World Academy of Science, Engineering and Technology

Authors: Essam A. Makky, Siti H. Mohd Rasdi, J. B. Al-Dabbagh, G. F. Najmuldeen

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The synthesis of silver nano particles (SNPs) extensively studied by using chemical and physical methods. Here, the biological methods were used and give benefits in research field in the aspect of very low cost (from waste to wealth) and safe time as well. The study aims to isolate and exploit the microbial power in the production of industrially important by-products in nano-size with high economic value, to extract highly valuable materials from hazardous waste, to quantify nano particle size, and characterization of SNPs by X-Ray Diffraction (XRD) analysis. Disposal X-ray films were used as substrate because it consumes about 1000 tons of total silver chemically produced worldwide annually. This silver is being wasted when these films are used and disposed. Different bacterial isolates were obtained from various sources. Silver was extracted as nano particles by microbial power degradation from disposal X-ray film as the sole carbon source for ten days incubation period in darkness. The protein content was done and all the samples were analyzed using XRD, to characterize of silver (Ag) nano particles size in the form of silver nitrite. Bacterial isolates CL4C showed the average size of SNPs about 19.53 nm, GL7 showed average size about 52.35 nm and JF Outer 2A (PDA) showed 13.52 nm. All bacterial isolates partially identified using Gram’s reaction and the results obtained exhibited that belonging to Bacillus sp.

Keywords: nanotechnology, bioremediation, disposal X-ray film, nanoparticle, waste, XRD

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Authors: M. S. Mahmoud

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In this paper, the adsorption performance of a novel environmental friendly material, calcium alginate gel beads as a non-conventional technique for the successful removal of copper ions from aqueous solution are reported on. Batch equilibrium studies were carried out to evaluate the adsorption capacity and process parameters such as pH, adsorbent dosages, initial metal ion concentrations, stirring rates and contact times. It was observed that the optimum pH for maximum copper ions adsorption was at pH 5.0. For all contact times, an increase in copper ions concentration resulted in decrease in the percent of copper ions removal. Langmuir and Freundlich's isothermal models were used to describe the experimental adsorption. Adsorbent was characterization using Fourier transform-infrared (FT-IR) spectroscopy and Transmission electron microscopy (TEM).

Keywords: adsorption, alginate polymer, isothermal models, equilibrium

Procedia PDF Downloads 441

Authors: Sharareh Mohseni

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Purpose: The objective of this study was to find a suitable solvent to produce saffron edible extract with improved chemical properties. Design/methodology/approach: Dried and pulverized stigmas of C. sativus L. (10g) was extracted with 300 ml of solvents including: distillated water (DW), ethanol/DW, methanol/DW, propylene glycol/DW, heptan/DW, and hexan/DW, for 3 days at 25°C and then centrifuged at 3000 rpm. Then the extracts were evaporated using rotary evaporator at 40°C. The fiber and solvent-free extracts were then analyzed by UV spectrophotometer to detect saffron quality parameters including crocin, picrocrocin and safranal. Findings: Distilled water/ethanol mixture as the extraction solvent, caused larger amounts of the plant constituents to diffuse out to the extract compared to other treatments and also control. Polar solvents including distilled water, ethanol, and propylene glycol (except methanol) were more effective in extracting crocin, picrocrocin, and saffranal than non-polar solvents. Social implications: Due to an enhancement of color and flavor, saffron extract is economical compared to natural saffron. Saffron Extract saves on preparation time and reduces the amount of saffron required for imparting the same flavor, as compared to dry saffron. Liquid extract is easier to use and standardize in food preparations compared to dry stamens and can be dosed precisely compared to natural saffron. Originality/value: No research had been done on production of saffron edible extract using the solvent studied in this survey. The novelty of this research is high and the results can be used industrially.

