Combined use of strain construction and affinity chromatography in the rapid, high-yield purification of 6-phosphogluconate dehydrogenase from Escherichia coli.
A rapid, high-yield method for purification of 6-phosphogluconate dehydrogenase from Escherichia coli K-12 is described. Sonic extracts prepared from heat-induced cultures of strain RW184, doubly lysogenic for the specialized transducing bacteriophage lambdacI857St68h80dgndhis and bearing a deletion of the gene for glucose 6-phosphate dehydrogenase, contained levels of 6-phosphogluconate dehydrogenase 15- to 20-fold higher than cultures of wild-type cells. Affinity chromatography on blue dextran-Sepharose with batchwise elution with 1 mM nicotinamide adenine dinucleotide phosphate affected a further 10-fold purification. Enzyme prepared in this manner was homogeneous according to electrophoresis on sodium dodecyl sulfate-polyacrylamide gels and immunoelectrophoresis using antiserum directed against it. Fructose 1,6-diphosphate is an inhibitor of enzyme activity.
14 references, click to expand
-
Medium for the differentiation of acid producing colonies of staphylococci.J Bacteriol. 1958 Oct;76(4):454-5 PMID: 13587488
-
Protein measurement with the Folin phenol reagent.J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
-
Blue dextran-sepharose: an affinity column for the dinucleotide fold in proteins.Proc Natl Acad Sci U S A. 1975 Feb;72(2):669-72 PMID: 164664
-
In vitro synthesis of a constitutive enzyme of Escherichia coli, 6-phosphogluconate dehydrogenase.Proc Natl Acad Sci U S A. 1975 Nov;72(11):4381-4 PMID: 172907
-
Binding of Cibacron blue F3GA to proteins containing the dinucleotide fold.Proc Natl Acad Sci U S A. 1976 Feb;73(2):361-5 PMID: 174104
-
Rapid purification of lactate dehydrogenase from rat liver and hepatoma: a new approach.Arch Biochem Biophys. 1974 Jan;160(1):279-84 PMID: 4364067
-
Mechanism for regulating the distribution of glucose carbon between the Embden-Meyerhof and hexose-monophosphate pathways in Streptococcus faecalis.J Bacteriol. 1971 May;106(2):456-67 PMID: 4396792
-
"Up-promoter" mutations of glucose 6-phosphate dehydrogenase in Escherichia coli.J Mol Biol. 1972 Oct 28;71(1):107-11 PMID: 4564298
-
Isolation of specialized transducing bacteriophages for gluconate 6-phosphate dehydrogenase (gnd) of Escherichia coli.J Bacteriol. 1974 Feb;117(2):468-76 PMID: 4590471
-
Affinity chromatography of kinases and dehydrogenases on Sephadex and Sepharose dye derivatives.Adv Exp Med Biol. 1974;42(0):123-33 PMID: 4602166
-
Genetic approaches to determination of enzyme quaternary structure.Biochemistry. 1971 Jan 19;10(2):204-7 PMID: 4923718
-
Glucose and gluconate metabolism in an Escherichia coli mutant lacking phosphoglucose isomerase.J Bacteriol. 1967 May;93(5):1571-8 PMID: 5337843
-
Isolation and properties of 6-phosphogluconate dehydrogenase from Escherichia coli. Some comparisons with the thermophilic enzyme from Bacillus stearothermophilus.Biochemistry. 1976 Sep 7;15(18):4026-33 PMID: 786365
-
Applications of blue dextran and Cibacron Blue F3GA in purification and structural studies of nucleotide-requiring enzymes.Biochem Biophys Res Commun. 1976 Oct 4;72(3):816-23 PMID: 791290
Contact
No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong
Qilu Normal University · Genelibs Bioinformatics Lab
750 Shunhua Rd, Jinan
2F, Bldg F, University Science Park
Tel: 0531-88819269
WeChat Official Account
Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.
Business Email
E-mail: product@genelibs.com
Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2
