IAL 21557
INSTITUTO DE AGROBIOTECNOLOGIA DEL LITORAL
Unidad Ejecutora - UE
artículos
Título:
Purification and Characterization of a Glutathione Reductase from Phaeodactylum tricornutum
Autor/es:
D.G. ARIAS; V.E. MÁRQUEZ; A.J. BECCARIA; S.A. GUERRERO; A.A. IGLESIAS
Revista:
PROTIST
Editorial:
ELSEVIER GMBH
Referencias:
Lugar: Frankfurt; Año: 2010 vol. 161 p. 91 - 101
ISSN:
1434-4610
Resumen:
Glutathionereductase(E.C.1.8.1.7)waspurifiedfrom Phaeodactylumtricornutum cells grownaxenicallyinanautotrophicmedium.Theoverallprocedurestartedwithpreparationofthecellextractandadditionofammoniumsulfateto20%saturation,followedbyanionexchangeandaffinityinteractionchromatography(Blue-A-and2,5-ADP-Sepharose).Completepurificationrequirednativepolyacrylamidegelelectrophoresisasthefinalstep.Theenzymewaspurifiedtohomogeneityandfunctionallycharacterized.Itsnativemolecularmasswasestimatedtobe118kDa;whichcorresponds toadimer.Theenzymeexhibitedaspecificactivityof190Umg1 withanoptimalactivityatpH8.0and32 1C.Wedetermined Km values of14 lM and60 lM forNADPHandoxidizedglutathione,respectively.Productsinhibitedtheenzymeaccordingtoahybridping-pongreactionmechanism.AfterMALDI-TOFanalysis,thepurifiedenzymewasunambiguouslyidentifiedasoneofthe twoproteinsannotatedasglutathionereductasesinthegenomeofthediatom.Thepropertiesofthe enzymehelptounderstandredoxmetabolicscenariosin P.tricornutum.