Keywords: Crocus sativus L., saffron extract, solvent extraction, distilled water

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Authors: I. Sani, A. Abdulhamid, F. Bello, Isah M. Fakai

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This research has focused on the application of green fluorescent protein (GFP) as a new technique for direct monitoring of fermentation processes involving cultured bacteria. To use GFP as a sensor for pH and oxygen, percentage ratio of red fluorescence to green (% R/G) was evaluated. Assessing the magnitude of the % R/G ratio in relation to low or high pH and oxygen concentration, the bacterial strains were cultivated under aerobic and anaerobic conditions. SCC1 strains of E. coli were grown in a 5 L laboratory fermenter, and during the fermentation, the pH and temperature were controlled at 7.0 and 370C respectively. Dissolved oxygen tension (DOT) was controlled between 15-100% by changing the agitation speed between 20-500 rpm respectively. Effect of reducing the DOT level from 100% to 15% was observed after 4.5 h fermentation. There was a growth arrest as indicated by the decrease in the OD650 at this time (4.5-5 h). The relative fluorescence (green) intensity was decreased from about 460 to 420 RFU. However, %R/G ratio was significantly increased from about 0.1% to about 0.25% when the DOT level was decreased to 15%. But when the DOT was changed to 100%, a little increase in the RF and decrease in the %R/G ratio were observed. Therefore, GFP can effectively detect and indicate any change in pH and oxygen level during fermentation processes.

Keywords: Escherichia coli SCC1, fermentation process, green fluorescent protein, red fluorescence

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Authors: J. B. Minari, O. B. Oloyede, A. A. Odutuga

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Myeloperoxidase is the most abundant enzyme found in the polymorphonuclear neutrophil and is known to play a central role in the host defense system of the leukocyte. The enzyme has been reported to interact with some drugs to generate free radical which inhibits its activity. This study investigated the effects of artesunate on the activity of the enzyme and the subsequent effect on the host immune system. In investigating the effects of the drugs on myeloperoxidase, the influence of concentration, pH, partition ratio estimation and kinetics of inhibition were studied. This study showed that artesunate is concentration-dependent inhibitor of myeloperoxidase with an IC50 of 0.078mM. Partition ratio estimation showed that 60 enzymatic turnover cycles are required for complete inhibition of myeloperoxidase in the presence of artesunate. The influence of pH on the effect of artesunate on the enzyme showed least activity of myeloperoxidase at physiological pH. The kinetic inhibition studies showed that artesunate caused a competitive inhibition with an increase in the Km value from 0.12mM to 0.26mM and no effect on the Vmax value. The Ki value was estimated to be 2.5mM. The results obtained from this study show that artesunate is a potent inhibitor of myeloperoxidase and it is capable of inactivating the enzyme. It is considered that the inhibition of myeloperoxidase in the presence of artesunate as revealed in this study may partly explain the impairment of polymorphonuclear neutrophil and consequent reduction of the strength of the host defense system against secondary infections.

Keywords: myeloperoxidase, artesunate, inhibition, nuetrophill

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Authors: Meriem Mansouri, Fatiha Bendali Saoudi, Noureddine Soltani

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Biological control presents a means of control for the protection of the environment. Bacillus thuringiensis israelensis Berliner 1915 is an inseticide of biological origin because it is a bacterium of the Bacillaceae family. This biocide has a biological importance, because of its specific larvicidal action against Culicidae, blood-sucking insects, responsible for several diseases to humans and animals through the world. As well as, its high specificity for these insects. Also, the freshwater mites, this necessarily parasitic group for aquatic species such as the Physidae, also have an effective biological control against the Culicidae, because of their voracious predation to the larvae of these insects. The present work aims to study the effects of the biocide Bacillus thuringiensis var israelinsis, against non-target adults of water mites Eylais hamata Koenike, 1897, as well as its associated host species Physa marmorata Fitzinger, 1833. After 12 days of oral treatment of adults with lethal concentration (LC50:0.08µg/ml), determined from essays on 4th instar larvae of Culex pipiens (hematophagous insects). No adverse effect has been recorded for adult individuals of Eylais hamata, contrary, snail Physa marmorata were sensitive for this dose of Bti. In parallel, after treatment at the Bti by LC50, the enzyme stress bio marker glutathione S-transferase, was measured after 24, 48 and 72 hours. The enzymatic activity of GST has increased after 24 and 48 hours following treatment.

