Isolation and Characterization of Mucinase Complex Secreted from Vibrio parahaemolyticus

  • Jun, In-Joon (Department of Microbiology, College of Natural Sciences, Pusan National University) ;
  • Kim, Yoon-Hee (Department of Microbiology, College of Natural Sciences, Pusan National University) ;
  • Kim, Min-Jeong (Department of Microbiology, College of Natural Sciences, Pusan National University) ;
  • Hwang, Ho-Soon (Department of Microbiology, College of Natural Sciences, Pusan National University) ;
  • Lee, Tae-Ho (Department of Microbiology, College of Natural Sciences, Pusan National University) ;
  • Cha, Jae-Ho (Department of Microbiology, College of Natural Sciences, Pusan National University)
  • Published : 2003.10.01

Abstract

Mucinase complex from V. parahaemolyticus ATCC 17802 was purified 6-fold with 0.4% yield by two sequential steps of Q-Sepharose and Superdex 200HR column chromatographies. Partially purified mucinase complex showed at least 8 times higher mucin-degrading activity than the culture filtrates. The mucinase complex also showed gelatin-and-casein-hydrolyzing activities, which demonstrates that the protein is a complex compound containing several proteases. The optimum pH and temperature of partially purified mucinase complex for mucin degradation was 8.0 and $35^{\circ}C$, respectively. The partially purified mucinase complex showed high cytotoxic activity on vero cells when examined by MTT assay and microscopic observations. Cytotoxicity was significantly increased in proportion to the concentration of the mucinase complex. Mouse experiments revealed that the jejunum, ileum. and large intestinal tissues were damaged by the injection of the mucinase complex. In particular, the reduction of the goblet cells in the large intestine was remarkable. Collectively, these data suggest that the mucinase complex partially purified from V. parahaemolyticus ATCC 17802 contributes to the adhesion and invasion of V. parahaemolyticus into the host intestinal tract.

Keywords

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