How to engineer glucose oxidase for mediated electron transfer

07/08/2018
 

Congratulations to Anne-Maria Wallraf on her recent publication!

Schematic view of the mediator QDM‐2 bound within the active site of GOx (left) with an enlarged view of the key amino acid residue Y414 (right). Copyright: Bio VI Figure. Schematic view of the mediator QDM‐2 bound within the active site of GOx (left) with an enlarged view of the key amino acid residue Y414 (right).

A joint protein engineering and computational study of glucose oxidase (GOx) for the development of improved glucose biosensors for diabetes care was performed. Position 414 in the active site was identified to modulate the electron shuttling from the FAD to two quinone-diimine mediators (QDM-1 and QDM-2) dependent on the polarity and size of residue 414 and the mediator. Variant GOx V7-I414Y showed increased mediator activity for the more polar QDM-2 with a simultaneously decreased activity for the less polar and smaller QDM-1 compared with parent V7 (QDM-1: 55.9 U/mg V7, 33.2 U/mg V7‐I414Y; QDM-2: 2.7 U/mg V7, 12.9 U/mg V7‐I414Y). The mediator binding model offers a promising possibility for the design of optimized enzyme-mediator couples.

This work was realized in the division Hybrid Catalysts and High Throughput Screening and it was possible through funding from Roche Diagnostics GmbH.

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E. Arango Gutierrez§, A. M. Wallraf§, A. Balaceanu, M. Bocola, M. D. Davari, T. Meier, H. Duefel, U. Schwaneberg, Biotechnol. Bioeng, 2018, DOI: 10.1002/bit.26785

§ These authors contributed equally