Biophysical characterization of a recombinant leucyl aminopeptidase from Bacillus kaustophilus.
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.
Chi
Meng-Chun
MC
Department of Applied Chemistry, National Chiayi University, Chiayi, Taiwan.
Chang
Hui-Ping
HP
Chang
Gu-Gang
GG
Wang
Tzu-Fan
TF
Huang
Hsien-Bin
HB
Lin
Long-Liu
LL
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
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D007931:Leucyl Aminopeptidase; D019791:Guanidine