Purification and characterization of heparinase that degrades both heparin and heparan sulfate from Bacillus circulans

• Published in: Bioscience, biotechnology, and biochemistry Vol. 66; no. 5; pp. 1181 - 1184
• Main Authors: Yoshida, E. (Shizuoka Univ. (Japan). Faculty of Agriculture), Sakai, K, Tokuyama, S, Miyazono, H, Maruyama, H, Morikawa, K, Yoshida, K, Tahara, Y
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 01.05.2002; Japan Society for Bioscience Biotechnology and Agrochemistry
• Subjects: Bacillus - enzymology; BACILLUS CIRCULANS; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; ELECTROPHORESIS; Electrophoresis, Polyacrylamide Gel; ENZYME ACTIVITY; Fundamental and applied biological sciences. Psychology; glycosaminoglycan; GLYCOSAMINOGLYCANS; heparan sulfate; HEPARIN; Heparin - metabolism; Heparin Lyase - chemistry; Heparin Lyase - isolation & purification; Heparin Lyase - metabolism; heparinase; Heparitin Sulfate - metabolism; HYDROLASES; Hydrolysis; Kinetics; Miscellaneous; Mission oriented research; Molecular Weight; MUCOPOLYSACCHARIDES; PURIFICATION; Substrate Specificity; SULPHATES; Polysaccharide-lyases; Purification; Bacillus circulans; Enzyme; Lyases; Bacillaceae; Bacillales; Enzymatic activity; Heparin lyase; Substrate specificity; Bacteria; Heparitin sulfate; Heparin; Carbon-oxygen lyases; Physicochemical properties
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.66.1181

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 45degC, 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.