CN115176959A - Application and method for degrading aflatoxin - Google Patents
Application and method for degrading aflatoxin Download PDFInfo
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Abstract
The invention relates to the technical field of microorganisms, in particular to application and a method for degrading aflatoxin, and particularly relates to application of hirsutella sinensis strains and paecilomyces hepiali in degrading aflatoxin, wherein the hirsutella sinensis strains and the paecilomyces hepiali effectively degrade various aflatoxins in a short time, and the method has very good application value in the fields needing to degrade aflatoxin, particularly in the fields of medicines, foods and feeds.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to application and a method for degrading aflatoxin, and especially relates to application and a method of Hirsutella sinensis (Hirsutella sinensis) and Paecilomyces hepiali (Paecilomyces hepiali) in degradation of aflatoxin.
Background
Aflatoxins were first discovered on aflatoxins isolated by british scientists in 1960 and were largely classified into two groups: b type of blue fluorescence and G type of green fluorescence, wherein B type comprises B1, B2 and B2 alpha, and G type comprises G1 and G2; also includes derivatives such as aflatoxins M1, M2, P1, Q, H, GM, toxol, etc. Aflatoxins, which are secondary metabolites produced by aspergillus flavus or aspergillus parasiticus, have been reported in 18 kinds, but the most studied are aflatoxins B1, B2, G1 and G2, wherein aflatoxin B1 has the highest toxicity, and is recognized as the first carcinogen in human by the cancer research center of the world health organization, and the toxicity of aflatoxins is 68, 75 and 10 times that of arsenicum, ammonium nitrite and potassium cyanide respectively.
Aflatoxins are relatively seriously polluted in grains and feeds, and have great threat to the health of human beings and livestock, while the traditional aflatoxin removal method mainly adopts a physical method and a chemical method. The physical methods mainly comprise a high-temperature method, a radiation method and a physical adsorption method, and the chemical methods comprise an alkali treatment method, an oxidation method and the like, but the methods easily cause secondary pollution to the environment, reduce the nutritional value of a sample, and have uncertain toxicity of degradation products and the like. The biological method has the characteristics of safety, high efficiency, non-reversibility, environmental friendliness and the like, and becomes a research hotspot for aflatoxin degradation. At present, some microorganisms have been reported to have the ability to remove aflatoxin, for example, concern, etc., screening identification of highly effective degradation strains of aflatoxin B1 and degradation thereof (proceedings of university of agriculture in china, vol. 25, no. 3/2016). In the patent publication No. CN101766273B, a mushroom polysaccharide composition capable of removing aflatoxin is disclosed, but the mushroom polysaccharide composition can only adsorb aflatoxin and can not degrade it.
However, how to effectively degrade aflatoxins using known bacteria still requires further investigation.
Disclosure of Invention
The invention provides an application of hirsutella sinensis and/or paecilomyces hepiali in aflatoxin degradation.
Specifically, the Aflatoxin may be any type of Aflatoxin (AF), such as B1, B2 α, G1, G2, or any derivative thereof, such as M1, M2, P1, Q, H1, GM, toxol, etc., preferably AFB1, AFB2, AFG1, AFG2, particularly preferably AFB1.
Specifically, the hirsutella sinensis can be any hirsutella sinensis strain, for example, different strains are obtained according to the prior art CN109082382A, CN105670937a, and the obtained sequences are numbered as strain # 1, strain # 2, strain # 3, strain # 4, strain # 5, strain # 6, strain # 7, strain # 8 and strain # 9, similarity analysis is performed on the obtained sequences and data stored in a GenBank (DNA sequence database), and the hirsutella sinensis is identified as hirsutella sinensis based on the 18S rDNA sequence homology of more than 95% according to the principle of microbial molecular genetics identification.
Specifically, the paecilomyces hepiali can be any paecilomyces hepiali strain, such as CGMCC3.15540, 3.17720 and 3.15421.
In an embodiment of the present invention, the application is an application of hirsutella sinensis in degrading aflatoxins of class B (e.g., AFB1, AFB2 α) and class G (e.g., AFG1, AFG 2).
