Reconstitution of Heme Enzymes with Artificial Metalloporphyrinoids

• Published in: Methods in enzymology Vol. 580; p. 439
• Main Authors: Oohora, K, Hayashi, T
• Format: Journal Article
• Language: English
• Published: United States 2016
• Subjects: Corrinoids - chemical synthesis; Corrinoids - chemistry; Heme - chemistry; Hemeproteins - chemical synthesis; Hemeproteins - chemistry; Kinetics; Metalloporphyrins - chemical synthesis; Metalloporphyrins - chemistry; Porphyrins - chemical synthesis; Porphyrins - chemistry; Protein Engineering - methods; Myoglobin; Reconstitution; Porphyrinoid; Heme enzyme; Heme
• ISSN: 1557-7988
• DOI: 10.1016/bs.mie.2016.05.049

An important strategy used in engineering of hemoproteins to generate artificial enzymes involves replacement of heme with an artificial cofactor after removal of the native heme cofactor under acidic conditions. Replacement of heme in an enzyme with a nonnatural metalloporphyrinoid can significantly alter the reactivity of the enzyme. This chapter describes the design and synthesis of three types of artificial metalloporphyrinoid cofactors consisting of mono-, di-, and tri-anionic ligands (tetradehydrocorrin, porphycene, and corrole, respectively). In addition, practical procedures for the preparation of apo-hemoproteins, incorporation of artificial cofactors, and characterization techniques are presented. Furthermore, the representative catalytic activities of artificial enzymes generated by reconstitution of hemoproteins are summarized.