6Y4J

Engineered Fructosyl Peptide Oxidase

6Y4J の概要

• エントリーDOI: 10.2210/pdb6y4j/pdb
• 分子名称: Fructosyl Peptide Oxidase, FLAVIN-ADENINE DINUCLEOTIDE, SULFATE ION, ... (5 entities in total)
• 機能のキーワード: oxidoreductase
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 49337.40

構造登録者

Donini, S.,Rigoldi, F.,Gautieri, A.,Parisini, E. (登録日: 2020-02-21, 公開日: 2021-01-13, 最終更新日: 2024-11-13)

主引用文献

Rigoldi, F.,Donini, S.,Torretta, A.,Carbone, A.,Redaelli, A.,Bandiera, T.,Parisini, E.,Gautieri, A.
Rational backbone redesign of a fructosyl peptide oxidase to widen its active site access tunnel.
Biotechnol.Bioeng., 117:3688-3698, 2020

PubMed Abstract: Fructosyl peptide oxidases (FPOXs) are enzymes currently used in enzymatic assays to measure the concentration of glycated hemoglobin and albumin in blood samples, which serve as biomarkers of diabetes. However, since FPOX are unable to work directly on glycated proteins, current enzymatic assays are based on a preliminary proteolytic digestion of the target proteins. Herein, to improve the speed and costs of the enzymatic assays for diabetes testing, we applied a rational design approach to engineer a novel enzyme with a wider access tunnel to the catalytic site, using a combination of Rosetta design and molecular dynamics simulations. Our final design, L3_35A, shows a significantly wider and shorter access tunnel, resulting from the deletion of five-amino acids lining the gate structures and from a total of 35 point mutations relative to the wild-type (WT) enzyme. Indeed, upon experimental testing, our engineered enzyme shows good structural stability and maintains significant activity relative to the WT.

PubMed: 32797625
DOI: 10.1002/bit.27535

実験手法

X-RAY DIFFRACTION (1.38 Å)