CN105255759B - One plant of aflatoxin and the difunctional degradation bacteria of ochratoxin and its application - Google Patents
One plant of aflatoxin and the difunctional degradation bacteria of ochratoxin and its application Download PDFInfo
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- CN105255759B CN105255759B CN201510681175.8A CN201510681175A CN105255759B CN 105255759 B CN105255759 B CN 105255759B CN 201510681175 A CN201510681175 A CN 201510681175A CN 105255759 B CN105255759 B CN 105255759B
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- aflatoxin
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- 230000015556 catabolic process Effects 0.000 title claims abstract description 63
- 238000006731 degradation reaction Methods 0.000 title claims abstract description 63
- 241000894006 Bacteria Species 0.000 title abstract description 28
- 229930195730 Aflatoxin Natural products 0.000 title description 8
- XWIYFDMXXLINPU-UHFFFAOYSA-N Aflatoxin G Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1C1C=COC1O2 XWIYFDMXXLINPU-UHFFFAOYSA-N 0.000 title description 8
- 239000005409 aflatoxin Substances 0.000 title description 8
- 229930183344 ochratoxin Natural products 0.000 title description 6
- OQIQSTLJSLGHID-WNWIJWBNSA-N aflatoxin B1 Chemical compound C=1([C@@H]2C=CO[C@@H]2OC=1C=C(C1=2)OC)C=2OC(=O)C2=C1CCC2=O OQIQSTLJSLGHID-WNWIJWBNSA-N 0.000 claims abstract description 55
- 239000002115 aflatoxin B1 Substances 0.000 claims abstract description 54
- 229930020125 aflatoxin-B1 Natural products 0.000 claims abstract description 54
- RWQKHEORZBHNRI-BMIGLBTASA-N ochratoxin A Chemical compound C([C@H](NC(=O)C1=CC(Cl)=C2C[C@H](OC(=O)C2=C1O)C)C(O)=O)C1=CC=CC=C1 RWQKHEORZBHNRI-BMIGLBTASA-N 0.000 claims abstract description 48
- VYLQGYLYRQKMFU-UHFFFAOYSA-N Ochratoxin A Natural products CC1Cc2c(Cl)cc(CNC(Cc3ccccc3)C(=O)O)cc2C(=O)O1 VYLQGYLYRQKMFU-UHFFFAOYSA-N 0.000 claims abstract description 47
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention provides one plant of Double function high efficient degradation bacteria (Stenotrophomonas sp.) CW117 and its application in degradation low stain concentration aflatoxin B1 and ochratoxin A.Compared with existing aspertoxin degradation bacteria, bacterial strain CW117 of the invention can reach splendid degradation effect under the conditions of low concentration endotoxin contamination.Under liquid fermentation condition, 72h is cultivated in the fermentation culture containing the final concentration of 20 μ g/L of aflatoxin B1 and ochratoxin A, bacterial strain CW117 is 99.4% to the degradation rate of ochratoxin A, and the degradation rate to aflatoxin B1 is 94.3%；The feed (20 μ g/kg of final concentration) of aflatoxin B1 and ochratoxin A combined pollution is handled with bacterial strain CW117,72h is 71.0% to the degradation rate of ochratoxin A, and the degradation rate to aflatoxin B1 is 54.5%.Bacterial strain CW117 has substantive application value and important meaning in food and feed biological detoxication application aspect.
Description
Technical field
The present invention relates to microbiology and biodegradable fields, specifically, being related to one plant of aflatoxin B1 and reddish brown song
The difunctional degradation bacteria of mould toxin A and its application.
