CN102067957A - Application of bacillus megaterium in degradation of peanut meal aflatoxin B1 - Google Patents
Application of bacillus megaterium in degradation of peanut meal aflatoxin B1 Download PDFInfo
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- CN102067957A CN102067957A CN2010105225427A CN201010522542A CN102067957A CN 102067957 A CN102067957 A CN 102067957A CN 2010105225427 A CN2010105225427 A CN 2010105225427A CN 201010522542 A CN201010522542 A CN 201010522542A CN 102067957 A CN102067957 A CN 102067957A
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- peanut meal
- afb
- bacillus megaterium
- aflatoxin
- fermentation
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Abstract
The invention provides application of bacillus megaterium in degradation of peanut meal aflatoxin B1, and belongs to the technical field of microbe application. A method comprises the following steps of: sterilizing crushed target peanut meal, adding a certain amount of aflatoxin B1, then adding bacillus megaterium liquid in an amount which is 5 to 10 percent of the inoculation amount, adding sterile water in a ratio of materials to water of 1:0.5-1.5, and fermenting for 48 to 72 hours at the temperature of between 30 and 40 DEG C; and adding a solvent into the peanut meal after fermentation is completed, shaking a table for 10 to 50 minutes, extracting the residual aflatoxin B1, and detecting the content of the aflatoxin B1 by using enzyme linked immunosorbent assay. The method has the advantages of capability of effectively reducing the aflatoxin B1 content of the peanut meal, low energy consumption, low environmental pollution and no need of wastewater treatment, strict sterile operation or continuous ventilation.
Description
Technical field
The present invention relates to a strain bacillus megaterium at degraded peanut meal AFB
1In application, belong to the using microbe technical field.
Background technology
Peanut meal is to be raw material with the shelling peanut, the byproduct after extracting grease.The nutritive value of peanut meal is higher, and metabolizable energy content surpasses soybean cake dregs, but peanut meal mainly is used as feed at present, and cheap.A main cause of restriction peanut meal price and extensive use is its easy infection Aspergillus flavus and produce aflatoxin, wherein with AFB
1The most representative.
AFB
1The removal method mainly contain physics, chemistry and biological method.Biological detoxication method treatment conditions gentleness, and can increase the nutritive value of product, be subjected to extensive concern.Bioanalysis removes AFB both at home and abroad
1Research mainly concentrate on microbial strains and directly act on AFB
1, and the enzyme that bacterial strain produces is made preparation act on AFB
1Related bacterial classification mainly contains the Armillariella tabescens of the above the average age for marriage research of domestic Liu, the slime bacteria of Wang Ning research and the stenotrophomonas maltophilia of Li Junxia research, and the lactic acid bacteria of foreign study, Bifidobacterium, saccharomycete, orange Flavobacterium and Rhodococcus erythropolis etc.Wherein Li Junxia adds the zymotic fluid of stenotrophomonas maltophilia in mouldy corn, the mouldy feed and Chinese herbal medicine, by liquid fermentation method degraded AFB wherein
1Do not see as yet in the domestic and international research report that at present bacillus megaterium can solid state fermentation peanut meal degraded AFB wherein
1
Solid state fermentation is at the nutritive value that improves oil meal and remove in the cottonseed dregs of rice aspect such as noxious material gossypol certain application has been arranged.A strain bacillus megaterium provided by the invention is at degraded peanut meal AFB
1In application, have that culture medium is simple, energy consumption is low, contaminated environment is less, do not need wastewater treatment, do not need strict aseptic manipulation, do not need advantages such as continuous ventilating.
Summary of the invention
The invention provides a strain bacillus megaterium at degraded peanut meal AFB
1In application, this application can effectively reduce AFB in the peanut meal
1Content.Its processing step is looked over so as to check after peanut meal is pulverized; Take by weighing a certain amount of peanut meal, put into plate, wrap, high pressure steam sterilization with brown paper; After the peanut meal sterilization cooling, add a certain amount of AFB in each culture dish
1Solution; With cultivating the bacillus megaterium seed liquor of 8～12h, be inoculated in the peanut meal by inoculum concentration 5～10%, mix; Sterilized water is pressed material-water ratio 1: 0.5～1.5, add in the peanut meal, mix; With postvaccinal peanut meal at the following heat insulating culture fermentation of 30～40 ℃ of temperature 48～72h; Fermented the back drying peanut dregs of rice to constant weight, added methanol-water (1+1) shaking table concussion 10～50min, extracted residual AFB in the peanut meal
1, detect its content with enzyme linked immunosorbent assay.
Bacillus megaterium used in this method obtains after the separation and purification screening from peanut soil with specificity method, and this bacterial strain has following character:
(1) morphological feature: bacterium colony is faint yellow, circle, and colony edge is neat, and surface wettability is smooth; Cellular morphology is short pole shape, and Gram is positive.
(2) physicochemical property
Advantage of the present invention is effectively to reduce AFB in the peanut meal
1Content, reduce the harm of toxin to people and animals, help the extensive use of peanut meal.And bacillus megaterium degraded peanut meal AFB
1Whole process, energy consumption is low, contaminated environment is less, do not need wastewater treatment, do not need strict aseptic manipulation, do not need continuous ventilating, has simple to operate, low cost and other advantages.
The specific embodiment
Embodiment 1.
