Kinetic characterization of a novel acid ectophosphatase from Enterobacter asburiae
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1007/s12275-015-5354-3 http://hdl.handle.net/11449/231367 |
Resumo: | Expression of acid ectophosphatase by Enterobacter asburiae, isolated from Cattleya walkeriana (Orchidaceae) roots and identified by the 16S rRNA gene sequencing analysis, was strictly regulated by phosphorus ions, with its optimal activity being observed at an inorganic phosphate concentration of 7 mM. At the optimum pH 3.5, intact cells released p-nitrophenol at a rate of 350.76 ± 13.53 nmol of p-nitrophenolate (pNP)/min/108 cells. The membrane-bound enzyme was obtained by centrifugation at 100,000 × g for 1 h at 4°C. p-Nitrophenylphosphate (pNPP) hydrolysis by the enzyme follows “Michaelis-Menten” kinetics with V = 61.2 U/mg and K0.5 = 60 μM, while ATP hydrolysis showed V = 19.7 U/mg, K0.5 = 110 μM, and nH = 1.6 and pyrophosphate hydrolysis showed V = 29.7 U/mg, K0.5 = 84 μM, and nH = 2.3. Arsenate and phosphate were competitive inhibitors with Ki = 0.6 mM and Ki = 1.8 mM, respectively. p-Nitrophenyl phosphatase (pNPPase) activity was inhibited by vanadate, while p-hydroxymercuribenzoate, EDTA, calcium, copper, and cobalt had no inhibitory effects. Magnesium ions were stimulatory (K0.5 = 2.2 mM and nH = 0.5). Production of an acid ectophosphatase can be a mechanism for the solubilization of mineral phosphates by microorganisms such as Enterobacter asburiae that are versatile in the solubilization of insoluble minerals, which, in turn, increases the availability of nutrients for plants, particularly in soils that are poor in phosphorus. |
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Repositório Institucional da UNESP |
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Kinetic characterization of a novel acid ectophosphatase from Enterobacter asburiaeacid ectophosphataseATPaseEnterobacter asburiaeinhibitionp-nitrophenylphosphatepyrophosphataseExpression of acid ectophosphatase by Enterobacter asburiae, isolated from Cattleya walkeriana (Orchidaceae) roots and identified by the 16S rRNA gene sequencing analysis, was strictly regulated by phosphorus ions, with its optimal activity being observed at an inorganic phosphate concentration of 7 mM. At the optimum pH 3.5, intact cells released p-nitrophenol at a rate of 350.76 ± 13.53 nmol of p-nitrophenolate (pNP)/min/108 cells. The membrane-bound enzyme was obtained by centrifugation at 100,000 × g for 1 h at 4°C. p-Nitrophenylphosphate (pNPP) hydrolysis by the enzyme follows “Michaelis-Menten” kinetics with V = 61.2 U/mg and K0.5 = 60 μM, while ATP hydrolysis showed V = 19.7 U/mg, K0.5 = 110 μM, and nH = 1.6 and pyrophosphate hydrolysis showed V = 29.7 U/mg, K0.5 = 84 μM, and nH = 2.3. Arsenate and phosphate were competitive inhibitors with Ki = 0.6 mM and Ki = 1.8 mM, respectively. p-Nitrophenyl phosphatase (pNPPase) activity was inhibited by vanadate, while p-hydroxymercuribenzoate, EDTA, calcium, copper, and cobalt had no inhibitory effects. Magnesium ions were stimulatory (K0.5 = 2.2 mM and nH = 0.5). Production of an acid ectophosphatase can be a mechanism for the solubilization of mineral phosphates by microorganisms such as Enterobacter asburiae that are versatile in the solubilization of insoluble minerals, which, in turn, increases the availability of nutrients for plants, particularly in soils that are poor in phosphorus.Faculdade de Ciências Agrárias e Veterinárias de Jaboticabal Departamento de TecnologiaFaculdade de Ciências Agrárias e Veterinárias de JaboticabalSato, Vanessa SayuriGaldiano Júnior, Renato F.Rodrigues, Gisele ReginaLemos, Eliana G. M.Junior, João Martins Pizauro2022-04-29T08:44:58Z2022-04-29T08:44:58Z2016-02-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article106-113http://dx.doi.org/10.1007/s12275-015-5354-3Journal of Microbiology, v. 54, n. 2, p. 106-113, 2016.1976-37941225-8873http://hdl.handle.net/11449/23136710.1007/s12275-015-5354-32-s2.0-84957580599Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengJournal of Microbiologyinfo:eu-repo/semantics/openAccess2024-06-07T15:32:22Zoai:repositorio.unesp.br:11449/231367Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T21:21:31.047911Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Kinetic characterization of a novel acid ectophosphatase from Enterobacter asburiae |
