Functional interfaces for biomimetic energy harvesting: CNTs-DNA matrix for enzyme assembly.

The development of 3D structures exploring the properties of nano-materials and biological molecules has been shown through the years as an effective path forward for the design of advanced bio-nano architectures for enzymatic fuel cells, photo-bio energy harvesting devices, nano-biosensors and bio-actuators and other bio-nano-interfacial architectures. In this study we demonstrate a scaffold design utilizing carbon nanotubes, deoxyribose nucleic acid (DNA) and a specific DNA binding transcription factor that allows for directed immobilization of a single enzyme. Functionalized carbon nanotubes were covalently bonded to a diazonium salt modified gold surface through carbodiimide chemistry creating a brush-type nanotube alignment. The aligned nanotubes created a highly ordered structure with high surface area that allowed for the attachment of a protein assembly through a designed DNA scaffold. The enzyme immobilization was controlled by a zinc finger (ZNF) protein domain that binds to a specific dsDNA sequence. ZNF 268 was genetically fused to the small laccase (SLAC) from Streptomyces coelicolor, an enzyme belonging to the family of multi-copper oxidases, and used to demonstrate the applicability of the developed approach. Analytical techniques such as X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and enzymatic activity analysis, allowed characterization at each stage of development of the bio-nano architecture. This article is part of a Special Issue entitled Biodesign for Bioenergetics--the design and engineering of electronic transfer cofactors, proteins and protein networks, edited by Ronald L. Koder and J.L. Ross Anderson.

[1]  Fei Wu,et al.  Fluorescence characterization of co-immobilization-induced multi-enzyme aggregation in a polymer matrix using Förster resonance energy transfer (FRET): toward the metabolon biomimic. , 2013, Biomacromolecules.

[2]  T. Tamaki,et al.  Evaluation of Immobilized Enzyme in a High-Surface-Area Biofuel Cell Electrode Made of Redox-Polymer-Grafted Carbon Black , 2010 .

[3]  Scott Banta,et al.  A chimeric fusion protein engineered with disparate functionalities-enzymatic activity and self-assembly. , 2009, Journal of molecular biology.

[4]  C. Pabo,et al.  Zif268 protein-DNA complex refined at 1.6 A: a model system for understanding zinc finger-DNA interactions. , 1996, Structure.

[5]  C. Pabo,et al.  High-resolution structures of variant Zif268-DNA complexes: implications for understanding zinc finger-DNA recognition. , 1998, Structure.

[6]  Plamen Atanassov,et al.  Glucose oxidase anode for biofuel cell based on direct electron transfer , 2006 .

[7]  Thomas P. McNamara,et al.  Bilirubin oxidase from Myrothecium verrucaria: X-ray determination of the complete crystal structure and a rational surface modification for enhanced electrocatalytic O2 reduction. , 2011, Dalton transactions.

[8]  N. Oyama,et al.  Incorporation of redox polymers to polyelectrolyte-coated electrode surfaces , 1983 .

[9]  A. Downard Potential-Dependence of Self-Limited Films Formed by Reduction of Aryldiazonium Salts at Glassy Carbon Electrodes , 2000 .

[10]  Pranab Goswami,et al.  Recent advances in material science for developing enzyme electrodes. , 2009, Biosensors & bioelectronics.

[11]  Henry Hess,et al.  Engineering enzymatic cascades on nanoscale scaffolds. , 2013, Current opinion in biotechnology.

[12]  Jennifer N Cha,et al.  Approaches for biological and biomimetic energy conversion. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Plamen Atanassov,et al.  Biological Fuel Cells: Cardinal Advances and Critical Challenges , 2014 .

[14]  Axel Kohlmeyer,et al.  Free energy landscape of a DNA-carbon nanotube hybrid using replica exchange molecular dynamics. , 2009, Nano letters.

[15]  Kateryna Artyushkova,et al.  Entrapment of enzymes and carbon nanotubes in biologically synthesized silica: glucose oxidase-catalyzed direct electron transfer. , 2008, Small.

[16]  David P. Hickey,et al.  Enzyme Cascade for Catalyzing Sucrose Oxidation in a Biofuel Cell , 2013 .

[17]  Merlin Crossley,et al.  Sticky fingers: zinc-fingers as protein-recognition motifs. , 2007, Trends in biochemical sciences.

[18]  J. Pinson,et al.  Attachment of Organic Layers to Materials Surfaces by Reduction of Diazonium Salts , 2012 .

[19]  Peijun Ji,et al.  Enzymes immobilized on carbon nanotubes. , 2011, Biotechnology advances.

[20]  Plamen Atanassov,et al.  Mechanistic study of direct electron transfer in bilirubin oxidase , 2012 .

[21]  G. Strack,et al.  Enzyme-Modified Buckypaper for Bioelectrocatalysis , 2013 .

[22]  I. Wheeldon,et al.  Bioactive proteinaceous hydrogels from designed bifunctional building blocks. , 2007, Biomacromolecules.

[23]  Engineering of a redox protein for DNA-directed assembly. , 2011, Chemical communications.

[24]  Nick L. Akers,et al.  Improving the environment for immobilized dehydrogenase enzymes by modifying Nafion with tetraalkylammonium bromides. , 2004, Biomacromolecules.

[25]  D. Bélanger,et al.  Direct Modification of a Gold Electrode with Aminophenyl Groups by Electrochemical Reduction of in Situ Generated Aminophenyl Monodiazonium Cations , 2006 .

[26]  N. Pavletich,et al.  Zinc finger-DNA recognition: crystal structure of a Zif268-DNA complex at 2.1 A , 1991, Science.

[27]  Fangfang Sun,et al.  A high-energy-density sugar biobattery based on a synthetic enzymatic pathway , 2014, Nature Communications.

[28]  S. Minteer,et al.  Metabolon Catalysts: An Efficient Model for Multi‐enzyme Cascades at Electrode Surfaces , 2011 .

[29]  D. Ivnitski,et al.  High electrocatalytic activity of tethered multicopper oxidase-carbon nanotube conjugates. , 2010, Chemical communications.

[30]  Itamar Willner,et al.  Enzyme cascades activated on topologically programmed DNA scaffolds. , 2009, Nature nanotechnology.

[31]  F. Rawson,et al.  Tailoring 3D Single-Walled Carbon Nanotubes Anchored to Indium Tin Oxide for Natural Cellular Uptake and Intracellular Sensing , 2012, Nano letters.

[32]  C. Pabo,et al.  Design and selection of novel Cys2His2 zinc finger proteins. , 2001, Annual review of biochemistry.

[33]  J. J. Monagle Carbodiimides. III. Conversion of Isocyanates to Carbodiimides. Catalyst Studies , 1962 .

[34]  Scott Calabrese Barton,et al.  Bioelectrocatalytic hydrogels from electron-conducting metallopolypeptides coassembled with bifunctional enzymatic building blocks , 2008, Proceedings of the National Academy of Sciences.

[35]  J. Yu,et al.  Complete oxidation of methanol in biobattery devices using a hydrogel created from three modified dehydrogenases. , 2013, Angewandte Chemie.

[36]  P. Wright,et al.  Zinc finger proteins: new insights into structural and functional diversity. , 2001, Current opinion in structural biology.

[37]  Nicolas Mano,et al.  Efficient direct electron transfer of PQQ-glucose dehydrogenase on carbon cryogel electrodes at neutral pH. , 2011, Analytical chemistry.