Cloning of the fabF gene in an expression vector and in vitro characterization of recombinant fabF and fabB encoded enzymes from Escherichia coli

P Edwards; J S Nelsen; J G Metz; K Dehesh

doi:10.1016/s0014-5793(96)01437-8

Cloning of the fabF gene in an expression vector and in vitro characterization of recombinant fabF and fabB encoded enzymes from Escherichia coli

FEBS Lett. 1997 Jan 27;402(1):62-6. doi: 10.1016/s0014-5793(96)01437-8.

Authors

P Edwards¹, J S Nelsen, J G Metz, K Dehesh

Affiliation

¹ Oils Division, Calgene Inc., Davis, CA 95616, USA.

PMID: 9013860
DOI: 10.1016/s0014-5793(96)01437-8

Abstract

Analysis of the beta-ketoacyl-ACP synthase (KAS) encoded by the fabF gene of Escherichia coli has been hampered by a reported instability of the cloned gene. Here we describe biochemical characterization of purified, active protein from the recombinant fabF gene. This enzyme has the properties ascribed to KAS II and not those of a putative KAS IV reported to be encoded by fabJ, a genomic clone with DNA sequence identical to that of fabF. We also characterize active protein from a recombinant fabB gene and suggest that this method may have a general utility for analysis of KAS enzymes.

MeSH terms

3-Oxoacyl-(Acyl-Carrier-Protein) Synthase / chemistry
3-Oxoacyl-(Acyl-Carrier-Protein) Synthase / genetics*
3-Oxoacyl-(Acyl-Carrier-Protein) Synthase / isolation & purification
3-Oxoacyl-(Acyl-Carrier-Protein) Synthase / metabolism*
Cerulenin / pharmacology
Cloning, Molecular
Escherichia coli / enzymology*
Escherichia coli / genetics
Genes, Bacterial
Genetic Vectors
Recombinant Proteins / chemistry
Recombinant Proteins / isolation & purification
Recombinant Proteins / metabolism
Substrate Specificity
Temperature

Substances

Recombinant Proteins
Cerulenin
3-Oxoacyl-(Acyl-Carrier-Protein) Synthase