title: Reconstituting redox active centers of heme-containing proteins with biomineralized gold toward peroxidase mimics with strong intrinsic catalysis and electrocatalysis for H2O2detection.
abstract: A facile and efficient enzymatic reconstitution methodology has been proposed for high-catalysis peroxidase mimics by remolding the redox active centers of heme-containing proteins with the in-site biomineralized gold using hemoglobin (Hb) as a model. Catalytic hemin (Hem) was extracted from the active centers of Hb for the gold biomineralization and then reconstituted into apoHb to yield the Hem-Au@apoHb nanocomposites showing dramatically improved intrinsic catalysis and electrocatalysis over natural Hb and Hem. The biomineralized gold, on the one hand, would act as "nanowires" to promote the electron transferring of the nanocomposites. On the other hand, it would create a reactivity pathway to pre-organize and accumulate more substrates towards the active sites of the peroxidase mimics. Steady-state kinetics studies indicate that Hem-Au@apoHb could present much higher substrate affinity (lower Michaelis constants) and intrinsic catalysis even than some natural peroxidases. Moreover, the application feasibility of the prepared artificial enzymes was demonstrated by colorimetric assays and direct electrocatalysis for H2O2sensing, showing a detection limitation low as 0.45μM. Importantly, such a catalysis active-center reconstitution protocol may circumvent the substantial improvement of the intrinsic catalysis and electrocatalysis of diverse heme-containing proteins or enyzmes toward the extensive applications in the chemical, enviromental, and biomedical catalysis fields.
authors.affiliation: Shandong Province Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu City, Shandong 273165, P.R. China.
authors.forename: Liyan
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Shandong Province Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu City, Shandong 273165, P.R. China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Li
authors.affiliation: Shandong Province Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu City, Shandong 273165, P.R. China.
authors.forename: Minmin
authors.initials: M
authors.lastname: Dong
authors.affiliation: Shandong Province Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu City, Shandong 273165, P.R. China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Shandong Province Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu City, Shandong 273165, P.R. China.
authors.forename: Ru
authors.initials: R
authors.lastname: Li
authors.affiliation: Shandong Province Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu City, Shandong 273165, P.R. China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Shandong Province Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu City, Shandong 273165, P.R. China.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Lv
authors.affiliation: Hospital of University, Qufu Normal University, Qufu City, Shandong Province 273165, P.R. China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Chen
authors.affiliation: Shandong Province Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu City, Shandong 273165, P.R. China. Electronic address: huawangqfnu@126.com.
authors.forename: Hua
authors.initials: H
authors.lastname: Wang
mesh_terms: D000818:Animals; D040761:Biomimetic Materials / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods; D002384:Catalysis; D002417:Cattle; D003124:Colorimetry / Q000379:methods; D006046:Gold / Q000737:chemistry*; D006418:Heme / Q000737:chemistry*; D006454:Hemoglobins / Q000737:chemistry*; D006861:Hydrogen Peroxide / Q000032:analysis*; D053761:Nanocomposites / Q000737:chemistry* / Q000648:ultrastructure; D010084:Oxidation-Reduction; D009195:Peroxidase / Q000737:chemistry*
chemical_list: D006454:Hemoglobins; D006418:Heme; D006046:Gold; D006861:Hydrogen Peroxide; D009195:Peroxidase
keywords: Enzyme reconstitution; Gold biomineralization; Intrinsic catalysis; Peroxidase mimics; Redox active centers