Keywords: biological control, Bacillus thuringiensis var israelinsis, culicidae, hydrachnidia, enzymatic activity

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Authors: Iram Siddique

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Synthetic seed technology is an alternative to traditional micropropagation for production and delivery of cloned plantlets. Synthetic seeds were produced by encapsulating nodal segments of C. angustifolia in calcium alginate gel. 3% (w/v) sodium alginate and 100 mM CaCl2. 2H2O were found most suitable for encapsulation of nodal segments. Synthetic seeds cultured on half strength Murashige and Skoog (MS) medium supplemented with thidiazuron (5.0 µM) + indole -3- acetic acid (1.0 µM) produced maximum number of shoots (10.9 ± 0.78) after 8 weeks of culture exhibiting (78%) in vitro conversion response. Encapsulated nodal segments demonstrated successful regeneration after different period (1-6 weeks) of cold storage at 4 °C. The synthetic seeds stored at 4 °C for a period of 4 weeks resulted in maximum conversion frequency (93%) after 8 weeks when placed back to regeneration medium. The isolated shoots when cultured on half strength MS medium supplemented with 1.0 µM indole -3- butyric acid (IBA), produced healthy roots and plantlets with well developed shoot and roots were successfully hardened off in plastic pots containing sterile soilrite inside the growth chamber and gradually transferred to greenhouse where they grew well with 85% survival rate. Changes in the content of photosynthetic pigments, net photosynthetic rate (PN), superoxide dismutase (SOD) and catalase (CAT) activity in C. angustifolia indicated the adaptation of micropropagated plants to ex vitro conditions.

Keywords: biochemical studies, nodal segments, rooting, synthetic seeds, thidiazuron

Procedia PDF Downloads 356

Authors: Khamael M. Abualnaja, Lidija Šiller, Benjamin R. Horrocks

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Metal-enhanced luminescence of silicon nano crystals (SiNCs) was determined using two different particle sizes of silver nano particles (AgNPs). SiNCs have been characterized by scanning electron microscopy (SEM), high resolution transmission electron microscopy (HRTEM), Fourier transform infrared spectroscopy (FTIR) and X-ray photo electron spectroscopy (XPS). It is found that the SiNCs are crystalline with an average diameter of 65 nm and FCC lattice. AgNPs were synthesized using photochemical reduction of AgNO3 with sodium dodecyl sulphate (SDS). The enhanced luminescence of SiNCs by AgNPs was evaluated by confocal Raman microspectroscopy. Enhancement up to ×9 and ×3 times were observed for SiNCs that mixed with AgNPs which have an average particle size of 100 nm and 30 nm, respectively. Silver NPs-enhanced luminescence of SiNCs occurs as a result of the coupling between the excitation laser light and the plasmon bands of AgNPs; thus this intense field at AgNPs surface couples strongly to SiNCs.

Keywords: silver nanoparticles, surface enhanced raman spectroscopy (SERS), silicon nanocrystals, luminescence

