@article{mbs:/content/journal/micro/10.1099/mic.0.26119-0, author = "Walker, Nicola D. and McEwan, Neil R. and Wallace, R. John", title = "Cloning and functional expression of dipeptidyl peptidase IV from the ruminal bacterium Prevotella albensis M384T", journal= "Microbiology", year = "2003", volume = "149", number = "8", pages = "2227-2234", doi = "https://doi.org/10.1099/mic.0.26119-0", url = "https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.26119-0", publisher = "Microbiology Society", issn = "1465-2080", type = "Journal Article", keywords = "MNA, 4-methoxynaphthylamide", keywords = "DPP, dipeptidyl peptidase", keywords = "pNA, p-nitroanilide", abstract = "Ruminal bacteria of the genus Prevotella play a crucial role in peptide breakdown in the rumen, a component of protein catabolism that leads to the inefficient use of dietary protein by ruminant animals. This is the first report of the cloning of a peptidase gene from a ruminal bacterium. Part of the dipeptidyl peptidase type IV (DPP-IV) gene from Prevotella albensis M384T was cloned using degenerate primers designed from conserved regions found within other known DPP-IV sequences. Flanking regions were determined by genomic walking. The DPP-IV gene was expressed in Escherichia coli. The cloned enzyme required a free N terminus and catalysed the removal of X-Pro dipeptide from proline-containing oligopeptides, where proline was the second residue from the N terminus. It was inhibited by serine protease inhibitors and the substrate analogue for mammalian DPP-IV, diprotin A. The properties of the cloned enzyme were similar to those of the native form in P. albensis and, in general, DPP-IVs from other organisms. The enzyme contained a conserved motif which is associated with the S9 class of prolyl oligopeptidases. The DPP-IV gene appeared not to be part of a contiguous operon. Regions with similarity to other putative promoters of Prevotella spp. were also identified. Construction of a phylogenetic tree demonstrated that the DPP-IV of P. albensis clusters with other DPP-IVs found in bacteria of the Cytophaga–Flexibacter–Bacteroidaceae (CFB) phylum, which are more closely related to eukaryotic DPP-IVs than the DPP-IV-like enzyme (PepX) of the lactic acid bacteria.", }