|
KMID : 0545119910010040266
|
|
Journal of Microbiology and Biotechnology
1991 Volume.1 No. 4 p.266 ~ p.273
|
|
|
Gene Expression and Secretion of the Anticoagulant Hirudin in Saccharomyces cerevisiae
|
|
Sohn, Jung Hoon
Lee, Sang kwon/Choi, Eui Sung/Rhee, Sang Ki
|
|
|
Abstract
|
|
|
|
Hirudin, a 65-amino acid protein isolated from the salivary gland of the bloodsucking leech, Hirudo medicinalis, is a potent thrombin-specific inhibitor and blocks the thrombin-mediated conversion of fibrinogen to fibrin in clot formation. We have studied the gene expression and secretion of hirudin in yeast, Saccharomyces cerevisiae. A gene coding for hirudin was synthesized based on the amino acid sequence and cloned into a yeast expression vector YEGα-1 containing the α-mating factor pre-pro leader sequence and galactose-indudble promoter, GAL10. Recombinant S. cerevisiae was found to secrete biologically active hirudin into the extracellular medium. The secreted recombinant hirudin was recovered from the culture medium and purified with ultrafiltration and reverse phase high performance liquid chromatography. Approximately 1 mg of hirudin per liter was produced under suboptimal culture conditions and brought to about 90% purity in two steps of purification.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
  
|