Cloning, expression, and purification of glutamine synthetase from Clostridum acetobutylicum

Journal Article · Mon Sep 01 00:00:00 EDT 1986 · Appl. Environ. Microbiol.; (United States)

OSTI ID:6934967

Usdin, K P; Zappe, H; Jones, D T; Woods, D R

A glutamine synthetase (GS) gene, glnA, from the gram-positive obligate anaerobe Clostridium acetobutylicum was cloned on recombinant plasmid pHZ200 and enabled Escherichia coli glnA deletion mutants to utilize (NH/sub 4/)/sub 2/ as a sole source of nitrogen. The cloned C. acetobutylicum gene was expressed from a regulatory region contained within the cloned DNA fragment. glnA expression was subject to nitrogen regulation in E. coli. This cloned glnA DNA did not enable an E. coli glnA ntrB ntrC deletion mutant to utilize arginine or low levels of glutamine as sole nitrogen sources, and failed to activate histidase activity in this strain which contained the Klebsiella aerogenes hut operon. The GS produced by pHZ200 was purified and had an apparent subunit molecular weight of approximately 59,000. There was no DNA or protein homology between the cloned C. acetobutylicum glnA gene and GS and the corresponding gene and GS from E. coli. The C. acetobutylicum GS was inhibited by Mg/sup 2 +/ in the ..gamma..-glutamyl transferase assay, but there was no evidence that the GS was adenylylated.

• Research Organization: Univ. of Cape Town, South Africa
• OSTI ID: 6934967
• Journal Information: Appl. Environ. Microbiol.; (United States), Journal Name: Appl. Environ. Microbiol.; (United States) Vol. 52:3; ISSN AEMID
• Country of Publication: United States
• Language: English

Related Subjects

550400 -- Genetics
550700* -- Microbiology
59 BASIC BIOLOGICAL SCIENCES
BACTERIA
CELL CONSTITUENTS
CLONING
CLOSTRIDIUM
CLOSTRIDIUM ACETOBUTYLICUM
DNA
ELEMENTS
ENZYME ACTIVITY
ENZYMES
GENE OPERONS
GENE RECOMBINATION
GENE REGULATION
LIGASES
METABOLISM
METHANOGENIC BACTERIA
MICROORGANISMS
MOLECULAR BIOLOGY
MUTANTS
NITROGEN
NONMETALS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PLASMIDS
PURIFICATION