Crystallization and preliminary X-ray diffraction analysis of fatty-acid hydroxylase cytochrome P450BSbeta from Bacillus subtilis

Acta Crystallographica. Section D, Biological Crystallography
Dong-Sun LeeYoshitsugu Shiro

Abstract

Cytochrome P450 isolated from Bacillus subtilis (P450BSbeta; MW 48 kDa) catalyzes the hydroxylation of long-chain fatty acids at the alpha and beta positions using H(2)O(2) as an oxidant. Crystals of the substrate-free form of P450BSbeta belonging to the trigonal space group P3(2)21 or P3(1)21 were obtained by the sitting-drop vapour-diffusion method using a precipitate solution consisting of 10%(w/v) PEG 4000 and 50 mM MES pH 6.8. Another crystal form, belonging to the rhombohedral space group R3 or R32, was obtained from precipitate solution consisting of 10% PEG 4000, 0.15 mM magnesium acetate and 50 mM MES pH 6.5 in the presence of 2 mM myristic acid (substrate). Using synchrotron radiation, both P450BSbeta crystals diffracted to 2.5 A resolution. Bijvoet and dispersive anomalous difference Patterson maps show a clear peak corresponding to the haem iron.

Citations

Dec 17, 2002·Current Opinion in Biotechnology·Vlada Urlacher, Rolf D Schmid
Feb 7, 2014·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Osami Shoji, Yoshihito Watanabe
Jul 20, 2004·The Journal of Biological Chemistry·Jason K YanoEric F Johnson
Nov 21, 2018·International Journal of Molecular Sciences·Bongumusa Comfort MthethwaKhajamohiddin Syed
Apr 4, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Ntombizethu Nokuphiwa MsomiKhajamohiddin Syed

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