title |
Kinetic characterization of a novel acid ectophosphatase from Enterobacter asburiae |
spellingShingle |
Kinetic characterization of a novel acid ectophosphatase from Enterobacter asburiae Sato, Vanessa Sayuri acid ectophosphatase ATPase Enterobacter asburiae inhibition p-nitrophenylphosphate pyrophosphatase |
title_short |
Kinetic characterization of a novel acid ectophosphatase from Enterobacter asburiae |
title_full |
Kinetic characterization of a novel acid ectophosphatase from Enterobacter asburiae |
title_fullStr |
Kinetic characterization of a novel acid ectophosphatase from Enterobacter asburiae |
title_full_unstemmed |
Kinetic characterization of a novel acid ectophosphatase from Enterobacter asburiae |
title_sort |
Kinetic characterization of a novel acid ectophosphatase from Enterobacter asburiae |
author |
Sato, Vanessa Sayuri |
author_facet |
Sato, Vanessa Sayuri Galdiano Júnior, Renato F. Rodrigues, Gisele Regina Lemos, Eliana G. M. Junior, João Martins Pizauro |
author_role |
author |
author2 |
Galdiano Júnior, Renato F. Rodrigues, Gisele Regina Lemos, Eliana G. M. Junior, João Martins Pizauro |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Faculdade de Ciências Agrárias e Veterinárias de Jaboticabal |
dc.contributor.author.fl_str_mv |
Sato, Vanessa Sayuri Galdiano Júnior, Renato F. Rodrigues, Gisele Regina Lemos, Eliana G. M. Junior, João Martins Pizauro |
dc.subject.por.fl_str_mv |
acid ectophosphatase ATPase Enterobacter asburiae inhibition p-nitrophenylphosphate pyrophosphatase |
topic |
acid ectophosphatase ATPase Enterobacter asburiae inhibition p-nitrophenylphosphate pyrophosphatase |
description |
Expression of acid ectophosphatase by Enterobacter asburiae, isolated from Cattleya walkeriana (Orchidaceae) roots and identified by the 16S rRNA gene sequencing analysis, was strictly regulated by phosphorus ions, with its optimal activity being observed at an inorganic phosphate concentration of 7 mM. At the optimum pH 3.5, intact cells released p-nitrophenol at a rate of 350.76 ± 13.53 nmol of p-nitrophenolate (pNP)/min/108 cells. The membrane-bound enzyme was obtained by centrifugation at 100,000 × g for 1 h at 4°C. p-Nitrophenylphosphate (pNPP) hydrolysis by the enzyme follows “Michaelis-Menten” kinetics with V = 61.2 U/mg and K0.5 = 60 μM, while ATP hydrolysis showed V = 19.7 U/mg, K0.5 = 110 μM, and nH = 1.6 and pyrophosphate hydrolysis showed V = 29.7 U/mg, K0.5 = 84 μM, and nH = 2.3. Arsenate and phosphate were competitive inhibitors with Ki = 0.6 mM and Ki = 1.8 mM, respectively. p-Nitrophenyl phosphatase (pNPPase) activity was inhibited by vanadate, while p-hydroxymercuribenzoate, EDTA, calcium, copper, and cobalt had no inhibitory effects. Magnesium ions were stimulatory (K0.5 = 2.2 mM and nH = 0.5). Production of an acid ectophosphatase can be a mechanism for the solubilization of mineral phosphates by microorganisms such as Enterobacter asburiae that are versatile in the solubilization of insoluble minerals, which, in turn, increases the availability of nutrients for plants, particularly in soils that are poor in phosphorus. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-02-01 2022-04-29T08:44:58Z 2022-04-29T08:44:58Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1007/s12275-015-5354-3 Journal of Microbiology, v. 54, n. 2, p. 106-113, 2016. 1976-3794 1225-8873 http://hdl.handle.net/11449/231367 10.1007/s12275-015-5354-3 2-s2.0-84957580599 |
url |
http://dx.doi.org/10.1007/s12275-015-5354-3 http://hdl.handle.net/11449/231367 |
identifier_str_mv |
Journal of Microbiology, v. 54, n. 2, p. 106-113, 2016. 1976-3794 1225-8873 10.1007/s12275-015-5354-3 2-s2.0-84957580599 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Journal of Microbiology |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
106-113 |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808129312745848832 |