In another embodiment of the present invention, the application is the application of paecilomyces hepiali in degrading aflatoxin B (such as AFB1, AFB2 and AFB2 α).
The invention also provides application of the biological composition of hirsutella sinensis in aflatoxin degradation.
The invention also provides application of the biological composition of the paecilomyces hepiali in aflatoxin degradation.
The invention also provides application of the biological composition of hirsutella sinensis and paecilomyces hepiali in aflatoxin degradation.
Specifically, the biological composition is a composition related to the above-mentioned strain, and comprises one or more of thalli such as viable bacteria or cell fragments of the above-mentioned strain, intracellular substances or extracellular metabolites of the above-mentioned strain, seed liquid, fermentation liquid, bacterial suspension, intracellular liquid, cell fragment liquid, seed liquid extract, fermentation liquid extract, and bacterial powder of the above-mentioned strain, and particularly one or more of seed liquid, fermentation liquid, and bacterial suspension.
In particular, the biological composition may be in any suitable form, such as a liquid, gel, powder, spray, and the like, preferably in liquid form.
Specifically, the seed liquid can be obtained by culturing the above-mentioned strain.
Specifically, the fermentation broth is obtained by further culturing the seed solution.
Specifically, the culture method comprises: the seed culture is cultured in a suitable medium, particularly a liquid medium.
Specifically, the above culture temperature may be any suitable culture temperature, for example, in the case of hirsutella sinensis strain, the culture temperature may be 4 to 25 ℃, particularly 10 to 20 ℃,13 to 16 ℃, e.g., 14 ℃,15 ℃; for paecilomyces hepialid strains, the cultivation temperature may be 15-37 ℃, particularly 20-30 ℃,24-26 ℃, e.g. 25 ℃.
Specifically, the above culture may be a shaking culture, and the shaking frequency may be 80 to 200rpm, particularly 100 to 150rpm, for example, 100, 110, 120, 130, 140, 150rpm.
Specifically, the above-mentioned culture time may be any suitable culture time, for example, for hirsutella sinensis strain, the culture time may be 5 to 20 days, particularly 10 to 15 days; for paecilomyces hepiali strain, the culture time can be 2-10 days, especially 4-6 days.
In one embodiment of the present invention, the culture medium may be a PDB culture medium.
Specifically, the above culture method further comprises secondary culture.
Specifically, the secondary culturing comprises: inoculating the culture solution into a proper culture medium for culturing.
Specifically, the amount of the inoculum in the secondary culture is 1 to 20% (v/v), particularly 5 to 10%.
Specifically, in the above secondary culture, the culture temperature may be any suitable culture temperature, for example, in the case of hirsutella sinensis strain, the culture temperature may be 4 to 25 ℃, particularly 10 to 20 ℃,13 to 16 ℃, e.g., 14 ℃,15 ℃; for paecilomyces hepialid strains, the cultivation temperature may be 15-37 ℃, particularly 20-30 ℃,24-26 ℃, e.g. 25 ℃.
Specifically, in the above secondary culture, the culture may be shaking culture, and the shaking frequency may be 80 to 200rpm, particularly 100 to 150rpm.
Specifically, the above-mentioned culture time may be any suitable culture time, for example, for hirsutella sinensis strain, the culture time may be 1 to 10 days, particularly 4 to 6 days; for paecilomyces hepiali strain, the culture time can be 1-10 days, especially 1-3 days.
Specifically, in the secondary culture, the culture medium may be a PDB medium.
In one embodiment of the present invention, the method includes: inoculating the strain to PDB culture medium, shaking for culturing, and transferring to PDB culture medium for culturing under the same conditions.
Specifically, the seed solution may be a secondary culture solution of the strain (as described above).
Specifically, the bacterial suspension can be prepared by suspending the cells of the above-mentioned strain in a buffer.
In one embodiment of the present invention, the bacterial suspension may be prepared by suspending bacterial cells obtained by separating the seed solution (for example, by centrifugation) in a buffer.
