A high-throughput screening for phosphatases using specific substrates

A high-throughput screening was developed for the detection of phosphatase activity in bacterial colonies. Unlike other methods, the current procedure can be applied to any phosphatase because it uses physiological substrates and detects the compelled product of all phosphatase reactions, that is, o...

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Autor principal: Senn, A.M
Otros Autores: Wolosiuk, R.A
Formato: Capítulo de libro
Lenguaje:Inglés
Publicado: Academic Press Inc. 2005
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Acceso en línea:Registro en Scopus
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024 7 |2 cas  |a fructose bisphosphatase, 9001-52-9; phosphatase, 9013-05-2 
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030 |a ANBCA 
100 1 |a Senn, A.M. 
245 1 2 |a A high-throughput screening for phosphatases using specific substrates 
260 |b Academic Press Inc.  |c 2005 
270 1 0 |m Senn, A.M.; Instituto Leloir, Fac. de Ciencias Exactas Y Naturales, Universidad de Buenos Aires, Patricias Argentinas 435, C1405BWE Buenos Aires, Argentina; email: asenn@leloir.org.ar 
506 |2 openaire  |e Política editorial 
504 |a Van Ommen Kloeke, F., Baty, A.M., Eastburn, C.C., Diwu, Z., Geesey, G.G., Novel method for screening bacterial colonies for phosphatase activity (1999) J. Microbiol. Methods, 38, pp. 25-31 
504 |a Boquet, P.L., Manoil, C., Beckwith, J., Use of TnphoA to detect genes for exported proteins in Escherichia coli: Identification of the plasmid-encoded gene for a periplasmic acid phosphatase (1987) J. Bacteriol., 169, pp. 1663-1669 
504 |a Pradel, E., Boquet, P.L., Acid phosphatases of Escherichia coli: Molecular cloning and analysis of agp, the structural gene for a periplasmic acid glucose phosphatase (1988) J. Bacteriol., 170, pp. 4916-4923 
504 |a Riccio, M.L., Rossolini, G.M., Lombardi, G., Chiesurin, A., Satta, G., Expression cloning of different bacterial phosphatase-encoding genes by histochemical screening of genomic libraries onto an indicator medium containing phenolphthalein diphosphate and methyl green (1997) J. Appl. Microbiol., 82, pp. 177-185 
504 |a Satta, G., Pompei, R., Grazi, G., Cornaglia, G., Phosphatase activity is a constant feature of all isolates of all major species of the family Enterobacteriaceae (1988) J. Clin. Microbiol., 26, pp. 2637-2641 
504 |a Manafi, M., Kneifel, W., Bascomb, S., Fluorogenic and chromogenic substrates used in bacterial diagnostics (1991) Microbiol. Rev., 55, pp. 335-348 
504 |a Cabib, E., Duran, A., Simple and sensitive procedure for screening yeast mutants that lyse at nonpermissive temperatures (1975) J. Bacteriol., 124, pp. 1604-1606 
504 |a Chaffin, D.O., Rubens, C.E., Blue/White screening of recombinant plasmids in gram-positive bacteria by interruption of alkaline phosphatase gene (phoZ) expression (1998) Gene, 219, pp. 91-99 
504 |a Thaller, M.C., Berlutti, F., Schippa, S., Selan, L., Rossolini, G.M., Bacterial acid phosphatase gene fusions useful as targets for cloning-dependent insertional inactivation (1998) Biotechnol. Prog., 14, pp. 241-247 
504 |a Satta, G., Pompei, R., Ingianni, A., The selective staining mechanism of phosphatase producing colonies in the diphosphatephenolphthalein-methyl green method for the detection of bacterial phosphatase activity (1984) Microbiologica, 7, pp. 159-170 
504 |a Satta, G., Grazi, G., Varaldo, P.E., Fontana, R., Detection of bacterial phosphatase activity by means of an original and simple test (1979) J. Clin. Pathol., 32, pp. 391-395 
504 |a Gee, K.R., Sun, W.C., Bhalgat, M.K., Upson, R.H., Klaubert, D.H., Latham, K.A., Haugland, R.P., Fluorogenic substrates based on fluorinated umbelliferones for continuous assays of phosphatases and beta-galactosidases (1999) Anal. Biochem., 273, pp. 41-48 
504 |a Guzman, L.M., Belin, D., Carson, M.J., Beckwith, J., Tight regulation, modulation, and high-level expression by vectors containing the arabinose pBAD promoter (1995) J. Bacteriol., 177, pp. 4121-4130 
504 |a Rodriguez-Suarez, R.J., Wolosiuk, R.A., High level expression in Escherichia coli: Purification and properties of chloroplast fructose-1,6-bisphosphatase from rapeseed (Brassica napus) leaves (1995) Photosynth. Res., 46, pp. 313-322 
