Tailoring FPOX enzymes for enhanced stability and expanded substrate recognition.

Autor: Estiri H; Department of Biotechnology, Latvian Institute of Organic Synthesis, Aizkraukles 21, Riga, 1006, Latvia., Bhattacharya S; Department of Biotechnology, Latvian Institute of Organic Synthesis, Aizkraukles 21, Riga, 1006, Latvia.; Faculty of Materials Science and Applied Chemistry, Riga Technical University, Paula Valdena 3, Riga, 1048, Latvia., Buitrago JAR; Department of Biotechnology, Latvian Institute of Organic Synthesis, Aizkraukles 21, Riga, 1006, Latvia., Castagna R; Department of Biotechnology, Latvian Institute of Organic Synthesis, Aizkraukles 21, Riga, 1006, Latvia.; Dipartimento di Chimica, Materiali e Ingegneria Chimica 'Giulio Natta', Politecnico di Milano, Piazza L. da Vinci 32, 20133, Milano, Italy., Legzdiņa L; Department of Biotechnology, Latvian Institute of Organic Synthesis, Aizkraukles 21, Riga, 1006, Latvia., Casucci G; Department of Biotechnology, Latvian Institute of Organic Synthesis, Aizkraukles 21, Riga, 1006, Latvia., Ricci A; Biomolecular Engineering Lab, Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133, Milano, Italy., Parisini E; Department of Biotechnology, Latvian Institute of Organic Synthesis, Aizkraukles 21, Riga, 1006, Latvia. emilio.parisini@osi.lv.; Department of Chemistry 'G. Ciamician', University of Bologna, Via Selmi 2, 40126, Bologna, Italy. emilio.parisini@osi.lv., Gautieri A; Biomolecular Engineering Lab, Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133, Milano, Italy. alfonso.gautieri@polimi.it.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2023 Oct 30; Vol. 13 (1), pp. 18610. Date of Electronic Publication: 2023 Oct 30.
DOI: 10.1038/s41598-023-45428-1
Abstrakt: Fructosyl peptide oxidases (FPOX) are deglycating enzymes that find application as key enzymatic components in diabetes monitoring devices. Indeed, their use with blood samples can provide a measurement of the concentration of glycated hemoglobin and glycated albumin, two well-known diabetes markers. However, the FPOX currently employed in enzymatic assays cannot directly detect whole glycated proteins, making it necessary to perform a preliminary proteolytic treatment of the target protein to generate small glycated peptides that can act as viable substrates for the enzyme. This is a costly and time consuming step. In this work, we used an in silico protein engineering approach to enhance the overall thermal stability of the enzyme and to improve its catalytic activity toward large substrates. The final design shows a marked improvement in thermal stability relative to the wild type enzyme, a distinct widening of its access tunnel and significant enzymatic activity towards a range of glycated substrates.
(© 2023. The Author(s).)
Databáze: MEDLINE
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