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荧光假单胞菌泛解酸酶的纯化及性质

Purification and properties of pantothenase from Pseudomonas fluorescens.

作者信息

Airas R K, Hietanen E A, Nurmikko V T

出版信息

Biochem J. 1976 Aug 1;157(2):409-13. doi: 10.1042/bj1570409.

DOI:10.1042/bj1570409
PMID:822843
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1163867/
Abstract

Pantothenase (EC 3.5.1.22) from Pseudomonas fluorescens UK-1 was purified to homogeneity as judged by disc-gel electrophoresis and isoelectric focusing. The purification procedure consisted of four steps: DEAE-Sephadex chromatography, (NH4)2SO4 precipitation, hydroxyapatite chromatography and preparative polyacrylamide-gel electrophoresis. Gel filtration on Ultrogel AcA 34 was used to determine the molecular weight, and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis to study the subunit molecular weight. The enzyme appeared to be composed of two subunits with mol.wts. of approx. 50000 each. The total mol.wt. of the enzyme was thus about 100000. The isoelectric point was 4.7 at 10 degrees C.

摘要

通过圆盘凝胶电泳和等电聚焦判断，荧光假单胞菌UK-1中的泛解酸酶（EC 3.5.1.22）被纯化至同质。纯化过程包括四个步骤：DEAE-葡聚糖凝胶色谱、硫酸铵沉淀、羟基磷灰石色谱和制备性聚丙烯酰胺凝胶电泳。使用Ultrogel AcA 34上的凝胶过滤来测定分子量，并用十二烷基硫酸钠/聚丙烯酰胺凝胶电泳研究亚基分子量。该酶似乎由两个亚基组成，每个亚基的分子量约为50000。因此，该酶的总分子量约为100000。在10℃时，等电点为4.7。

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Kinetic studies on pantothenase from Pseudomonas fluorescens. Effects of pH on substrate and inhibitor binding.荧光假单胞菌泛解酸酶的动力学研究。pH对底物和抑制剂结合的影响。
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本文引用的文献

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DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS.圆盘电泳。II. 方法及其在人血清蛋白中的应用。
Ann N Y Acad Sci. 1964 Dec 28;121:404-27. doi: 10.1111/j.1749-6632.1964.tb14213.x.
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Spectroscopic determination of tryptophan and tyrosine in proteins.蛋白质中色氨酸和酪氨酸的光谱测定
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The aerobic pseudomonads: a taxonomic study.需氧假单胞菌属：一项分类学研究。
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The bacterial degradation of pantothenic acid. I. Over-all nature of the reaction.泛酸的细菌降解。I. 反应的整体性质。
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Measurement of molecular weights by electrophoresis on SDS-acrylamide gel.通过在SDS-聚丙烯酰胺凝胶上进行电泳来测定分子量。
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Thermal inactivation and reactivation of an enzyme in vivo. Pantothenate hydrolase of Pseudomonas fluorescens.体内一种酶的热失活与再激活。荧光假单胞菌的泛酸水解酶
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Kinetic studies on pantothenase from Pseudomonas fluorescens. Effects of pH on substrate and inhibitor binding.荧光假单胞菌泛解酸酶的动力学研究。pH对底物和抑制剂结合的影响。
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