In one embodiment of the present invention, the buffer is PBS buffer.
Specifically, the intracellular fluid can be prepared as follows: breaking cell wall of the above strain (such as by grinding), and separating to obtain supernatant as intracellular fluid.
Specifically, the cell debris solution can be prepared by: the cells of the above strain are broken into walls (for example, by grinding), separated, and the obtained solid is suspended in a buffer.
In one embodiment of the present invention, the buffer is PBS buffer.
Specifically, the seed liquid extract and the fermentation liquid extract can be obtained by appropriately extracting the seed liquid or the fermentation liquid.
Specifically, the bacterial powder is bacterial strain powder, especially live bacterial powder, such as live bacterial lyophilized powder.
The invention also provides the application of hirsutella sinensis and/or paecilomyces hepiali in medicines, foods and feeds.
The invention also provides application of the biological composition of hirsutella sinensis in medicines, foods and feeds.
The invention also provides application of the biological composition of the paecilomyces hepialid in medicines, foods and feeds.
The invention also provides application of the biological composition of hirsutella sinensis and paecilomyces hepiali in medicines, foods and feeds.
Specifically, hirsutella sinensis or paecilomyces hepiali, and biological compositions thereof have the corresponding definitions of the invention.
Specifically, the application can be the application of hirsutella sinensis and/or paecilomyces hepiali in the production, processing, storage, transportation and other processes of medicines, foods and feeds.
In particular, the medicament, food and feed may be any medicament, food or feed that may be contaminated with aflatoxins, which may be suspected of containing aflatoxins and/or having unknown levels of aflatoxins. In particular, the above-mentioned applications are the use for degrading (even completely eliminating) aflatoxins in pharmaceuticals, foods and feeds.
Specifically, the medicine can be a traditional Chinese medicine (the medicine can be polluted by aflatoxin) and a traditional Chinese medicine product prepared from the traditional Chinese medicine, or a medicine produced by microbial fermentation (wherein a microbial fermentation raw material can be polluted by aflatoxin, so that the produced medicine can be possibly polluted by aflatoxin).
Specifically, the food may be a food which is daily ingested by human beings, and may include, for example, foodstuffs, nuts, and products thereof.
Specifically, the above-mentioned grain may be a cereal, for example, one or more of corn, wheat, barley, rye, rice, sorghum and millet; legumes, such as one or more of soybeans, peas, and peanuts; oilseeds, for example, rape, soybean, sunflower and cotton. The above-described foodstuffs may also be milled, for example, wet or dry milled grains, including milled fractions containing gluten, protein, starch, bran and/or oil.
Specifically, the nut can be peanut, walnut, melon seed, etc.
Specifically, the product may be a milk product, a seasoning (a fermented seasoning such as fermented bean curd, fermented soybean, soy sauce, soybean paste, etc.), an edible oil (such as an oil extracted from the grain, for example, peanut oil, soybean oil, sunflower seed oil, cottonseed oil, etc.), and the like.
In particular, the feed described above can be any compound, mixture or composition suitable or intended for ingestion by an animal, e.g., can include foodstuffs, animal proteins, and the like.
Specifically, the animal protein may be, for example, fish meal, skim milk, whey, silkworm pupa powder, bone meal, or the like.
The invention also provides an aflatoxin degradation agent (or an aflatoxin antidote) which comprises hirsutella sinensis and/or paecilomyces hepiali and a biological composition containing any one or two of the bacteria.
Specifically, hirsutella sinensis, paecilomyces hepiali and the biological composition have the corresponding definitions of the invention.
The invention also provides application of the biological composition of hirsutella sinensis and/or paecilomyces hepiali, and hirsutella sinensis and/or paecilomyces hepiali in preparation of an aflatoxin degradation agent (or an aflatoxin antidote).
The present invention also provides a method of degrading aflatoxins (reducing or even eliminating aflatoxin toxicity) which comprises the step of using hirsutella sinensis and/or paecilomyces hepiali, or using a biological composition of hirsutella sinensis and/or paecilomyces hepiali.
