Isolation, Purification, and Characterization of a Laccase-Degrading Aflatoxin B1 from <i>Bacillus amyloliquefaciens</i> B10
Aflatoxins, widely found in feed and foodstuffs, are potentially harmful to human and animal health because of their high toxicity. In this study, a strain of <i>Bacillus amyloliquefaciens</i> B10 with a strong ability to degrade aflatoxin B1 (AFB1) was screened; it could degrade 2.5 μg/...
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MDPI AG
2022-03-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/14/4/250 |
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| author | Dongwei Xiong Jun Wen Gen Lu Tianxi Li Miao Long |
| author_facet | Dongwei Xiong Jun Wen Gen Lu Tianxi Li Miao Long |
| author_sort | Dongwei Xiong |
| collection | DOAJ |
| description | Aflatoxins, widely found in feed and foodstuffs, are potentially harmful to human and animal health because of their high toxicity. In this study, a strain of <i>Bacillus amyloliquefaciens</i> B10 with a strong ability to degrade aflatoxin B1 (AFB1) was screened; it could degrade 2.5 μg/mL of AFB1 within 96 h. The active substances of <i>Bacillus amyloliquefaciens</i> B10 for the degradation of AFB1 mainly existed in the culture supernatant. A new laccase with AFB1-degrading activity was separated by ammonium sulfate precipitation, diethylaminoethyl (DEAE) and gel filtration chromatography. The results of molecular docking showed that B10 laccase and aflatoxin had a high docking score. The coding sequence of the laccase was successfully amplified from cDNA by PCR and cloned into <i>E. coli</i>. The purified laccase could degrade 79.3% of AFB1 within 36 h. The optimum temperature for AFB1 degradation was 40 °C, and the optimum pH was 6.0–8.0. Notably, Mg<sup>2+</sup> and dimethyl sulfoxide (DMSO) could enhance the AFB1-degrading activity of B10 laccase. Mutation of the three key metal combined sites of B10 laccase resulted in the loss of AFB1-degrading activity, indicating that these three metal combined sites of B10 laccase play an essential role in the catalytic degradation of AFB1. |
| first_indexed | 2024-03-09T10:28:01Z |
| format | Article |
| id | doaj.art-459cb6e4f1984fb09b79afc49d667248 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T10:28:01Z |
| publishDate | 2022-03-01 |
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| record_format | Article |
| series | Toxins |
| spelling | doaj.art-459cb6e4f1984fb09b79afc49d6672482023-12-01T21:29:41ZengMDPI AGToxins2072-66512022-03-0114425010.3390/toxins14040250Isolation, Purification, and Characterization of a Laccase-Degrading Aflatoxin B1 from <i>Bacillus amyloliquefaciens</i> B10Dongwei Xiong0Jun Wen1Gen Lu2Tianxi Li3Miao Long4Key Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science & Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, ChinaKey Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science & Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, ChinaKey Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science & Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, ChinaKey Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science & Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, ChinaKey Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science & Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, ChinaAflatoxins, widely found in feed and foodstuffs, are potentially harmful to human and animal health because of their high toxicity. In this study, a strain of <i>Bacillus amyloliquefaciens</i> B10 with a strong ability to degrade aflatoxin B1 (AFB1) was screened; it could degrade 2.5 μg/mL of AFB1 within 96 h. The active substances of <i>Bacillus amyloliquefaciens</i> B10 for the degradation of AFB1 mainly existed in the culture supernatant. A new laccase with AFB1-degrading activity was separated by ammonium sulfate precipitation, diethylaminoethyl (DEAE) and gel filtration chromatography. The results of molecular docking showed that B10 laccase and aflatoxin had a high docking score. The coding sequence of the laccase was successfully amplified from cDNA by PCR and cloned into <i>E. coli</i>. The purified laccase could degrade 79.3% of AFB1 within 36 h. The optimum temperature for AFB1 degradation was 40 °C, and the optimum pH was 6.0–8.0. Notably, Mg<sup>2+</sup> and dimethyl sulfoxide (DMSO) could enhance the AFB1-degrading activity of B10 laccase. Mutation of the three key metal combined sites of B10 laccase resulted in the loss of AFB1-degrading activity, indicating that these three metal combined sites of B10 laccase play an essential role in the catalytic degradation of AFB1.https://www.mdpi.com/2072-6651/14/4/250aflatoxin<i>Bacillus amyloliquefaciens</i>laccasedegradationmolecular dockingmutagenesis |
| spellingShingle | Dongwei Xiong Jun Wen Gen Lu Tianxi Li Miao Long Isolation, Purification, and Characterization of a Laccase-Degrading Aflatoxin B1 from <i>Bacillus amyloliquefaciens</i> B10 Toxins aflatoxin <i>Bacillus amyloliquefaciens</i> laccase degradation molecular docking mutagenesis |
| title | Isolation, Purification, and Characterization of a Laccase-Degrading Aflatoxin B1 from <i>Bacillus amyloliquefaciens</i> B10 |
| title_full | Isolation, Purification, and Characterization of a Laccase-Degrading Aflatoxin B1 from <i>Bacillus amyloliquefaciens</i> B10 |
| title_fullStr | Isolation, Purification, and Characterization of a Laccase-Degrading Aflatoxin B1 from <i>Bacillus amyloliquefaciens</i> B10 |
| title_full_unstemmed | Isolation, Purification, and Characterization of a Laccase-Degrading Aflatoxin B1 from <i>Bacillus amyloliquefaciens</i> B10 |
| title_short | Isolation, Purification, and Characterization of a Laccase-Degrading Aflatoxin B1 from <i>Bacillus amyloliquefaciens</i> B10 |
| title_sort | isolation purification and characterization of a laccase degrading aflatoxin b1 from i bacillus amyloliquefaciens i b10 |
| topic | aflatoxin <i>Bacillus amyloliquefaciens</i> laccase degradation molecular docking mutagenesis |
| url | https://www.mdpi.com/2072-6651/14/4/250 |
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