Cloning and functional expression of zeta-carotene desaturase, a novel carotenoid biosynthesis gene from Ficus carica

Carotene desaturation, an essential step in the carotenoid biosynthesis pathway, is catalyzed by two enzymes, phytoene desaturase (PDS) and ζ-carotene desaturase (zeta carotene desaturase, ZDS). Here we describe cloning and E. Coli expression of zds-Fc, a novel Ficus carica ζ-carotene desaturase catalyzing dehydrogenation of ζ-carotene into neurosporene and finally lycopene. The ζ-carotene desaturase (ZDS) gene was amplified from the fig tree by rapid amplification of cDNA ends (RACE) and spanned a 1746 bp open reading frame (ORF), encoding a protein of 582 amino acid residues with a predicted molecular weight of 64kD. The N-terminal region of this polypeptide contained a putative transit sequence for plastid targeting. By phylogenetic and sequence analyses, zds-Fc showed high homology with previously described ζ-carotene desaturases from higher plant species (Al-Babili et al. 1998; Cong et al. 2009; Matthews et al. 2003; Yan et al. 2011). Additionally, sequence analysis revealed a high degree of conservation among plant ZDSs. The deduced ZDS protein, designated FcZDS, also contains an N-terminus dinucleotide-binding, followed by a conserved region identified in other carotene desaturase sequences. These data, taken together, confirm our cloned zds-Fc as an integral part of the ZDS family of protei

Cloning and functional expression of zeta-carotene desaturase, a novel carotenoid biosynthesis gene from Ficus carica

Carotene desaturation, an essential step in the carotenoid biosynthesis pathway, is catalyzed by two enzymes, phytoene desaturase (PDS) and ζ-carotene desaturase (zeta carotene desaturase, ZDS). Here we describe cloning and E. Coli expression of zds-Fc, a novel Ficus carica ζ-carotene desaturase catalyzing dehydrogenation of ζ-carotene into neurosporene and finally lycopene. The ζ-carotene desaturase (ZDS) gene was amplified from the fig tree by rapid amplification of cDNA ends (RACE) and spanned a 1746 bp open reading frame (ORF), encoding a protein of 582 amino acid residues with a predicted molecular weight of 64kD. The N-terminal region of this polypeptide contained a putative transit sequence for plastid targeting. By phylogenetic and sequence analyses, zds-Fc showed high homology with previously described ζ-carotene desaturases from higher plant species (Al-Babili et al. 1998; Cong et al. 2009; Matthews et al. 2003; Yan et al. 2011). Additionally, sequence analysis revealed a high degree of conservation among plant ZDSs. The deduced ZDS protein, designated FcZDS, also contains an N-terminus dinucleotide-binding, followed by a conserved region identified in other carotene desaturase sequences. These data, taken together, confirm our cloned zds-Fc as an integral part of the ZDS family of proteins

Potential use of plant proteolytic enzymes in hemostasis

The aim of this chapter is to review the development and state of the art in the application of plant proteases, and their effects on hemostasis. Reviewing the proteases that inhibit or enhance platelet aggregation, blood coagulation and fibrinolysis. The ultimate goal is the use of these plant proteases to improve current therapies and overcome drawbacks and deficiencies of the current drugs associated mainly with bleeding disorders. Different methods of extraction and identification have been investigated. Some effects of the identified proteases have been evaluated. We give here an overview of the latest advances in the identification and application of proteases in hemostasis.Fil: Pepe, Alfonso. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Tito, Florencia Rocio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Guevara, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentin