Purification and characterization of a novel heparinase

L. H. Bohmer*, M. J. Pitout, P. L. Steyn, L. Visser

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

Abstract

A unique heparinase was isolated from a recently discovered Gram-negative soil bacterium. The enzyme (heparinase III) was purified by hydroxylapatite chromatography, chromatofocusing, and gel permeation chromatography. The enrichment was 48x, and the specific activity of catalytically pure heparinase was 127 IU/mg of protein. Similar to the heparinase I from Flavobacterium heparinum, heparinase III also degrades heparin to mainly disaccharide fragments. It is specific for heparin and also breaks down heparan sulfate, but not hyaluronic acid and chondroitin sulfate. Heparinase III, however differs markedly from heparinase I in several other aspects: it has a higher molecular mass (94 versus 43 kDa), pI (9.2 versus 8.5), its K(m) and k(cat) are different, and it has a higher energy of activation (15.6 versus 6.3 kcal/mol). Optimal activity was also found at higher pH (7.6 versus 6.5) and temperature (45 versus 37°C). Furthermore, the amino acid composition of heparinase III is quite different from that of heparinase I.

Original languageEnglish
Pages (from-to)13609-13617
Number of pages9
JournalJournal of Biological Chemistry
Volume265
Issue number23
Publication statusPublished - 1990
Externally publishedYes

Fingerprint

Dive into the research topics of 'Purification and characterization of a novel heparinase'. Together they form a unique fingerprint.

Cite this