Specifically, the aflatoxin, hirsutella sinensis, paecilomyces hepiali and the biological composition have the corresponding definitions of the invention.
In particular, the method may comprise the step of mixing the biological composition with a material to be treated, optionally subjected to a biodegradation process, wherein the material to be treated is a material suspected of containing aflatoxins and/or having an unknown level of aflatoxins (e.g. pharmaceuticals, food, feed).
The inventor discovers through a large number of experiments that hirsutella sinensis and paecilomyces hepiali can effectively degrade a plurality of aflatoxins in a short time, and the strain has very good application value in the field needing to degrade the aflatoxins, particularly in the fields of medicines, foods and feeds.
Drawings
FIG. 1 shows the kinetics of AFB1 degradation of strain # 8.
Detailed Description
Unless defined otherwise, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.
Hirsutella sinensis (Hirsutella sinensis), paecilomyces hepiali (Paecilomyces hepiali Chen).
Aflatoxins (AF) are difuranosiclotoxins produced by some strains of aspergillus flavus and aspergillus parasiticus. Aflatoxins can be mainly classified into two groups: b type of blue fluorescence and G type of green fluorescence, wherein the B type comprises B1, B2 and B2 alpha, and the G type comprises G1 and G2; also included are derivatives such as aflatoxins M1, M2, P1, Q, H, GM, toxol (Aflatoxicol), and the like.
In the present invention, "biological composition of hirsutella sinensis" refers to a composition related to hirsutella sinensis, "biological composition of paecilomyces hepiali" refers to a composition related to paecilomyces hepiali (e.g. china common microorganism culture collection and management center (CGMCC) numbers 3.15540, 3.17720, 3.15421), for example, a preparation obtained by treating the thallus thereof, for example, a seed liquid, a fermentation liquid, a bacterial suspension, an intracellular liquid, a cell debris liquid, etc., and a preparation obtained by treating the seed liquid thereof, for example, a seed liquid supernatant, a fermentation liquid extract, a seed liquid extract, etc. Thus, a "biological composition of hirsutella sinensis or paecilomyces hepiali" comprises hirsutella sinensis or paecilomyces hepiali (e.g. viable bacteria or cell debris thereof), intracellular material or extracellular metabolites.
The disclosures of the various publications, patents, and published patent specifications cited herein are hereby incorporated by reference in their entirety.
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood 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 1: degradation of aflatoxin by different strains
The method comprises the steps of obtaining strains 1# to 9# of hirsutella sinensis different from each other according to CN109082382A, CN105670937A in the prior art (wherein the strain 2# is the strain L0106 described in CN 109082382A), obtaining 9 strains in total, carrying out similarity analysis on obtained sequences and data stored in GenBank (DNA sequence database), and finding that the homology based on 18S rDNA sequences is higher than 95% according to the principle of microbial molecular genetics identification, so that nine strains obtained by screening are all hirsutella sinensis. Wherein the specific detection base sequences of the strain No. 1, the strain No. 2, the strain No. 3, the strain No. 4, the strain No. 7, the strain No. 8 and the strain No. 9 are respectively shown in SEQ ID NO. 1-7.
The paecilomyces hepiali strain is purchased from China general microbiological culture Collection center (CGMCC) and belongs to the institute of microbiology of China academy of sciences, and the CGMCC is 3.15540, 3.17720 and 3.15421 in serial number.
1. Preparation of seed liquid of hirsutella sinensis strains 1# to 9#, paecilomyces hepiali CGMCC3.15540, 3.17720 and 3.15421
(1) Preparation of seed liquid of hirsutella sinensis strain
Taking a hirsutella sinensis strain slant, scraping 1 slant hypha of which the number is 4, into a PDB shake flask culture medium (200 mL/500 mL), carrying out shake culture at 120rpm, wherein the culture temperature is 15 ℃, and the culture time is 12 days. Transferring the strain to a PDB culture medium according to the inoculation amount of 10 percent, and continuously culturing for 4 days to obtain seed liquid.