Bacillus megaterium degraded peanut meal AFB
1The concrete operations flow process be: bacillus megaterium is cultivated 24h 37 ℃ of LB solid mediums, and picking list bacterium colony inserts in the LB fluid nutrient medium, and 37 ℃, the concussion of 180r/min shaking table is cultivated; 10mg is purchased in the AFB of Fermentek company
1(98: 2, V/V) in the solution, concentration was 200 μ g/mL, i.e. AFBs to be dissolved in 50mL benzene-acetonitrile
1Storing solution is got a certain amount of AFB
1Storing solution is used N
2Blow, vapor away solvent benzol and acetonitrile, (PBS pH7.4), makes AFB to add a certain proportion of phosphate buffer
1Ultimate density is 2000ng/mL; Peanut meal is pulverized the back cross 20 mesh sieves; Take by weighing 5g peanut meal (being accurate to 0.01g) in culture dish, wrap, 121 ℃ of high pressure steam sterilization 20min with brown paper; After the peanut meal sterilization cooling, adding 250 μ l concentration in each culture dish is the AFB of 2000ng/mL
1Solution; With cultivating the bacillus megaterium seed liquor of 12h, be inoculated in the peanut meal by inoculum concentration 10%, mix; Sterilized water was pressed material-water ratio 1: 1, add in the peanut meal, mix; With postvaccinal peanut meal at the following heat insulating culture fermentation of 35 ℃ of temperature 60h; Fermented back 50 ℃ of drying peanut dregs of rice to constant weight, added the methanol-water (1+1) of 10 times of volumes of peanut meal quality, shaking table concussion 30min extracts residual AFB in the peanut meal
1, detect its content with enzyme linked immunosorbent assay, obtain AFB in the peanut meal
1Degradation rate can reach 69.34%.
Claims (4)
1. a strain bacillus megaterium is at degraded peanut meal AFB
1In application, it is characterized in that this strain bacterium is bacillus megaterium (Bacillus megaterium), the screening from peanut soil, be preserved in Chinese typical culture collection center, deposit number is CCTCC M2010272.
2. a strain bacillus megaterium according to claim 1 is at degraded peanut meal AFB
1In application, it is characterized in that to contain AFB
1Peanut meal as solid-state fermentation culture medium, the described bacterial strain of inoculation claim 1, control material-water ratio (the ratio of the quality of peanut meal and the volume of sterilized water, g/mL), seed liquor incubation time, bacterium liquid inoculum concentration (the long-pending percentage that accounts for the peanut meal quality of bacteria liquid, %), fermentation temperature and fermentation time, carry out solid state fermentation.With residual AFB in the peanut meal of solvent extraction fermentation back
1, detect its content with enzyme linked immunosorbent assay.
According to claim 1 and the described strain bacillus megaterium of claim 2 at degraded peanut meal AFB
1In application, it is characterized in that described material-water ratio is 1: 0.5～1.5, the seed liquor incubation time is 8～12h, bacterium liquid inoculum concentration is 5～10%, fermentation temperature is 30～40 ℃, fermentation time is 48～72h.
According to claim 1 and the described strain bacillus megaterium of claim 2 at degraded peanut meal AFB
1In application, it is characterized in that add methanol-water (1+1) behind the solid state fermentation in peanut meal, shaking table concussion 10～50min extracts wherein residual AFB
1
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105225427A CN102067957A (en) | 2010-10-28 | 2010-10-28 | Application of bacillus megaterium in degradation of peanut meal aflatoxin B1 |
Applications Claiming Priority (1)
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|---|---|---|---|
| CN2010105225427A CN102067957A (en) | 2010-10-28 | 2010-10-28 | Application of bacillus megaterium in degradation of peanut meal aflatoxin B1 |
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| CN102067957A true CN102067957A (en) | 2011-05-25 |
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| CN2010105225427A Pending CN102067957A (en) | 2010-10-28 | 2010-10-28 | Application of bacillus megaterium in degradation of peanut meal aflatoxin B1 |
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Cited By (6)
| 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 |
| CN105166602A (en) * | 2015-08-07 | 2015-12-23 | 山东省花生研究所 | Method for degrading aflatoxins in peanut meal by utilizing solid fermentation of fructificatio amaurodermatis rudae |
| CN106065396A (en) * | 2015-11-24 | 2016-11-02 | 沈阳农业大学 | A kind of vegetable bacillus cereus and cultural method thereof and application |
| CN108456648A (en) * | 2018-03-04 | 2018-08-28 | 河南亿万中元生物技术有限公司 | A kind of method of aflatoxin B1 in degradation peanut meal |
| CN110129229A (en) * | 2019-05-19 | 2019-08-16 | 河南工业大学 | A kind of probiotics preparation method of protect liver and degrading aflatoxin B 1 |
| CN110583964A (en) * | 2019-09-23 | 2019-12-20 | 江南大学 | Biological removal method for efficiently removing four aflatoxins in peanut meal |
-
2010
- 2010-10-28 CN CN2010105225427A patent/CN102067957A/en active Pending
Cited By (6)
| 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 |
| CN105166602A (en) * | 2015-08-07 | 2015-12-23 | 山东省花生研究所 | Method for degrading aflatoxins in peanut meal by utilizing solid fermentation of fructificatio amaurodermatis rudae |
| CN106065396A (en) * | 2015-11-24 | 2016-11-02 | 沈阳农业大学 | A kind of vegetable bacillus cereus and cultural method thereof and application |
| CN108456648A (en) * | 2018-03-04 | 2018-08-28 | 河南亿万中元生物技术有限公司 | A kind of method of aflatoxin B1 in degradation peanut meal |
| CN110129229A (en) * | 2019-05-19 | 2019-08-16 | 河南工业大学 | A kind of probiotics preparation method of protect liver and degrading aflatoxin B 1 |
| CN110583964A (en) * | 2019-09-23 | 2019-12-20 | 江南大学 | Biological removal method for efficiently removing four aflatoxins in peanut meal |
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Application publication date: 20110525 |