5OC3
Crystal structure of Ser67Cys/Pro121Cys Amadoriase I mutant from Aspergillus Fumigatus
Summary for 5OC3
| Entry DOI | 10.2210/pdb5oc3/pdb |
| Descriptor | Fructosyl amine:oxygen oxidoreductase, FLAVIN-ADENINE DINUCLEOTIDE, GLYCEROL, ... (4 entities in total) |
| Functional Keywords | thermoresistance, flavin dependant enzyme, glycated aminoacid, oxidoreductase |
| Biological source | Neosartorya fumigata |
| Total number of polymer chains | 2 |
| Total formula weight | 105393.92 |
| Authors | Rigoldi, F.,Donini, S.,Gautieri, A.,Parisini, E. (deposition date: 2017-06-29, release date: 2018-02-28, Last modification date: 2024-11-06) |
| Primary citation | 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 Cited by 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: 29445091DOI: 10.1038/s41598-018-19991-x PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.153 Å) |
Structure validation
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