Background technology
Aspergillus class mycotoxin is mainly produced by the secondary metabolism that the Aspergillus such as aspergillus flavus, aspergillus parasiticus and Aspergillus ochraceus generate
Object, it is separated to identify 20 various structures analogs, wherein with aflatoxin B1 (AFB1) and ochratoxin A (OTA)
Toxicity is maximum, is polluted in agricultural production and food industry the most extensively.Aflatoxin is by the World Health Organization within 1993
(WHO) it is I class carcinogenic substance that Agency for Research on Cancer, which delimited, because it has strongly carcinogenic, teratogenesis and mutagenicity, has been increasingly becoming
Publilc health and scientific research field focus of attention.Aflatoxin and ochratoxin Polluted grains and oil crops have become entirely
Ball sex chromosome mosaicism, in animal husbandry, two class aspertoxins can endanger animal health by polluting feed, cause Animal husbandry production rate low
Under, serious economic loss is caused, and by direct or indirect (animal product propagation) approach contaminated food products, influence the mankind's
Health.Animal takes in AFB1 or AFB2 from feed, through being converted into AFM1 or AFM2 in vivo, is present in milk, meat and birds, beasts and eggs
In, pollute human foods chain.The end of the year 2011, the exceeded event of aflatoxin that China's Dairy Industry occurs are AFB1 pollutions
The typical case that feed causes dairy produce AFM1 exceeded has caused the great attention of the Chinese government and food security department.With
Gradually intensification of the people to mycotoxin recognizing dangers, for the detection of aspergillus class mycotoxin, pollution prevention and detoxification technology
The research work of aspect is also deepening continuously, of close concern to each other with people's quality of the life, health and economic interests.
In recent years, it was found that multiple-microorganism has significant degradation or suction-operated to aflatoxin.For example, butyric acid
Vibrio (Butyrivibrio sp.), Bacillus acidi lactici (Lactobacillus sp.), streptococcus (Streptococcus
Sp.), Bifidobacterium (Bifidobacterium sp.) and acinetobacter (Acinetobacter sp.) etc..Report at present
Most virus-free strains exist in practical applications it is following of both common problem：First, part virus-free strain is de-
Malicious mechanism belongs to physical absorption, and the biodegradation of unsubstantiality, is adsorbed on the toxin of somatic cells wall, in animal or human body
Desorption may occur under different physicochemical environments, without substance detoxification meaning；Second, detoxification of the report strain to toxin
Rate majority measures under 100 μ g/kg concentration conditions higher above, bent for low concentration in the complex matrices such as raw-food material and feed
The practical detoxification capability study of mould toxoid (being less than 20 μ g/kg) is rarely reported.However, low dosage, combined pollution and high pollution
Rate is the universals of aspergillus class mycotoxin pollution again, and the low concentration aspergillus class mycotoxin of 20 μ g/kg can be right
People and animals' internal organs damage, and especially by the agricultural product of a variety of toxin combined pollutions, toxicity has more significant synergistic effect.
Therefore, excavating has under degradation low consistency conditions, the virus-free strain of a variety of aspergillus class mycotoxin combined pollutions, in agricultural production
There is great practice significance and application value with field of food safety.
Invention content
The object of the present invention is to provide one plant of aflatoxin B1 and ochratoxin A Double function high efficient degradation bacteria and its answer
With.
In order to realize the object of the invention, the present invention is from by the serious contaminated soil of polycyclic aromatic hydrocarbon (oil plant, Auto repair shop
Near) and isolated in by the mildew and rot food samples that mycotoxin seriously pollutes one plant can efficient degradation aflatoxin B1 and
The bacterial strain CW117 of ochratoxin A.The Main Morphology of the bacterium is characterized as：Gram-negative, no gemma are rod-shaped to curved bar shape
Feature has single polar flagella, can move (Fig. 1).Bacterial strain CW117 is strictly aerobic, and optimum growth temperature is 35 DEG C, most adaptability
Long pH is 7.0-7.5.
Morphological feature and 16S rDNA sequences (SEQ ID No.1) based on bacterial strain CW117, oligotrophy is accredited as by the bacterial strain
Monad (Stenotrophomonas sp.).The bacterial strain is preserved on June 14th, 2015 in China typical culture collection
The heart (abbreviation CCTCC, address：Wuhan, China, Wuhan University, postcode 430072), deposit number is CCTCC NO：M 2015371.