504 |a Chen, P.S., Toribara, T.Y., Warner, H., Microdetermination of phosphorus (1956) Anal. Chem., 28, pp. 1756-1758 
504 |a Lin-Goerke, J.L., Robbins, D.J., Burczak, J.D., PCR-based random mutagenesis using manganese and reduced dNTP concentration (1997) BioTechniques, 23, pp. 409-412 
504 |a Sambrook, J., Fritsch, E.F., Maniatis, T., (1989) Molecular Cloning: A Laboratory Manual, , Cold Spring Harbor Laboratory Press Cold Spring Harbor, NY 
504 |a Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J., Protein measurement with the Folin phenol reagent (1951) J. Biol. Chem., 193, pp. 265-275 
504 |a Wolosiuk, R.A., Ballícora, M.A., Hagelin, K., The reductive pentose phosphate cycle for photosynthetic CO2 assimilation: Enzyme modulation (1993) FASEB J., pp. 622-637 
504 |a Fiske, C.H., Subbarow, Y., The colorimetric determination of phosphorus (1925) J. Biol. Chem., 66, pp. 375-400 
504 |a Chifflet, S., Torriglia, A., Chiesa, R., Tolosa, S., A method for the determination of inorganic phosphate in the presence of labile organic phosphate and high concentrations of protein: Application to lens ATPases (1988) Anal. Biochem., 168, pp. 1-4 
504 |a Cariani, L., Thomas, L., Brito, J., Del Castillo, J.R., Bismuth citrate in the quantification of inorganic phosphate and its utility in the determination of membrane-bound phosphatases (2004) Anal. Biochem., 324, pp. 79-83 
504 |a Wahler, D., Reymond, J.-L., High-throughput screening for biocatalysts (2001) Curr. Opin. Biotechnol., 12, pp. 535-544 
504 |a Goddard, J.-P., Reymond, J.-L., Enzyme assays for high-throughput screening (2004) Curr. Opin. Biotechnol., 15, pp. 314-322 
520 3 |a A high-throughput screening was developed for the detection of phosphatase activity in bacterial colonies. Unlike other methods, the current procedure can be applied to any phosphatase because it uses physiological substrates and detects the compelled product of all phosphatase reactions, that is, orthophosphate. In this method, substrates diffuse from a filter paper across a nitrocellulose membrane to bacterial colonies situated on the opposite face, and then reaction products flow back to the paper. Finally, a colorimetric reagent discloses the presence of orthophosphate in the filter paper. We validated the performance of this assay with several substrates and experimental conditions and with different phosphatases, including a library of randomly mutagenized rapeseed chloroplast fructose-1,6-bisphosphatase. This procedure could be extended to other enzymatic activities provided that an appropriate detection of reaction products is available. © 2004 Elsevier Inc. All rights reserved.  |l eng 
536 |a Detalles de la financiación: Agencia Nacional de Promoción Científica y Tecnológica 
536 |a Detalles de la financiación: Universidad de Buenos Aires 
536 |a Detalles de la financiación: Consejo Nacional de Investigaciones Científicas y Técnicas 
536 |a Detalles de la financiación: We are indebted to M. Berkmen and J. Beckwith (Harvard Medical School) for the generous gift of MB260 (agp) and MB65 (phoA) E. coli strains. This work was supported by a grant from the Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT) and the Universidad de Buenos Aires. Alejandro Senn holds a fellowship and Ricardo Wolosiuk is an investigator of the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). 
593 |a Instituto Leloir, Fac. de Ciencias Exactas Y Naturales, Universidad de Buenos Aires, Patricias Argentinas 435, C1405BWE Buenos Aires, Argentina 
690 1 0 |a ENZYME ACTIVITY 
690 1 0 |a HIGH-THROUGHPUT SCREENING 
690 1 0 |a PHOSPHATASES 
690 1 0 |a TRANSFORMED BACTERIA 
690 1 0 |a FRUCTOSE BISPHOSPHATASE 
690 1 0 |a PHOSPHATASE 
690 1 0 |a ARTICLE 
690 1 0 |a BACTERIUM COLONY 
690 1 0 |a BIOFILTER 
690 1 0 |a CONTROLLED STUDY 
690 1 0 |a ENZYME ACTIVITY 
690 1 0 |a ENZYME MECHANISM 
690 1 0 |a ENZYME SPECIFICITY 
690 1 0 |a HIGH THROUGHPUT SCREENING 
690 1 0 |a NONHUMAN 
690 1 0 |a PHYSIOLOGY 
690 1 0 |a PRIORITY JOURNAL 
690 1 0 |a RAPESEED 
690 1 0 |a BACTERIA (MICROORGANISMS) 
690 1 0 |a SINAPIS ARVENSIS 
650 1 7 |2 spines  |a MUTAGENESIS 
700 1 |a Wolosiuk, R.A. 
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