(2) Preparation of seed liquid of paecilomyces hepiali
Collecting paecilomyces hepiali strain slant, scraping a 1.5cm x 1.5cm strain block in PDB shake flask culture medium (200 mL/500 mL), shaking at 150rpm, culturing at 25 deg.C for 5 days. Inoculating to PDB culture medium according to the inoculation amount of 5%, and culturing for 2 days to obtain seed liquid.
HPLC detection (this method was used for all examples of the invention)
Extracting dichloromethane: centrifuging the sample at 12000rpm for 10min, taking 1mL of supernatant, adding 1mL of methanol, mixing, adding 1mL of dichloromethane, shaking, mixing uniformly, extracting, carefully sucking the lower organic phase liquid, extracting repeatedly for three times, and combining the extracts. Blow-drying by a nitrogen blowing instrument, dissolving by using 1mL of methanol after dichloromethane is completely volatilized, fixing the volume, filtering by using a 0.22 mu m filter membrane, and detecting by HPLC.
The chromatographic column is Pickering MYCOTOX TM C18 column (4.6 x 250mm,5 μm), mobile phase methanol: acetonitrile: water (volume ratio 22.
3. Degradation experiments
And respectively taking the strains to prepare seed solutions in a PDB culture medium, respectively adding 30 mu l of AFB1 standard substance (acetonitrile is used as a solvent) into 5mL of prepared strain seed solutions until the initial concentration of the system is 18ng/mL, carrying out shake culture at 14 ℃, carrying out 120rpm, and detecting the content of AFB1 by HPLC-light derivatization.
AFB1 degradation rate = (starting sample AFB1 concentration-sample AFB1 concentration to be measured)/starting sample AFB1 concentration 100%.
The experimental results are shown in the following table 1, the hirsutella sinensis strains 1# to 9#, the paecilomyces hepiali CGMCC3.15540, 3.17720 and 3.15421 can all degrade AFB1:
TABLE 1 degradation of AFB1 by different strains
Example 2: strain No. 8 degrading aflatoxin
5mL of strain No. 8 seed solution is taken, 30 mu L of aflatoxin AFB1 standard solution (acetonitrile is used as a solvent) is added, and PDB and the seed solution with the same volume are respectively added with the standard solution and the acetonitrile to be used as blank control, which is specifically shown in the following table 2. Both blank and experimental groups were cultured with shaking at 120rpm at 15 ℃ and 1mL was sampled at 0, 48, 72, 96, 120h, respectively.
TABLE 2 Experimental design
| Control 1 | 5mL of blank Medium + 30. Mu.L of Standard sample |
| Control 2 | 5mL seed liquid + 30. Mu.L acetonitrile |
| Low concentration test group 1 | 5mL seed solution + 30. Mu.L standard sample |
| High concentration test group 2 | 5mL seed liquid + 50. Mu.L standard sample |
Note: the original concentration of the standard sample is 3ppm, the concentration of AFB1 in the low-concentration experimental group 1 is 17.89ppb, and the concentration of AFB1 in the high-concentration experimental group 2 is 29.70ppb.
The detection of the content of the aflatoxin is carried out according to the method in the example 1, and the result shows that the hirsutella sinensis seed liquid can degrade an aflatoxin standard sample, the degradation rate of 90ng of a low-concentration sample in 96 hours is 98%, and the degradation rate of a high-concentration sample in 120 hours is 100%. The kinetics results of aflatoxin degradation of high concentration samples are shown in figure 1.
Distinguishing between degradation and desorption
Taking samples of low-concentration and high-concentration experimental groups, centrifuging at 12000rpm for 10min, taking supernatant, and extracting with chloroform; meanwhile, PBS solution (pH 7.4) with the same volume as the supernatant is added into the thallus sediment to a centrifuge tube, the thallus is resuspended, the centrifugation is carried out for 10min at 12000rpm, and the supernatant is discarded.
