Degrading Ochratoxin A and Zearalenone Mycotoxins Using a Multifunctional Recombinant Enzyme
Zearalenone (ZEA) is an estrogenic and ochratoxin A (OTA) is a hepatotoxic <i>Fusarium</i> mycotoxin commonly seen in cereals and fruits products. No previous investigation has studied on a single platform for the multi degradation mycotoxin. The current study aimed to investigate the bi...
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MDPI AG
2019-05-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/11/5/301 |
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| author | Md Shofiul Azam Dianzhen Yu Na Liu Aibo Wu |
| author_facet | Md Shofiul Azam Dianzhen Yu Na Liu Aibo Wu |
| author_sort | Md Shofiul Azam |
| collection | DOAJ |
| description | Zearalenone (ZEA) is an estrogenic and ochratoxin A (OTA) is a hepatotoxic <i>Fusarium</i> mycotoxin commonly seen in cereals and fruits products. No previous investigation has studied on a single platform for the multi degradation mycotoxin. The current study aimed to investigate the bifunctional activity of a novel fusion recombinant. We have generated a recombinant fusion enzyme (ZHDCP) by combining two single genes named zearalenone hydrolase (ZHD) and carboxypeptidase (CP) in frame deletion by crossover polymerase chain reaction (PCR). We identified enzymatic properties and cell cytotoxicity assay of ZHDCP enzyme. Our findings have demonstrated that ZEA was completely degraded to the non-toxic product in 2 h by ZHDCP enzyme at an optimum pH of 7 and a temperature of 35 °C. For the first time, it was found out that ZEA 60% was degraded by CP degrades in 48 h. Fusion ZHDCP and CP enzyme were able to degrade 100% OTA in 30 min at pH 7 and temperature 30 °C. ZEA- and OTA-induced cell death and increased cell apoptosis rate and regulated mRNA expression of Sirt1, Bax, Bcl2, Caspase3, TNFα, and IL6 genes. Our novel findings demonstrated that the fusion enzyme ZHDCP possess bifunctional activity (degrade OTA and ZEA), and it could be used to degrade more mycotoxins. |
| first_indexed | 2024-12-10T07:41:24Z |
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| id | doaj.art-7e78c2db1ed042b6bb497748f6edf58f |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-12-10T07:41:24Z |
| publishDate | 2019-05-01 |
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| spelling | doaj.art-7e78c2db1ed042b6bb497748f6edf58f2022-12-22T01:57:18ZengMDPI AGToxins2072-66512019-05-0111530110.3390/toxins11050301toxins11050301Degrading Ochratoxin A and Zearalenone Mycotoxins Using a Multifunctional Recombinant EnzymeMd Shofiul Azam0Dianzhen Yu1Na Liu2Aibo Wu3SIBS-UGENT-SJTU Joint Laboratory of Mycotoxin Research, CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, ChinaSIBS-UGENT-SJTU Joint Laboratory of Mycotoxin Research, CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, ChinaSIBS-UGENT-SJTU Joint Laboratory of Mycotoxin Research, CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, ChinaSIBS-UGENT-SJTU Joint Laboratory of Mycotoxin Research, CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, ChinaZearalenone (ZEA) is an estrogenic and ochratoxin A (OTA) is a hepatotoxic <i>Fusarium</i> mycotoxin commonly seen in cereals and fruits products. No previous investigation has studied on a single platform for the multi degradation mycotoxin. The current study aimed to investigate the bifunctional activity of a novel fusion recombinant. We have generated a recombinant fusion enzyme (ZHDCP) by combining two single genes named zearalenone hydrolase (ZHD) and carboxypeptidase (CP) in frame deletion by crossover polymerase chain reaction (PCR). We identified enzymatic properties and cell cytotoxicity assay of ZHDCP enzyme. Our findings have demonstrated that ZEA was completely degraded to the non-toxic product in 2 h by ZHDCP enzyme at an optimum pH of 7 and a temperature of 35 °C. For the first time, it was found out that ZEA 60% was degraded by CP degrades in 48 h. Fusion ZHDCP and CP enzyme were able to degrade 100% OTA in 30 min at pH 7 and temperature 30 °C. ZEA- and OTA-induced cell death and increased cell apoptosis rate and regulated mRNA expression of Sirt1, Bax, Bcl2, Caspase3, TNFα, and IL6 genes. Our novel findings demonstrated that the fusion enzyme ZHDCP possess bifunctional activity (degrade OTA and ZEA), and it could be used to degrade more mycotoxins.https://www.mdpi.com/2072-6651/11/5/301fusion enzymeochratoxin AcarboxypeptidasezearalenoneZHD101HK293 cellLX2 cell |
| spellingShingle | Md Shofiul Azam Dianzhen Yu Na Liu Aibo Wu Degrading Ochratoxin A and Zearalenone Mycotoxins Using a Multifunctional Recombinant Enzyme Toxins fusion enzyme ochratoxin A carboxypeptidase zearalenone ZHD101 HK293 cell LX2 cell |
| title | Degrading Ochratoxin A and Zearalenone Mycotoxins Using a Multifunctional Recombinant Enzyme |
| title_full | Degrading Ochratoxin A and Zearalenone Mycotoxins Using a Multifunctional Recombinant Enzyme |
| title_fullStr | Degrading Ochratoxin A and Zearalenone Mycotoxins Using a Multifunctional Recombinant Enzyme |
| title_full_unstemmed | Degrading Ochratoxin A and Zearalenone Mycotoxins Using a Multifunctional Recombinant Enzyme |
| title_short | Degrading Ochratoxin A and Zearalenone Mycotoxins Using a Multifunctional Recombinant Enzyme |
| title_sort | degrading ochratoxin a and zearalenone mycotoxins using a multifunctional recombinant enzyme |
| topic | fusion enzyme ochratoxin A carboxypeptidase zearalenone ZHD101 HK293 cell LX2 cell |
| url | https://www.mdpi.com/2072-6651/11/5/301 |
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