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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Isolation and Characterization of Four Carboxypeptidases in Canavalia lineata Cotyledons

  • Yang, Jong-Moon (Department of Biology, College of Natural Sciences, Seoul National University) ;
  • Rhew, Tae-Hyong (Department of Biology, College of Natural Sciences, Pusan National University) ;
  • Koh, Suck-Chan (Department of Biology, College of Natural Sciences, Cheju National University) ;
  • Kwon, Young-Myung (Department of Biology, College of Natural Sciences, Seoul National University)
  • Received : 1995.05.19
  • Published : 1995.09.30
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Abstract

Four carboxypeptidases, CP1, CP2, CP3, and CP4 were isolated from the cotyledons of germinating seedlings of Canavalia lineata by sequential chromatography on the following four columns: 1) CM-cellulose, 2) Sephacryl 5-300, 3) Procion red dye, and 4) Sephacryl S-200. A number of properties of the enzymes, such as substrate specificity, molecular weight, optimum pH, thermal stability, have been determined. Enzyme activities were measured using the Cbz(carbobenzoxy)-dipeptides containing phenylalanine at the penultimate position. The $K_m$ values of four carboxypeptidases for Cbz-Phe-Ala were 0.50, 0.65, 1.30, and 1.35 mM, respectively. The inhibition studies indicated that the four carboxypeptidases were all serine type. Each of the carboxypeptidases with molecular weights of 145, 114, 105, and 104 kDa, respectively, had the optimum enzyme activity at pH 5.0~6.0. And they were sensitive to high temperature.

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