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KMID : 0377519810060030387
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Chung-Ang Journal of Medicine
1981 Volume.6 No. 3 p.387 ~ p.396
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Purification and Characterization of Bound Form Trehalase from Rabbit Kidney
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Kim Sung-Hwan
Lee Dong-Wook
Lee Hi-Sung
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Abstract
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The distribution and same properties of trehalase (α, α-trehalose 1-D-glucohydrolase, EC 3.2.1.28) in rabbit kidney was investigated in this study. The bound form of trehalase was solubilized from the mitochondrial and nucleic fraction with a mixture of 0.5% Triton X-100 and 0.2% sodium deo-xycholate. Solubilized enzyme was purified by ammonium sulfate precipitation, Sephadex G-200 gel filtration and DEAE-cellulose column chromatography. The activity of trehalase was measured by Lapp and Mason. The results are summarized as follows; 1. The activity of trehalase in rabbit kidney was 232 units/g. The distribution of trehalase in the subcellular organells indicated that 25% of the activity was in cytosol, 45% in mitochoridrial and about 30% in nucleic fraction. The specific activity were 0.92, 4.78, and 3.72, respectively. 2. The bound form of trehalase was purified approximately 82-fold. 3, The molecular weight of the purified enzyme was estimated to be 103,000 daltons by Sephadex G-200 gel filtration. 4. The optimum pH of the enzyme was 6.0 in phosphate buffer and 4.8 in acetate buffer. 5. The purified enzyme was optimally active at 55℃ in phosphate buffer at pH 6.0. 6. The enzyme was highly specific for trehalose.
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