ESTERASE FROM THE OIL-DEGRADING ACINETOBACTER-LWOFFII RAG-1 - SEQUENCE-ANALYSIS AND OVER-EXPRESSION IN ESCHERICHIA-COLI

The est gene encoding an esterase from Acinetobacter lwoffii RAG-1 was cloned into E. coli under the control of the P(L) promoter of the pha ge lambda. The N-terminal sequence of the first 20 amino acids of the heterolgous expressed esterase corresponded to that obtained from the nucleotide sequence. Antibodies prepared against the over-expressed re combinant esterase in E. coli were used to locate the enzyme primarily in the membrane fractions of A lwoffii RAG-1. Comparison with homolog ous proteins from both eukaryotic and prokaryotic organisms suggest th at the RAG-1 esterase exhibits sequence motifs characteristic of both serine proteases and of lipases.