Purification and characterization of a fibrinolytic enzyme and identification of fibrinogen clotting enzyme in a marine green alga, Codium divaricatum.
A fibrinolytic enzyme was isolated from a marine green alga, Codium divaricatum, and designated C. divaricatum protease (CDP). This protease effectively hydrolyzed fibrinogen A alpha chain, while it had very low hydrolyzing efficiency for B beta and gamma chains. This property was similar to that of alpha-fibrinogenase isolated from snake venom. Protease activity peaked at pH 9, and was completely inhibited by diisopropyl fluorophosphate (DFP) and phenylmethylsulfonyl fluoride (PMSF), identifying it as a serine protease. Its molecular form was single polypeptide structure and molecular weight was estimated as 31,000 by SDS-PAGE. Fibrinogen clotting enzyme was also identified in a fraction by ion-exchange chromatography. Analysis of clots formed by the enzyme and by thrombin by SDS-PAGE showed that the fibrinogen clotting enzyme would act like thrombin and have high substrate specificity.
Matsubara
K
K
Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Japan. kmatsuba@fhw.oka-pu.ac.jp
Hori
K
K
Matsuura
Y
Y
Miyazawa
K
K
D001777:Blood Coagulation; D056890:Eukaryota / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism*; D005343:Fibrinolytic Agents / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D010940:Plant Proteins / Q000302:isolation & purification / Q000378:metabolism*
D005343:Fibrinolytic Agents; D010940:Plant Proteins; D005340:Fibrinogen