In vitro Synthesis of a Constitutive Enzyme of Escherichia coli, 6-phosphogluconate Dehydrogenase

Enzymes of central intermediary metabolism are among the constitutive proteins of Escherichia coli. 6-Phosphogluconate dehydrogenase [6-phospho-D-gluconate: nicotinamide adenine dinucleotide phosphate 2-oxidoreductase (decarboxylating), EC 1.1.1.44], an enzyme of the hexose monophosphate shunt, was...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1975-11, Vol.72 (11), p.4381-4384
Main Authors: Isturiz, Tomas, Wolf, Richard E.
Format: Article
Language:English
Subjects:
Bacteriophages
Cell-Free System
Cyclic AMP - pharmacology
Cyclic GMP - pharmacology
Dehydrogenases
DNA
DNA, Bacterial - metabolism
Enzymes
Escherichia coli
Escherichia coli - enzymology
Gene expression regulation
Genes
Genes, Regulator - drug effects
Kinetics
Messenger RNA
Phosphogluconate Dehydrogenase - biosynthesis
Protein Biosynthesis - drug effects
Protein synthesis
RNA
RNA, Bacterial - biosynthesis
RNA, Messenger - biosynthesis
Time Factors
Transcription, Genetic - drug effects
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Summary:Enzymes of central intermediary metabolism are among the constitutive proteins of Escherichia coli. 6-Phosphogluconate dehydrogenase [6-phospho-D-gluconate: nicotinamide adenine dinucleotide phosphate 2-oxidoreductase (decarboxylating), EC 1.1.1.44], an enzyme of the hexose monophosphate shunt, was synthesized in vitro in a coupled transcription-translation system directed by DNA from a specialized transducing phage carrying gnd, the structural gene. Enzyme synthesized in vitro was detected by radio-chemical assay of its activity. Kinetic experiments using inhibitors of transcription and translation suggest that synthesis of the mRNA for the enzyme was initiated at the gnd promoter. Neither cyclic AMP nor cyclic GMP had any effect on the amount of enzyme synthesized in vitro.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.72.11.4381