JOURNAL OF BIOTECHNOLOGY
Year:2019 | Volume:10 | Issue:2 |Papers Count:20

Hsp70 family members are central components of the cellular network of molecular chaperones and folding catalysts. The gene encoding a protein related to Hsp70 or DnaK in the domain bacteria is called dnaK. DnaK proteins are involved in de novo protein folding, formation, and disassembly of protein complexes and degradation of misfolded proteins. The gene dnaJ which codes for Hsp40 in bacteria, modulate the activities of DnaK by acting as co-chaperone. In the present study, we cloned and expressed DnaK from Bacillus halodurans Guj1 were identified. The dnaK gene of B. halodurans was successfully expressed in E. coli BL21 (DE3) using pET-28a+ expression system. The open reading frame of the cloned gene contained 1839bp and encoded 612 amino acid residues. Calculated molecular weight and pI of the protein were 66. 18kDa and 4. 55 respectively. The deduced amino acid sequence of B. halodurans Guj1 showed about 60% identity with the E. coli counterpart. The 3D structure of dnaK from B. halodurans was constructed using the crystal structure of human HSP70 chaperone BiP as the template, which showed an identity of 88. 8% together. Partially purified recombinant DnaK by heat treatment showed a band at approximately 70kDa on SDS-PAGE. Our findings showed that the recombinant DnaK improved the refolding efficiency of the carbonic anhydrase by 27% after 60min at 54° C. According to the results obtained, DnaK from B. halodurans can potentially be used for improving the functional properties of enzymes and proteins in various applications.

The production of double haploid plants can be used as an effective method for plant breeding. In this research, in order to produce chickpea haploid plants, 1mm-long anthers of Bivanij cultivar containing the microspores at uni-nucleate stage were isolated from suitable buds (3mm in length) and exposed to different centrifuge (150g, 300g and 600g each for 3, 6 and 10 minutes) and electrical shock pretreatments (0, 100, 150 and 200V) in the 2ml microtubes containing 1. 5ml of RM-IK medium. The treated anthers were then cultured in an EDM culture medium containing 10mg/l 2, 4-D and 10mg/l silver nitrate to induce callus and embryos. Results showed significant differences between different levels of centrifugation, different levels of electric shock and their interactions for the studied traits. The highest percentage of embryogenesis was observed in centrifuge pretreatments of 150g for 6 minutes, 300g for 3 minutes, 150g for 3 minutes in combination with 150V electrical shock, 300g for 6 and 10 minutes and combination of 150g centrifuge pretreatment for 3 minutes with electric shock of 200V, while the highest percentage of plant regeneration was obtained from centrifuge pretreatment of 300g for 6 minutes and also the combination of centrifuge pretreatment of 150g for 3 minutes with electric shock of 150V.

Effluent from dairy industry has a high amount of nutrients such as nitrate and phosphate. In this work nitrate and phosphate removal from treated dairy wastewater in the presence of organic load was investigated. For this purpose, 400ml of synthetic wastewater was inoculated with 2ml of seed culture of microalgae Chlorella salina. During the growth period, nitrate and phosphate concentration in synthetic wastewater was measured for 1, 3, 5 and 7 days with the standard method (APHA). Results showed that removal of nitrate and phosphate by the microalgae from synthetic wastewater was 100% and 95%, respectively. Also, maximum biomass production in 7 days of experiment was about 0. 7g/L. These values showed that Chlorella Salina could be potential candidates by showing their intrinsic merit for removal of phosphate and nitrate from dairy wastewater and can be used in treated outlet refinement from the dairy treatment plant to be used before entering to the environment.

