无花果拟盘多毛孢pestheic acid生物合成中PtaE参与的苯氧化偶联反应

PtaE catalyzes the phenol oxidative reaction of pestheic acid biosynthesis in Pestalotiopsis fici

ptaE是pestheic acid生物合成基因簇中的漆酶编码基因,前期的体内敲除实验表明其可能参与pestheic acid生物合成中的苯氧化偶联反应。本实验通过构建带有His标签的pta E互补株,从中分离纯化了PtaE蛋白。体外酶活测定证实了PtaE参与pestheic acid生物合成中的苯氧化偶联反应,能够催化氯代二苯酮chloroisosulochrin和非氯代二苯酮isosulochrin分别生成pestheic acid及其非氯代形式RES-1214-1。对PtaE的底物专一性分析表明,PtaE具有较广泛的底物专一性,能够降解有毒物质邻苯二酚、对苯二酚、双酚A。

One laccase coding gene, pta E, is located in the pestheic acid biosynthetic gene cluster and predicted to be involved in the phenol oxidative coupling reaction of pestheic acid biosynthesis through gene disruption. To confirm its function both in vivo and in vitro, the pta E complemented strain was constructed and PtaE was isolated as the His-tagged protein from it. Enzymatic reaction analysis showed that PtaE could catalyze the conversion of chloroisosulochrin to pestheic acid and isosulochrin to RES-1214-1. Moreover, the substrate specificity of PtaE was determined. The results demonstrated that PtaE could catalyze catechol, hydroquinone and bisphenol A, indicating that it has a wide range of substrate catalytic activity.