Isolation and Characterization of Hieronymain II, Another Peptidase Isolated from Fruits of <i>Bromelia hieronymi</i> Mez (Bromeliaceae)

From unripe fruits of <i>Bromelia hieronymi</i> Mez (Bromeliaceae), a partially purified protease preparation was obtained by acetone fractionation of the crude extract. Purification was achieved by anionic exchange chromatography (FPLC) on Q-Sepharose HP followed by cationic exchange ch...

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Detalles Bibliográficos
Autores principales: Bruno, Mariela Anahí, Trejo, Sebastián Alejandro, Avilés, Xavier F., Caffini, Néstor Oscar, López, Laura María Isabel
Formato: Articulo
Lenguaje:Inglés
Publicado: 2006
Materias:
Ciencias Exactas
Biología
Bromelia hieronymi
Bromeliaceae
cysteine proteinase
plant peptidases
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/131725
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-131725
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Exactas
Biología
Bromelia hieronymi
Bromeliaceae
cysteine proteinase
plant peptidases
spellingShingle Ciencias Exactas
Biología
Bromelia hieronymi
Bromeliaceae
cysteine proteinase
plant peptidases
Bruno, Mariela Anahí
Trejo, Sebastián Alejandro
Avilés, Xavier F.
Caffini, Néstor Oscar
López, Laura María Isabel
Isolation and Characterization of Hieronymain II, Another Peptidase Isolated from Fruits of <i>Bromelia hieronymi</i> Mez (Bromeliaceae)
topic_facet Ciencias Exactas
Biología
Bromelia hieronymi
Bromeliaceae
cysteine proteinase
plant peptidases
description From unripe fruits of <i>Bromelia hieronymi</i> Mez (Bromeliaceae), a partially purified protease preparation was obtained by acetone fractionation of the crude extract. Purification was achieved by anionic exchange chromatography (FPLC) on Q-Sepharose HP followed by cationic exchange chromatography (SP-Sepharose HP). Homogeneity of the new enzyme, named hieronymain II, was confirmed by SDS-PAGE and mass spectroscopy (MALDI-TOF-TOF). The molecular mass of was 23,411 Da, and maximum proteolytic activity (more than 90% of maximum activity) was achieved at pH 7.5-9.0 on casein and at pH 7.30-8.3 on Z-Phe-Arg-p-nitroanilide. The enzyme was completely inhibited by E-64 and iodoacetic acid and activated by the addition of cysteine. The N-terminal sequence of hieronymain II (AVPQSIDWRVYGAV) was compared with those of 12 plant cysteine proteases which showed more than 70% of identity. Kinetic enzymatic assays were made on Z-Phe-Arg-p-nitroanilide (K<sub>m</sub> = 0.72mM, k<sub>cat</sub> = 1.82 seg⁻¹, k<sub>cat</sub>/K<sub>m</sub> = 2.54seg⁻¹ mM⁻¹). No detectable activity could be found on PFLNA or Z-Arg-Arg-p-nitroanilide.
format Articulo
Articulo
author Bruno, Mariela Anahí
Trejo, Sebastián Alejandro
Avilés, Xavier F.
Caffini, Néstor Oscar
López, Laura María Isabel
author_facet Bruno, Mariela Anahí
Trejo, Sebastián Alejandro
Avilés, Xavier F.
Caffini, Néstor Oscar
López, Laura María Isabel
author_sort Bruno, Mariela Anahí
title Isolation and Characterization of Hieronymain II, Another Peptidase Isolated from Fruits of <i>Bromelia hieronymi</i> Mez (Bromeliaceae)
title_short Isolation and Characterization of Hieronymain II, Another Peptidase Isolated from Fruits of <i>Bromelia hieronymi</i> Mez (Bromeliaceae)
title_full Isolation and Characterization of Hieronymain II, Another Peptidase Isolated from Fruits of <i>Bromelia hieronymi</i> Mez (Bromeliaceae)
title_fullStr Isolation and Characterization of Hieronymain II, Another Peptidase Isolated from Fruits of <i>Bromelia hieronymi</i> Mez (Bromeliaceae)
title_full_unstemmed Isolation and Characterization of Hieronymain II, Another Peptidase Isolated from Fruits of <i>Bromelia hieronymi</i> Mez (Bromeliaceae)
title_sort isolation and characterization of hieronymain ii, another peptidase isolated from fruits of <i>bromelia hieronymi</i> mez (bromeliaceae)
publishDate 2006
url http://sedici.unlp.edu.ar/handle/10915/131725
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