Expression and purification of plasmid-encoded Thermoplasma acidophilum citrate synthase from Escherichia coli

• Published in: FEBS letters Vol. 282; no. 1; pp. 132 - 134
• Main Authors: Sutherland, Katharine J., Danson, Michael J., Hough, David W., Towner, Paul
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 22-04-1991; Elsevier
• Subjects: Analytical, structural and metabolic biochemistry; Archaebacterium; Base Sequence; Binding Sites; Biological and medical sciences; Citrate (si)-Synthase - biosynthesis; Citrate (si)-Synthase - genetics; Citrate (si)-Synthase - isolation & purification; Citrate synthase; cloning; Cloning, Molecular; Electrophoresis, Polyacrylamide Gel; Enzymes and enzyme inhibitors; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Gene expression, thermophilicity; genes; Ligases; Molecular Sequence Data; Oligonucleotides; Plasmids; Thermoplasma - enzymology; Thermoplasma - genetics; Thermoplasma acidophilum; Gene expression, thermophilicity; Thermoplasma acidophilum; Citrate synthase; Archaebacterium; Purification; Ribosome; Archaeobacteria; Enzyme; Escherichia coli; Binding site; Gene expression; Citrate (si)-synthase; Bacteria; Genetic engineering; Enterobacteriaceae; Chromatofocusing; Ultraviolet visible spectrometry
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(91)80461-B

The citrate synthase gene from the thermophilic archaebacterium.
Thermoplasma acidophilum was expressed in
Escherichia coli, yielding an active product of the expected molecular weight. Manipulation of the citrate synthase gene in a series of pUC19 constructs showed that the presumed
Thermoplasma ribosome binding site is recognized by the
E. coli ribosome. A rapid purification of the expression of the expression product to homogeneity was achieved, based on the thermostability of
Thermoplasma citrate synthase.