ABSTRACT
Aims: Ribulose 1,5-bisphosphate (RuBP) carboxylase/oxygenase (RubisCO) catalyses a key reaction by which inorganic carbon is assimilated into organic carbon found in the biosphere. The present study was aimed to isolate this enzyme from leaves of Phytolacca dioica L. Study Design: In this work, first crude extracts from leaves at different stages of development were assayed to isolate this enzyme, then young leaves of P. dioica were used considering high value of specific activity. Methodology: Classical methods for protein isolation have been used to characterise RubisCO from P. dioica leaves. Results: RubisCO was isolated from young leaves by gel-filtration. The pure RubisCO showed two predominant bands (56- and 15-kDa) by SDS-PAGE. N-terminal sequences data on large (56 kDa) and Small (15 kDa) subunits obtained by automatic Edman degradation show a high percentage of identity with large and small subunit of other RubisCo enzymes. Moreover, the N-terminal amino acid sequence obtained by Edman degradation of the expressed large subunit (56 kDa) corresponds to the traduced one found by the analysis of the chloroplast genome of P. dioica (access number AFU65422). Conclusion: The data on RubisCO from young leaves of Phytolacca dioica L. (RubisCO-Pd), obtained in the present work, could be used as the starting point for biological characterization of this enzyme.
ABSTRACT
The introduction of generics drugs has brought the need for more control of their quality and purity. In Italy from June 23, 2013 the Pfizer® has no longer the patent for the Viagra’s production and other industries produce equivalent products containing Sildenafil citrate. Study design: Thus, in this work, the chemical profiles of both Viagra Italian Pfizer® and 3 Italian commercial Sildenafil citrate tables (generic pharmaceutical manufacturers) for male erectile dysfunction were obtained by using UltraPerformanceLiquid Chromatography (UPLC). Methodology: UPLC methodology was successfully used for the assay of Sildenafil citrate in different products in Italy which are under the cover of alternative systems of medicine. Results: The results show that: i) the chromatographic profiles obtained from Italian Sildenafil citrate tablets are identical and not present other active pharmaceutical ingredients; ii) the commercial samples have a quantity of Sildenafil citrate comparable with the corresponding labelled amounts. Conclusion: The UPLC method can be used for determination of Sildenafil citrate tables marketed by generic pharmaceutical manufacturers in Italy.