Degrading Ochratoxin A and Zearalenone Mycotoxins Using a Multifunctional Recombinant Enzyme
Degrading Ochratoxin A and Zearalenone Mycotoxins Using a Multifunctional Recombinant Enzyme
复制标题
使用多功能重组酶降解赭曲霉毒素 A 和玉米赤霉烯酮霉菌毒素
DOI：
10.3390/toxins11050301
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发表时间：
2019-05-01
期刊：
影响因子：
4.2
通讯作者：
Wu, Aibo
中科院分区：
文献类型：
--
作者：
Azam, Md Shofiul;Yu, Dianzhen;Wu, Aibo
玉米赤霉烯酮 (ZEA) 是一种雌激素，赭曲霉毒素 A (OTA) 是一种肝毒性镰刀菌毒素，常见于谷物和水果产品中。此前还没有对多重降解霉菌毒素的单一平台进行过研究。当前的研究旨在研究新型融合重组体的双功能活性。我们通过交叉聚合酶链式反应 (PCR) 将玉米赤霉烯酮水解酶 (ZHD) 和羧肽酶 (CP) 两个单基因组合在框缺失中，产生了重组融合酶 (ZHDCP)。我们鉴定了 ZHDCP 酶的酶学特性和细胞毒性测定。我们的研究结果表明，ZHDCP 酶在最适 pH 值 7 和温度 35 °C 下，在 2 小时内将 ZEA 完全降解为无毒产物。首次发现CP在48小时内降解了ZEA 60%。融合 ZHDCP 和 CP 酶在 pH 7 和温度 30 °C 下能够在 30 分钟内降解 100% OTA。 ZEA 和 OTA 诱导细胞死亡，增加细胞凋亡率，并调节 Sirt1、Bax、Bcl2、Caspase3、TNFα 和 IL6 基因的 mRNA 表达。我们的新发现表明融合酶 ZHDCP 具有双功能活性（降解 OTA 和 ZEA），可用于降解更多的霉菌毒素。
Zearalenone (ZEA) is an estrogenic and ochratoxin A (OTA) is a hepatotoxic Fusarium 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.