Iranian Journal of Biotechnology
Volume:12 Issue: 3, Summer 2014

Curcumin as an antibacterial agent is a yellow natural compound extracted from turmeric root. Nanoparticles can decrease the defects of usual drug delivery systems. Chitosan is a low toxic, biodegradable, biocompatible and safe polymer which is used in production of nanoparticles. Nanoparticles including chitosan-tripolyphosphate (TPP) can increase antibacterial properties of curcumin. Curcumin loaded chitosan-TPP nanoparticles were synthesized by ionotropic gelation method from chitosan, curcumin and TPP salt. The skin of anesthetized mice was inoculated with staphylococcus aureus and pseudomonas aeruginosa. The infected mice were treated with curcumin loaded chitosan-TPP nanoparticles during 3 days. Antibacterial characteristics of curcumin-nanoparticles treated mice were evaluated by bacterial culture of infected mice. Our results showed the size of 160 ± 10 nm and the charge of +7 ± 2 mV in curcumin loaded chitosan-TPP nanoparticles. These nanoparticles were also spiral shape. The encapsulation efficiency of curcumin loaded chitosan-TPP nanoparticles was 75 ± 2%. Bacterial culture showed that curcumin loaded chitosan-TPP nanoparticles inhibited staphylococcus aureus and pseudomonas aeruginosa growth. Our study demonstrated that curcumin loaded chitosan-TPP nanoparticles can be utilized as a potent agent in treatment of Staphylococcus aureus and Pseudomonas aeruginosa infections.

Epidermal growth factor receptor (EGFR) has been shown to play a critical role in tumor cell growth and its overexpression has been observed in many epithelial tumors. In the field of cancer vaccine research, displaying the peptide mimotope on the surface of phage particles has shown promising results. In this study using m13-PVIII phage display system, two constructs were prepared: triple tandem repeat of EGFR mimotpe displaying particles (3M) and single EGFR mimotope displaying phage particle (1M). To investigate the anti-tumor properties of phage vaccine, C57BL/6 mice Lewis lung carcinoma xenograft model was established and treated with 3M phage vaccine, 1M phage vaccine and control agents. Immunization of mice with these phage-based vaccines showed strong immune response against phage-mimotope. 3M phage vaccine showed more potency against tumor in comparison with control groups. Also the survival time was extended in phage vaccine treated tumor-bearing mice compared with untreated mice. Our findings suggest that mimotope-displaying phage vaccine can induce specific antibodies with antitumoral activity, which its potential as a candidate vaccine for EGFR-specific cancer immunotherapy needs to be more investigated in future studies.

Proteolytic enzymes have an important role in variety of physiological and pathological functions. They have been used in therapeutic and pharmaceutical applications. Characterizations of extracellular proteases from various strains of S. marcescens indicate that most strains produce a very similar major metalloprotease. This metalloprotease (serrapeptidase, serrapeptase) is an important pharmaceutical agent. Serrapeptase has been used in Asian and European countries for the treatment of inflammatory diseases, cardiovascular disorders, and bacterial infections. Purification and Characterization of Extracellular Metalloprotease from Serratia sp. ZF03 for therapeutic purposes. In this study the protease gene encoding a zinc-metalloprotease was isolated from the previously isolated red-pigmented Serratia sp. ZF03. The gene was sequenced and submitted to the GenBank. Proteolytic activity was detected by skim milk agar plate method and zymography. This fragment was found to encode an extracellular zinc-metalloendopeptidase with a molecular weight of approximately 50 kDa. The metalloprotease was purified by ammonium sulfate precipitation and dialysis, and then characterized. The effects of various inhibitors and reagents on protease activity and its kinetic parameters were also determined. The nucleotide sequence demonstrated that deduced amino acid sequence has high identity with those of metalloprotease from serralysin family. Production of metalloprotease was highest at 48th h of cultivation. Optimum protease activity occurred at a temperature range of 50˚C-55˚C and a pH range of 8.0-10. EDTA as a metal chelator, significantly inhibited protease activity. Zymography and inhibition assays showed that metalloprotease is the major secreted protease of Serratia sp. ZF03. The kinetic parameters, Km and Vm, were 0.00105 mg/ml and 0.0531 mM/min, respectively. Since the metalloprotease of this strain has strong proteolytic properties and good stability, it can be suitable candidate to be used as an effective drug in the medicine and pharmaceutical industries.