In vitro test shows that bacterial strain CW117 has significant ochratoxin (OTA) and aflatoxin in the short time
The degradation effect of B1 (AFB1).In the fluid nutrient medium of the final concentration of 20 μ g/L of OTA, bacterial strain CW117 is handled for 24 hours, to OTA
Degradation rate to reach the degradation rate of 79.9%, the 72h bacterial strains be 99.4%；In the Liquid Culture of the final concentration of 20 μ g/L of AFB1
In base, for 24 hours, the degradation rate to AFB1 is that 59.0%, 72h degradation rates reach 94.3% for bacterial strain CW117 processing.With bacterial strain CW117
The feed (20 μ g/kg of final concentration) of endotoxin contamination is handled, it is 54.5% that the OTA degradation rates of 72h, which are 71.0%, AFB1 degradation rates,.
The zymetology feature testing result of degradation bacteria strains shows that bacterial strain CW117 is mainly extracellular enzymatic to the mechanism of degradation of OTA
Degradation, extracellular crude enzyme liquid 72h are 83.1% to the degradation rate of OTA (20 μ g/L of final concentration), and bacterial strain CW117 is to AFB1's
Mechanism of degradation is mainly intracellular enzymatic degradation, and extracellular crude enzyme liquid 72h is only to the degradation rate of AFB1 (20 μ g/L of final concentration)
29.7%.
The present invention also provides microbial inoculums and its complex micro organism fungicide containing Stenotrophomonas CW117.
The present invention also provides the aflatoxin B1 prepared by Stenotrophomonas CW117 or described microbial inoculums and ochratoxins
The difunctional biodegradation agents of A.
Preferably, active constituent is the crude enzyme liquid prepared by the microbial inoculum in the biodegradation agent.
It is highly preferred that active constituent is zymotic fluid, the bacterial strain CW117 zymotic fluid of bacterial strain CW117 in the biodegradation agent
Through centrifugation gained supernatant or cellular lysate liquid.
Wherein, the culture medium for being used to prepare bacterial strain CW117 zymotic fluids is：Tryptone 17.0 g/L, soya peptone 3.0g/L,
Glucose 2.5g/L, NaCl 5.0g/L, K2HPO42.5g/L, pH value 7.0-7.5；Fermentation condition is：35 DEG C, 180r/min shakes
Swing culture 48h.
The present invention also provides Stenotrophomonas CW117, the microbial inoculum and the biodegradation agents in aflatoxin B1 and
Application in ochratoxin A biodegradation.
The present invention also provides Stenotrophomonas CW117, the microbial inoculum and the biodegradation agent go mouldy raw-food material or
Application in feed detoxification treatment.
Wherein, aflatoxin B1 and the content of ochratoxin A are respectively 20 in go mouldy raw-food material or the feed
μ g/kg and hereinafter, but it is not limited to 20 μ g/kg and following.
The present invention further provides Stenotrophomonas CW117 or described microbial inoculums to prepare aflatoxin B1 and Aspergillus ochraceus poison
Application in plain A Double function high efficients biodegradation agent.
Compared with existing aspertoxin degradation bacteria, Stenotrophomonas CW117 of the invention can be in low concentration aspergillus flavus
Reach splendid degradation effect under toxin B1 and ochratoxin A pollutional condition.It is advised according to state food and forage health standard
Fixed, cereal agricultural products aflatoxin content has to be lower than 20 μ g/kg, and ochratoxin content must not exceed 5 μ g/kg, raises
Ochratoxin A in material also must not exceed 100 μ g/kg (GB 2761-2011, GB13078.2-2006).In fact, animal
Toxicological experiment shows continuously to feed the feed containing 20 μ g/kg toxin, so that it may cause serious wound with the internal organs to rat
Evil (Wei et al., 2014DOI 10.1002/jsfa.6649).Therefore, bacterial strain CW117 has low concentration AFB1 and OTA
Efficient degradation-detoxification ability, has substantive application value and important meaning in foods/feeds biological detoxication application aspect.