Luciferase from firefly Photinus pyralis (P. py) is a peroxisomal enzyme that converts a heterocyclic substrate luciferin to an excited state oxyluciferin in the presence of Mg+2-ATP and O2. Excited oxyluciferin with the emission of visible light is changed to its ground state. The combination of rapidity, sensitivity, and convenience has led to the development of a broad range of luminescence applications. In spite of wide ranges applications, firefly luciferase is unstable against changes in chemical and physical conditions, thereby reduce its precision and sensitivity. The most undesirable instability of the luciferase is low thermostability and high susceptibility to proteolytic degradation. According to previous studies, limited proteolysis by trypsin of P. py luciferase indicated six cleavage sites on two accessible regions: 206-220 (Including K206, R213, and R218) and 329-341 (Including K329, R330, and R337) on N-terminal domain. In this study, we used site-directed mutagenesis to introduce one point mutation on the 329-341 accessible regions of P. py luciferase, in order to investigate the role of R330 on the enzyme structure and function which R330 changed to Q. Based on limited proteolysis data, R330Q mutant didn’ t significantly change compared to wild type, but this mutation caused several alterations in enzymatic properties including shifting the pH optimum from 7. 5 to 8 and increasing the thermal inactivation. Based on the results, it can be concluded that whilst Arg330 is a conserved residue but not effects on trypsinolysis stability.

Chronic myeloid leukemia (CML) is a malignant blood disease with a particular chromosomal aberration and it is known as a common form of leukemia. Chromene family exhibit strong anti-cancer effects. Therefore, in this study, the effect of two derivatives of dihydro-pyrano [2, 3-g] chromene family is investigated on cell toxicity and apoptosis induction in K562 cancer cells and compared them with Peripheral Blood Mononuclear (PBMC) normal cells. The K562 cell line was cultured in the presence of the mentioned chromene derivatives at a concentration of 40-200μ M for 24-72 hours. The effect of these compounds on growth and viability of K562 cell line and PBMC cells were studied via MTT assay and apoptosis induction was investigated by flow cytometry. The results showed that these chromene derivatives inhibit K562 cell line growth. Moreover, increasing the chromene concentration and the time of exposure to it increase the cell toxicity. Among these compounds, 4-No2pgC was known as a compound with high toxicity (IC50=129± 2. 75) and 4-MePgC recognized as a compound with low toxicity (IC50=214± 3. 42) after 72 hours exposure to the K562 cell line. Furthermore, the results of flow cytometry demonstrated the effect of apoptosis induction of these compounds on the K562 cell line. According to the obtained results from this research, chromene derivatives can induce apoptosis in the K562 cell line and these compounds have a less toxic effect on normal cells than cancer cells. In conclusion, these derivatives can be considered as a proper candidate for the treatment of leukemia.

Immunotoxins, as a critical approach for cancer therapy, have ability to induction death in cancerous cells based on specific ligands for targeting cancer-specific antigens and toxin domains in its context. Bearing in mind, discovery the cancer-specific antigens, as well as immunotoxin characterization for modeling based on linkers application, is a critical step for drugs design, which is considered in this study for ovarian cancer based on in-silico biology. The results of this study, led to the detection of 29 antigens with expression capacity on the surface of ovarian cancer cells, with the highest and specific expression associated with MAGE4 and CA125 antigens. Moreover, the 3D structure of MAGE4 was performed, and the pattern of its expression was determined to rely on HLA proteins. On the other hand, among connecting proteins to this antigen TRIM69 selected as the most effective ligand. Subsequently, the assembling between the domain of Corynebacterium diphtheria and this ligand with (GGGGS) 3 linker in 5 positions led to the creation of 50 models, with different quality and structure. However, among these models, S4 drug showed the best structure and function including binding affinity and immunogenicity after simulation in physiological condition. Generally, this result led to present the MAGE4 as a suitable candidate for immunotoxin development for ovarian cancer, as well as an effective immunotoxin which should be considered in an experimental condition.

