Preliminary functional characterization, cloning and primary sequence of Fastuosain, a cysteine peptidase isolated from fruits of Bromelia fastuosa.
The present work reports the characterization of Fastuosain, a novel cysteine protease of 25kDa, purified from the unripe fruits of Bromelia fastuosa, a wild South American Bromeliaceae. Proteolytic activity, measured using casein and synthetic substrates, was dependent on the presence of thiol reagents, having maximum activity at pH 7.0. The present work reports cDNA cloning of Fastuosain; cDNA was amplified by PCR using specific primers. The product was 1096pb long. Mature fastuosain has 217 residues, and with the proregion has a total length of 324 residues. Its primary sequence showed high homology with ananain(74%), stem bromelain (66%) and papain (44%).
Cabral
Hamilton
H
Departamento de Química e Ciências Ambientais,IBILCE-UNESP, State University of São Paulo, São José do Rio Preto SP, Brazil.
Leopoldino
Andréia M
AM
Tajara
Eloiza H
EH
Greene
Lewis J
LJ
Faça
Vitor M
VM
Mateus
Rogério P
RP
Ceron
Carlos R
CR
de Souza Judice
Wagner A
WA
Julianod
Luiz
L
Bonilla-Rodriguez
Gustavo O
GO
D000595:Amino Acid Sequence; D001483:Base Sequence; D031231:Bromelia / Q000201:enzymology*; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D017931:DNA Primers; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D018076:DNA, Complementary; D003546:Cysteine Endopeptidases; C509400:fastuosain, Bromelia fastuosa