Cloning and nucleotide sequence of the gene coding for citrate synthase from a thermotolerant Bacillus sp.

F. J. Schendel, P. R. August, C. R. Anderson, R. S. Hanson, M. C. Flickinger

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

The structural gene coding for citrate synthase from the gram-positive soil isolate Bacillus sp. strain C4 (ATCC 55182) capable of secreting acetic acid at pH 5.0 to 7.0 in the presence of dolime has been cloned from a genomic library by complementation of an Escherichia coli auxotrophic mutant lacking citrate synthase. The nucleotide sequence of the entire 3.1-kb HindIII fragment has been determined, and one major open reading frame was found coding for citrate synthase (ctsA). Citrate synthase from Bacillus sp. strain C4 was found to be a dimer (M(r), 84,500) with a subunit with an M(r) of 42,000. The N-terminal sequence was found to be identical with that predicted from the gene sequence. The kinetics were best fit to a bisubstrate enzyme with an ordered mechanism. Bacillus sp. strain C4 citrate synthase was not activated by potassium chloride and was not inhibited by NADH, ATP, ADP, or AMP at levels up to 1 mM. The predicted amino acid sequence was compared with that of the E. coli, Acinetobacter anitratum, Pseudomonas aeruginosa, Rickettsia prowazekii, porcine heart, and Saccharomyces cerevisiae cytoplasmic and mitochondrial enzymes.

Original languageEnglish (US)
Pages (from-to)335-345
Number of pages11
JournalApplied and environmental microbiology
Volume58
Issue number1
DOIs
StatePublished - 1992

Fingerprint Dive into the research topics of 'Cloning and nucleotide sequence of the gene coding for citrate synthase from a thermotolerant Bacillus sp.'. Together they form a unique fingerprint.

Cite this