Adding methanol solution with equal volume to the precipitate, suspending, mixing, performing 40KHZ ultrasonic treatment for 30min, and filtering with 0.22 μm filter membrane to obtain precipitate.
The content of aflatoxin in the thalli precipitation extract and the sample of the supernatant is respectively detected, and the result shows that aflatoxin is not detected in the seed solution supernatant and the thalli extract, namely the strain No. 8 has the degradation effect on aflatoxin, but not the adsorption effect.
Example 3: strain No. 8 degrades aflatoxins AFG2, AFG1, AFB2 and AFB1
Seed solutions were prepared in the same manner as in example 1, and 4.95mL of the seed solution was taken and added with 50. Mu.L of each of the mixed standard AFB1 (1000 ppb), AFB2 (300 ppb), AFG1 (1000 ppb) and AFG2 (300 ppb) to initial concentrations of 10ppb, 3ppb, 10ppb and 3ppb, respectively, at 14 ℃ and 120rpm, and a PDB medium was used as a control, followed by shaking reaction for 144 hours, extraction with methylene chloride, and HPLC-light derivatization detection.
The result shows that the strain No. 8 can effectively degrade AFG2, AFG1, AFB2 and AFB1 at 144h, and the residual rates are 23%, 0, 10% and 0 respectively.
Example 4: degradation experiment of different components in strain 8# seed solution on AFB1
Preparing a seed solution by the same method as the example 1;
1. supernatant fluid
54mL of seed liquid was collected and centrifuged to obtain a supernatant.
2. Bacterial suspension:
the pellet was washed twice with PBS (pH7.4), and then suspended in the same volume of PBS to obtain a bacterial suspension.
3. Preparation of intracellular fluid and cell debris:
15mL of the suspension was ground, the slurry centrifuged, the supernatant was counted as the intracellular fluid, and the pellet was resuspended in 15mL of PBS and counted as cell debris.
4. Degrading AFB1 by different components:
respectively taking 5mL of the prepared seed liquid, supernatant, bacterial suspension, intracellular fluid and cell debris, adding 30 μ L of AFB1 standard substance one by one until the AFB1 initial concentration is 18ng/mL, carrying out shaking culture at 15 ℃ and 120rpm, respectively sampling for 96h, and carrying out HPLC-light derivatization detection.
The results of the experiment are shown in table 3 below.
TABLE 3 degradation test results of different components on AFB1 at 96h
The degradation effect of intracellular fluid without live bacteria and supernatant with basically removed bacteria is very low, i.e. hirsutella sinensis itself has degradation effect, and cell debris may contain live bacteria, thus having certain degradation effect.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and the like that are within the spirit and principle of the present invention are included in the present invention.
The foregoing embodiments and methods described in this disclosure may vary based on the abilities, experience, and preferences of those skilled in the art.
The mere order in which the steps of a method are listed in the present invention does not constitute any limitation on the order of the steps of the method.