Microalgae are microscopic algae found in a wide range of habitats including freshwater and marine systems. Over the last decades, biotechnological processes based on microalgae have been receiving increasing interest due to their potential to produce large quantities of valuable products used as human food supplements, pharmaceuticals and animal feed. Microalgae have also been proved as an efficient and cost-effective platform for recombinant protein production. Most progress in this field has been achieved using Chlamydomonas reinhardtii, a photosynthetic unicellular alga which has been mostly considered as a model organism in different studies. High growth rate, ease of cultivation, well-established genetics and the ability to perform post-translational modifications are the most beneficial attributes that have made C. reinhardtii an attractive system for the expression of recombinant proteins. In this review, we focus on C. reinhardtii as a novel platform for the development of advanced recombinant products and compare them with other commonly used expression systems. We also present a comprehensive overview of its structure, life cycle, culture systems, and media in detail and then discuss the strategies for engineering its three different genomes to produce recombinant cells. Finally, algal culture collections in the world are introduced.

Biosurfactants are surface tension reducing compounds produced by a wide range of microorganisms. These compounds are caused to facilitate the absorption insoluble substrate by microbial cells. The aim of this study was to investigate the effects of nanoparticles of Fe/SDS on the biosurfactant production by Pseudomonas aeruginosa in culture is molasses. For this purpose were used different concentrations of nanoparticles 1, 500 and 1000 mg/L. As a result the concentration of 1mg /L of Fe/SDS nanoparticles has the best effect on the growth of bacteria and biosurfactant production. This concentration increased 23. 21% cell growth and 20. 73% biosurfactant production compared with control samples. By increasing the concentration of nanoparticles reduced growth rate and biosurfactant production was observed. This indicates that the nanoparticles having negative effects of higher concentrations. The results showed that low concentrations of nanoparticles Fe/SDS has positive effects on bacterial biosurfactant production and therefore a good alternative to chemical surfactants for use in the petroleum industry.

Accumulation of polyethylene (PE) wastes has become a major environmental problem. The objective of this research was to assess the potential for microbial degradation of sun-treated low-density PE as a natural way to eliminate PE wastes in semi-industrial condition. Lowdensity polyethylene (LDPE) films were exposed to one month of sun radiation treatment and then cultured with two PE-degrading bacteria (Sphigobacterium moltivorum IRN11 and Delftia tsuruhatensis IRN27) in aerobic bioreactors over 100 days. Weight loss percentage of the PE and the culture pH were measured. Also, Changes in the chemical structure of the LDPE were assessed by FT-IR and surface erosion and microbial layer formation by bacterial activity was observed by Scanning Electron Microscopy. Partial increases in the culture pH were recorded during the incubation period. The weight loss percentage for T-LDPE samples cultured with Sphigobacterium moltivorum IRN11 and Delftia tsuruhatensis IRN27 was 3. 31%± 0. 013 and 3. 98%± 0. 025 in TLDPE samples, respectively, and functional carbonyl-groups in the TLDPE samples decreased significantly due to bacterial hydrolysis. SEM images showed the different microbial layer formation on sun-treated low-density polyethylene (T-LDPE) for both bacteria. Our results suggest that exposure of LDPE to sun radiation had a significant effect on biodegradation of Ld-PE films and that the two bacteria tested were able to enhance the biodegradation the T-LDPE.

Liposomes or biological vesicles are formed from cholesterol, phospholipids, and water. Also, sometimes other biological and non-biological molecules imported in the structure of liposome. The stability of the liposomes in the treatment of diseases and drug delivery, it is vitally important and can be influenced by the composition of phospholipid. In addition, the presence or absence of cholesterol may also affect the stability of liposome. Also, the formation of liposomes is affected by the presence or absence of cholesterol. In this study, we are seeking to affect the presence or absence of cholesterol on the stability and the formation of the liposome. For this purpose, the molecular dynamics simulation method is used. Liposomes that they are simulated was of two types of liposomes type I and liposome type II. The formation analyzes including radial distribution function and solvent accessible surface area showed that each of liposomes created. The type I liposome created one nanodisc structure and type II liposome created two nanodisc structures. Also, energy analysis including total energy, van der Waals interaction energy, and electrostatic interaction energy showed that type I liposome is more stable. Because the cholesterol molecules are the presence of in this liposome structure, that ability to gives hydrogen bonding with side lipids and an increase of stability. In addition, hydrophobic interactions between cholesterol and phospholipids as well as distribution and proper orientation of these parts play a major stake in the stability of the structure.

