Secretory expression and characterization of a novel peroxiredoxin for zearalenone detoxification in Saccharomyces cerevisiae.
Secretory expression and characterization of a novel peroxiredoxin for zearalenone detoxification in Saccharomyces cerevisiae.
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DOI：
10.1016/j.micres.2012.08.002
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发表时间：
2013-01
影响因子：
6.7
通讯作者：
Yuqian Tang;J. Xiao;Yi Chen;Yigang Yu;Xinglong Xiao;Yuanshan Yu;Hui Wu
中科院分区：
文献类型：
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作者：
Yuqian Tang;J. Xiao;Yi Chen;Yigang Yu;Xinglong Xiao;Yuanshan Yu;Hui Wu
玉米赤霉烯酮（ZEN）是一种镰刀菌毒素，被认为是一种雌激素性内分泌干扰物，可引起家畜生殖器官的严重形态和功能障碍。越来越多的注意力已经支付到一个有效的战略ZEN去污的发展。ZEN被一种从不动杆菌SM 04中分离的新型过氧化物氧还蛋白（Prx）氧化成较小的雌激素代谢产物。将Prx编码基因克隆到分泌型载体pYES 2-alpha（pYα）中，并将α（α）信号肽基因插入pYES 2的多克隆位点。经2%（w/v）半乳糖诱导72 h后，酿酒酵母INVSc 1表达了重组Prx，经12% SDS-PAGE分析，其分子量约为20 kDa。重组Prx在细胞外上清中的表达量为0.24mg/mL。重组Prx在ZEN降解开始时显示出梯度增加。12 h后，每单位重组Prx对ZEN的最终降解量为0.43μg。重组Prx酶活最高的温度、H_2O_2浓度和pH分别为80 ℃、20 mM和9.0。与其他过氧化物酶相比，重组Prx酶的热稳定性和耐碱性都有很大提高。结果表明，重组Prx在S.啤酒。
Zearalenone (ZEN) is a Fusarium mycotoxin, which is considered to be an oestrogenic endocrine disruptor found to cause severe morphological and functional disorders of reproductive organs in livestock. Increasing attention has been paid to the development of an effective strategy for ZEN decontamination. ZEN is oxidized into smaller estrogenic metabolites by a novel peroxiredoxin (Prx) isolated from Acinetobacter sp. SM04. The Prx coding gene was cloned in a secretory vector pYES2-alpha (pYα) with an alpha (α) signal peptide gene inserted into the multiple cloning site of pYES2. The recombinant Prx was secreted from Saccharomyces cerevisiae INVSc1 after inducing with 2% (w/v) galactose for 72h, and was found to be nearly 20kDa through 12% SDS-PAGE. The expressed amount of recombinant Prx was 0.24mg/mL in the extracellular supernatant. Recombinant Prx showed a gradient increase at the beginning of ZEN degradation. The final ZEN degradation amount was 0.43μg by one unit recombinant Prx after 12h. Furthermore, the temperature, H2O2concentration, and pH for highest peroxidase activity of recombinant Prx were 80°C, 20mM and 9.0, respectively. When compared with other peroxidases, the thermal stability and alkali resistance of recombinant Prx were much better. The results suggest that recombinant Prx is successfully expressed in S. cerevisiae.