Description of the drawings
Fig. 1 is the electron micrograph of bacterial strain CW117 of the present invention.
Fig. 2 is bacterial strain CW117 in the embodiment of the present invention 2 and 3 to the dynamic degradation curve of aspertoxin.
Fig. 3 is bacterial strain CW117 crude enzyme liquids in the embodiment of the present invention 2 to the dynamic degradation curve of aspertoxin.
Specific implementation mode
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..Unless otherwise specified, embodiment
Used in the conventional means that are well known to those skilled in the art of technological means, raw materials used is commercial goods.
Screening, identification and the culture of 1 aflatoxin B1 of embodiment and ochratoxin A degradation bacteria
1, the screening of bacterial strain
Sample collection is from by the serious contaminated soil of polycyclic aromatic hydrocarbon (near oil plant, Auto repair shop) and by mycotoxin
The mildew and rot food seriously polluted.Taking acquisition sample 0.5g to be added in 150mL enriched mediums, (OTA and AFB1 contents are respectively 20 μ
G/L or so), enrichment culture 7 days in 30 DEG C of constant incubators.After first time enrichment culture, 5mL enrichment culture liquid is taken to connect
In kind to OTA and AFB1 enriched mediums fresh 150mL, the concentration of OTA and AFB1 is increased to 30 μ g/L, in the same terms
Lower continuation enrichment culture 7 days.The experiment of third time enrichment culture is completed with identical enrichment method, improves the concentration of OTA and AFB1
To 50 μ g/L.After enrichment culture, enrichment culture liquid is made into gradient dilution with sterile water, gradient dilution liquid is respectively coated on
OTA or AFB1 solid separation culture mediums are cultivated 3 days under the conditions of 30 DEG C.After culture, picking culture dish single bacterium colony, which is inoculated in, to be contained
Degradation test cultures 48h is carried out in the enriched medium for there are each 20 μ g/L of OTA and AFB1, while setting does not connect the negative right of bacterium
According to.After degradation culture, culture solution is extracted through dichloromethane, according to national standards High Performance Liquid Chromatography with Fluorescence Detection
(HPLC-FLD is carried out with reference to National Standard of the People's Republic of China GB/T 25220-2010 and GB/T 18979-2003) inspection
Two toxoid residual concentrations in culture solution are surveyed, calculate degradation rate compared with the control.Finally, separation obtains one plant in low concentration OTA
The bacterial strain CW117 of efficient degradation rate is still kept with (OTA and AFB1 contents≤20 μ g/L) under the conditions of AFB1.
Wherein, enriched medium is：Glucose 5.0g/L, peptone 5.0g/L, 2.5 g/L of yeast powder, ammonium citrate
1.0g/L, NaAc3H2O 2.5g/L, MnSO4·4H2O 0.05g/L, K2HPO41.0g/L, MgSO4·7H2O 0.2g/L,
Tween-80 0.5mL, OTA add concentration 20-50 μ g/L, and AFB1 adds concentration 20-50 μ g/L, adjusts pH to 6.2-6.5.
Isolation medium is：Glucose 10.0g/L, peptone 10.0g/L, yeast powder 5.0g/L, ammonium citrate 2.0g/L,
MgSO4·7H2O 0.58g/L, NaAc3H2O 5.0g/L, MnSO4·4H2O 0.28g/L, Tween-80 1.0mL adjust pH extremely
6.2-6.5。
Above-mentioned culture medium high pressure sterilization 15min under the conditions of 120 DEG C uses the preceding sterile OTA and AFB1 toxin standard of addition
Solution is to normal concentration, and solid medium is by the above formula plus 2.0% agar powder.
2, the identification of bacterial strain CW117
The Main Morphology of bacterial strain CW117 is characterized as：Gram-negative, no gemma, rod-shaped to curved rod like features have single
Root polar flagella can move (Fig. 1).The bacterium is strictly aerobic, and optimum growth temperature is 35 DEG C, and the most suitable growth pH is 7.0-7.5.
