Direct bioelectrocatalysis at the interfaces by genetically engineered dehydrogenase

Abstract

There is an emerging interest in developing bio-functionalisation routes serving as platforms for assembling diverse enzymes onto material surfaces. Specifically, the fabrication of next-generation, laboratory-on-a-chip-based sensing and energy-harvesting systems requires controlled orientation and organisation of the proteins at the inorganic interfaces. Herein, the authors take the initial steps towards designing multifunctional, enzyme-based platforms by genetically integrating the engineered materialselective peptide tags for tethering redox enzymes onto electrode surfaces. The authors engineered a fusion protein that genetically conjugates gold-binding peptide to formate dehydrogenase derived from Candida methylica. The expressed proteins were tested for both enzyme activity and self-directed gold-surface functionalisation ability. Their findings demonstrate the successful self-immobilisation of the engineered enzyme onto different gold electrodes while retaining the catalytic activity.Building on the functionalisation by the peptides, a fusion enzyme-integrated circuit-based biosensor system was designed. The catalytic conversion of the formate by the engineered dehydrogenase was successfully monitored on the electrode surface at subsequent intervals. The engineered peptide-mediated self-integrated electrode systems can be extended to develop a wide range of biosensing and energy-harvesting platforms using different combinations of materials and biomolecules. This paper contains supporting information that will be made available online once the issue is published. In the meantime, if you wish to get a copy of the supplementary file, please contact the Journals Editor, Sarah Brown, at sarah.brown@icepublishing.com.

Editorial

“…The paper by Yucesoy et al 10 relates to the emerging interest in developing bio-functionalisation routes. The work introduces a new platform that could serve as a bioengineering platform for assembling diverse enzymes onto material surfaces for laboratoryon-a-chip-based sensing and energy-harvesting systems.…”

Probing Selective Self-Assembly of Putrescine Oxidase with Controlled Orientation Using a Genetically Engineered Peptide Tag

“…Another approach that has been developed recently by our groups and others is the use of genetically engineered affinity peptides for the selective, specific immobilization of biomolecules at particular material interfaces. − These peptides have been developed using cell surface display and phage display to have a high affinity for a specific material (e.g., Au, Ag, silica, graphite). − While a number of peptides with high affinity for different materials have been developed and demonstrated, detailed investigation of the immobilization of active enzymes using these peptides is relatively unexplored. ,,,− We and others studied solid binding peptides to control orientation of the proteins and demonstrated self-immobilization of several enzymes, including alkaline phosphatase, glutathione S-transferase, and lactate dehydrogenase on specific materials, − but details including information regarding the correlation of orientation and coverage with activity are still lacking …”

Deep Directed Evolution of Solid Binding Peptides for Quantitative Big-data Generation

“…Despite well-established phage display selection procedures and subsequent improvements towards increasing the winner peptides' potential, the methods are still far from leveraging the huge combinatorial potential since the diversity of the libraries is in the order of 10 9 variants. 2,7,15,18,19 The drawback is primarily due to the limited scalability of clone selection techniques and characterization. Moreover, in low-throughput Sanger sequencing workflows traditionally used, the number of peptides isolated in a reasonable time frame after several rounds of the biopanning process is limited to 10-100 clones.…”