Molecular analysis of an esterase-encoding gene from a lipolytic psychrotrophic pseudomonad.
An esterase gene (estA) from a lipolytic psychotroph (Pseudomonas sp. LS107d2), was cloned in Escherichia coli and its nucleotide sequence was determined, revealing an ORF encoding a polypeptide of 389 amino acid residues, with a molecular mass of 42276 Da. Labelling of plasmid-encoded proteins with [35S]methionine, using the maxicell procedure, gave a single polypeptide of molecular mass 42 kDa, consistent with that calculated from the ORF. Colonies of E. coli cells containing estA produced a clear halo when grown on solid media containing tributyrin; no clearance was produced when cells were grown on media containing triolein. Extracts of cells containing estA also hydrolysed water-soluble nitrophenol esters, but were unable to cleave water-insoluble substrates. The preference for water-soluble substrates indicates that the gene product is an esterase.
McKay
D B
DB
Division of Science and Technology, Griffith University, Brisbane, Queensland, Australia.
Jennings
M P
MP
Godfrey
E A
EA
MacRae
I C
IC
Rogers
P J
PJ
Beacham
I R
IR
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D004950:Esterases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D005798:Genes, Bacterial; D006041:Goats; D008066:Lipolysis / Q000235:genetics*; D008892:Milk / Q000032:analysis; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics*; D013379:Substrate Specificity
D001426:Bacterial Proteins; D004950:Esterases