Rapid identification of enterococci by pyrrolidonyl aminopeptidase activity (PYRase)

Mitchell, M.J.; Conville, Patricia S.; Gill, Vee J.

doi:10.1016/0732-8893(87)90176-3

Cited by 8 publications

(9 citation statements)

References 5 publications

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“…The principal value of PYRase as a diagnostic marker is in the differentiation of S. pyogenes and enterococci from most other Grampositive cocci. 9,14 A range of selective culture media have been designed for detection of enterococci and these have traditionally relied upon chromogenic substrates for detection of b-glucosidase activity, which is a less specific marker than PYRase. One reason for this is likely to be the lack of available chromogenic substrates for PYRase that are suitable for use in culture media.…”

Section: Evaluation Of Substrates 8-10mentioning

confidence: 99%

“…The organic layer was dried (MgSO 4 ) and evaporated giving the product. 7 Compound 16c was synthesized from compound 11c (0.30 g, 1.00 mmol), Boc-Lalanine (0.20 g, 1.05 mmol), N-methylmorpholine (0.10 g, 1.00 mmol) and IBCF (0.14 g, 1.00 mmol) in dry THF (20 mL 9.86 (1H, s, NH), 9.63 (1H, s, NH), 9.23 The solvent was evaporated to giving compound 9a (0. 16 9.97 (1H, s, NH), 9.31 (1H, s, br, OH), 7.98 (3H, s, br, NH 3 + ), 7.32-7.20 (2H, m, Ar-H), 6.73 7.1.5.1.…”

Section: -(2-hydroxy-5-nitrophenyl)-n-[2-[3-(2-hydroxy-5-nitrophenylmentioning

confidence: 99%

“…b-Alanyl aminopeptidase has been detected in Pseudomonas aeruginosa, a common respiratory pathogen in cystic fibrosis patients. 7 L-Pyroglutamyl aminopeptidase activity is useful for differentiation within the family Enterobacteriaceae 3,8 and also for detection of enterococci 9 and Streptococcus pyogenes. 10 A diverse range of chromogenic aminopeptidase substrates have previously been described and some relevant examples (structures 1-5, AA = amino acid) are shown in Figure 1.…”

Section: Introductionmentioning

confidence: 99%

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Novel chromogenic aminopeptidase substrates for the detection and identification of clinically important microorganisms

Cellier

James

Orenga

et al. 2014

Bioorganic & Medicinal Chemistry

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A series of amino acid derivatives 8-10, 42 and 43 have been prepared as chromogenic enzyme substrates in order to detect aminopeptidase activity in clinically important Gram-negative and Gram-positive bacteria. Enzymatic hydrolysis liberates the amino acid moiety and either a 4-aminophenol or a 4-dialkylaminoaniline derivative which undergoes oxidative coupling with 1-naphthol or a substituted 1-naphthol giving an indophenol dye. Substrates and 1-naphthols were incorporated into an agar-based culture medium and this allowed growth of intensely coloured bacterial colonies based on hydrolysis by specific enzymes. Red/pink coloured colonies were produced by the substrates 8-10 and blue coloured colonies were formed by the substrates 42 and 43. The L-alanyl aminopeptidase substrates 8 targeted L-alanyl aminopeptidase activity and gave coloured colonies with a range of Gram-negative bacteria. Substrates 9 targeted β-alanyl aminopeptidase activity and generated coloured colonies with selected Gram-negative species including Pseudomonas aeruginosa. Three substrates for L-pyroglutamyl acid aminopeptidase (10a, 10c and 43) were hydrolysed by enterococci and Streptococcus pyogenes to generate coloured colonies. Two yeasts were also included in the study, but they did not produce coloured colonies with any of the substrates examined.

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Section: Evaluation Of Substrates 8-10mentioning

confidence: 99%

Section: -(2-hydroxy-5-nitrophenyl)-n-[2-[3-(2-hydroxy-5-nitrophenylmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Novel chromogenic aminopeptidase substrates for the detection and identification of clinically important microorganisms

Cellier

James

Orenga

et al. 2014

Bioorganic & Medicinal Chemistry

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“…In the 1980s, detection methods were developed in bacterial diagnosis based on the use of chromogenic and fluorogenic substrates. 138 Beyond the detection of PYRase activity in bacterial diagnosis, the characterization of PYRase genes could also lead to promising a p p 1 i~a t i o n s . l~~ For example, group A streptococci have been identified using DNA probes to the PYRase gene.lsg With this approach, it may be possible to replace the PYRase activity test with a molecular probe test for a more definitive identification of bacterial species.…”

