Isolation and Characterization of Fuji Apple Peroxidase

사과 Peroxidase의 분리 및 특성

  • Jee, Wan-Jung (Department of Food Science, Seoul Woman's University) ;
  • Cho, Nam-Sook (Department of Food Science, Seoul Woman's University) ;
  • Kim, In-Cheol (Department of Food Science and Technology, Seoul National University) ;
  • Park, Kwan-Hwa (Department of Food Science and Technology, Seoul National University) ;
  • Choi, Eon-Ho (Department of Food Science, Seoul Woman's University)
  • 지완정 (서울여자대학교 식품과학과) ;
  • 조남숙 (서울여자대학교 식품과학과) ;
  • 김인철 (서울대학교 식품공학과) ;
  • 박관화 (서울대학교 식품공학과) ;
  • 최언호 (서울여자대학교 식품과학과)
  • Published : 1991.08.01

Abstract

Three peroxidase fractions (peak I, II, III) were isolated from Fuji apples using CM-cellulose chromatography. The homogeneity of the isolated peroxidase isozymes was established by isoelectric focusing and electrophoresis. Isoelectric points of the isozymes were 3.80, 3.82, and 3.85, respectively. The optimum pH of peroxidase isozymes were pH 5.0(peak I) or 5.5(peak II, III), and optimum temperature was $40^{\circ}C$ when assayed by using guaiacol and $H_{2}O_{2}$ as substrates. Inactivation rate of three peroxidase isozymes were different at temperature of $70^{\circ}C$ and at pH of 5.5. The isozyme of peak II was found to be more heat stable than those of peak I and III. D values at $70^{\circ}C$ of peroxidase isozymes (peak I, II, III) were estimated to be 660 sec, 1,320 sec, and 600 sec, respectively. The thermal stability of Fuji apple peroxidase was not influenced in the presence of 0.032 M sucrose or lactose. However, the thermal stability of the enzyme was decreased by fructose and glucose.

Fuji 사과로부터 acetone 처리 및 CM-cellulose column chromatography를 통하여 3개의 peroxidase isozyme(peak I, II, III)을 분리하고 isoelectric focusing에 의하여 pI값이 각각 3.80, 8.82, 8.35임을 확인하였다. Peroxidase isozyme의 최적 pH는 guaiacol과 $H_{2}O_{2}$를 기질로 하였을 때 peak I이 pH5.0, peak II, III은 pH5.5 이었으며 최적온도는 peak I, II, III 모두 $40^{\circ}C$이었다. $70^{\circ}C$에서 각 isozyme의 D값은 660초, 1,320초, 600초로서 peak II isozyme이 가장 열에 안정하였다. 당(0.032 M)의 존재하에서는 sucrose와 lactose는 효소의 열안정성에 영향을 미치지 않았으나 fructose와 glucose는 열안정성을 현저하게 감소시켰다. 이는 환원당과 효소단백질의 상호작용에 기인한 것으로 생각된다.

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