production of pharmaceutical proteins via genetic engineering of green plant is a promising field in biotechnology. Long time required to generate transgenic lines is major drawback of stable transformation techniques. Transient gene expression is a quick method to produce recombinant proteins in plants. The main goal of the present study was to evaluate agroinfiltration as an efficient and rapid method for production of recombinant antigen of FMDV. tobacco leaves were transformed via agroinfiltration using a needle-free syringe. Presence of the gene cassette was verified by polymerase chain reaction (PCR). Expression of the foreign gene was evaluated using Real Time PCR, protein dot blot and enzyme linked immunosorbent assay (ELISA). PCR analysis confirmed successful transformation of plant leaves. Expression of foreign protein was confirmed at both transcription and translation levels. Results of Real Time PCR assay indicated that the foreign gene was transcribed in transformed leaves. ELISA results showed that the foreign gene was expressed in transformed leaves in high level. In this investigation, we demonstrated the efficacy of agroinfiltration for transient expression of FMDV coat protein in tobacco plants. Based on the results obtained in this investigation, we believe that this method can be used as an effective and quick way for expression of recombinant antigens in plant tissues.

Interleukie-8 is a proinflammatory cytokine and a potent chemotactic and activation factor for neutrophils. It has been reported to be expressed in high levels by various cell types after immune stimulation and mediates neutrophil recruitment in host. The aim of this study was to analysis of the relationship between single nucleotide polymorphisms (SNPs) at position -180 G/A in the promoter region of bovine IL8 gene with milk production traits and somatic cell score (SCS). The part of promoter region in the bovine IL8 gene containing -180 G/A SNP was screened by single strand conformation polymorphism (SSCP) and DNA sequencing in Holstein cattle of Iran. The association analysis between different genotypes of IL8-180 SNP and performance traits of Holstein cattle was carried out by Mixed procedure of SAS 9.1 program. A total of 3 distinct SSCP patterns were observed. Sequence analysis indicated a reported polymorphism at position -180 G/A relative to the start codon. This SNP created a putative binding site for Oct-1 transcription factor which associated with lower SCS. The association of IL8-180 genotypes was studied with milk production traits and SCS. The IL8-180 associated (P)

In Algeria the date wastes production is estimated to be 85.000 tons. Date wastes are an economical source of carbohydrates for conversion to industrial enzymes because it is readily available and relatively low priced. The aim of the present study was to investigate the potential of using date wastes as a substrate for the production of α-amylase and invertase. Thirty strains of A. niger were isolated from the saline soils collected from five arid locations in Algeria. The process parameters; time, temperature, sugar content, initial pH, nitrogen source, nitrogen and phosphorus content, on the production of these enzymes were optimized. The obtained results show that the strain of Aspergillus niger ANSS-B5 produced the highest level of these enzymes. For α-amylase production, the cumulative effect of fermentation period of 96 h, temperature of 30°C, sugar content of 20 g/L, initial pH=5.5, supplemented with yeast extract as nitrogen source, and yeast extract and potassium phosphate at 5.0 g/L, content during the fermentation process of date wastes syrup produced α-amylase levels up to 285.6 U/ml. Invertase to 195.56 U/ml were produced under optimum conditions of 96h, temperature of 30°C, initial pH=6.0, sugars content of 40.0 g/L and utilisation of yeast extract and potassium phosphate at concentrations of 11.0 and 3.5 g/L, respectively. It is concluded from these results, using date wastes product as carbon source is very interesting for industrial scale application for the production of these enzymes.

Many plant growth-promoting bacteria including Rhizobia contain the 1-aminocyclopropane-1-carboxylate (ACC) deaminase enzyme that can cleave ACC, and thereby lower the level of ethylene in stressed plants. Drought and salinity are the most common environmental stress factors for plants in Iran. The main aim of this research was development of biofertilizers containing ACC deaminase enzyme which is very important in drought and salinity stressed conditions.In this research 168 isolates of native Sinorhizobium meliloti were evaluated for ACC deaminase activity. These isolates were classified in four groups based on growth rate on ACC containing medium and enzyme activity. One isolate from each group was selected for molecular characterization. The nucleotide sequence of 16S rRNA gene of selected isolates was determined. The ACC deaminase genes (acdS) on total and chromosomal DNA of S. meliloti KYA40, and KYA71 strains were isolated. The ACC deaminase gene was cloned in pTZ57R/T plasmid. Plasmids were extracted from E.coli strain DH5 harboring recombinant plasmid and sequenced. The sequence of acdS genes from strains KYA71 and KYA40 and corresponding proteins were analyzed with respect to available sequences in NCBI database. The 16S rRNA gene sequences of S. meliloti strains submitted to the GeneBank/NCBI database. The acdS gene of KYA71 may be located on chromosomal DNA and in KYA40 is on one of mega plasmids. These two genes had 99% similarity with three nucleotide differences which only lead to change in one amino acid 48, threonine in KYA40 acdS gene and methionine in KYA71.The comparison of amino acid sequences of KYA40 and KYA71 with other sequences in database showed that the amino acids 37 to 58 in almost all strains are similar. Therefore, it is concluded that it may be a conserve region in this location of acdS genes and any changes in this region may cause change in ACC deaminase activity.

