title: Biophysical characterization of a recombinant leucyl aminopeptidase from Bacillus kaustophilus.
abstract: The biophysical properties of Bacillus kaustophilus leucyl aminopeptidase (BkLAP) were examined in terms of analytical ultracentrifugation, fluorescence spectroscopy, and circular dichroism. By using the analytical ultracentrifuge, we demonstrated that tetrameric BkLAP exists as the major form in solution at protein concentration of 1.5 mg/ml at pH 8.0. The native enzyme started to unfold beyond ~1 M GdnHCl and reached an unfolded intermediate with [GdnHCl](1/2) at 1.8 M. Thermal unfolding of BkLAP was found to be highly irreversible and led to a marked formation of aggregates.
authors.affiliation: Department of Applied Chemistry, National Chiayi University, Chiayi, Taiwan.
authors.forename: Meng-Chun
authors.initials: MC
authors.lastname: Chi
authors.forename: Hui-Ping
authors.initials: HP
authors.lastname: Chang
authors.forename: Gu-Gang
authors.initials: GG
authors.lastname: Chang
authors.forename: Tzu-Fan
authors.initials: TF
authors.lastname: Wang
authors.forename: Hsien-Bin
authors.initials: HB
authors.lastname: Huang
authors.forename: Long-Liu
authors.initials: LL
authors.lastname: Lin
mesh_terms: D001407:Bacillus / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002942:Circular Dichroism; D019791:Guanidine / Q000737:chemistry; D007931:Leucyl Aminopeptidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D014461:Ultracentrifugation
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D007931:Leucyl Aminopeptidase; D019791:Guanidine