Journal of Applied Biotechnology Reports
Volume:1 Issue: 1, Winter 2014

Organophosphorus chemicals are compounds which have been used as pesticides and insecticides in agriculture. They’re also used as nervous agents and have raised many problems for human and environment. Among the most important methods of decontamination from these compounds are biodegradation methods. Using OPH enzyme in degradation the mentioned compounds is seen as one of the desirable ways, but low activity and specification and low thermostability are among factors significantly decreasing the optimal application of this enzyme. Using methods of protein engineering based on the alteration of specific protein positions in order to improve the activity, specification and thermostability are some common ways used currently. Numerous studies have been done to increase activity and thermostability of OPH enzyme with alteration of some special amino acids the result of which was an increase against different substrates. OPH enzyme active site connected to substrates that consisted of three large, small and releasing packets were one of the goal areas of changing amino acids used by researchers to improve engineered activities. Among other ways of making enzymes more rigid and stable were bending loops by replacing Proline, creating disulfide bonds, ionic bonds by replacing charged amino acids.

Mammalian ∆-(1)-Pyrroline-5-carboxylate synthase (P5CS) enzyme catalyzes the coupled phosphorylation and reduction-conversion of glutamate to ∆-(1)-pyrroline-5-carboxylate (P5C), a critical step in the proline, ornithine, citrulline and arginine biosynthesis. In plants and mammals, P5CS consists of two separate enzymatic domains: N-terminal γ-glutamyl kinase (γ-GK) and C-terminal γ-glutamyl phosphate reductase (γ–GPR). Hyperammonemia has been reported as a new inborn disorder, with a range of clinical symptoms which is associated with a reduced synthesis of proline, ornithine, citruline and arginine. A missense mutation, R84Q, which alters the conserved residue in γ-GK domain, is responsible for this disorder. In this study using in-silico approaches as a new bioinformatics method, sequence analysis was performed and the tertiary structure of γ-GK domain of human P5CS, which includes the R84Q missense mutation, was predicted and the mutation effects on structural and functional features of P5CS enzyme were analyzed. Our analysis showed that this substitution has an affect on the molecular surface accessibility and total energy of the modeled structure. We conclude that this mutation results in a reduced activity of P5CS enzyme and an impaired synthesis of these amino acids.

Androgenesis via anther culture or microspore culture is one of the current methods for producing haploid and double haploid plants in maize. To use of this method in maize breeding program it should be able to regenerate enough plantlets. Anthers culture usually is carried out indirectly via callus induction and regeneration on at least two different media. In this study a responsive genotype, ETH-M82, using a new single culture medium was used for embryogenesis and regeneration. We tested different growth regulators (2,4-D; kinetin; NAA & IAA) in modified YP medium. After6-week, direct regeneration on some treatments was observed. The highest frequency of direct formation of plantlets (in 100 anthers) occurred on medium supplemented with 2mgl-1 IAA and 2mgl-1 kinetin (4%).Best results with an average of 3.1plantlets in replication were obtained with the medium solidified with agar, while in difcobactoagar only 1.4 of plantlets in replication was produced. This experiment suggested that agar and plant growth regulators in the medium were beneficial for producing embryo and plantlet from maize anthers.

DNA microarrays consist of collection of DNA microscopic spots that In order to form an array attached to a solid surface such as glass, plastic or silicon chip. The pieces of fixed DNA considered as a searcher. In this technology it is possible to test sample against thousands probes for specific genes. With this ability, arrays accelerate the biological investigations, gene finding, molecular detection and disease diagnosis. Microarray technology can be seen as a continued development of southern blotting. The most important stage in this technology is data analysis. To analysis such large data whit high degree of confidence and reliability needs reliable bioinformatics tools. Infectious diseases still is major problem for human. One of the most important application of microarray technology is the possibility of testing for the presence of thousands micro-organism in environmental and clinical samples only in a single excrement. Thereby we take an important step in rapid and accurate detection of infectious diseases. Here, we present E-Predict algorithm and DetectiV package that is based on species identification in microarray. We demonstrate the application of E-Predict and DetectiV for viral detection in a large publicly available dataset and show that DetectiV performs better than E-Predict. DetectiV is implemented as a package for R - powerful, open source software for statistical programming - that containing visualization, normalization and significance testing functions.s

The effects of different factors on growth and bio-oxidation efficiency of a native strain of Acidithiobacillus ferrooxidans have been evaluated by the utilization of response surface methodology, RSM. Medium pH and iron concentration were found to be the most significant factors while temperature and ammonia concentrations had the least weight within the ranges investigated. Optimum operational conditions for maximizing Fe2+ oxidation were found to be 31 oC, 7 g/Liron concentration, 4.5 g/Ltotal ammonium salt concentration and medium pH 1.85. Maximum recovery of 98% of Zinc is the main outcome of results as observed at 7 g/Lof Fe2+, under optimized experimental conditions. The response of a bacterial strain to metals toxicity also studied. The isolate showed good resistance to most of the toxic metals. The proteomics approach was used to identify the differentially expressed proteins under heavy metal stress. Four of the differentially expressed proteins were identified as major outer membrane protein of A. ferrooxidans, ribulose large bisphosphate carboxylase subunit, and holo synthase.

T-2 toxin is the most poisonous trichothecene produced by Fusarium species especiallyF.sporotrichioides. T-2 toxin is a biological contaminant in a number of agricultural commodities that can cause severe diseases among humans and animals and even lead to death. The aim of the current study is the analysis of T-2 mycotoxin in Fusarium species by high-performance liquid chromatography (HPLC) combined with florescence detection and derivatization with 1-antroylnitrile (1-AN). Totally, 11Fusariumisolates and reference strains were studied. The isolates were tested for the T-2 toxin production after growing on rice substrate followed by using specific “Multisep 225 Trich Clean up columns” purification. In this study, T-2 toxin production was ranged from 197.05ug/kg to 8503.07ug/kg. This is the first study of T-2 toxin analysis by HPLC-F in Iran.

Non-beneficial and harmful weeds are plants that are unwanted, outside their home farms are growing and have the potential to exceed. This study was done in order to screening fungai and isolating NLP phytotoxine from them for selective biocontrol of Amaranthus retroflexus and Rumes crispus as a dicot, common and chemical herbicide resistance weeds. NLPs are effective just on dicot plants. Contaminated soil and dicotyledons plants were Collected from different regions of Iran. after collecting and culturing them, The effect of Supernatant from fungal cultures, was asseyd by spraying of 5 µl /cm3 of it mid 20 µl tween-20 on leaves of Amaranthus retroflexus, Rumes crispus and wheat as negative control that were cultured in MS media and pots in 3 replications with completely randomized design in laboratory and research green house of baqiatallah university. The effects were assessed according to numbering method. Finally, the QAT5and G7-1strains was selected from 9 top strains, because was more destructive than others on Amaranthus retroflexus and Rumes crispus respectively from necrosis to cell death with number 4 according to numbring method and has non-harmful effect on the wheat (Triticum aestivum). SDS-page results showed phytotoxine that was produced by QAT5 strain was a protein and this from G7-1 was non-protein. For better result on SDS-page protein was concentrated using by ammonium sulfat method, but about G7-1 again this outcomewas repeated. The protein purification of QAT5strain using FPLC showed the presence of a protein with about 24 kDa like other family members of this protein. Considering this fact that these phytotoxines according to the result had similarity features to what founded befor about NLPs, they are recommended as biocontrol factor of these weeds insteade of chemical herbicides.