Partial purification and characterization of glycylprolyl dipeptidyl aminopeptidase in porcine pancreas |
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Authors: | Makoto Ichinose Renju Maeda Takaaki Fukuda Bunji Watanabe Tadahiko Ishimaru Motomori Izumi Seibei Miyake Masaharu Takamori |
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Institution: | 1. Department of Clinical Research, Nagasaki Chuo National Hospital, 1001, Kubara, Omura City, Nagasaki 856, Japan;2. The 1st Department of Internal Medicine, Nagasaki University School of Medicine, 7-1, Sakamoto-Cho, Nagasaki, 852 Japan |
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Abstract: | This study reports the presence of glycylprolyl dipeptidyl aminopeptidase in porcine pancreas, and its partial purification and some properties. Crude enzyme preparation was obtained by extraction from acetone-dried powder of the pancreas at pH 7.6. For solubilization of enzyme, freezing and thawing were carried out. Crude enzyme extract was fractionated with ammonium sulfate precipitation, gel filtration on Sephadex G-200 column and ion-exchange chromatography on DEAE-cellulose. Partially purified enzyme showed 2897-folds purification. The enzyme activity on polyacrylamide gel electrophoresis showed good agreement with a main protein band stained with Coomassie brilliant blue. Molecular weight of this enzyme from the pancreas was estimated to be 300 000 by gel filtration on Sephacryl S-300 column. Optimum pH was between 8.5 and 9.0, and Km value for glycylproline-p-nitroanilide tosilate was 0.33 mM. This enzyme from the pancreas was a serine enzyme and was relatively stable to heat at 60°C for 10 min. |
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Keywords: | Dipeptidyl aminopeptidase Peptidase (Porcine pancreas) |
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