Overexpression, crystallization and preliminary X-ray crystallographic analysis of the ectoine hydroxylase from Sphingopyxis alaskensis.
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 O2-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.
Crystal Farm and X-ray Facility, Heinrich-Heine-Universität, Universitätsstrasse 1, 40225 Düsseldorf, Germany.
Astrid
A
Hoeppner
Department of Biology, Laboratory for Microbiology, Philipps-University Marburg, Marburg, Germany.
Nils
N
Widderich
Department of Biology, Laboratory for Microbiology, Philipps-University Marburg, Marburg, Germany.
Erhard
E
Bremer
Institut für Biochemie, Heinrich-Heine-Universität, Universitätsstrasse 1, 40225 Düsseldorf, Germany.
Sander H J
SH
Smits
D000599:Amino Acids, Diamino / Q000378:metabolism*; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray / Q000379:methods*; D007501:Iron / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D042301:Sphingomonadaceae / Q000145:classification / Q000201:enzymology*
D000599:Amino Acids, Diamino; C045628:ectoine; D007501:Iron; D006899:Mixed Function Oxygenases
Sphingopyxis alaskensis; ectoine hydroxylase