CN105039199A - Deep-sea Bacillus circulans and application thereof in suppression of aflatoxin - Google Patents
Deep-sea Bacillus circulans and application thereof in suppression of aflatoxin Download PDFInfo
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- CN105039199A CN105039199A CN201510314566.6A CN201510314566A CN105039199A CN 105039199 A CN105039199 A CN 105039199A CN 201510314566 A CN201510314566 A CN 201510314566A CN 105039199 A CN105039199 A CN 105039199A
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Abstract
The invention provides a deep-sea bacterial strain capable of preventing and controlling aflatoxin, and particularly relates to a Bacillus circulans FA13 bacterial strain which is formed by separating light-yellow ooze sediment in water of 3203m in depth in South Atlantic, collected in the China General Microbiological Culture Collection Center and numbered as CGMCC No.10663. After the FA13 bacterial strain is fermented under optimized conditions and obtained fermentation supernate is diluted by 30 times, the fermentation supernate can still kill spores of aflatoxin producing bacteria and suppress producing of aflatoxin 100%, in other words, the fermentation supernate has capability of suppressing production of aflatoxin from a growing source, has no suppression effect on intestinal flora and unicellular fungus saccharomyces cerevisia and has higher using safety; active metabolite in fermentation liquid is stable in property and resistant to high temperature, acid and alkali and degrading of pepsin, trypsin and chymotrypsin. The FA13 bacterial strain and a fermentation product thereof can be used for effectively preventing and controlling aflatoxin pollution of feed, grain and field crops.
Description
Technical field
The present invention relates to Deep-Sea Microorganisms technology and the biological Control Technology field of mycotoxins, the specifically application of a strain deep-sea Bacillus circulans and suppression aflatoxin thereof.
Background technology
Aflatoxin is the mycotoxins that the toxicity that found at occurring in nature is at present the strongest, and has carcinogenic, teratogenesis and inhibitive ability of immunity by force, is classified as a class carcinogenic substance by international cancer mechanism.Produce primarily of the filamentous fungus such as flavus, Aspergillus parasiticus, extensively can pollute grain, feed etc., after human and animal eats aflatoxin-contaminated food and feed, can the health of serious harm human and animal, therefore, how effectively prevention and control aflatoxin contamination is key subjects global urgently to be resolved hurrily.
The prefered method of effective prevention and control aflatoxin contamination feed and grain presses down poison, namely the generation of aflatoxin in feed and grain is suppressed, instead of except poison, namely the aflatoxin produced is removed, because except poison, not only cost is large, effect is also undesirable, is difficult to eradicate, and can cause the loss of nutrition.
Summary of the invention
Technical problem to be solved by this invention overcomes above-mentioned the deficiencies in the prior art, and the technical scheme that the present invention solves the problems of the technologies described above employing is: the application of a strain deep-sea Bacillus circulans and suppression aflatoxin thereof
,its feature comprises the steps:
(1) the present invention said deep-sea Bacillus circulans be Bacillus circulans FA13 bacterial strain (
bacilluscirculansfA13), and be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on March 24th, 2015, deposit number is CGMCCNo.10663;
(2) the present invention said FA13 strains separation is from the light yellow ooze settling of the 3203 meters of depth of waters in South Atlantic, be separated acquisition by the following method: fresh light yellow ooze 10 times of serial dilutions are after 55 DEG C of water-bath 6min, coat Gause I substratum, 28 degree of lower constant temperature culture, timely picking list bacterium colony, and through purifying of ruling further, obtain pure strain FA13 bacterial strain;
(3) the said FA13 bacterial strain of the present invention is accredited as a new strains of Bacillus circulans through 16SrRNA gene molecule, and thalline is shaft-like, can produce gemma, Gram-positive;
(4) by deep-sea of the present invention Bacillus circulans FA13 inoculation in Optimal Medium; at 20-36 DEG C of temperature; shaking culture 4-12 days; the fermented liquid containing high-efficiency activated material can be obtained; even if after this fermented liquid dilution 30 times; still 100% can suppress the spore germination of aflatoxin producing strains and the synthesis of aflatoxin, after dilute 40 times, still can the synthesis of 100% suppression aflatoxin.
(5) the active substance stable in properties in the fermented liquid of FA13 bacterial strain of the present invention, process 30 minutes under 121 DEG C of High Temperature High Pressure after, activity is not lost; Process 6 hours in the scope of pH2-12 after, activity is not lost; With stomach en-, Chymotrypsin and trypsinase etc. under optimum conditions enzymolysis after 6 hours activity still do not lose;
(6) active substance in the fermented liquid of FA13 bacterial strain of the present invention has good biological safety, to the intestinal microflora such as intestinal bacteria, gas bacillus unrestraint effect, does not also have restraining effect to unicellular fungi yeast saccharomyces cerevisiae;
(7) deep-sea of the present invention Bacillus circulans FA13 bacterial strain and fermented liquid thereof can be applicable to effective prevention and control of the aflatoxin contamination of feed, grain, field crop etc.
