Purification and characterization of heparinase that degrades both heparin and heparan sulfate from Bacillus circulans.
A heparinase that degrades both heparin and heparan sulfate (HS) was purified to homogeneity from the cell-free extract of Bacillus circulans HpT298. The purified enzyme had a single band on SDS-polyacrylamide gel electrophoresis with an estimated molecular mass of 111,000. The enzyme showed optimal activity at pH 7.5 and 45 degrees C, and its activity was stimulated in the presence of 5 mM CaCl2, BaCl2, or MgCl2. Analysis of substrate specificity and degraded disaccharides demonstrated that the enzyme acts on both heparin and HS, similar to heparinase II from Flavobacterium heparinum.
Yoshida
Eiichi
E
Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, Japan.
Sakai
Kazuya
K
Tokuyama
Shinji
S
Miyazono
Hirofumi
H
Maruyama
Hiroshi
H
Morikawa
Kiyoshi
K
Yoshida
Keiichi
K
Tahara
Yasutaka
Y
D001407:Bacillus / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D006493:Heparin / Q000378:metabolism*; D019763:Heparin Lyase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism*; D006868:Hydrolysis; D007700:Kinetics; D008970:Molecular Weight; D013379:Substrate Specificity
D006493:Heparin; D019763:Heparin Lyase; D006497:Heparan Sulfate