The DmpA aminopeptidase from Ochrobactrum anthropi LMG7991 is the prototype of a new terminal nucleophile hydrolase family

Fanuel, Laurence; Goffin, Colette; CHEGGOUR, Abdellatif; Devreese, Bart; DRIESSCHE, Gonzales VAN; Joris, Bernard; Beeumen, Jozef Van; Frère, Jean-Marie

doi:10.1042/bj3410147

Cited by 39 publications

(41 citation statements)

References 30 publications

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“…A blast search indicated that the amino acid sequence deduced from bapA was similar to those of hypothetical protein PA1486 from Pseudomonas aeruginosa PAO1 [17], putative d-aminopeptidase PP3844 from Pseudomonas putida KT2440 [18], hypothetical protein PLU2258 similar to d-aminopeptidase from Photorhabdus luminescens (ssp. laumondii) TTO1 [19], aminopeptidase Atu5242 from Agrobacterium tumefaciens C58 plasmid AT [20] and DmpA (l-aminopeptidase d-Ala-esterase ⁄ amidase) from O. anthropi LMG7991 [15] (Table 1). Figure 3 shows the alignment of the primary structures of BapA from Pseudomonas sp.…”

Section: Resultsmentioning

confidence: 99%

“…DmpA has been reported to be a homotetrameric enzyme, each subunit of which is composed of two polypeptides generated by a possible autocatalytic cleavage of a precursor polypeptide. Interestingly, DmpA changes its stereoselectivity depending on the substrate, catalyzing the l-stereoselective hydrolysis of peptides and d-stereoselective hydrolysis of amino acid amides and esters [15]. The gel-filtration chromatography and SDS ⁄ PAGE analysis of the purified BapA revealed that the number of subunits in the native form and the polypeptide composition of each subunit were significantly similar to those for DmpA.…”

Section: Discussionmentioning

confidence: 95%

“…, D-Ala-Gly, D-Ala-(Gly) 2 (D-Ala) 2 , D-Ala-L-Ala (D-Ala)3 (D-Ala)4 , L-Ala-Gly, L-Ala-(Gly) 2 (L-Ala) 2 , L-Ala-D-Ala, L-Ala-D-Ala-L-Ala, DL-Ala-DL-Asn, DL-Ala-DL-Ile, DL-Ala-DL-Leu, DL-Ala-DL-Met, DL-Ala-DL-Phe, DL-Ala-DL-Ser, DL-Ala-DL-Val, L-Asp-D-Ala, L-Pro-Gly, L-Pro-L-Phe, c-Aminobutyryl-L-His (homocarnosine), Gly-NH 2 , D-Phe-NH 2 , D-Asp-NH 2 , D-Glu-NH 2 , D-Gln-NH 2 , D-Pro-NH 2 , L-Ala-NH 2 , L-Leu-NH 2 , L-Phe-NH 2 , L-Tyr-NH 2 , L-Trp-NH 2 , L-Ser-NH 2 , L-Thr-NH 2 , L-Lys-NH 2 and L-Pro-NH 2 . a Data from reference[15]; N.D. not detectable; N.I. -Xaa dipeptidase from Pseudomonas sp.…”

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confidence: 99%

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A DmpA‐homologous protein from Pseudomonas sp. is a dipeptidase specific for β‐alanyl dipeptides

Komeda

Asano

2005

The FEBS Journal

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Many different kinds of microbial hydrolases acting d-stereoselectively on amino acid amides or peptides have been characterized, and some of them have been applied to the production of optically active d-amino acids from the corresponding racemic amino acid amides [1]. The d-stereoselective amidases and peptidases known to date can be classified into four groups based on their primary structures. We have determined the nucleotide sequence of a DNA fragment covering the flanking region of the R-stereoselective amidase gene, ramA, from the Pseudomonas sp. MCI3434 genome and found an additional gene, bapA, coding for a protein showing sequence similarity to DmpA aminopeptidase from Ochrobactrum anthropi LMG7991 (43% identity). The DmpA (called l-aminopeptidase d-Ala-esterase ⁄ amidase) hydrolyzes alanine-p-nitroanilide, alaninamide, and alanine methylester with a preference for the d-configuration of the alanine, whereas the enzyme acts as an l-stereoselective aminopeptidase on a tripeptide Ala- (Gly) A recombinant BapA exhibiting hydrolytic activity toward d-alaninep-nitroanilide was purified from the cell-free extract of an Escherichia coli transformant overexpressing the bapA gene and characterized. The purified enzyme contained two polypeptides corresponding to residues 1-238 (a-peptide) and 239-366 (b-peptide) of the precursor as observed for DmpA. On gel-filtration chromatography, BapA in the native form appeared to be a tetramer. It had maximal activity at 60°C and pH 9.0-10.0, and was inactivated in the presence of p-chloromercuribenzoate, N-ethylmaleimide, dithiothreitol, Zn 2+ , Ag + , Cd 2+ or Hg 2+ . The enzyme hydrolyzed d-alanine-p-nitroanilide more efficiently than l-alanine-p-nitroanilide the same as DmpA. Furthermore, BapA was found to hydrolyze peptide bonds of b-alanyl dipeptides including b-Ala-l-Ala, b-Ala-Gly, b-Ala-l-His (carnosine), b-Ala-l-Leu, and (b-Ala) 2 with high efficiency compared to d-alanine-p-nitroanilide. b-Alaninamide was also efficiently hydrolyzed, but the enzyme did not act on the peptides containing proteinogenic amino acids or their d-counterparts for N-terminal residues. Based on its unique substrate specificity, the enzyme should not be called l-aminopeptidase d-Ala-esterase ⁄ amidase but b-Ala-Xaa dipeptidase.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 95%

mentioning

confidence: 99%

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A DmpA‐homologous protein from Pseudomonas sp. is a dipeptidase specific for β‐alanyl dipeptides