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Authors: Fellah Sihem

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The optimal use of soil moisture by culture, is related to the leaf area index, which stood in the cycle and its modulation according to the prevailing stress intensity. For a given stock of water in the soil, cultivar adapted and saving water is one that is no luxury consumption during the preanthesis. It modulates the leaf area index to regulate sweating in the degree of its water supply. In plants water saving, avoidance of dehydration is related to the reduction of water loss by cuticular and stomatal pathways. Muchow and Sinclair reported that the test of relative water content (TRE) is considered the best indicator of leaf water status. The search for indicators of the ability of the plant to make good use of the water, under water stress is a prerequisite for progress in improving performance under water stress. This experiment aims to characterize a set of durum wheat varieties, tested jars and vegetation under different levels of water stress to the surface of the leaf, relative water content, cell integrity, the accumulated biomass and efficiency of water use. The experiment was conducted during the 2005/2006 academic year, at the Agricultural Research Station of the Field Crop Institute of Setif, under semi-controlled conditions. Five genotypes of durum wheat (Triticum durum Desf) were evaluated for their ability to tolerate moderate and severe water stress. The results showed that geno types respond differently to water stress. Dry matter accumulation and growth rate varied among geno types and were significantly reduced. At severe water stress biomass accumulated by Boussalam was the least affected.

Keywords: water stress, triticum durum, biomass, cell membrane integrity, relative water content

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Authors: Nassima Behidj-Benyounes, Thoraya Dahmene, Khaled Benyounes Nadjiba Chebouti1and F/Zohra Bissaad

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Essential oils have a significant antimicrobial activity. These oils can successfully replace the antibiotics. So, the microorganisms show their inefficiencies resistant for the antibiotics. For this reason, we study the physicochemical analysis and antimicrobial activity of the essential oil of Daucus carota. The extraction is done by steam distillation of water which brought us a very significant return of 4.65%. The analysis of the essential oil is performed by GC/MS and has allowed us to identify 32 compounds in the oil of D. carota flowering tops of Bouira. Three of which are in the majority are the α-pinene (22.3%), the carotol (21.7%) and the limonene (15.8%).

Keywords: Daucus carota, essential oil, α-pinene, carotol, limonene

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Authors: Dalila Bouamra, Chekib Arslane Baki, Abdelhamid Bouchebour, Fatiha Koussa, Amel Benamara, Seoussen Kada

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The activity of methanolic extract from Anacyclus pyrethrum was evaluated using λ-carrageenan 1% induced paw edema in Wistar Albinos rats. The oral administration of 200 mg/kg, 400 mg/kg and 600 mg/kg, body weight of methanolic extract, one hour before induction of inflammation, exerted a significant inhibition effect of 47%, 57% and 62% respectively after 4h λ-carrageenan treatment and highly significant inhibition effect of 57%, 66% and 75% respectively after 8h λ-carrageenan treatment, compared to non treated group (100%) and that treated with aspirin, a standard anti-inflammatory drug. On the other hand, the effect of the plant extract on stomach was macroscopically and microscopically studied. The plant extract has an impact on the loss ratio of granulocytes that have invaded the stomach after a period of inflammation at a dose of 600 mg/kg body weight.

Keywords: inflammation, Anacyclus pyrethrum, gastritis, Wistar Albinos rats

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Authors: T. Bettaieb, S. Soufi, S. Arbaoui

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Stevia rebaudiana Bert. is a natural sweet plant. The leaves contain diterpene glycosides stevioside, rebaudiosides A-F, steviolbioside and dulcoside, which are responsible for its sweet taste and have commercial value all over the world as sugar substitute in foods and medicines. Stevia rebaudiana Bert. is sensitive temperature lower than 9°C. The possibility of its outdoor culture in Tunisian conditions demand genotypes tolerant to low temperatures. In order to evaluate the low temperature tolerance of eight genotypes of Stevia rebaudiana, the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalases (CAT) were measured. Before carrying out the analyses, three genotypes of Stevia were exposed for 1 month at a temperature regime of 18°C during the day and 7°C at night similar to winter conditions in Tunisia. In response to the stress generated by low temperature, antioxidant enzymes activity revealed on native gel and quantified by spectrophotometry showed variable levels according to their degree of tolerance to low temperatures.

