Expression, purification and preliminary X-ray diffraction studies of RebC.
The flavin-dependent monooxygenase RebC is a key enzyme in the biosynthesis of the indolocarbazole rebeccamycin. The synthesis of rebeccamycin is of great interest as it has been shown to be a natural antitumour agent. The enzyme has been recombinantly expressed in Escherichia coli and purified to homogeneity. Hanging-drop vapour diffusion in combination with microseeding was used to obtain suitable crystals for X-ray diffraction. Data were collected to 2.4 A; the crystals belonged to space group P2(1), with unit-cell parameters a = 63.08, b = 77.85, c = 63.94 A, alpha = gamma = 90, beta = 108.11 degrees .
van Staalduinen
Laura M
LM
Department of Biochemistry, Queen's University, Kingston K7L 3N6, Canada.
Bhattacharya
Anupam
A
Groom
Katherine
K
Zechel
David L
DL
Jia
Zongchao
Z
D000192:Actinomycetales / Q000201:enzymology; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification
D006899:Mixed Function Oxygenases; C525129:RebC protein, Lechevalieria aerocolonigenes