Purification and characterization of a collagenolytic protease from the filefish, Novoden modestrus.
A serine collagenolytic protease was purified from the internal organs of filefish, Novoden modestrus, by ammonium sulfate, ion-exchange chromatography on a DEAE-Sephadex A-50, ion-exchange rechromatography on a DEAE-Sephadex A-50, and gel filtration on a Sephadex G- 150 column. The molecular mass of the filefish serine collagenase was estimated to be 27.0 kDa by gel filtration and SDS-PAGE. The purified collagenase was optimally active at pH 7.0-8.0 and 55 degrees C. The purified enzyme was rich in Ala, Ser, Leu, and Ile, but poor in Trp, Pro, Tyr, and Met. In addition, the purified collagenolytic enzyme was strongly inhibited by N-P-toluenesulfonyl-L-lysine chloromethyl ketone (TLCK), diisopropylfluorophosphate (DFP), and soybean trypsin inhibitor.
Kim
Se-Kwon
SK
Department of Chemistry, Pukyong National University, Pusan 608-737, Korea. sknkim@mail.pknu.ac.kr
Park
Pyo-Jam
PJ
Kim
Jong-Bae
JB
Shahidi
Fereidoon
F
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D017364:Collagenases / Q000737:chemistry* / Q000302:isolation & purification*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005399:Fishes / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D061965:Matrix Metalloproteinase Inhibitors; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification*; D013696:Temperature; D014107:Tosyllysine Chloromethyl Ketone / Q000494:pharmacology
D000596:Amino Acids; D004791:Enzyme Inhibitors; D061965:Matrix Metalloproteinase Inhibitors; D008670:Metals; D014107:Tosyllysine Chloromethyl Ketone; D012697:Serine Endopeptidases; D017364:Collagenases