J. Abendroth et al., Purification of a D-hydantoinase using a laboratory-scale Streamline phenyl column as the initial step, J CHROMAT B, 737(1-2), 2000, pp. 187-194
A D-hydantoinase from Thermus sp. was overexpressed in Escherichia coli and
purified to homogeneity for subsequent crystallization. The purification w
as performed with hydrophobic interaction chromatography as the capture ste
p followed by anion-exchange chromatography and gel permeation chromatograp
hy as intermediate purification and polishing steps, respectively. The hydr
ophobic interaction step was done in fluidized bed mode in a laboratory-sca
le Streamline column made from conventional laboratory equipment. The whole
purification protocol could be finished within one day. The purified enzym
e crystallizes. The crystals are suitable for X-ray protein structure analy
sis and diffract to at least 2.3 Angstrom resolution. Complete data sets ha
ve been measured up to 2.6 Angstrom resolution. The X-ray structure is curr
ently being solved. (C) 2000 Published by Elsevier Science BN. All rights r
eserved.