Phenoxazinone Synthase-like Activity of Rationally Designed Heme Enzymes Based on Myoglobin
文献类型： 外文期刊
作者： Sun, Li-Juan 1 ; Yuan, Hong 2 ; Xu, Jia-Kun 4 ; Luo, Jie 1 ; Lang, Jia-Jia 1 ; Wen, Ge-Bo 1 ; Tan, Xiangshi 2 ; Lin, Ying-Wu 1 ;
作者机构： 1.Univ South China, Hengyang Med Sch, Hengyang 421001, Peoples R China
2.Fudan Univ, Dept Chem, Shanghai 200433, Peoples R China
3.Fudan Univ, Inst Biomed Sci, Shanghai 200433, Peoples R China
4.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Key Lab Sustainable Dev Polar Fisheries, Qingdao 266071, Peoples R China
5.Univ South China, Lab Prot Struct & Funct, Hengyang 421001, Peoples R China
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.
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