Abstract
A gene encoding a so far uncharacterized β-peptidyl aminopeptidase from the opportunistic human pathogen Pseudomonas aeruginosa PAO1 was cloned and actively expressed in the heterologue host Escherichia coli. The gene was identified in the genome sequence by its homology to the S58 family of peptidases. The sequence revealed an open reading frame of 1,101 bp with a deduced amino acid sequence of 366 amino acids. The gene was amplified by PCR, ligated into pET22b(+) and was successfully expressed in E. coli BL21 (DE3). It was shown that the enzyme consists of two polypeptides (α- and β-subunit), which are processed from the precursor. The enzyme is specific for N-terminal β-alanyl dipeptides (β-Ala-Xaa). BapF hydrolyses efficiently β-alanine at the N-terminal position, including H-β3hAla-pNA, H–D-β3hAla-pNA and β-Ala-l-His (l-carnosine). d- and l-alaninamide were also hydrolysed by the enzyme.
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Fuchs, V., Jaeger, KE., Wilhelm, S. et al. The BapF protein from Pseudomonas aeruginosa is a β-peptidyl aminopeptidase. World J Microbiol Biotechnol 27, 713–718 (2011). https://doi.org/10.1007/s11274-010-0484-6
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DOI: https://doi.org/10.1007/s11274-010-0484-6