Aims: The study of the behavior of active suspended fluid introduced a new topic for a wide range of applications such as reactors, microfluidic pumps, cultivating surfaces and antibacterial surfaces. The motility of bacterial suspension has generated a shear force (Hydrodynamic interaction) across the cells, which has an effect on the fluid viscosity. In this study, shear rheometer was used to evaluate the viscosity behavior of Escherichia coli (E. coli) in a water/ polymer environment as a function of bacteria concentration and shear rate. Materials and Methods: In the experimental study, the activity of E. coli was evaluated by choosing the constant concentration (0. 01g/ml) and molecular weight of polyvinylpyrrolidone (360KDa). Also, the surface tensions of E. coli, Acetobacter xylinum, and S. aureus solutions were calculated using Wilhelmy test. Findings: Activity of E. coli suspension results the lower viscosity comparing with the bacteriafree solution. The relative viscosities of the solutions were analyzed in a wide range of shear rates and bacterial concentrations. At a low shear rate up to1S-1, the relative viscosity was found to be less than a unit value (Less than water). Also, due to the collective motility break up to smaller parts at high shear rates, the viscosity increased. The critical volume fraction was defined in determined bacterial concentration (0. 8v/v) to analyze the collective movement of bacteria. The interfacial tension was reduced by bacteria presence of three different stain types that confirmed the effect of bacteria activity on the flow behavior. Conclusion: The activity of E. coli bacteria makes it easy for fluid to flow at low shear rates. The viscosity reduction of active particle has a potential to demonstrate a variety of novel applications when a reduced energy level is needed.

Cancer stem cells are responsible for the formation the resistance to treatment, tumor relapse, and metastasis. miRNAs play an important role in the regulation of biological processes. Therefore, the purpose of this review is to candidate miRNAs that are involved in the regulation of all three properties including stemness, metastasis, and drug resistance and find their target genes and signaling pathways by using literature learning and data mining. The present systematic review is done to identify stemness-regulating miRNAs. By using CORMINE database, metastasis and drug resistance regulating miRNAs collected. Finally, we compared these three lists of miRNAs and found common miRNAs in these three properties. ONCO. IO database and KEGG pathway have been done to obtain the interaction between miRNA-miRNA target and cancer-related signaling pathway respectively. We collected 191 stemness-regulating miRNAs from 21 excluded studies. Based on CORMINE database, 161 miRNAs and 57 miRNAs had metastasis and stemness features respectively. We obtained 7 common miRNAs that 4 of them including has-miR-34a, has-miR-23a, has-miR-30a, has-miR-100 has a significant role for targeting signaling pathways involved in cancer and their most important targets regulate many processes of cells. These data suggest that three important properties can regulate by common miRNAs. Therefore, target these miRNAs or their targets can be helpful to stop tumor growth and metastasis and may be useful biomarkers for early detection of gastric cancer.

Genome editing using targetable nucleases is an emerging technology for precise genome modification in many organisms with hight ability and capability. All targeted genome engineering relies on the introduction of a site-specific doublestrand break (DSB) in a pre-determined genomic locus by a rare-cutting DNA endonuclease. Subsequent repair of this DSB by non-homologous end joining (NHEJ) or homology-directed repair (HDR) generates the desired genetic modifications such as gene disruption, gene insertion, gene correction, etc. Three types of endonucleases, namely ZFNs (zinc finger nucleases), TALENs (transcription activator-like effector nucleases), and the CRISPR (clustered regularly interspersed short palindromic regions) associated (Cas9) system have been predominantly utilized for gene editing. Targeted genome engineering or editing enables researchers to modify genomic loci of interest in a precise manner, which has a turning point in medicine, biological research, and biotechnology. Treatment of human immunodeficiency virus (HIV) infection with ZFN-mediated CCR5 gene disruption is one of the indicator examples of the ability of ZFNs in genome editing. The emergence of TALENs in 2010 has enabled the genome modification of non-model organisms, while the emergence of the CRISPR/Cas9 system in 2013 as a revolutionary genome-editing tool has allowed us to anticipate the forthcoming new era of genome editing research. Soon, it is likely that tgenome editing also will provide the possibility of treating genetic diseases. Genome editing is also hoped to be available for use in the generation of crops and livestock with useful traits. An example would be the production of edible fungi resistant to browning by inactivation of the genes encoding polyphenol oxidase in 2016 under the non-GMO genetically edited crop plants and production of herbicide-resistant rice and rapeseed using CRISPR/Cas9 systems. In this article, we review essential genome editing tools, summarize their applications in crop improvement, as well as, next-generation crop breeding and their computational resources will be discussed.

