Premium
Overexpression, crystallization and preliminary X‐ray crystallographic analysis of the ectoine hydroxylase from Sphingopyxis alaskensis
Author(s) -
Hoeppner Astrid,
Widderich Nils,
Bremer Erhard,
Smits Sander H. J.
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14004798
Subject(s) - ectoine , dioxygenase , enzyme , chemistry , substrate (aquarium) , crystallization , yield (engineering) , biochemistry , decarboxylation , stereochemistry , crystallography , biology , organic chemistry , amino acid , proline , catalysis , materials science , osmoprotectant , ecology , metallurgy
The ectoine hydroxylase (EctD) is a member of the non‐haem‐containing iron(II)‐ and 2‐oxoglutarate‐dependent dioxygenase superfamily. Its mononuclear iron centre is a prerequisite for the activity of this enzyme and promotes the O 2 ‐dependent oxidative decarboxylation of 2‐oxoglutarate, which is coupled to a two‐electron oxidation of the substrate ectoine to yield 5‐hydroxyectoine. An expression and purification protocol for the EctD enzyme from Sphingopyxis alaskensis was developed and the protein was crystallized using the sitting‐drop vapour‐diffusion method. This resulted in two different crystal forms, representing the apo and iron‐bound forms of the enzyme.