蜡样芽孢杆菌的脱氧核糖5-磷酸醛缩酶：纯化及性质

Deoxyribose 5-phosphate aldolase of Bacillus cereus: purification and properties.

作者信息

Sgarrella F, Del Corso A, Tozzi M G, Camici M

机构信息

Istituto di Chimica Biologica, Facoltà di Farmacia, Università di Sassari, Italy.

出版信息

Biochim Biophys Acta. 1992 Jan 9;1118(2):130-3. doi: 10.1016/0167-4838(92)90139-5.

DOI:10.1016/0167-4838(92)90139-5

PMID:1730028

Abstract

Deoxyribose 5-phosphate aldolase was purified 41 times from Bacillus cereus induced by growth on deoxyribonucleosides. The purification procedure includes ammonium sulphate fractionation, gel filtration on Sephadex G-100, ion-exchange chromatography on DEAE-Sephacel and preparative electrophoresis on 10% polyacrylamide gel. The enzyme is stable above pH 6.5, but is rapidly inactivated by sulfhydryl reagents. Being insensitive to EDTA, it may be considered as a Class I aldolase. Among a number of compounds tested (including some carboxylic acids, free and phosphorylated pentoses, nucleotides and nucleosides), none has been found to affect the enzyme activity. The enzyme appears to be dimeric, with a subunit Mr of 23,600. A Km of 4.4 x 10(-4) M was calculated for dRib 5-P.

摘要

通过在脱氧核苷上生长诱导的蜡样芽孢杆菌中，磷酸脱氧核糖醛缩酶被纯化了41倍。纯化步骤包括硫酸铵分级分离、Sephadex G - 100凝胶过滤、DEAE - Sephacel离子交换色谱以及10%聚丙烯酰胺凝胶制备电泳。该酶在pH 6.5以上稳定，但会被巯基试剂迅速灭活。它对EDTA不敏感，可被视为I类醛缩酶。在测试的许多化合物（包括一些羧酸、游离和磷酸化的戊糖、核苷酸和核苷）中，未发现任何一种会影响酶活性。该酶似乎是二聚体，亚基分子量为23,600。计算得出dRib 5 - P的Km为4.4×10⁻⁴ M。