Journal of Applied Biotechnology Reports
Volume:2 Issue: 4, Autumn 2015

Endoplasmic reticulum has a critical role in the synthesis and folding of secretory and membrane proteins. High accumulation of proteins in ER activates the unfolded protein response and glucose regulated protein 78 or GPR78 plays an essential role in this pathway. Unfolded protein response is activated in cancerous cells due to their adverse condition to survive and it has been shown that GRP78 can be expressed in tumor cell membrane. Overexpresion and localization of GRP78 makes it a suitable target for the treatment of cancer. This review describes cellular localization, biological function, and role of GRP78 in cancer induction. Methods for tumor inhibition via GRP78 are also discussed.

Cotton is one of the most important crops in the world and increasing cotton yield is the main goal in the cotton industry. One important factor for crop improvement is increasing produced fibers by each developing seed. It is very interesting to manipulate fiber properties such as length, micronaire, color and strength which necessitate cotton ovule culture. Ovule culture is used as a tool to study physiology and biochemistry of secondary cell wall synthesis, effects of plant growth regulators, nutrition and environmental conditions on fiber and ovule development, inter-specific hybridization and embryo recue. This technique can be used for analysis of functional genes in fibers as transient expression systems. In this regard, fiber specific promoters should be identified in developing fibers. Optimum growth regulator combinations can increase fiber yield and uniformity in vitro conditions. Despite mature ovules exogenous Indole-3-acetic acid and gibberellic acid are required for fiber development of unfertilized ovules in vitro condition even though these two hormones also induce fiber production in fertilized ovules. On the whole cotton ovule culture can be used as a model before permanent cotton transformation and field trials.

In this report, we describe a procedure for in-silico design of a novel haloalkane dehalogenase protein that exhibits luciferase property which can be potentially used in biosensor applications. From a PDB BLAST search, the selected haloalkane dehalogenase (PDB code: 1EDE) had a close structural homology with a lucifearse (PDB code: 2PSJ chain A) sharing an identity of 33%. Initially, the amino acids in luciferase interacting with chromophore colentrizine were analyzed by a 3 ns molecular dynamics simulation. Later, Colentrizine was docked to the haloalkane dehalogenase followed by 5 ns molecular dynamics simulation to find out the frequently interacting residues with the ligand and amino acid Asp170, Lys192, Arg193, Lys259, Lys261 were selected in haloalkane dehalogenase structure for substitution purpose. Four mutants were generated by substituting these positions with Phe, Tyr, Trp respectively as they have comparable molecular weight. Following several selection strategies based on energy minimization, structural accuracy, ligand binding score, % of hydrophobicity and aromaticity it was observed that the protein substituted with Phe in all the positions is the best one which was further validated by a 10 ns molecular dynamics simulation by Gromacs 4.5.0 software. The selected designed protein is further analysed for their substrate specificity towards 10 selected haloalkane ligands in comparison to the unmutated counterpart by Autodock 4.2 tool. The result shows, that the designed protein has also improved the substrate specificity towards four toxic pollutants.

An efficient method for producing doubled haploid plants of date palm (Phoenix dactylifera L.) was established using in vitro colchicine treatment of anthers. To this study male inflorescences were harvested in the appropriate developmental stage. Separated anthers after one day cold (4°C) pretreatment, in uninucleate stage of microspore were transferred to the modified Y3 medium containing different concentrations of colchicine (125- 250- 500- 1000 mgl-1) for one of four treatment durations (12- 24- 36- 48 hours) and were compared to control anthers (without colchicine treatment). Based on results, different concentrations of colchicine, different exposure times and their interaction had a significant different effect on callus induction. No callus and embryo were produced from control anthers; however, the callus and embryo were induced from colchicine-treated anthers. Among used concentration in 1000 mgl-1 and 125 mgl-1, the range of callus was very low, and also the 500 mgl-1 concentration and duration of 12 hours had the highest range of callus. In concentration of 500 mgl-1 and duration of 36 hours some direct embryo was observed. These results indicate that the colchicine treatment of date palm anthers can induce callus and embryo for the production of doubled haploid lines.

Copper sulfate was used as a precursor to prepare CuO nanoparticles in reverse micelles (o/w microemulsion). This is a technique which allows the preparation of ultrafine metal oxide nanoparticles within the size ranging from 50 to 60 nm. The preparation of nano copper (II) oxide studied was investigated in the inverse microemulsion system. Therefore the nucleation of metal particles proceeds in the water capsules of the microemulsion. Tween 80 was added as surfactant. The products were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The study provides a simple and efficient route to synthesize CuO nanoparticles at room temperature.

Proteases catalyze the hydrolysis of peptide bonds in proteins. They are widelydistributed in nature, nearly in all plants, animals and microorganisms. Plant latex is a rich source of proteases. Latex production is unique property of Euphorbian plant, which could be a potential source of proteolytic enzymes. The highest proteolytic activity was observed in the latex of Pedilanthus tithymaloides Linn., which is followed by lattices of Euphorbia tirucalli Linn., Euphorbia nivulia Buch.-Ham., and Euphorbia nerifolia Linn. Therefore, P. tithymaloides, E. tirucalli, E. nivulia, and E. nerifolia lattices were selected as potential sources of proteases. These proteases were effectively used to remove hair from goat skin indicating its potential in leather processing industry. These proteolytic enzymes showed potential environmental waste management applications such as degradation of chicken feather waste. Additionally, the crude enzymes of the selected plants exhibited potentgelatinolytic activity almost correlated to release of silver from waste X–ray film. They also showed remarkable destaining property, indicating their importance in detergent industry. These enzymes have remarkable silk degumming property demonstrating their use in textile industries.

The use of organophosphorus hydrolase (OPH) enzyme to degrade Chemical Warfare Agents is one of the most frequently used decontamination methods. OPH is a ~36 kDa homodimeric metalloprotein that is found in the membrane of Flavobacterium sp. strain ATCC 27551 and Brevundimonas diminuta MG and is capable of hydrolyzing a wide range of oxon and thion , such as paraoxon and parathion. OPH gene (opd) has been expressed in many hosts, such as bacteria, insect cells, fungi, and Streptomyces spp. High level and soluble expression and correct folding of each protein are of important factors. Fusion proteins, including TRX, Gb1 and MBP, are commonly used to increase solubility, folding and in some cases, stability. The present study evaluated thioredoxin (TRX) role in OPH expression level, solubility and stability by cloning the opd gene into pET32a and pET21a and expressing the resulting vectors in E. coli shuffle T7. The pET32a vector encodes a fusion protein containing TRX that is not present in the pET21a. The results revealed an increased expression level, solubility and stability in OPH produced by the pET32a-opd construct compared to the pET21a vector due to the presence of the TRX fusion in pET32a vector.