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ONLINEISSN:1347-6947
PRINTISSN:0916-8451
Bioscience, Biotechnology, and Biochemistry
Vol. 66 (2002) , No. 9 pp.1873-1879
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Cloning, Sequencing, and Expression of the Gene from Bacillus circulans That Codes for a Heparinase That Degrades Both Heparin and Heparan Sulfate
Eiichi YOSHIDA1), Shinji ARAKAWA1), Taizo MATSUNAGA1), Shigeki TORIUMI1), Shinji TOKUYAMA1), Kiyoshi MORIKAWA2) and Yasutaka TAHARA1)
1) Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University
2) Central Research Laboratories, Seikagaku Corp.
(Received February 28, 2002)
(Accepted May 8, 2002)
  The gene, designated hep, coding for a heparinase that degrades both heparin and heparan sulfate, was cloned from Bacillus circulans HpT298. Nucleotide sequence analysis showed that the open reading frame of the hep gene consists of 3,150 bp, encoding a precursor protein of 1,050 amino acids with a molecular mass of 116.5 kDa. A homology search found that the deduced amino acid sequence has partial similarity with enzymes belonging to the family of acidic polysaccharide lyases that degrade chondroitin sulfate and hyaluronic acid. Recombinant mature heparinase (111.2 kDa) was produced by the addition of IPTG from Escherichia coli harboring pETHEP with an open reading frame of the mature hep gene and was purified to homogeneity by SDS-polyacrylamide gel electrophoresis. Analyses of substrate specificity and degraded disaccharides indicated that the recombinant enzyme acts on both heparin and HS, as does heparinase purified from the wild-type strain.
Key words:heparin; heparan sulfate; heparinase gene; Bacillus circulans

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To cite this article:
Eiichi YOSHIDA, Shinji ARAKAWA, Taizo MATSUNAGA, Shigeki TORIUMI, Shinji TOKUYAMA, Kiyoshi MORIKAWA and Yasutaka TAHARA, “Cloning, Sequencing, and Expression of the Gene from Bacillus circulans That Codes for a Heparinase That Degrades Both Heparin and Heparan Sulfate”, Biosci. Biotechnol. Biochem., Vol. 66, 1873-1879 (2002) .
doi:10.1271/bbb.66.1873
JOI  JST.JSTAGE/bbb/66.1873
Copyright (c) 2003 by Japan Society for Bioscience, Biotechnology, and Agrochemistry

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