Keywords: chilling tolerance, enzymatic activity, stevia rebaudiana bert, low thermal stress

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Authors: Shilpi Malik, Ramneek Kaur, Archita Gupta, Deepshikha Yadav, Ashwani Mathur, Manisha Singh

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Glucosamine (GS) is the most abundant naturally occurring amino monosaccharide and is normally produced in human body via cellular glucose metabolism. It is regarded as the building block of cartilage matrix and is also an essential component of cartilage matrix repair mechanism. Besides that, it can also be explored for its prebiotic potential as many bacterial species are known to utilize the amino sugar by acquiring them to form peptidoglycans and lipopolysaccharides in the bacterial cell wall. Glucosamine can therefore be considered for its fermentation by bacterial species present in the gut. Current study is focused on exploring the potential of glucosamine as prebiotic. The studies were done to optimize considerable concentration of GS to reach GI tract and being fermented by the complex gut microbiota and food grade GS was added to various Simulated Fluids of Gastro-Intestinal Tract (GIT) such as Simulated Saliva, Gastric Fluid (Fast and Fed State), Colonic fluid, etc. to detect its degradation. Since it was showing increase in microbial growth (CFU) with time, GS was Further, encapsulated to increase its residential time in the gut, which exhibited improved resistance to the simulated Gut conditions. Moreover, prepared microspehres were optimized and characterized for their encapsulation efficiency and toxicity. To further substantiate the prebiotic activity of Glucosamine, studies were also performed to determine the effect of Glucosamine on the known probiotic bacterial species, i.e. Lactobacillus delbrueckii (MTCC 911) and Bifidobacteriumbifidum (MTCC 5398). Culture conditions for glucosamine will be added in MRS media in anaerobic tube at 0.20%, 0.40%, 0.60%, 0.80%, and 1.0%, respectively. MRS media without GS was included in this experiment as the control. All samples were autoclaved at 118° C for 15 min. Active culture was added at 5% (v/v) to each anaerobic tube after cooling to room temperature and incubated at 37° C then determined biomass and pH and viable count at incubation 18h. The experiment was completed in triplicate and the results were presented as Mean ± SE (Standard error).The experimental results are conclusive and suggest Glucosamine to hold prebiotic properties.

Keywords: gastro intestinal tract, microspheres, peptidoglycans, simulated fluid

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Authors: Huyen T. Pham, Nhue P. Nguyen, Tien Q. Phi, Phuong T. Dang, Hy G. Le

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Since the marine environmental conditions are extremely different from the other ones, so that marine actinomycetes might produce novel bioactive compounds. Therefore, actinomycete strains were screened from marine water and sediment samples collected from the coastal areas of Northern Vietnam. Ninety-nine actinomycete strains were obtained on starch-casein agar media by dilution technique, only seven strains, named HP112, HP12, HP411, HPN11, HP 11, HPT13 and HPX12, showed significant antibacterial activity against both gram-positive and gram-negative bacteria (Bacillus subtilis ATCC 6633, Staphylococcus epidemidis ATCC 12228, Escherichia coli ATCC 11105). Further studies were carried out with the most active HP411strain against Candida albicans ATCC 10231. This strain could grow rapidly on starch casein agar and other media with high salt containing 7-10% NaCl at 28-30oC. Spore-chain of HP411 showed an elongated and circular shape with 10 to 30 spores/chain. Identification of the strain was carried out by employing the taxonomical studies including the 16S rRNA sequence. Based on phylogenetic and phenotypic evidence it is proposed that HP411 to be belongs to species Streptomyces variabilis. The potent of the crude extract of fermentation broth of HP411that are effective against wide range of pathogens: both gram-positive, gram-negative and fungi. Further studies revealed that the crude extract HP411 could obtain the anticancer activity for cancer cell lines: Hep-G2 (liver cancer cell line); RD (cardiac and skeletal muscle letters cell line); FL (membrane of the uterus cancer cell line). However, the actinomycetes from marine ecosystem will be useful for the discovery of new drugs in the furture.

