Purification and Characterization of Heparinase That Degrades Both Heparin and Heparan Sulfate from Bacillus circulans

Eiichi YOSHIDA; Kazuya SAKAI; Shinji TOKUYAMA; Hirofumi MIYAZONO; Hiroshi MARUYAMA; Kiyoshi MORIKAWA; Keiichi YOSHIDA; Yasutaka TAHARA

doi:10.1271/bbb.66.1181

Microbiology & Fermentation Technology Notes

Purification and Characterization of Heparinase That Degrades Both Heparin and Heparan Sulfate from Bacillus circulans

Eiichi YOSHIDA, Kazuya SAKAI, Shinji TOKUYAMA, Hirofumi MIYAZONO, Hiroshi MARUYAMA, Kiyoshi MORIKAWA, Keiichi YOSHIDA, Yasutaka TAHARA

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Keywords: heparinase, heparin, heparan sulfate, Bacillus circulans, glycosaminoglycan

JOURNAL FREE ACCESS

2002 Volume 66 Issue 5 Pages 1181-1184

DOI https://doi.org/10.1271/bbb.66.1181

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Abstract

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°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 haparin and HS, similar to heparinase II from Flavobacterium heparinum.

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