Komeda

Asano

2005

The FEBS Journal

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“…Finally, DmpA, an N-terminal nucleophile hydrolase, exhibits different stereospecificities according to the nature of the substrates. It hydrolyses D-alanyl-amide,p-nitroanilide and -methylester more efficiently than their L-counterparts but is a strict L-aminopeptidase with peptides (Fanuel et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

The dppA gene of Bacillus subtilis encodes a new d‐aminopeptidase

Cheggour

Fanuel

Duez

et al. 2000

Molecular Microbiology

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SummaryDifferent strains of Bacillus were screened for their ability to hydrolyse D-alanyl-p-nitroanilide. Activity was detected in Bacillus pumilus, Bacillus brevis, Bacillus licheniformis 749I and Bacillus subtilis 168. The last strain was the best producer and was selected for the production and purification of the enzyme. The determination of the N-terminal sequence identified the enzyme as the product of the dppA gene (previously named dciAA) belonging to the dipeptide ABC transport (dpp) operon expressed early during sporulation. Open reading frames (ORFs) encoding putative related proteins were found in the genomes of a variety of Archaea and both sporulating and non-sporulating bacteria. The enzyme behaves as a D-aminopeptidase and represents the prototype of a new peptidase family. Among the tested substrates, the highest activities were found with D-Ala-D-Ala and D-Ala-Gly-Gly. The active enzyme behaves as an octamer of identical 30 kDa subunits. It exhibits a broad pH optimum, extending between pH 9 and 11. It is reversibly inhibited in the presence of Zn 21 chelators, and the sequence comparisons highlight the conservation of potential Zn-binding residues. As it has been shown by others that null mutations in the dpp operon do not inhibit spore formation, the physiological role of DppA is probably an adaptation to nutrient deficiency.

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“…This may be the case for d-aminopeptidase from Ochrobactrum anthropi in particular, as it is capable of hydrolysing not only d-amino acid amides and d-amino acid p-nitroanilides but also l-amino acid p-nitroanilides (Asano et al 1989). Furthermore, the second d-aminopeptidase from O. anthropi, DmpA, exhibits different stereospecificity as a function of substrate type (Fanuel et al 1999). It hydrolyses d-alanyl-amide, -pNa,methyl-ester more efficiently than their l-counterparts but is a strict l-aminopeptidase with peptides.…”

Section: The Role Of Peps In Nitrogen Supply and Other Cellular Functmentioning

confidence: 99%

The role of aminopeptidase PepS in the growth of Streptococcus thermophilus is not restricted to nitrogen nutrition

Thomas

Besset

Courtin

et al. 2010

Journal of Applied Microbiology

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Aims: To investigate the effect of an absence of aminopeptidase PepS on the growth of Streptococcus thermophilus on different media and at different temperatures. Methods and Results: Using gene interruption, a negative mutant of the Strep. thermophilus CNRZ385 strain was constructed for the aminopeptidase PepS (strain ΔpepS). Checks were first of all made using biochemical assays that the ΔpepS strain lacks the peptide hydrolase activity of aminopeptidase PepS. It was demonstrated that the absence of the aminopeptidase PepS exerted a negative effect on growth whatever the culture medium (M17, chemically defined medium, milk). The role of aminopeptidase PepS in growth was enhanced at a high temperature (45°C vs 37°C). The ΔpepS strain was more resistant to lysozyme than the wild‐type strain. Conclusions: We were able to demonstrate that aminopeptidase PepS probably plays a pleiotropic role through its involvement in growth via nitrogen nutrition, as well as via other cellular functions/metabolisms (such as peptidoglycane metabolism). Significance and Impact of the Study: This study constitutes the first report on the role of a member of the M29 MEROPS family of metallopeptidases (http://merops.sanger.ac.uk/).

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The DmpA aminopeptidase from Ochrobactrum anthropi LMG7991 is the prototype of a new terminal nucleophile hydrolase family

Cited by 39 publications

References 30 publications

A DmpA‐homologous protein from Pseudomonas sp. is a dipeptidase specific for β‐alanyl dipeptides

A DmpA‐homologous protein from Pseudomonas sp. is a dipeptidase specific for β‐alanyl dipeptides

The dppA gene of Bacillus subtilis encodes a new d‐aminopeptidase

The role of aminopeptidase PepS in the growth of Streptococcus thermophilus is not restricted to nitrogen nutrition

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