Keywords: marine actinomycetes, antibacterial, anticancer, Streptomyces variabilis

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Authors: Faheema Khan

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To investigate the ability of sensitive and tolerant genotype of Glycine max to adapt to a saline environment in a field, we examined the growth performance, water relation and activities of antioxidant enzymes in relation to photosynthetic rate, chlorophyll a fluorescence, photosynthetic pigment concentration, protein and proline in plants exposed to salt stress. Ten soybean genotypes (Pusa-20, Pusa-40, Pusa-37, Pusa-16, Pusa-24, Pusa-22, BRAGG, PK-416, PK-1042, and DS-9712) were selected and grown hydroponically. After 3 days of proper germination, the seedlings were transferred to Hoagland’s solution (Hoagland and Arnon 1950). The growth chamber was maintained at a photosynthetic photon flux density of 430 μmol m−2 s−1, 14 h of light, 10 h of dark and a relative humidity of 60%. The nutrient solution was bubbled with sterile air and changed on alternate days. Ten-day-old seedlings were given seven levels of salt in the form of NaCl viz., T1 = 0 mM NaCl, T2=25 mM NaCl, T3=50 mM NaCl, T4=75 mM NaCl, T5=100 mM NaCl, T6=125 mM NaCl, T7=150 mM NaCl. The investigation showed that genotype Pusa-24, PK-416 and Pusa-20 appeared to be the most salt-sensitive. genotypes as inferred from their significantly reduced length, fresh weight and dry weight in response to the NaCl exposure. Pusa-37 appeared to be the most tolerant genotype since no significant effect of NaCl treatment on growth was found. We observed a greater decline in the photosynthetic variables like photosynthetic rate, chlorophyll fluorescence and chlorophyll content, in salt-sensitive (Pusa-24) genotype than in salt-tolerant Pusa-37 under high salinity. Numerous primers were verified on ten soybean genotypes obtained from Operon technologies among which 30 RAPD primers shown high polymorphism and genetic variation. The Jaccard’s similarity coefficient values for each pairwise comparison between cultivars were calculated and similarity coefficient matrix was constructed. The closer varieties in the cluster behaved similar in their response to salinity tolerance. Intra-clustering within the two clusters precisely grouped the 10 genotypes in sub-cluster as expected from their physiological findings.Salt tolerant genotype Pusa-37, was further analysed by 2-Dimensional gel electrophoresis to analyse the differential expression of proteins at high salt stress. In the Present study, 173 protein spots were identified. Of these, 40 proteins responsive to salinity were either up- or down-regulated in Pusa-37. Proteomic analysis in salt-tolerant genotype (Pusa-37) led to the detection of proteins involved in a variety of biological processes, such as protein synthesis (12 %), redox regulation (19 %), primary and secondary metabolism (25 %), or disease- and defence-related processes (32 %). In conclusion, the soybean plants in our study responded to salt stress by changing their protein expression pattern. The photosynthetic, biochemical and molecular study showed that there is variability in salt tolerance behaviour in soybean genotypes. Pusa-24 is the salt-sensitive and Pusa-37 is the salt-tolerant genotype. Moreover this study gives new insights into the salt-stress response in soybean and demonstrates the power of genomic and proteomic approach in plant biology studies which finally could help us in identifying the possible regulatory switches (gene/s) controlling the salt tolerant genotype of the crop plants and their possible role in defence mechanism.

