Phenoxazinone Synthase-like Activity of Rationally Designed Heme Enzymes Based on Myoglobin

文献类型： 外文期刊

第一作者： Sun, Li-Juan

作者： Sun, Li-Juan;Lang, Jia-Jia;Wen, Ge-Bo;Yuan, Hong;Tan, Xiangshi;Yuan, Hong;Tan, Xiangshi;Xu, Jia-Kun;Lin, Ying-Wu;Lin, Ying-Wu

作者机构：

期刊名称：BIOCHEMISTRY （ 影响因子：2.9； 五年影响因子：2.9 ）

ISSN： 0006-2960

年卷期： 2023 年 62 卷 2 期

页码：

收录情况： SCI

摘要： The design of functional metalloenzymes is attractive for the biosynthesis of biologically important compounds, such as phenoxazinones and phenazines catalyzed by native phenoxazinone synthase (PHS). To design functional heme enzymes, we used myoglobin (Mb) as a model protein and introduced an artificial CXXC motif into the heme distal pocket by F46C and L49C mutations, which forms a de novo disulfide bond, as confirmed by the X-ray crystal structure. We further introduced a catalytic Tyr43 into the heme distal pocket and found that the F43Y/F46C/L49C Mb triple mutant and the previously designed F43Y/F46S Mb exhibit PHS-like activity (80-98% yields in 5-15 min), with the catalytic efficiency exceeding those of natural metalloenzymes, including o-aminophenol oxidase, laccase, and dye-decolorizing peroxidase. Moreover, we showed that the oxidative coupling product of 1,6-disulfonic-2,7-diaminophenazine is a potential pH indicator, with the orange-magenta color change at pH 4-5 (pKa = 4.40). Therefore, this study indicates that functional heme enzymes can be rationally designed by structural modifications of Mb, exhibiting the functionality of the native PHS for green biosynthesis.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序