Bioinformatics Evaluation of Plant Chlorophyllase, the Key Enzyme in Chlorophyll Degradation

Background and Objective: Chlorophyllase catalyzes the hydrolysis of chlorophylls to chlorophyllide and phytol. Recently, several applications including removal of chlorophylls from vegetable oils, use in laundry detergents and production of chlorophyllides have been described for chlorophyllase. However, there is little information about the biochemical characteristics of chlorophyllases.Material and Methods: 35 chlorophyllase protein sequences were obtained from the National Centre for Biotechnology Information database. All of the sequences were analyzed using bioinformatics tools for their conserved domain, phylogenetic relationships and biochemical characteristics.Results and Conclusion: The overall domain architecture of chlorophyllases consisted of the esterases/lipases superfamily domain over their full length and the alpha/beta hydrolase family domain over the middle part of their sequences. Plant chlorophyllases could be classified into 4 clades. Molecular weight and pI of the chlorophyllases ranged 32.65-37.77 kDa and 4.80-8.97, respectively. The most stable chlorophyllase is probably obtained from Malus domestica. Chlorophyllases form Solanum pennellii, Triticum aestivum, Triticum urartu, Arabidopsis lyrata, Pachira macrocarpa, Prunus mume and Malus domestica were predicted to be soluble upon overexpression in Escherichia coli, Beta vulgaris and Chenopodium album chlorophyllases were predicted to form no disulfide bond. Chlorophyllases from Jatropha curcas, Amborella trichopod, Setaria italica, Piper betle, Triticum urartu and Arabidopsis thaliana were predicted to be in non-N-glycosylated form.Conflict of interest: The authors declare no conflict of interest

Impact of different culture media on hairy roots growth of Valeriana officinalis L.

Transformed hairy root cultures of Valeriana officinalis were established by infection with Agrobacterium rhizogenes strain ATCC 15834. To determine the effect of different media on the growth of V. officinalis hairy roots, MS, B5 media (1.0X and 0.5X strength), N6 medium and a modified MS medium without phytohormones were used. In addition, different NH₄⁺ to NO₃^- ratios in MS medium were studied. The effects of these treatments were evaluated after 21 days of culture in relation to hairy root growth. B₅ and ½ B₅ media were the best basal media for hairy root growth. MS medium supplemented with a 20:20 ratio (mM) of NH₄⁺ to NO₃^- displayed highest growth rates and biomass yield in hairy root cultures. The present study demonstrated that the composition of culture medium and the ratio of different nitrogen sources have significant impact on the growth of V. officinalis hairy roots

Antibacterial effect of cerium oxide nanoparticle against Pseudomonas aeruginosa

Background Antibiotics have been widely used for the treatment of bacterial infections for decades. However, the rapid emergence of antibiotic-resistant bacteria has created many problems with a heavy burden for the medical community. Therefore, the use of nanoparticles as an alternative for antibacterial activity has been explored. In this context, metal nanoparticles have demonstrated broad-spectrum antimicrobial activity. This study investigated the antimicrobial activity of naked cerium oxide nanoparticles dispersed in aqueous solution (CNPs) and surface-stabilized using Pseudomonas aeruginosa as a bacterial model. Methods Gelatin-polycaprolactone nanofibers containing CNPs (Scaffold@CNPs) were synthesized, and their effect on P. aeruginosa was investigated. The minimum inhibitory and bactericidal concentrations of the nanoparticls were determined in an ATCC reference strain and a clinical isolate strain. To determine whether the exposure to the nanocomposites might change the expression of antibiotic resistance, the expression of the genes shv, kpc, and imp was also investigated. Moreover, the cytotoxicity of the CNPs was assessed on fibroblast using flow cytometry. Results Minimum bactericidal concentrations for the ATCC and the clinical isolate of 50 µg/mL and 200 µg/mL were measured, respectively, when the CNPs were used. In the case of the Scaffold@CNPs, the bactericidal effect was 50 µg/mL and 100 µg/mL for the ATCC and clinical isolate, respectively. Interestingly, the exposure to the Scaffold@CNPs significantly decreased the expression of the genes shv, kpc, and imp. Conclusions A concentration of CNPs and scaffold@CNPs higher than 50 μg/mL can be used to inhibit the growth of P. aeruginosa. The fact that the scaffold@CNPs significantly reduced the expression of resistance genes, it has the potential to be used for medical applications such as wound dressings.Medicine, Faculty ofNon UBCInfectious Diseases, Division ofMedicine, Department ofReviewedFacultyResearche