Abstract
Peptidases of Prevotella spp. play an important role in the breakdown of protein to ammonia in the rumen. This study describes a peptidase cloned from Prevotella albensis M384. DNA from P. albensis was used to complement a peptidase-deficient strain of Escherichia coli, CM107. A cloned fragment, Pep581, which enabled growth of E. coli CM107, contained an ORF of 1452 bp, encoding a 484 amino acid residue protein with a calculated molecular weight of 53.2 kDa and a theoretical pI of 4.90. Pep581 shared similar sequence identity of 47% with PepD from E coli, and it was also a metallo-aminopeptidase. A putative catalytic metal binding region was identified in Pep581, similar to that found in the related PepT (a tripeptidase) and PepA (an oligopeptidase). Gel filtration indicated Pep581 was a dimer in its native state, similar to PepD of E. coli. PepD is a broad specificity dipeptidase that has been found in several prokaryotes. The enzyme expressed from Pep581 differed from PepD enzymes previously characterised in that it hydrolysed tri- and oligopeptides in addition to dipeptides, cleaving single amino acids from the N terminus. (C) 2005 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.
Original language | English |
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Pages (from-to) | 399-404 |
Number of pages | 6 |
Journal | FEMS Microbiology Letters |
Volume | 243 |
Issue number | 2 |
DOIs | |
Publication status | Published - 15 Feb 2005 |
Keywords
- aminopeptidase
- Prevotella albensis
- peptidase
- rumen
- Escherichia coli
- rumen bacteria
- physicochemical parameters
- enzyme activity
- ruminicola
- microorganisms
- protein
- hydrolysis
- metabolism
- breakdown