Catecholamines are active plant-based drug compounds in Pisum sativum, Phaseolus vulgaris and Vicia faba Species

Introduction: Catecholamines (L-DOPA and dopamine) are the key metabolites found in nervous system and their endogenous deficiency is associated with different patho-physiological disorders. Therefore, it is important to screen the new herbal sources of catecholamines for drug preparation. In this study, the amount of L-DOPA and dopamine were investigated in the leaves and roots of three species from legume family such as Pisum sativum (garden pea), Phaseolus vulgaris (haricot bean) and Vicia faba (broad bean); using TLC and HPLC. Methods: The seeds of P. sativum, P. Vulgaris and V. faba were treated and cultured under the glasshouse conditions. The extraction from 1 gram of each plant sample was obtained and assayed for L-DOPA and dopamine using thin layer chromatography (TLC) and reversed-phase HPLC. Results: The results indicated that all cultivars accumulated different levels of L-DOPA and dopamine in leaves and roots. The quantitative results showed that the metabolites concentrations were high in the leaves of P. Sativum and V. faba compared to that in roots. Conclusion: The present study may provide a new avenue for preparation and estimation of L-DOPA and dopamine from plant sources and may be used for further analysis and therapeutic studies.</p

A r c h i v e o f S I D Iranian Chemical Society Molecular Cloning, Expression and Sequence Analysis of DNA Polymerase I from an Iranian Thermophilic Bacterium, Bacillus sp. G (2006)

Thermostable DNA polymerases are widely used in DNA amplification reactions such as the Polymerase Chain Reaction (PCR), requiring the activity of the enzymes at high temperatures. The aim of the present study was to assess the potential biotechnological capabilities of Iranian thermostable DNA polymerases. To this end, we cloned the gene encoding a DNA polymerase from a novel thermophilic eubacterium, Bacillus sp. G (2006). Phylogentic analysis of this gene revealed that the new isolate belongs to the genera Bacillus. Sequence analysis of the fragment produced by degenerate primers also showed that it consists of 2,631 bp encoding an 876 amino acid protein, and subsequent amino acid sequence analysis of this DNA polymerase showed that it belongs to family A-type DNA polymerases. The expression vector pET28a (+) was chosen for expression of the gene fragment in the mesophilic host bacterium E. coli BL21. This expression vector has some advantages such as attachment of a Poly-His tag to the N-terminus of the protein for the ease of purification and a powerful promoter of lac-Z induced by IPTG. The band corresponding to the protein product was observed in the molecular weight range of about 100KDa on the SDS-PAGE gel after heat and Ni +2 -NTA column chromatography. Using the dot blot technique, the polymerase activity of the enzyme was qualitatively confirmed at 70 °C. Therefore, it is suggested that optimizations of this activity could make this enzyme appropriate for PCR processes in future

Renilla luciferase-labeled Annexin V: a new probe for detection of apoptotic cells.

The Ca2+-dependent binding of Annexin V to phosphatidylserine on cell surfaces is a reliable marker for apoptosis that is widely used in flow cytometry based apoptosis assays. In this paper, we report a new class of Annexin V-based probes for apoptosis. Luciferase from Renilla reniformis (RLuc) was linked to Annexin V and expressed successfully in a soluble form in Escherichia coli BL21 (DE3). The new probe, Rluc/Annexin V, was purified and functionally assayed for detection of apoptosis in actinomycin D-induced apoptotic Jurkat cells. Moreover, the spontaneous apoptosis in neutrophils was shown using the new probe. The results indicate that Rluc/Annexin V can bind to the apoptotic cells, and the signal of Renilla luciferase can be detected by luminometric measurements. The availability of Rluc/Annexin V may be of potential commercial interest for improving current apoptosis assays