RNA plays a key role in many aspects of biological processes and its tertiary structure is critical for its biological function. RNA secondary structure represents various significant portions of RNA tertiary structure. Since the biological function of RNA is concluded indirectly from its primary structure, it would be important to analyze the relations between the RNA sequences and their structures. One important tool to perform this kind of analysis is the neutral network which is a collection of RNA sequences, all coding the same secondary structure, where each RNA sequence is distinguished from the others by no more than a single base mutation. Another high level and useful representation of an RNA secondary structure is the RNA shape, where it is holding the vicinity and nesting of structural components and reducing their lengths to one unit. This allows us to analyze the huge structural space corresponding to the larger RNA sequences. In this study, a new concept, entitled Variation Network, over the set of all RNA shapes is introduced. Based on this concept, the potential relations between random and natural RNA sequences, as well as their corresponding structures are analyzed. To explore the relations between random and natural RNA sequences and their corresponding structures, different properties including frequency, normalized frequency, shape energy average, variation rate, normalized variation rate, neighborhood energy average, and stability are obtained and analyzed. The correlations among these properties of random and natural Variation Networks are presented. Base on the obtained correlations, all the employed datasets are highly correlated to each other from the frequency point of view, whereas they are not well correlated from the thermodynamic energy point of view. Since the thermodynamic energy value of an RNA sequence over its secondary structure plays a key role in its function, this research conclude that the natural RNA sequences are not generated randomly.

Populations of Magnaporthe, the causal agent of rice blast disease, are pathotypically and genetically diverse, and therefore their interaction with different rice cultivars and also antagonistic microorganisms is very complicate. The objectives of the present study were to characterize phylogenetic relationships of 114 native Magnaporthe strains, isolated from rice and different weeds in the North region of Iran and study their interaction with the fungal and bacterial antagonists. Phylogenetic studies (lineage structure, cluster analysis and gene flow) were performed using AFLP DNA fingerprinting. Antagonistic effects of the native fungal (Trichoderma harzianum) and bacterial (Bacillus subtilis and Pseudomonas fluorescens) against Magnaporthe strains were assayed at In vitro levels using factorial experiments based on completely randomized designs (CRD) and mean comparison tests. Totally, 39 clonal lineages including 48 haplotypes were identified among the strains of M. grisea and designated here as A–Z. AFLP marker could finely differentiate the strains isolated from various hosts. The strains isolated from Setaria sp. were much close to those from rice (Oryza sativa L.). Magnaporthe strains isolated from Digitaria sp. showed higher genetic variation than other strains. Genetic distances revealed by AFLP markers could finely differentiate M. grisea and M. salvinii. The rate of gene flow was an evidence of low gene transferring among Magnaporthe populations and existence of a complex species for Magnaporthe strains. The fungal and bacterial antagonists showed different reactions against different Magnaporthe strains. These results confirmed high genetic diversity between the Magnaporthe strains which also previously determined by AFLP experiments. It is concluded that the Magnaporthe populations in Iran have complex genetic diversity, and therefore, to achieve an efficient control of different strains and pathotypes of Magnaporthe sp, it is necessary to use different bacterial and fungal biocontrol agents as a dynamic and integrated control system.

There are varieties of purification techniques for separation of human plasma proteins such as salting out, ion exchange chromatography, and ethanol fractionation. There are limitations for each method, for example in salting out method, the salt has to be removed in an additional step. Ion exchange chromatography is difficult for scaling up, and plasma fractionation is a time consuming method and it needs machinery and plant. In the present study the fractionation of human plasma by polyethylene glycol was investigated. The purpose of this study was to investigate the possibility of the fractionation of human plasma by polyethylene glycol. Human plasma fractionation was carried out by using polyethylene glycol at different concentrations from five to twenty percent, and it was followed by centrifugation. After each step of addition of polyethylene glycol the supernatant was removed for further fractionation by addition of higher concentration of polyethylene glycol. Suitable intermediate sources for protein purification were obtained by fractionation of human plasma by polyethylene glycol. Fibrinogen in fraction 5%, IgG and IgM in fraction 10%, IgA in fraction 20%, and finally albumin and α1-Antitrypsin in supernatant 20% of polyethylene glycol were achieved. By our study we could obtain four different fractions as intermediate sources for protein purification which cannot be easily obtained from plasma fractionation by cold ethanol fractionation.