Purification of glutamine synthetase from a variety of bacteria.
We have developed two procedures which allow the very rapid purification of glutamine synthetase (GS) from a diverse variety of bacteria. The first procedure, based upon differential sedimentation, depends upon the association of GS with deoxyribonucleic acid in cell extracts. The second procedure, derived from the method of C. Gross et al (J. Bacteriol. 128:382-389, 1976) for purifying ribonucleic acid polymerase by polyethylene glycol (PEG) precipitation, enabled us to obtain high yields of GS from either small or large quantities of cells. We used the PEG procedure to purify GS from Klebsiella aerogenes, K. pneumoniae, Escherichia coli, Salmonella typhimurium, Rhizobium sp. strain 32H1, R. meliloti, Azotobacter vinelandii, Pseudomonas putida, Caulobacter crescentus, and Rhodopseudomonas capsulata. The purity of the GS obtained, judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was high, and in many instances only a single protein band was detected.
D001395:Azotobacter / Q000201:enzymology; D001419:Bacteria / Q000201:enzymology*; D002498:Centrifugation; D011232:Chemical Precipitation; D004755:Enterobacteriaceae / Q000201:enzymology*; D005974:Glutamate-Ammonia Ligase / Q000302:isolation & purification*; D008722:Methods; D011092:Polyethylene Glycols; D011549:Pseudomonas / Q000201:enzymology; D012231:Rhizobium / Q000201:enzymology; D012241:Rhodopseudomonas / Q000201:enzymology
D011092:Polyethylene Glycols; D005974:Glutamate-Ammonia Ligase