16S rDNA identifications：Designed for expanding a pair of of universal primer of bacterial strain CW11716S rDNA sequences：27F 5'-
AGAGTTTGATCCTGGCTCAG-3' and 1492R 5'-GGTTACCTTGTTACGACTT-3'.
The genomic DNA of bacterial strain CW117 is extracted as template, carries out PCR amplification, and amplified production is sequenced.Bacterium
The 16S rDNA sequences of strain CW117 are as shown in SEQ ID No.1.
3, the culture of bacterial strain
Most suitable solid medium is：Tryptone 17.0g/L, soya peptone 3.0g/L, glucose 2.5 g/L, NaCl
5.0g/L, K2HPO42.5g/L, agar 20.0g/L, pH value 7.0-75.Most suitable fluid nutrient medium is that fine jade is free of in above-mentioned formula
Fat.
Optimal culture conditions are：35 DEG C, 180r/min shaken cultivations 48h.
2 high performance liquid chromatography of embodiment detects the degradation characteristic of bacterial strain CW117
Degradation bacteria strains degradation dynamic detects：Degradation bacteria strains CW117 was continuously activated into for 2 generations on most suitable solid medium, is connect
Kind is incubated overnight in 4mL fluid nutrient mediums, obtains fresh bacterium solution.The 50 fresh bacterium solutions of μ L are inoculated in 4mL OTA test trainings respectively
Base (containing 20.0 μ g/L OTA) and AFB1 test medias (containing 20.0 μ g/L AFB1) are supported, by the test tube shaken cultivation after inoculation
0,3 repetitions are arranged in 24,48 and 72h, each degradation experiment.With e. coli k12 (E.coli K12) for negative control bacterium
Strain.Mixing after culture after 8000r/min centrifuges 10min, collects supernatant and bacterial sediment respectively.
Supernatant of degrading passes through OTA immune affinity columns and AFB1 immune affinity columns respectively, methanol elution, collect eluent in
In glass sample injection bottle, according to national standards detection method (GB/T 25220-2010 and GB/T 18979-2003) carry out OTA and
The high performance liquid chromatography fluoroscopic examination (HPLC-FLD) of AFB1 residual quantities.Bacterial sediment after centrifugation, using the methanol of chromatographically pure
Resuspension mixing is carried out, the residual toxin of 10min extraction cell absorption is acutely shaken, precipitation is abandoned after centrifugation, supernatant crosses 0.22 μm
The residual toxin that cell absorption is carried out by HPLC-FLD after filter membrane detects.
Degradation bacteria strains zymetology feature detects：Degradation bacteria strains CW117 was continuously activated into for 2 generations on most suitable solid medium, it will
Inoculation after activation is incubated overnight in 50mL fluid nutrient mediums, obtains OD600=0.8 fresh bacterium solution.Bacterium solution is mixed
It is even, it is sub-packed in 50mL centrifuge tubes, 10000r/min centrifuges 15min, abandons precipitation.The accurate culture supernatant 40mL that measures is in nothing
In the clean triangular flask of bacterium, ochratoxin A/aflatoxin B1 is added respectively, its final concentration is made to reach 20 μ g/L.It will prepare
30 DEG C good of triangular flask is protected from light stationary culture, in the residual concentration of 0,24,48 and 72h sample detections OTA/AFB1.Each degradation
3 repetitions of experimental setup, and using E.coli K12 as negative control bacterial strain.The sample of taking-up is sub-packed in 2mL centrifuge tubes,
Centrifugation, abandons precipitation.Supernatant passes through OTA immune affinity columns and AFB1 immune affinity columns respectively, methanol elution, collect eluent in
In glass sample injection bottle, OTA and AFB1 residues detections are carried out respectively.
