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毛竹笋中脱氧核糖核酸酶的提纯和特性(PDF)

《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

Issue:
1991年01期
Page:
15-22
Column:
研究论文
publishdate:
1900-01-01

Article Info:/Info

Title:
PURIFICATION AND PROPERTIES OF DEOXYRIBONUCLEASE FROM PHYLLOSTACHYS PUBESCENS SHOOTS
Article ID:
1000-2006(1991)01-0015-08
Author(s):
Lu Xianhui
Department of Forestry
Keywords:
Phyllostachys Bamboo shoot Deoxyribonuclease
Classification number :
-
DOI:
10.3969/j.jssn.1000-2006.1991.01.003
Document Code:
A
Abstract:
The deoxyribonuclease (DNase) from Phyllostachys pubescebs shoots was purified by heat treatment, ammonium sulfate fractionation, Sephadex G-100 and DEAE-cellulose column chromatography. The data show that final purification obtained was 27.2 fold, photographs of SDS-polyacrylamide dise gels stained with Coomassie blue R-250 after electrophoresis of this DNase. preparation showed single band. The MW of the enzyme was 5.4×104. The enzyme gave pH optima of 5.0-5.2 and temperature optima of 65℃ . The purified enzyme preparation hydrolyzes denatured DNA and the 5’-AME at 1 : 1 : 0.1 activities. The absolute requirement for a sulfhydryl compound for maintenance of activity at pH 4. 8 and the marked inhibition of the enzyme activity by the same sulfhydryl compounds at pH 8.0 Except for KH2po4at lower concentration (1 x 10~5M) , there several ative effects on the enzymeactivity, approximately 45% and 42% inhibition and sodium fluoride of 1 x 10~3M. The addition of 10~2M one valent cation K+, Na+and NH+4 to the incubative mixture resulted in a 10%-20% stimulation of the DNase activity, Of a number cf two valent Cations tested, only ca++and Mg++produce a slightly stimulating effect on activity (5%-8% stimulation) , other cation found to exert markedly inhibiting effect on the activity includes Co++, Cu++, Zn++, Hg++, and Mn++. Metal chelating agent EDTA (10~3M) resulted in marked inhibition of the DNase activity, which implies that this DNase contains metal ions for maintained activity.

References

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Last Update: 1900-01-01