Sequence listing
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<212> DNA
<213> Hirsutella sinensis (Hirsutella sinensis)
<400> 3
cctccgctta ttgatatgct taagttcagc gggtatccct acctgatccg aggtcaactg 60
gagggtgtgg tggtttcacg gcgtgaccgc ctccgcgctc ccggtgcgag gttctcagcg 120
agctactgcg caggggagcc gcggcggggt cgccactgcg tttagggggc ggcgcggggc 180
cgtgaccccc aaggccgggc cccgccgccg cgaggcgggg ggctcgaggg ttgagatgac 240
gctcggacag gcatgcccgc cagagtgctg gcgggcgcaa tgtgcgttca aagattcgat 300
ggttcactga attctgcaat tcacattact tatcgcattt cgctgcgttc ttcatcgatg 360
ccagaaccaa gagatccgtt gttgaaagtt ttgattcctt tgcttgcttc ttgactgaga 420
gatgccactg cgacaggagg gtctgggggt cctccggcgg gcgccctggt ccgggccctg 480
gggcgccggg gcggtcccgc cgaggcaact gctgtggtgt tcgcaggggg tttgggagtg 540
gtgactcgat aatgatccct ccgcaggtca ccctacgg 578
<210> 4
<211> 564
<212> DNA
<213> Hirsutella sinensis (Hirsutella sinensis)
<400> 4
cgtagggggg cctgcggagg gatcattatc gagtcaccac tcccaaaccc cctgcgaaca 60
ccacagcagt tgcctcggcg ggaccgcccc ggcgccccag ggcccggacc agggcgcccg 120
ccggaggacc cccagaccct cctgtcgcag tggcatctct cagtcaagaa gcaagcaaat 180
gaatcaaaac tttcaacaac ggatctcttg gttctggcat cgatgaagaa cgcagcgaaa 240
tgcgataagt aatgtgaatt gcagaattca gtgaaccatc gaatctttga acgcacattg 300
cgcccgccag cactctggcg ggcatgcctg tccgagcgtc atctcaaccc tcgagccccc 360
cgcctcgcgg cggcggggcc cggccttggg ggtcacggcc ccgcgccgcc ccctaaacgc 420
agtggcgacc ccgccgcggc tcccctgcgc agtagctcgc tgagaacctc gcaccgggag 480
cgcggaggcg gtcacgccgt gaaaccacca caccctccag ttgacctcgg atcaggtagg 540
gatacccgct gaacttaagc atat 564
<210> 5
<211> 561
<212> DNA
<213> Hirsutella sinensis (Hirsutella sinensis)
<400> 5
acctgcggag ggatcattat cgagtcacca ctcccaaacc ccctgcgaac accacagcag 60
ttgcctcggc gggaccgccc cggcgcccca gggcccggac cagggcgccc gccggaggac 120
ccccagaccc tcctgtcgca gtggcatctc tcagtcaaga agcaagcaaa tgaatcaaaa 180
ctttcaacaa cggatctctt ggttctggca tcgatgaaga acgcagcgaa atgcgataag 240
taatgtgaat tgcagaattc agtgaaccat cgaatctttg aacgcacatt gcgcccgcca 300
gcactctggc gggcatgcct gtccgagcgt catctcaacc ctcgagcccc ccgcctcgcg 360
gcggcggggc ccggccttgg gggtcacggc cccgcgccgc cccctaaacg cagtggcgac 420
cccgccgcgg ctcccctgcg cagtagctcg ctgagaacct cgcaccggga gcgcggaggc 480
ggtcacgccg tgaaaccacc acaccctcca gttgacctcg gatcaggtag ggatacccgc 540
tgaacttaag catatcaata a 561
<210> 6
<211> 554
<212> DNA
<213> Hirsutella sinensis (Hirsutella sinensis)
<400> 6
cctgcggagg gatcattatc gagtcaccac tcccaaaccc cctgcgaaca ccacagcagt 60
tgcctcggcg ggaccgcccc ggcgccccag ggcccggacc agggcgcccg ccggaggacc 120
cccagaccct cctgtcgcag tggcatctct cagtcaagaa gcaagcaaat gaatcaaaac 180
tttcaacaac ggatctcttg gttctggcat cgatgaagaa cgcagcgaaa tgcgataagt 240
aatgtgaatt gcagaattca gtgaaccatc gaatctttga acgcacattg cgcccgccag 300
cactctggcg ggcatgcctg tccgagcgtc atctcaaccc tcgagccccc cgcctcgcgg 360
cggcggggcc cggccttggg ggtcacggcc ccgcgccgcc ccctaaacgc agtggcgacc 420
ccgccgcggc tcccctgcgc agtagctcgc tgagaacctc gcaccgggag cgcggaggcg 480
gtcacgccgt gaaaccacca caccctccag ttgacctcgg atcaggtagg gatacccgct 540
gaacttaagc atat 554
<210> 7
<211> 552
<212> DNA
<213> Hirsutella sinensis (Hirsutella sinensis)
<400> 7
tgcggaggga tcattatcga gtcaccactc ccaaaccccc tgcgaacacc acagcagttg 60
cctcggcggg accgccccgg cgccccaggg cccggaccag ggcgcccgcc ggaggacccc 120
cagaccctcc tgtcgcagtg gcatctctca gtcaagaagc aagcaaatga atcaaaactt 180
tcaacaacgg atctcttggt tctggcatcg atgaagaacg cagcgaaatg cgataagtaa 240
tgtgaattgc agaattcagt gaaccatcga atctttgaac gcacattgcg cccgccagca 300
ctctggcggg catgcctgtc cgagcgtcat ctcaaccctc gagccccccg cctcgcggcg 360
gcggggcccg gccttggggg tcacggcccc gcgccgcccc ctaaacgcag tggcgacccc 420
gccgcggctc ccctgcgcag tagctcgctg agaacctcgc accgggagcg cggaggcggt 480
cacgccgtga aaccaccaca ccctccagtt gacctcggat caggtaggga tacccgctga 540
acttaagcat at 552
Claims (10)
