterilized and cooled juice, wherein the initial concentration is 107～108CFU/mL, 28-33 ℃, 150-180 rpm, fermenting for 48-60 hours to make the concentration of the saccharomyces cerevisiae bacterial liquid reach 108～109CFU/mL。
Filtering the fermented juice residue to remove residue, and adding the filtrate of Bacillus amyloliquefaciens fermented juice residue into the filtrateAdding 0.2-0.7% of cinnamaldehyde, 0.04-0.4% of immobilized laccase and 0.04-0.1% of tween into the juice after yeast fermentation, wherein the tween comprises the following types: mixing Tween-20, Tween-40, Tween-60, Tween-80, and Tween-85, and making into AFB1And the enzymatic activity of the immobilized laccase is 48-56U/mL, and the purity of the Tween is more than 99%.
Sterilizing 7d mildewed peanut powder 10g at 115 deg.C for 10min, and collecting the AFB10.5g of degradation agent, 20mL of acetic acid-sodium acetate buffer solution with pH 4.5 was added, and the mixture was reacted at 40 ℃ and 180rpm in the dark for 18 hours. After the reaction is finished, adding 10mL of methanol into the peanut powder, and extracting AFB in the peanuts by ultrasonic treatment for 20min1Centrifuging at 12000rpm for 10min after ultrasonic treatment, filtering the supernatant with 0.22 μm filter membrane, and detecting AFB in the peanut powder by HPLC1Of (c) is added. Without addition of AFB1Moldy peanut powder of degradant was used as a blank control. The results are shown in FIG. 2.

Claims (6)

1. Preparation of aflatoxin B1（AFB1) The preparation method of the degradation agent is characterized by comprising the following steps: firstly, juicing fresh aloe and fresh lemon together, filtering, crushing juice residues, adding an inorganic salt solution, adding glucose into juice, then respectively inoculating bacillus amyloliquefaciens and saccharomyces cerevisiae into the sterilized and cooled juice residues and juice, finally filtering the fermented juice residues to remove residues, adding the filtrate into the fermented juice, and adding cinnamaldehyde, immobilized laccase and tween to prepare AFB (active fragment binding protein)1A degradation agent.
2. As claimed in claim 1, fresh aloe vera includes the following species: aloe vera, aloe arborescens, aloe vera, aloe saponaria, aloe vera, aloe barbadensis, added in an amount of 32-57% by mass of the degrading agent, wherein the aloe polysaccharide content is 0.60-1.15% by mass of the degrading agent; the fresh lemons include the following species: the lemon is lemon, perfume lemon, Taiwan lime and Beijing lime, and the addition amount is 42-67 percent of the mass of the degradation agent; juicing fresh aloe and fresh lemon, and filtering to obtain juice residues and juice: crushing the juice residues to 40-80 meshes, adding inorganic salt accounting for 0.1-0.3 percent of the mass of the degradation agent, adding glucose accounting for 1-2 percent of the mass of the degradation agent, and uniformly stirring; sterilizing the juice residues and the juice respectively at the low temperature of 80-100 ℃ for 15-20 min by using steam, and cooling to room temperature; tweens include the following classes: tween-20, Tween-40, Tween-60, Tween-80 and Tween-85, wherein the addition amount is 0.04-0.1 percent of the mass of the degradation agent (mass fraction).
3. As stated in claim 1, the formulation of inorganic salts in claim 2 is (in mass fraction of the degradation agent): (NH)4)2SO4 0.03~0.09%、KH2PO4 0.03~0.09%、MgSO4 0.01~0.03%、NaCl 0.01~0.03%、CuSO4 0.01~0.03%、CdCl2 0.001~0.01%。
4. As described in claim 1, the sterilized and cooled juice residue is added with Bacillus amyloliquefaciens (CGMCC NO: 1.8719) bacterial liquid with initial concentration of 109~1010CFU/mL, culturing at 30-37 ℃ for 36-48 h to make the concentration of the bacillus amyloliquefaciens liquid reach 1010~1011CFU/mL; adding Saccharomyces cerevisiae (preservation number of CGMCC NO: 2.3889) bacterial liquid into the sterilized and cooled juice, wherein the initial concentration is 107~108CFU/mL, 28-33 ℃, 150-180 rpm, fermenting for 48-60 hours to make the concentration of the saccharomyces cerevisiae bacterial liquid reach 108~109 CFU/mL。
5. The method as claimed in claim 1, wherein the fermented juice residue is filtered to remove residue, the filtrate obtained by fermenting the juice residue with Bacillus amyloliquefaciens is added to the juice fermented with Saccharomyces cerevisiae, and then 0.2-0.7% (by mass) of cinnamaldehyde, 0.04-0.4% (by mass) of immobilized laccase, and 0.04-0.1% (by mass) of Tween are added to the mixture to obtain AFB1And the degrading agent, wherein the enzyme activity of the aldehyde immobilized laccase is 48-56U/mL, and the purity of the Tween is more than 99%.
6. The AFB of claim 11The degradation agent can be applied to grain and oil processing.
CN202111206577.4A 2021-10-17 2021-10-17 Aflatoxin B1Preparation of the degradation agent Pending CN113995082A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202111206577.4A CN113995082A (en) 2021-10-17 2021-10-17 Aflatoxin B1Preparation of the degradation agent

