title: Purification, biochemical characterization and cloning of a new cationic peroxidase isoenzyme from artichoke.
abstract: A cationic soluble peroxidase isoenzyme (CysPrx) has been purified and characterized from artichoke (Cynara cardunculus subsp. scolymus (L.) Hegi) leaves by combination of aqueous two phase extraction, ion exchange chromatography, and gel filtration. The purification fold was 149 and the activity recovery 5.5%. CysPrx was stable from 5 to 45 °C with a pH optimum around 5.5; the pI was 8.3 and the MW of 37.7 ± 1.5 kDa. MALDI-TOF MS analysis provided partial peptide sequences and resolved CysPrx isoenzyme into two putative isoforms. The presence of these isoforms was confirmed by the isolation of full-length cDNA encoding CysPrx that generate two slightly different sequences coding for two putative CysPrx: CysPrx1 and CysPrx2. The obtained MS peptides showed a 35% coverage with 100% identity with the two CysPrx deduced protein sequences. A molecular modeling analysis was carried out to predict in silico the protein structure and compare it with other plant Prx structures. Considering that CysPrx is quite stable, the study carried out in this paper will offer new insights for the production of the recombinant protein for utilization of CysPrx as an alternative Prx for food technology, biomedical analysis and bioremediation.
authors.affiliation: Institute of Sciences of Food Production-CNR, Bari, Italy. angela.cardinali@ispa.cnr.it
authors.forename: Angela
authors.initials: A
authors.lastname: Cardinali
authors.forename: Nunzia
authors.initials: N
authors.lastname: Tursi
authors.forename: Angela
authors.initials: A
authors.lastname: Ligorio
authors.forename: Maria Gabriella
authors.initials: MG
authors.lastname: Giuffrida
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Napolitano
authors.forename: Rocco
authors.initials: R
authors.lastname: Caliandro
authors.forename: Lucrezia
authors.initials: L
authors.lastname: Sergio
authors.forename: Donato
authors.initials: D
authors.lastname: Di Venere
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Lattanzio
authors.forename: Gabriella
authors.initials: G
authors.lastname: Sonnante
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D031943:Cynara scolymus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D018076:DNA, Complementary / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010455:Peptides / Q000032:analysis*; D010544:Peroxidases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D010802:Phylogeny; D018515:Plant Leaves / Q000201:enzymology*; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D018076:DNA, Complementary; D007527:Isoenzymes; D010455:Peptides; D010940:Plant Proteins; D010544:Peroxidases