HPLC detects OTA conditions：Mobile phase：Acetonitrile：Water：Acetic acid (volume ratio 96:102:4), fluorescence detector：Excitation
Wavelength 333nm, launch wavelength 460nm, flow velocity：1.0mL/min, sample size：20 μ l, chromatographic column：C18Chromatographic column (4.6 ×
250mm, 5 μm of Waters of filler diameter)；HPLC detects AFB1 conditions：Mobile phase：Methanol：Water (volume ratio 45:55), glimmering
Photodetector：Excitation wavelength 360nm, launch wavelength 420nm, flow velocity：0.8mL/min, sample size：20 μ l, chromatographic column：C18Color
Compose column (4.6 × 250mm, 5 μm of Waters of filler diameter), 0.05% iodine solution post-column derivation method.
Bacterial strain CW117 to the dynamic degradation curve of aspertoxin as shown in Fig. 2, the experimental results showed that, CW117 strain fermentations
Liquid processing reaches 79.9% for 24 hours, to the degradation rate of OTA, and the degradation rate of the processing 72h bacterial strains is 99.4%；Processing is right for 24 hours
The degradation rate of AFB1 is 59.0%, and degradation rate is more than 94.3% after handling 72h.In addition, thalline to toxin without suction-operated.
Bacterial strain CW117 crude enzyme liquids are to the dynamic degradation curve of aspertoxin as shown in figure 3, degradation bacteria strains zymetology feature detects
The results show that bacterial strain CW117 is mainly extracellular enzymatic degradation to the mechanism of degradation of OTA, extracellular crude enzyme liquid 72h is (whole to OTA
20 μ g/L of concentration) degradation rate be 83.1%, bacterial strain CW117 is mainly intracellular enzymatic degradation to the mechanism of degradation of AFB1,
Extracellular crude enzyme liquid 72h is only 29.7% to the degradation rate of AFB1 (20 μ g/L of final concentration).
Applications of the 3 bacterial strain CW117 of embodiment in corn bean pulp type feed detoxification treatment
1, experiment material
Strain activation and culture base I：Tryptone 17.0g/L, soya peptone 3.0g/L, glucose 2.5g/L, NaCl 5.0g/
L, K2HPO42.5g/L, agar 20.0g/L.
Strain activation and culture base II：Peptone 5.0g/L, beef extract 30.0g/L, NaCl 5.0g/L, pH 7.0-7.2.
Above-mentioned culture medium high pressure sterilization 15min at 120 DEG C, experiment feed are corn bean pulp type daily ration.
2, experimental method
Appropriate OTA and AFB1 standard reserving solutions are added into 50mL phosphate buffers, 100g is poured into immediately after mixing
It in the Feed Sample crushed, stirs evenly, so that the final concentration of OTA and AFB1 is respectively reached 40.0 μ g/kg, feed is in the moon
It dries for use cool ventilation.2 generation of bacteria to be tested is continuously activated on solid activation medium I, and the strain after activation is inoculated in
150mL liquid activation medium II cultivates 48h, obtains OD600=1.0 fresh bacterium solution.Take the fresh bacterium solutions of 100mL and 100g systems
The combined pollution feed got ready is uniformly mixed, and prepared sample is cultivated under the conditions of 35 DEG C, in 0h, for 24 hours, 48h and 72h
3 repetitions are arranged in sampling, each degradation experiment.Meanwhile the pollution toxin prepared with the fresh blank cultures of 100mL and 100g
Mixing is uniformly mixed to handle as negative control.Processing group and blank control group carry out OTA and AFB1 immediately per sub-sampling 15g
Extraction and analysis.
The extraction and purification of toxin.The sample after 10g cultures is weighed in 150mL conical flask with stopper, be added 1g NaCl and
20mL methanol-water mixtures (volume ratio 4:1), room temperature concussion extraction 30min removes bottle stopper and is filtered in clean through Cao Wen filter paper
Cup in (or at 3000r/min room temperature centrifuge 5min), accurately pipette 5.0mL supernatants and that the dilution of 20mL pure water is added be mixed
It is even, it filters 1-2 times through glass fiber filter paper and is clarified to filtrate, carry out immune affinity column purification process.