In recent years, many studies have been performed on magnetic nanoparticles synthesis for use in various field of sciences. The design and synthesis of these nanoparticles have a direct impact on their physico-chemical properties and their applications, especially in the field of biological sciences. One of the simplest and most efficient methods for synthesis of magnetic nanoparticles is a chemical co-precipitation method, which is mainly associated with aggregation of nanoparticles. In this study, the synthesis of magnetic nanoparticles by co-precipitation method was performed in the presence of sodium hydroxide, ammonium hydroxide, and sodium citrate, and then coated with tetraethyl ortho-silicate. The size, surface charge, superparamagnetic properties, crystalline structure, and surface functional groups of nanoparticles were characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), zeta potential, vibrating sample magnetometer (VSM), X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FTIR). The results showed that the synthesis of magnetic nanoparticles in the presence of citrate or acidic acid and silica coating increase zeta potential of nanoparticles up to-70. 8 and-79. 2mV and leads to increase their stability and dispersion and prevents aggregation of magnetic nanoparticles without significant effect on their superparamagnetic properties.

Maize (Zea mays L. ) is one of the most cultivated crops worldwide, owing to its versatility and wide adaptability, and serves as food, animal feed, and raw material for various industrial products. The purpose of the current research was the classification of maize inbred lines in order to produce hybrid seeds based on agro-morphological traits. Each of 100 maize inbred lines was planted in 6 pots as 6 replications and arranged in completely randomized design in an open area near to greenhouse in 2015. The result of the analysis of variance revealed significant differences among lines for all studied traits. The highest correlation was seen between cob’ s length and cob’ s weight. Stepwise regression analysis revealed that 66. 4% of seed yield per plant variation was determined by cob’ s length and cob’ s weight. Cluster analysis divided inbred lines into 4 groups. The highest Mahalanobis distance (28. 07) was observed between cluster 2 and 4. The result of principal component analysis confirmed the calcification by cluster analysis. The genotypes from groups 2 and 4 can be potentially used as parental lines in hybrid varieties production and development of segregating populations.

Some diseases such as gout, the formation of kidney stones, Lesch-Nyhan syndrome, Heart disease, diabetes type II and metabolic syndrome are caused due to the high concentration of uric acid. Within drugs, uricase significantly decreases the level of uric acid in plasma. The production, formulation and preservation proteins need special conditions so that there was no alteration in their structure and highest activity and response, at the same time the lowest immunogenicity can be achieved. In this study, uricase from Aspergillus flavus was cloned and expressed in Escherichia coli BL21. The protein was then purified using affinity chromatography. The enzyme activity and stability were compared with the common industrial Rasburicase. Results showed higher activity and stability at different temperatures (50, 37, 25, 4, and-20° C). Since uricase has an important role in the prevention and cure of mentioned diseases, therefore, the stable form of this enzyme could be a potential candidate for drug development.

