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Biotechnology and bioprocess engineering v.15 no.3, 2010년, pp.467 - 475   SCIE
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Overexpression of Acid Protease of Saccharomycopsis fibuligera in Yarrowia lipolytica and Characterization of the Recombinant Acid Protease for Skimmed Milk Clotting

Yu, Xin-Jun    (Unesco Chinese Center of Marine Biotechnology, Ocean University of China   ); Li, Hui-Juan    (College of Chemical and Environmental Engineering, Shandong University of Science & Technology   ); Li, Jing    (Unesco Chinese Center of Marine Biotechnology, Ocean University of China   ); Chi, Zhen-Ming    (Unesco Chinese Center of Marine Biotechnology, Ocean University of China  );
  • 초록

    The gene encoding an acid protease natively produced by Saccharomycopsis fibuligera was cloned and overexpressed in Yarrowia lipolytica and the resultant recombinant acid protease was purified and characterized. The molecular mass of the purified enzyme was estimated as 94.8 kDa by gel filtration chromatography. The optimal pH and temperature of the purified acid protease were 3.5 and $33^{\circ}C$ , respectively, and the enzyme was very stable over a pH range of 1.0 ~ 3.0. The recombinant acid protease was activated by $Zn^{2+}$ , but was inhibited by $Hg^{2+}$ , $Fe^{2+}$ , $Fe^{3+}$ , and $Mg^{2+}$ , EDTA, EGTA, iodoacetic acid, and pepstatin. The purified recombinant acid protease from the positive transformant 71 had high milk clotting activity, suggesting that it may be used as a rennet substitute in the cheese industry.


  • 주제어

    Saccharomycopsis fibuligera .   Yarrowia lipolytica .   acid protease .   milk clotting activity.  

  • 참고문헌 (30)

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