Accurate to pipette 10.0mL (representing 1.0g samples) above-mentioned clear filtrate, in implantation glass syringe, adjusting pressure makes
Solution slow transits through immune affinity column with 1-2 drops/sec of flow velocity, until there is partial air by cylinder, is eluted with 10mL pure water
Pillar (is repeated once), discards whole effluxes and partial air is made to pass through cylinder.It is accurate that the elution of 1.5mL methanol, flow velocity is added
It is 1 drop/sec, collects eluent in glass sample injection bottle, carry out HPLC-FLD detections.
Bacterial strain CW117 to the dynamic degradation curve of aspertoxin as shown in Figure 1, the experimental results showed that, bacterial strain CW117 processing
The feed (20 μ g/Kg of final concentration) of toxin combined pollution, 72h are 71.0% to OTA degradation rates, are to AFB1 degradation rates
54.5%.Illustrate drops of the bacterial strain CW117 with extracellular enzymatic degradation capability to the aspergillus toxoids A of low concentration in complex matrices
It is preferable to solve effect, takes second place to the degradation effect of aflatoxin B1.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (10)
- Stenotrophomonas 1. (Stenotrophomonas sp.) CW117, deposit number is CCTCC NO：M 2015371.
- 2. the microbial inoculum containing Stenotrophomonas CW117 described in claim 1.
- 3. aflatoxin B1 and reddish brown prepared by microbial inoculum described in Stenotrophomonas CW117 or claim 2 described in claim 1 The difunctional biodegradation agents of aspertoxin A.
- 4. biodegradation agent according to claim 3, which is characterized in that its active constituent is thick to be prepared by the microbial inoculum Enzyme solution.
- 5. biodegradation agent according to claim 3, which is characterized in that its active constituent be bacterial strain CW117 zymotic fluid, Bacterial strain CW117 zymotic fluids are through centrifugation gained supernatant or cellular lysate liquid.
- 6. biodegradation agent according to claim 5, which is characterized in that be used to prepare the culture of bacterial strain CW117 zymotic fluids Base is：Tryptone 17.0g/L, soya peptone 3.0g/L, glucose 2.5g/L, NaCl 5.0g/L, K2HPO42.5g/L, pH value 7.0-7.5；Fermentation condition is：35 DEG C, 180r/min shaken cultivations 48h.
- 7. any one of microbial inoculum, claim 3-6 life described in Stenotrophomonas CW117, claim 2 described in claim 1 Application of the object degradation agent in aflatoxin B1 and ochratoxin A biodegradation.
- 8. any one of microbial inoculum, claim 3-6 life described in Stenotrophomonas CW117, claim 2 described in claim 1 Application of the object degradation agent in go mouldy raw-food material or feed detoxification treatment.
- 9. application according to claim 8, which is characterized in that aflatoxin B1 in go mouldy raw-food material or the feed Content with ochratoxin A is respectively 20 μ g/kg and hereinafter, but is not limited to 20 μ g/kg and following.
- 10. microbial inoculum described in Stenotrophomonas CW117 or claim 2 described in claim 1 is preparing aflatoxin B1 and reddish brown Application in the difunctional biodegradation agents of aspertoxin A.
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| CN108396004B (en) * | 2018-04-10 | 2020-08-07 | 华南农业大学 | Aflatoxin B capable of efficiently degrading1Escherichia coli CG1061 |
| CN109762761B (en) * | 2019-01-10 | 2022-01-07 | 河南农业大学 | Stenotrophomonas acidiphila capable of efficiently degrading aflatoxin B1 and application thereof |
| CN111607579B (en) * | 2019-02-22 | 2021-10-08 | 安徽农业大学 | Ochratoxin hydrolase, encoding gene, recombinant vector and application thereof |
| CN116286384A (en) * | 2022-12-15 | 2023-06-23 | 中国农业科学院农产品加工研究所 | Destruxin of destruxin-OT 3 of Metarrhizium anisopliae, and detoxification enzyme produced by destruxin-OT 3 and application of destruxin |
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