Embodiment
Below in conjunction with embodiment, the present invention will be further described, but therefore do not limit the present invention among described scope of embodiments.
Embodiment 1: the liquid fermenting of deep-sea Bacillus circulans FA13 bacterial strain
By the liquid seeds of the FA13 bacterial strain of incubated overnight, be inoculated in the liquid nutrient medium containing 0.5-1.5% yeast extract paste, 0.1-0.3% maltose and 1.0-3.0% glucose by 1% inoculum size, 6-12 days is cultivated under 20-36 degree, 100-120 rev/min, terminate fermentation, carry out centrifugal to fermented liquid, namely the cell-free supernatants obtained is containing the efficient fermented liquid suppressing the active substance of aflatoxin, the pollution of prevention and control aflatoxin can be directly used in, or be processed into the pollution of dry powder for prevention and control aflatoxin.
Embodiment 2: the fermented liquid of deep-sea Bacillus circulans FA13 bacterial strain and diluent thereof suppress the activity of aflatoxin
Add in the cell free fermentation supernatant liquor that embodiment 1 is obtained with the nutritive ingredient of fermention medium equivalent after as the nutrient solution cultivating aflatoxin producing strains, and cell free fermentation supernatant liquor fermention medium embodiment 1 to be obtained dilutes after 30 times and 40 times respectively as the nutrient solution cultivating aflatoxin producing strains, contrast is fermention medium, inoculate the spore suspension of aflatoxin producing strains of 1% respectively in above-mentioned substratum after, 28 degree of cultivations 6 days, collected by centrifugation mycelium, after weighing, the aflatoxin intermediate product in mycelium is extracted by methanol-hydrogen sodium hydroxide solution, light absorption value is measured at 560nm place, calculate bacteriostasis rate and press down malicious rate, result shows, the spore of the aflatoxin producing strains inoculated in the cell free fermentation supernatant liquor of undiluted cell free fermentation supernatant liquor and dilution 30 times is not sprouted, in the substratum that its renewed vaccination is grown to suitable spore germination after 6 days, still without spore germination, show that the spore of aflatoxin producing strains is killed after cultivating in the cell free fermentation supernatant liquor of FA13 bacterial strain, thus, the cell free fermentation supernatant liquor of FA13 bacterial strain and the cell free fermentation supernatant liquor of dilution 30 times have the ability of killing spore completely and suppressing aflatoxin to produce, show to reach more than 80% to the inhibiting rate of mycelial growth to the detection of the FA13 cell free fermentation supernatant liquor of dilution 40 times, 100% is reached to the inhibiting rate that aflatoxin produces.
Embodiment 3: the temperature stability of the active substance in the fermented liquid of deep-sea Bacillus circulans FA13 bacterial strain
Cell free fermentation supernatant liquor embodiment 1 obtained is 60,80
oat C temperature, heat treated naturally cools to room temperature and 121 after 2 hours
ounder C, autoclaving naturally cooled to room temperature after 30 minutes, by the fermented liquid of above-mentioned process, to inoculate 1% respectively together with blank cultures without the fermented liquid of Temperature Treatment the spore suspension of aflatoxin producing strains after, 28 degree of cultivations 6 days, observe spore germination situation, even if result shows 121
ounder C, the autoclaving fermented liquid of 30 minutes also can be the same with the fermented liquid without Temperature Treatment, suppresses the spore germination of aflatoxin producing strains and the generation of aflatoxin completely, shows that the antibacterial cytotoxic activity material that presses down in fermented liquid has temperature stability.
Claims (1)
1. a strain is separated from the light yellow ooze sedimental deep-sea bacterial strain of the 3203 meters of depth of waters in South Atlantic, for Bacillus circulans FA13 bacterial strain (
bacilluscirculansfA13), be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on March 24th, 2015, deposit number is CGMCCNo.10663.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112021461A (en) * | 2020-09-16 | 2020-12-04 | 沈阳师范大学 | Method for inhibiting aflatoxin in feed of weaned pigs |
| CN113186111A (en) * | 2020-12-15 | 2021-07-30 | 哈尔滨工业大学（威海） | Deep sea actinomycete rhodococcus erythropolis and application thereof in inhibiting aflatoxin |
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| CN104293882A (en) * | 2014-09-29 | 2015-01-21 | 哈尔滨工业大学（威海） | Method for quantitatively and rapidly screening active materials for inhibiting production of aflatoxin |
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| CN104328161A (en) * | 2014-09-29 | 2015-02-04 | 哈尔滨工业大学（威海） | Method of intuitively screening microorganism strains for inhibiting aflatoxin |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112021461A (en) * | 2020-09-16 | 2020-12-04 | 沈阳师范大学 | Method for inhibiting aflatoxin in feed of weaned pigs |
| CN113186111A (en) * | 2020-12-15 | 2021-07-30 | 哈尔滨工业大学（威海） | Deep sea actinomycete rhodococcus erythropolis and application thereof in inhibiting aflatoxin |
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| CN105039199B (en) | 2019-08-16 |
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