Keywords: glycine max, salt stress, RAPD, genomic and proteomic variability

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Authors: K.Narasimhulu, Y. Pydi Setty

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The objective of this present study is the optimization of process parameters in biosorption of Ni(II) and Cd(II) ions by Pseudomonas putida using Response Surface Methodology in a Packed bed bioreactor. The experimental data were also tested with theoretical models to find the best fit model. The present paper elucidates RSM as an efficient approach for predictive model building and optimization of Ni(II) and Cd(II) ions using Pseudomonas putida. In packed bed biosorption studies, comparison of the breakthrough curves of Ni(II) and Cd(II) for Agar immobilized and PAA immobilized Pseudomonas putida at optimum conditions of flow rate of 300 mL/h, initial metal ion concentration of 100 mg/L and bed height of 20 cm with weight of biosorbent of 12 g, it was found that the Agar immobilized Pseudomonas putida showed maximum percent biosorption and bed saturation occurred at 20 minutes. Optimization results of Ni(II) and Cd(II) by Pseudomonas putida from the Design Expert software were obtained as bed height of 19.93 cm, initial metal ion concentration of 103.85 mg/L, and flow rate of 310.57 mL/h. The percent biosorption of Ni(II) and Cd(II) is 87.2% and 88.2% respectively. The predicted optimized parameters are in agreement with the experimental results.

Keywords: packed bed bioreactor, response surface mthodology, pseudomonas putida, biosorption, waste water

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Authors: Noura El-Ahmady El-Naggar

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L-asparaginase is an important enzyme as therapeutic agents used in combination therapy with other drugs in the treatment of acute lymphoblastic leukemia in children. L-asparaginase producing actinomycete strain, NEAE-K, was isolated from soil sample and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as new subsp. Streptomyces rochei subsp. chromatogenes NEAE-K and sequencing product (1532 bp) was deposited in the GenBank database under accession number KJ200343. The study was conducted to screen parameters affecting the production of L-asparaginase by Streptomyces rochei subsp. chromatogenes NEAE-K on solid state fermentation using Plackett–Burman experimental design. Sixteen different independent variables including incubation time, moisture content, inoculum size, temperature, pH, soybean meal+ wheat bran, dextrose, fructose, L-asparagine, yeast extract, KNO3, K2HPO4, MgSO4.7H2O, NaCl, FeSO4. 7H2O, CaCl2, and three dummy variables were screened in Plackett–Burman experimental design of 20 trials. The most significant independent variables affecting enzyme production (dextrose, L-asparagine and K2HPO4) were further optimized by the central composite design. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase by Streptomyces rochei subsp. chromatogenes NEAE-K from solid state fermentation: g/L (soybean meal+ wheat bran 15, dextrose 3, fructose 4, L-asparagine 8, yeast extract 2, KNO3 1, K2HPO4 2, MgSO4.7H2O 0.5, NaCl 0.1, FeSO4. 7H2O 0.02, CaCl2 0.01), incubation time 7 days, moisture content 50%, inoculum size 3 mL, temperature 30°C, pH 8.5.

Keywords: streptomyces rochei subsp. chromatogenes neae-k, 16s rrna, identification, solid state fermentation, l-asparaginase production, plackett-burman design, central composite design

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Authors: Amir Wahid, Fazal Hadi, Amin Ullah Jan

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Tomato and Cauliflower seedlings were grown in-vitro under salt concentrations (0, 2, 4, 8, and 10 dSm-1) with objectives to investigate; (1) The effect of salinity on seedling growth and free proline production, (2) the correlation between seedling growth and proline contents, (3) comparative salt tolerance of both species. Different concentrations of salt showed considerable effect on percent (%) germination of seeds, length and biomass of shoot and root and also showed effect on percent water content of both plants seedlings. Germination rate in cauliflower was two times higher than tomato even at highest salt concentration (10 dSm-1). Seedling growth of both species was less effected at low salt concentrations (2 and 4 dSm-1) but at high concentrations (6 and 8 dSm-1) the seedling growth of both species was significantly decreased. Particularly the tomato root was highly significantly reduced. The proline level linearly increased in both species with increasing salt concentrations up-to 4 dSm-1 and then declined. The cauliflower showed higher free proline level than tomato under all salt treatments. Overall, the cauliflower seedlings showed better growth response along with higher proline contents on comparison with tomato seedlings.

