Phenoxazinone Synthase-like Activity of Rationally Designed Heme Enzymes Based on Myoglobin
| العنوان: | Phenoxazinone Synthase-like Activity of Rationally Designed Heme Enzymes Based on Myoglobin |
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| المؤلفون: | Li-Juan Sun, Ge-Bo Wen, Ying-Wu Lin, Xiangshi Tan, Hong Yuan, Jia-Kun Xu, Jia-Jia Lang, Jie Luo |
| المصدر: | Biochemistry. 62:369-377 |
| بيانات النشر: | American Chemical Society (ACS), 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | chemistry.chemical_classification, Oxidase test, ATP synthase, biology, Stereochemistry, Biochemistry, chemistry.chemical_compound, Enzyme, Biosynthesis, chemistry, Myoglobin, pH indicator, biology.protein, Heme, Peroxidase |
| الوصف: | 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. |
| تدمد: | 1520-4995 0006-2960 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::1a3010954d0d29c2ea308eefa92b1a46 https://doi.org/10.1021/acs.biochem.1c00554 |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi.dedup.....1a3010954d0d29c2ea308eefa92b1a46 |
| قاعدة البيانات: | OpenAIRE |
Find this issue from the American Chemical Society | تدمد: | 15204995 00062960 |
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