MicroRNAs (miRNAs) are single-stranded RNAs that play key roles in cellular disorders or disease diagnosis. Thus the method for sensitive and selective detection of miRNAs is imperative to clinical diagnosis. Recently it has witnessed the rapid development of Metal Nanocluster-Based fluorescent probe design and its successful applications in detecting various targets, such as ssDNA, miRNA and Metal Ions. The DNA scaffolded Metal nanoclusters display excellent photostability, subnanometer size, nontoxicity, biocompatibility and thus well-suited as a fluorescent probe for biochemical applications. Here we develop a DNA/Metal Nanoclusters (MNCs)-based turn-on fluorescence method in the presence of target microRNAs as a potential biomarker for screening cancer. DNA scaffold Metal nanocluster was fabricated by a one-pot wet-chemical strategy and characterized by TEM and DLS techniques. This nanobisensor had a detection limit of 0. 64pM. Conclusion, this nanobiosensors could become a potential alternative tools for detection of miRNAs in biological samples and useful in biomedical research and early clinical diagnosis.

Saccharomyces yeast genus has a wide application in biotechnology that several studies on this subject are performed. They have also known as a dietary supplement in breeding all kinds of creatures in particular for aquatics. Species of this genus are considered as probiotics, as well as a lot of beneficial effects on growth are creatures and Yeasts can be cultured in a variety of waste and cheap substrates, including hydrocarbons and petroleum. So finding an Appropriate and inexpensive culture media for optimal growth of yeast is important. In the present study, along with the study of Saccharomyces species diversity as intestinal flora of domestic rainbow trout flora, optimum growth conditions the strains of Saccharomyces cerevisiae was studied in different culture media as YPD, YPAD, YPG, YPAC, and DM. This research not only emphasized the diversity of Saccharomyces cerevisiae strains as an intestinal flora in domestic fishes, but also revealed the optimum growth of yeasts in YPAD media.

A biological network represents the interaction between a set of macromolecules to drive a particular biological process. In a biological environment, abnormalities happen not only in one molecule but also through a biological network. One of the most effective methods to detect anomaly is the comparison between healthy and diseased networks. In this regard, biological network alignment is one of the most efficient ways to find the difference between healthy and diseased cells. This problem, protein-protein interaction network alignment, has been raised in two main types: Local network alignment and Global network alignment. According to the NP-completeness of this problem, different non-deterministic approaches have been proposed to tackle the Global network alignment problem. Recently, NetAl has been introduced as a common algorithm to align two networks. Although this algorithm can align two networks at the appropriate time, it does not consider biological features. In this study, we present a new framework called PRAF to improve the results of network alignment algorithms such as NetAl by considering some biological features like gene ontology (GO).

Conventional wastewater treatment systems are not capable of removing phosphorus effectively. The entry of phosphorus into water resources leads to the formation of an Eutrophication phenomenon. One of the methods for phosphorus removal is the use of microalgae. In this way, besides helping with advanced sewage treatment, it can produce algae with many applications. The purpose of this study was to determine the simultaneous and to compare the phosphorus removal (rate of phosphorus) and Spirulina biomass production in a photobioreactor, using two kinds of treated sewage. The experiments were carried out with the manufacture of a photobioreactor and air injection by means of a fine bubble diffuser into sewage-containing reactors. The light source in this test was designed as fluorescent light bulbs and alternating radiation. Urban wastewater effluent and refined sewage were used as a culture medium in a photobioreactor. The amount of phosphorus in the purified sewage was measured by spectrophotometry at a wavelength of 690nm. The phosphorus removal and algal biomass production were measured in different culture medium containing wastewater with various concentrations of phosphorus. The initial concentrations of phosphorus in refined urban sewage and sanitary sewage were 1. 96 and 0. 4mg L-1, respectively. Phosphorus removal during microalgae cultivation with municipal wastewater and sanitary sewerage (removal of phosphorus) for 8 days, was 71. 9% and 37%, respectively. Biomass production in this time were 0. 18 and 0. 025g/l, respectively. By decreasing the concentration of phosphorus in the wastewater, the amount of biomass production and (removal of) phosphorus removal decreased. Treated domestic and sanitary sewage can be injected directly, without prior treatment, in photobioreactor and it is possible to remove phosphorus and to produce algal biomass.