Premium
Molecular characterization of an Escherichia coli mutant with a temperature‐sensitive malonyl coenzyme A‐acyl carrier protein transacylase
Author(s) -
Verwoert Ira I.G.S.,
Verhagen Etienne F.,
van der Linden Karin H.,
Verbree Elizabeth C.,
Nijkamp H.John J.,
Stuitje Antoine R.
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)00630-x
Subject(s) - escherichia coli , mutant , mutagenesis , enzyme , biochemistry , mutation , coenzyme a , chemistry , strain (injury) , biology , amino acid , acyl carrier protein , microbiology and biotechnology , gene , anatomy , reductase
The temperature‐sensitive malonyl CoA‐ACP transacylase found in the Escherichia coli strain LA2‐89, carrying the fabD89 allele, was shown to result from the presence of an amber mutation in the fabD gene, at codon position 257, in combination with the supE44 genotype of this strain. The truncated form of the protein produced as the result of the amber mutation was demonstrated to be enzymatically inactive, whereas amber suppression rendered the resulting enzyme temperature labile. Site‐directed mutagenesis of codon 257 revealed a requirement for an aromatic amino acid at this position in the polypeptide chain, to assure temperature stability of the enzyme.