Section: Applications Involving Pyrasementioning

confidence: 99%

Pyrrolidone carboxyl peptidase (Pcp): An enzyme that removes pyroglutamic acid (pGlu) from pGlu‐peptides and pGlu‐proteins

Awade¹,

Cleuziat²,

Gonzalès³

et al. 1994

Proteins

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Pyrrolidone carboxyl peptidase (EC 3.4.11.8) is an exopeptidase commonly called PYRase, which hydrolytically removes the pGlu-proteins. pGlu also known as pyrrolidone carboxylic acid may occur naturally by an enzymatic procedure or may occur as an artifact in proteins or peptides. The enzymatic synthesis of pGlu suggests that this residue may have important biological and physiological functions. Several studies are consistent with this supposition. PYRase has been found in a variety of bacteria, and in plant, animal, and human tissues. For over two decades, biochemical and enzymatic properties of PYRase have been investigated. At least two classes of PYRase have been characterized. The first one includes the bacterial and animal type I PYRases and the second one the animal type II and serum PYRases. Enzymes from these two classes present differences in their molecular weight and in their enzymatic properties. Recently, the genes of PYRases from four bacteria have been cloned and characterized, allowing the study of the primary structure of these enzymes, and their over-expression in heterelogous organisms. Comparison of the primary structure of these enzymes revealed striking homologies. Type I PYRases and bacterial PYRases are generally soluble enzymes, whereas type II PYRases are membrane-bound enzymes. PYRase II appears to play as important a physiological role as other neuropeptide degrading enzymes. However, the role of type I and bacterial PYRases remains unclear. The primary application of PYRase has been its utilization for some protein or peptide sequencing. Development of chromogenic substrates for this enzyme has allowed its use in bacterial diagnosis.

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“…The PYR-positive isolates could be further distinguished by using a bile-esculin test, for which only enterococci will give a positive result. A number of systems incorporating substrates for the PYR test on paper strips (65,88,96,120,167,234) and in liquid (28,93) and agar (74) media have been devised for the detection of PYRase (EC 3.4.11.8) and reviewed (51,70,152,166,234,242). Commercially available chromogenic tests based on the pyrrolidonyl carboxylic acid or pyroglutamic acid conjugates of ␤-naphthylamine, a colorimetric test based on a proprietary substrate (65), and one fluorogenic test based on a proprietary substrate (88,234), as well as their performances, have been summarized (152).…”

Section: Streptococcaceae and Related Organismsmentioning

confidence: 99%

Use of Enzyme Tests in Characterization and Identification of Aerobic and Facultatively Anaerobic Gram-Positive Cocci

Bascomb¹,

Manafi

1998

Clin Microbiol Rev

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SUMMARY The contribution of enzyme tests to the accurate and rapid routine identification of gram-positive cocci is introduced. The current taxonomy of the genera of aerobic and facultatively anaerobic cocci based on genotypic and phenotypic characterization is reviewed. The clinical and economic importance of members of these taxa is briefly summarized. Tables summarizing test schemes and kits available for the identification of staphylococci, enterococci, and streptococci on the basis of general requirements, number of tests, number of taxa, test classes, and completion times are discussed. Enzyme tests included in each scheme are compared on the basis of their synthetic moiety. The current understanding of the activity of enzymes important for classification and identification of the major groups, methods of testing, and relevance to the ease and speed of identification are reviewed. Publications describing the use of different identification kits are listed, and overall identification successes and problems are discussed. The relationships between the results of conventional biochemical and rapid enzyme tests are described and considered. The use of synthetic substrates for the detection of glycosidases and peptidases is reviewed, and the advantages of fluorogenic synthetic moieties are discussed. The relevance of enzyme tests to accurate and meaningful rapid routine identification is discussed.

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Rapid identification of enterococci by pyrrolidonyl aminopeptidase activity (PYRase)

Cited by 8 publications

References 5 publications

Novel chromogenic aminopeptidase substrates for the detection and identification of clinically important microorganisms

Novel chromogenic aminopeptidase substrates for the detection and identification of clinically important microorganisms

Pyrrolidone carboxyl peptidase (Pcp): An enzyme that removes pyroglutamic acid (pGlu) from pGlu‐peptides and pGlu‐proteins

Use of Enzyme Tests in Characterization and Identification of Aerobic and Facultatively Anaerobic Gram-Positive Cocci

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