Purification, crystallization and preliminary X-ray diffraction experiment of nattokinase from Bacillus subtilis natto.
Nattokinase is a single polypeptide chain composed of 275 amino acids (molecular weight 27,724) which displays strong fibrinolytic activity. Moreover, it can activate other fibrinolytic enzymes such as pro-urokinase and tissue plasminogen activator. In the present study, native nattokinase from Bacillus subtilis natto was purified using gel-filtration chromatography and crystallized to give needle-like crystals which could be used for X-ray diffraction experiments. The crystals belonged to space group C2, with unit-cell parameters a=74.3, b=49.9, c=56.3 Å, β=95.2°. Diffraction images were processed to a resolution of 1.74 Å with an Rmerge of 5.2% (15.3% in the highest resolution shell) and a completeness of 69.8% (30.0% in the highest resolution shell). This study reports the first X-ray diffraction analysis of nattokinase.
Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba 288-0025, Japan.
Yasuhide
Y
Yanagisawa
Toshiyuki
T
Chatake
Kaori
K
Chiba-Kamoshida
Sawa
S
Naito
Tadanori
T
Ohsugi
Hiroyuki
H
Sumi
Ichiro
I
Yasuda
Yukio
Y
Morimoto
D001412:Bacillus subtilis / Q000201:enzymology*; D002850:Chromatography, Gel; D003460:Crystallization; D018360:Crystallography, X-Ray; D055672:Static Electricity; D013381:Subtilisins / Q000737:chemistry* / Q000302:isolation & purification*; D014961:X-Ray Diffraction
D013381:Subtilisins; C053771:nattokinase