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202111206577.4A CN113995082A (en) 2021-10-17 2021-10-17 Aflatoxin B1Preparation of the degradation agent

Publications (1)

Publication Number Publication Date
CN113995082A true CN113995082A (en) 2022-02-01

Family

ID=79923310

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202111206577.4A Pending CN113995082A (en) 2021-10-17 2021-10-17 Aflatoxin B1Preparation of the degradation agent

Country Status (1)

Country Link
CN (1) CN113995082A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114457063A (en) * 2022-03-11 2022-05-10 安徽黑娃食品科技有限公司 Aspergillus flavus toxin B in degradation peanut1Preparation method of immobilized enzyme
CN115501910A (en) * 2022-10-25 2022-12-23 陕西中医药大学 Organic-inorganic photocatalyst AE/FeOOH and preparation method and application thereof

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103966147A (en) * 2014-05-29 2014-08-06 江南大学 Bacillus amyloliquefacien for degrading aflatoxin B1 in peanut meal
CN106119179A (en) * 2016-09-18 2016-11-16 天津北洋百川生物技术有限公司 A kind of preparation method of the complex micro organism fungicide for water body purification
CN108251458A (en) * 2018-01-25 2018-07-06 青岛农业大学 A kind of degradation agent of aflatoxin, preparation method and application
CN108893434A (en) * 2018-07-21 2018-11-27 泸州正泰生物工程有限公司 A kind of microbiological garbage treatment agent and the preparation method and application thereof of degrading aspergillus flavus element B1

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103966147A (en) * 2014-05-29 2014-08-06 江南大学 Bacillus amyloliquefacien for degrading aflatoxin B1 in peanut meal
CN106119179A (en) * 2016-09-18 2016-11-16 天津北洋百川生物技术有限公司 A kind of preparation method of the complex micro organism fungicide for water body purification
CN108251458A (en) * 2018-01-25 2018-07-06 青岛农业大学 A kind of degradation agent of aflatoxin, preparation method and application
CN108893434A (en) * 2018-07-21 2018-11-27 泸州正泰生物工程有限公司 A kind of microbiological garbage treatment agent and the preparation method and application thereof of degrading aspergillus flavus element B1

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114457063A (en) * 2022-03-11 2022-05-10 安徽黑娃食品科技有限公司 Aspergillus flavus toxin B in degradation peanut1Preparation method of immobilized enzyme
CN115501910A (en) * 2022-10-25 2022-12-23 陕西中医药大学 Organic-inorganic photocatalyst AE/FeOOH and preparation method and application thereof
CN115501910B (en) * 2022-10-25 2024-02-09 陕西中医药大学 Organic-inorganic photocatalyst AE/FeOOH, and preparation method and application thereof

Similar Documents

Publication Publication Date Title
Mussatto Biotechnological potential of brewing industry by-products
CN107937292B (en) Saccharomyces cerevisiae culture and fermentation process thereof
CN113995082A (en) Aflatoxin B1Preparation of the degradation agent
CN106755188A (en) The preparation method and brown alga oligose of a kind of brown alga oligose monomer
Varelas et al. An evaluation study of different methods for the production of β‐D‐glucan from yeast biomass
TWI706036B (en) Method of concentrating protein in grain powder
Taskin et al. Biomass and exopolysaccharide production by Morchella esculenta in submerged culture using the extract from waste loquat (Eriobotrya japonica L.) kernels
CN1231593C (en) Technology for producing glucomannan using neutral beta-mannase to degradate fine konjaku flour
CN105586286A (en) Method for producing heavy metal absorbent through microbiological treatment on lignite and related compound microbial agent
CN116058432B (en) Citric acid mycelium residue microbial feed additive and preparation method and application thereof
CN112592914A (en) Special green alga polysaccharide lyase and production process thereof
CN108570422A (en) One plant of lactobacillus buchneri bacterial strain and its ensilage fermentation agent
CN114431454B (en) Preparation method of bee pollen ferment
KR20160074300A (en) Method of Making Material for Health Food by Fermentation of Medical Herbs
CN113564215B (en) Preparation method of biosurfactant with carbon dioxide and/or lignocellulose as substrate
CN107348107A (en) A kind of bran polysaccharide feed addictive with antioxidation activity and preparation method and application
CN109652479B (en) Method for improving antioxidant capacity of dendrobe polysaccharide
CN111620965A (en) Preparation method of flammulina velutipes chitin
CN111909822A (en) Method for fermenting pineapple vinegar by cell immobilization method
CN102719511B (en) Method for extracting L-arabinose from crop byproduct by utilizing multi-strain mixed fermentation
CN111820362A (en) Biodegradation method for aflatoxin in DDGS
CN108841894B (en) Method for preparing arabinoxylo-oligosaccharide through probiotic transformation
Yakimova et al. Alpha-galactosidase and invertase from Penicillium chrysogenum sp. 23: purification, characteristics and hydrolysis of raffinose
CN113121714B (en) Ganoderma lucidum oligosaccharide with probiotic activity and preparation method and application thereof
CN112293580B (en) Feed and preparation process thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination