CN113215014A - Deep sea Kocuria rosea for degrading aflatoxin - Google Patents
Deep sea Kocuria rosea for degrading aflatoxin Download PDFInfo
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- CN113215014A CN113215014A CN202011510102.XA CN202011510102A CN113215014A CN 113215014 A CN113215014 A CN 113215014A CN 202011510102 A CN202011510102 A CN 202011510102A CN 113215014 A CN113215014 A CN 113215014A
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- Prior art keywords
- m2pmix1
- m2mix5
- aflatoxin
- strain
- rosea
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- 241000191948 Kocuria rosea Species 0.000 title claims abstract description 10
- 229930195730 Aflatoxin Natural products 0.000 title abstract description 15
- XWIYFDMXXLINPU-UHFFFAOYSA-N Aflatoxin G Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1C1C=COC1O2 XWIYFDMXXLINPU-UHFFFAOYSA-N 0.000 title abstract description 15
- 239000005409 aflatoxin Substances 0.000 title abstract description 15
- 230000000593 degrading effect Effects 0.000 title abstract description 8
- 238000011065 in-situ storage Methods 0.000 claims abstract description 3
- 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 abstract description 26
- 229930020125 aflatoxin-B1 Natural products 0.000 abstract description 26
- 238000000855 fermentation Methods 0.000 abstract description 14
- 230000004151 fermentation Effects 0.000 abstract description 14
- 239000006228 supernatant Substances 0.000 abstract description 11
- 230000015556 catabolic process Effects 0.000 abstract description 10
- 238000006731 degradation reaction Methods 0.000 abstract description 10
- 238000001784 detoxification Methods 0.000 abstract description 9
- 239000002115 aflatoxin B1 Substances 0.000 abstract description 6
- 239000001963 growth medium Substances 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 3
- 235000013305 food Nutrition 0.000 abstract description 3
- 244000005700 microbiome Species 0.000 abstract 1
- 239000000047 product Substances 0.000 abstract 1
- 101100449517 Arabidopsis thaliana GRH1 gene Proteins 0.000 description 20
- 101100434479 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) AFB1 gene Proteins 0.000 description 20
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- 241000544061 Cuculus canorus Species 0.000 description 4
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- 244000178231 Rosmarinus officinalis Species 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- 241001137883 Cucumaria Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 231100000678 Mycotoxin Toxicity 0.000 description 1
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002636 mycotoxin Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- LJCNRYVRMXRIQR-OLXYHTOASA-L potassium sodium L-tartrate Chemical compound [Na+].[K+].[O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O LJCNRYVRMXRIQR-OLXYHTOASA-L 0.000 description 1
- 229940074439 potassium sodium tartrate Drugs 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- 235000011006 sodium potassium tartrate Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
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
- A23L5/28—Removal of unwanted matter, e.g. deodorisation or detoxification using microorganisms
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Microbiology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a deep sea strain capable of degrading aflatoxin B1 and having a detoxification effect, which is Kocuria rosea M2PMix1-M2Mix5-13 strain (a)Kocuria rosea M2PMix1-M2Mix5-13), a culture isolated from 1617M depth of mountain top of the Western Pacific dock land, which was subjected to in situ directional enrichment culture for 1 year, and has been deposited in Guangdong province collection center for microorganism strains with the deposit number GDMCC No. 61240. Fermenting the M2PMix1-M2Mix5-13 strain in an M2 culture medium, wherein the degradation rate of the aflatoxin B1 in 1 day at 37 ℃ of the obtained fermented cell-free supernatant reaches 94.01 +/-1.45%; the degradation rate of the fermented cell-free supernatant obtained after 4 days of fermentation on the aflatoxin B1 reaches 99.89 +/-0.02 percent at 37 ℃ for 1 day. The Cokucuria rosea M2PMix1-M2Mix5-13 strain and the fermentation product thereof can be used for related food and drinksAnd removing and detoxifying the polluted aflatoxin in feed and the like.
Description
The technical field is as follows:
the invention relates to the technical field of deep sea microbial technology and mycotoxin biological prevention and control, in particular to a deep sea Kocuria rosea for degrading aflatoxin.
Background art:
the aflatoxin pollution of grains and feeds is a worldwide problem, and the grains and feeds polluted by the aflatoxin not only bring serious harm to the health of human beings and animals, but also cause huge economic loss in industries such as agriculture, food processing industry, livestock breeding industry, aquaculture industry and the like, so that the removal and detoxification of the aflatoxin is a focus and a research focus of attention of domestic and foreign scientists and industries.
The removal and detoxification of aflatoxin comprises physical detoxification, chemical detoxification and biological detoxification, wherein physical detoxification methods such as high temperature and radiation are relatively complex in actual operation, chemical detoxification methods such as ozone and ammoniation pollute the environment, are high in cost and long in consumed time, and are difficult to use on a large scale.
The invention content is as follows:
the technical problem to be solved by the invention is to overcome the defects of the prior art, and the technical scheme adopted by the invention for solving the technical problem is as follows: the deep sea Kocuria rosea for degrading aflatoxin is characterized by comprising the following steps:
(1) the deep sea Kuck rosea of the invention is Kuck rosea M2PMix1-M2Mix5-13 strain(s) ((R))KocuriaroseaM2PMix1-M2Mix5-13) and has been deposited at the Guangdong province culture Collection in 26/10 of 2020,the address of the preservation unit is No. 59 building No. 5 building of No. 100 college of the Pieli Zhonglu, Guangzhou city, and the preservation number is GDMCC No. 61240;
(2) the deep sea Cuckoo rosea M2PMix1-M2Mix5-13 strain is separated from a culture obtained by in-situ directional enrichment culture for 1 year at 1617M depth of the mountain top of the Western Pacific dock Lang flat-topped sea mountain, diluted by 10 times with sterile water, coated on an M2 culture medium containing 1 mu g/ml aflatoxin B1, cultured at constant temperature of 20 ℃, picked in time and subjected to single colony selection and further streaking purification to obtain a pure strain M2PMix1-M2Mix 5-13;
(3) the M2PMix1-M2Mix5-13 strain is identified as Kukelia rosea by 16S rRNA gene molecules (K, KKocuria rosea ) A new strain of (a);
(4) inoculating the deep sea Kocuria rosea M2PMix1-M2Mix5-13 strain in an M2 culture medium, and performing shaking culture at the temperature of 18-28 ℃ for 1-5 days to obtain a fermentation broth containing active substances for efficiently degrading aflatoxin;
(5) after the fermentation liquid cell-free centrifugal supernatant is mixed with the aflatoxin B1, the mixture is cultured for 1 day at 37 ℃, and the degradation rate of the aflatoxin B1 can reach 94.01 +/-1.45-99.89 +/-0.02%;
(6) the deep sea Cucumaria rosea M2PMix1-M2Mix5-13 strain and the fermentation liquor thereof can be used for degradation and detoxification of related aflatoxin pollution of food, drinks, feeds and the like.
Detailed Description
The present invention is further illustrated by the following examples, which are not intended to limit the scope of the invention.
Example 1: fermentation of deep sea Cuckoo roseus M2PMix1-M2Mix5-13 strain
Inoculating liquid seeds of Kocuria rosea M2PMix1-M2Mix5-13 strain into a liquid culture medium containing 0.5% of sodium acetate, 0.05% of peptone, 0.05% of yeast extract powder, 0.05% of beef extract, 0.05% of glucose, 0.05% of sucrose, 0.05% of soluble starch, 0.005% of trisodium citrate, 0.005% of malic acid, 0.005% of potassium sodium tartrate, 0.1% of ammonium nitrate, 0.02% of ammonium chloride and 100ml of water, culturing for 1-5 days at 18-28 ℃ and 250 rpm of 150-.
