5OC3

Crystal structure of Ser67Cys/Pro121Cys Amadoriase I mutant from Aspergillus Fumigatus

5OC3 の概要

• エントリーDOI: 10.2210/pdb5oc3/pdb
• 分子名称: Fructosyl amine:oxygen oxidoreductase, FLAVIN-ADENINE DINUCLEOTIDE, GLYCEROL, ... (4 entities in total)
• 機能のキーワード: thermoresistance, flavin dependant enzyme, glycated aminoacid, oxidoreductase
• 由来する生物種: Neosartorya fumigata
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 105393.92

構造登録者

Rigoldi, F.,Donini, S.,Gautieri, A.,Parisini, E. (登録日: 2017-06-29, 公開日: 2018-02-28, 最終更新日: 2024-11-06)

主引用文献

Rigoldi, F.,Donini, S.,Giacomina, F.,Sorana, F.,Redaelli, A.,Bandiera, T.,Parisini, E.,Gautieri, A.
Thermal stabilization of the deglycating enzyme Amadoriase I by rational design.
Sci Rep, 8:3042-3042, 2018

PubMed Abstract: Amadoriases are a class of FAD-dependent enzymes that are found in fungi, yeast and bacteria and that are able to hydrolyze glycated amino acids, cleaving the sugar moiety from the amino acidic portion. So far, engineered Amadoriases have mostly found practical application in the measurement of the concentration of glycated albumin in blood samples. However, these engineered forms of Amadoriases show relatively low absolute activity and stability levels, which affect their conditions of use. Therefore, enzyme stabilization is desirable prior to function-altering molecular engineering. In this work, we describe a rational design strategy based on a computational screening method to evaluate a library of potentially stabilizing disulfide bonds. Our approach allowed the identification of two thermostable Amadoriase I mutants (SS03 and SS17) featuring a significantly higher T (55.3 °C and 60.6 °C, respectively) compared to the wild-type enzyme (52.4 °C). Moreover, SS17 shows clear hyperstabilization, with residual activity up to 95 °C, whereas the wild-type enzyme is fully inactive at 55 °C. Our computational screening method can therefore be considered as a promising approach to expedite the design of thermostable enzymes.

PubMed: 29445091
DOI: 10.1038/s41598-018-19991-x

実験手法

X-RAY DIFFRACTION (2.153 Å)