title: Purification and biochemical characterization of a 17 kDa fibrinolytic enzyme from Schizophyllum commune.
abstract: A fibrinolytic enzyme of the mushroom, Schizophyllum commune was purified with chromatographic methods, including a DEAE-Sephadex A-50 ion-exchange column and gel filtrations with Sephadex G-75 and Sephadex G-50 columns. The analysis of fibrin-zymography and SDS-PAGE showed that the enzyme was a monomeric subunit that was estimated to be approximately 17 kDa in size. The fibrinolytic activity of the enzyme in plasminogen-rich and plasminogen-free fibrin plates was 1.25 and 0.44 U/ml, respectively. The N-terminal amino acid sequence of the purified enzyme was identified as HYNIXNSWSSFID, which was highly distinguished from known fibrinolytic enzymes. The relative activity of the purified enzyme with an addition of 5 mM EDTA, Phosphoramidon, and Bestatin was about 76, 64, and 52%, respectively, indicating that it is a metalloprotease. The optimum temperature for the purified enzyme was approximately 45°C, and over 87% of the enzymatic activity was maintained as a stable state in a pH range from 4.0 to 6.0. Therefore, our results suggest that the potential thrombolytic agent from S. commune is a unique type of fibrinolytic enzyme.
authors.affiliation: Department of Biotechnology, Dong-A University, Busan, 604-714, Republic of Korea.
authors.forename: In Suk
authors.initials: IS
authors.lastname: Park
authors.forename: Jeong Uck
authors.initials: JU
authors.lastname: Park
authors.forename: Min Jeong
authors.initials: MJ
authors.lastname: Seo
authors.forename: Min Jeong
authors.initials: MJ
authors.lastname: Kim
authors.forename: Hye Hyeon
authors.initials: HH
authors.lastname: Lee
authors.forename: Sung Ryeal
authors.initials: SR
authors.lastname: Kim
authors.forename: Byoung Won
authors.initials: BW
authors.lastname: Kang
authors.forename: Yung Hyun
authors.initials: YH
authors.lastname: Choi
authors.forename: Woo Hong
authors.initials: WH
authors.lastname: Joo
authors.forename: Yong Kee
authors.initials: YK
authors.lastname: Jeong
mesh_terms: D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000378:metabolism; D004795:Enzyme Stability; D005341:Fibrinolysin / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D012567:Schizophyllum / Q000201:enzymology*; D020539:Sequence Analysis, Protein; D013696:Temperature
chemical_list: D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D005341:Fibrinolysin