title: Addressable self-immobilization of lactate dehydrogenase across multiple length scales.
abstract: Successful nanobiotechnology implementation largely depends on control over the interfaces between inorganic materials and biological molecules. Controlling the orientations of biomolecules and their spatial arrangements on the surface may transform many technologies including sensors, to energy. Here, we demonstrate the self-organization of L-lactate dehydrogenase (LDH), which exhibits enhanced enzymatic activity and stability on a variety of gold surfaces ranging from nanoparticles to electrodes, by incorporating a gold-binding peptide tag (AuBP2) as the fusion partner for Bacillus stearothermophilus LDH (bsLDH). Binding kinetics and enzymatic assays verified orientation control of the enzyme on the gold surface through the genetically incorporated peptide tag. Finally, redox catalysis efficiency of the immobilized enzyme was detected using cyclic voltammetry analysis in enzyme-based biosensors for lactate detection as well as in biofuel cell energy systems as the anodic counterpart. Our results demonstrate that the LDH enzyme can be self-immobilized onto different gold substrates using the short peptide tag under a biologically friendly environment. Depending on the desired inorganic surface, the proposed peptide-mediated path could be extended to any surface to achieve single-step oriented enzyme immobilization for a wide range of applications.
authors: Materials Science and Engineering, University of Washington, Seattle, WA 98195, USA.
authors: Sibel
authors: S
authors: Cetinel
authors: H Burak
authors: HB
authors: Caliskan
authors: Deniz T
authors: DT
authors: Yucesoy
authors: A Senem
authors: AS
authors: Donatan
authors: Esra
authors: E
authors: Yuca
authors: Mustafa
authors: M
authors: Urgen
authors: Nevin G
authors: NG
authors: Karaguler
authors: Candan
authors: C
authors: Tamerler
mesh_terms: D001674:Bioelectric Energy Sources; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods; D002384:Catalysis; D003001:Cloning, Molecular; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D001411:Geobacillus stearothermophilus / Q000201:enzymology; D006046:Gold / Q000737:chemistry; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification; D053768:Metal Nanoparticles / Q000737:chemistry; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation
chemical_list: D004800:Enzymes, Immobilized; D010455:Peptides; D006046:Gold; D007770:L-Lactate Dehydrogenase