Example 2: activity of cell-free supernatant of deep sea Cuckoo rosea M2PMix1-M2Mix5-13 strain fermented for 1 day for degrading aflatoxin
To the cell-free fermentation supernatant obtained by 1 day fermentation in example 1, AFB1 was added to make the final concentration of AFB1 4 μ g/ml, AFB1 was added to the corresponding medium to make the final concentration of AFB1 4 μ g/ml as a control, after 1 day stationary culture at 37 degrees, chloroform was added to extract AFB1 to the control and treatment, the volume of extracted AFB1 was determined by chromatographic methanol, AFB1 was determined by HPLC, the monitoring wavelength was 203nm, the peak area of AFB1 was recorded, the degradation rate of AFB1 was calculated, the degradation rate (%) = (peak area of control AFB 1-peak area of cell-free fermentation supernatant AFB 1)/peak area x 100 of control AFB1, and the degradation rate of aflatoxin by the fermentation cell-free supernatant of custara rosmarin M2PMix1-M2Mix5-13 strain was determined and calculated to be 94.01 ± 1.45%.
Example 3: activity of deep sea Cuckoo rosea M2PMix1-M2Mix5-13 strain fermented cell-free supernatant for degrading aflatoxin for 4 days
To the cell-free fermentation supernatant obtained by 4 days of fermentation in example 1, AFB1 was added to make the final concentration of AFB1 4 μ g/ml, AFB1 was added to the corresponding medium to make the final concentration of AFB1 4 μ g/ml as a control, after 1 day of stationary culture at 37 degrees, chloroform was added to extract AFB1 to the control and treatment, the volume of extracted AFB1 was determined by chromatographic methanol, AFB1 was determined by HPLC, the monitoring wavelength was 203nm, the peak area of AFB1 was recorded, the degradation rate of AFB1 was calculated, the degradation rate (%) = (peak area of control AFB 1-peak area of cell-free fermentation supernatant AFB 1)/peak area x 100 of control AFB1, and the degradation rate of aflatoxin by the fermentation cell-free supernatant of custara rosmarin M2PMix1-M2Mix5-13 strain was determined and calculated to be 99.89 ± 0.02%.
Claims (1)
1. A culture separated from the crest 1617m of the Taiwan beast of Western Pacific ocean for in-situ directional enrichment culture for 1 yearThe deep sea strain of (A) is Kocuria rosea M2PMix1-M2Mix5-13 strain (a)Kocuria rosea M2PMix1-M2Mix5-13), which has been deposited at 26.10.2020 in Guangdong province culture Collection with the deposit number GDMCC No. 61240.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011510102.XA CN113215014A (en) | 2020-12-18 | 2020-12-18 | Deep sea Kocuria rosea for degrading aflatoxin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011510102.XA CN113215014A (en) | 2020-12-18 | 2020-12-18 | Deep sea Kocuria rosea for degrading aflatoxin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113215014A true CN113215014A (en) | 2021-08-06 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011510102.XA Pending CN113215014A (en) | 2020-12-18 | 2020-12-18 | Deep sea Kocuria rosea for degrading aflatoxin |
Country Status (1)
| Country | Link |
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| CN (1) | CN113215014A (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754561A (en) * | 2017-02-13 | 2017-05-31 | 浙江大学 | A kind of thermophilic salt rosiness Cook bacteria strain and its application |
-
2020
- 2020-12-18 CN CN202011510102.XA patent/CN113215014A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754561A (en) * | 2017-02-13 | 2017-05-31 | 浙江大学 | A kind of thermophilic salt rosiness Cook bacteria strain and its application |
Non-Patent Citations (2)
| Title |
|---|
| 侯玉煌;丁宏标;: "角蛋白酶研究进展及其在饲料工业中的应用", 中国畜牧杂志, no. 01, 10 January 2018 (2018-01-10) * |
| 杨锐;郭玉荣;魏晓红;孔玉洁;朱丽林;: "86株库克菌的鉴定与抗菌药物敏感性试验", 国际检验医学杂志, no. 09, 15 May 2016 (2016-05-15) * |
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