6K3D
Structure of multicopper oxidase mutant
Summary for 6K3D
| Entry DOI | 10.2210/pdb6k3d/pdb |
| Related | 3aw5 |
| Descriptor | Multicopper oxidase, COPPER (II) ION, CU-O-CU LINKAGE, ... (4 entities in total) |
| Functional Keywords | cupredoxin-like domains, oxidoreductase |
| Biological source | Pyrobaculum aerophilum (strain ATCC 51768 / IM2 / DSM 7523 / JCM 9630 / NBRC 100827) |
| Total number of polymer chains | 1 |
| Total formula weight | 49956.10 |
| Authors | Sakuraba, H.,Ohshida, T.,Satomura, T.,Yoneda, K.,Ohshima, T. (deposition date: 2019-05-17, release date: 2020-05-20, Last modification date: 2023-11-22) |
| Primary citation | Satomura, T.,Hirano, T.,Inagaki, K.,Horinaga, K.,Takamura, E.,Sakamoto, H.,Ohshida, T.,Ohshima, T.,Sakuraba, H.,Suye, S.I. Activity enhancement of multicopper oxidase from a hyperthermophile via directed evolution, and its application as the element of a high performance biocathode. J.Biotechnol., 325:226-232, 2021 Cited by PubMed Abstract: Although multicopper oxidase from the hyperthermophilic archaeon Pyrobaculum aerophilum (McoP) can be particularly useful in biotechnological applications, e.g., as a specific catalyst at the biocathode of biofuel cells (BFCs), owing to its high stability against extremely high temperatures and across a wide range of pH values, this application potential remains limited due to the enzyme's low catalytic activity. A directed evolution strategy was conducted to improve McoP catalytic activity, and the No. 571 mutant containing four amino acid substitutions was identified, with specific activity approximately 9-fold higher than that of the wild type enzyme. Among the substitutions, the single amino acid mutant F290I was essential in enhancing catalytic activity, with a specific activity approximately 12-fold higher than that of the wild type enzyme. F290I thermostability and pH stability were notably comparable with values obtained for the wild type. Crystal structure analysis suggested that the F290I mutant increased loop flexibility near the T1 Cu center, and affected electron transfer between the enzyme and substrate. Additionally, electric current density of the F290I mutant-immobilized electrode was 7-fold higher than that of the wild type-immobilized one. These results indicated that F290I mutant was a superior catalyst with potential in practical biotechnological applications. PubMed: 33164755DOI: 10.1016/j.jbiotec.2020.10.019 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.919 Å) |
Structure validation
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