1. Application of hirsutella sinensis and/or paecilomyces hepiali in degrading aflatoxin.
2. The application of a biological composition of a strain in degrading aflatoxin is disclosed, wherein the strain is hirsutella sinensis and/or paecilomyces hepiali.
3. Use according to claim 2, wherein the composition comprises a biomass of hirsutella sinensis or paecilomyces hepiali, preferably a live biomass.
4. The use according to claim 2, wherein the composition comprises one or more of seed liquor, fermentation liquor, bacterial suspension, supernatant, intracellular fluid, cell debris fluid, seed liquor extract, fermentation liquor extract, bacterial powder of the strain, preferably one or more of seed liquor, fermentation liquor, bacterial suspension.
5. The use of any one of claims 1 to 4, wherein the aflatoxin is selected from the group consisting of: aflatoxins B1, B2 α, G1, G2, aflatoxin derivatives M1, M2, P1, Q, H, GM, aflatoxins, especially AFB1, AFB2, AFG1, AFG2.
6. Application of hirsutella sinensis and/or paecilomyces hepiali in medicine, food or feed.
7. An aflatoxin-degrading agent comprising hirsutella sinensis and/or paecilomyces hepiali.
8. The use of claim 7, wherein the aflatoxin degradation agent comprises one or more of a seed liquor, a fermentation liquor, a bacterial suspension, a supernatant, an intracellular fluid, a cell debris fluid, a seed liquor extract, a fermentation liquor extract, a bacterial powder of the strain, preferably one or more of a seed liquor, a fermentation liquor, a bacterial suspension.
9. A method for degrading aflatoxins, which comprises the step of using hirsutella sinensis and/or paecilomyces hepiali.
10. The method of claim 9, wherein the method comprises the step of using one or more of seed liquor, fermentation liquor, bacterial suspension, supernatant, intracellular fluid, cell debris fluid, seed liquor extract, bacterial powder of hirsutella sinensis and/or paecilomyces hepiali.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105670937A (en) * | 2016-01-07 | 2016-06-15 | 广东东阳光药业有限公司 | Method for acquiring high quality Hirsutella sinensis strain |
| CN109082382A (en) * | 2018-06-15 | 2018-12-25 | 浙江工业大学 | Cordyceps sinensis Hirsutella sinensis ZJB18002 and its application |
| CN109161497A (en) * | 2018-08-21 | 2019-01-08 | 山东省花生研究所 | A kind of microorganism formulation of aflatoxin degradation and application |
-
2021
- 2021-04-07 CN CN202110376570.0A patent/CN115176959A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105670937A (en) * | 2016-01-07 | 2016-06-15 | 广东东阳光药业有限公司 | Method for acquiring high quality Hirsutella sinensis strain |
| CN109082382A (en) * | 2018-06-15 | 2018-12-25 | 浙江工业大学 | Cordyceps sinensis Hirsutella sinensis ZJB18002 and its application |
| CN109161497A (en) * | 2018-08-21 | 2019-01-08 | 山东省花生研究所 | A kind of microorganism formulation of aflatoxin degradation and application |
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