Keywords: NaCl (Sodium Chloride), EC (Electrical Conductivity), MS (Murashig and Skoog), ANOVA (Analysis of Variance), LSD (Least Significant Differences)

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Authors: Furqan M. Kadhum, Asmaa A. Hussein, Maysaa Ch. Hatem

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Hyaluronidase enzyme was purified from the clinical isolate Staphyloccus aureus in three purification steps, first by precipitation with 90% saturated ammonium sulfate, ion exchange chromatography on DEAE-Cellulose, and gel filtration chromatography throughout Sephacryl S-300. Specific activity of the purified enzyme was reached 930 U/mg protein with 7.4 folds of purification and 46.5% recovery. The enzyme has an average molecular weight of about 69 kDa, with an optimum pH of enzyme activity and stability at pH 7, also the optimum temperature for activity was 37oC. The enzyme was stable with full activity at a temperature ranged between 30-40 oC. Metal ions showed variable inhibitory degree with the strongest effect for Fe+3, however, the chelating and reducing agents had no or little effects. Cytotoxic studies for purified and crude hyaluronidase against cancer cell Hep G2 type at different enzyme concentrations and exposure times showed that the inhibition effect of both crude and purified enzyme increased by increasing the enzyme concentration with no change was observed at 24hr, while at 48 and 72 hrs the same inhibition rate were observed for purified enzyme and differ for the crude filtrate.

Keywords: hyaluronidase, S. aureus, metal ions, cytotoxicity

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Authors: Francielo Vendruscolo

Abstract:

This review summarizes the potential of starch agroindustrial residues as substrate for biohydrogen production. Types of potential starch agroindustrial residues, recent developments and bio-processing conditions for biohydrogen production will be discussed. Biohydrogen is a clean energy source with great potential to be an alternative fuel, because it releases energy explosively in heat engines or generates electricity in fuel cells producing water as only by-product. Anaerobic hydrogen fermentation or dark fermentation seems to be more favorable, since hydrogen is yielded at high rates and various organic waste enriched with carbohydrates as substrate result in low cost for hydrogen production. Abundant biomass from various industries could be source for biohydrogen production where combination of waste treatment and energy production would be an advantage. Carbohydrate-rich nitrogen-deficient solid wastes such as starch residues can be used for hydrogen production by using suitable bioprocess technologies. Alternatively, converting biomass into gaseous fuels, such as biohydrogen is possibly the most efficient way to use these agroindustrial residues.

Keywords: biofuel, dark fermentation, starch residues, food waste

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Authors: Rashi Rajput, Manisha Singh

Abstract:

Escitalopram oxalate (ETP), an FDA approved antidepressant drug from the category of SSRI (selective serotonin reuptake inhibitor) and is used in treatment of general anxiety disorder (GAD), major depressive disorder (MDD).When taken orally, it is metabolized to S-demethylcitalopram (S-DCT) and S-didemethylcitalopram (S-DDCT) in the liver with the help of enzymes CYP2C19, CYP3A4 and CYP2D6. Hence, causing side effects such as dizziness, fast or irregular heartbeat, headache, nausea etc. Therefore, targeted and sustained drug delivery will be a helpful tool for increasing its efficacy and reducing side effects. The present study is designed for formulating mucoadhesive nanoparticle formulation for the same Escitalopram loaded polymeric nanoparticles were prepared by ionic gelation method and characterization of the optimised formulation was done by zeta average particle size (93.63nm), zeta potential (-1.89mV), TEM (range of 60nm to 115nm) analysis also confirms nanometric size range of the drug loaded nanoparticles along with polydispersibility index of 0.117. In this research, we have studied the in vitro drug release profile for ETP nanoparticles, through a semi permeable dialysis membrane. The three important characteristics affecting the drug release behaviour were – particle size, ionic strength and morphology of the optimised nanoparticles. The data showed that on increasing the particle size of the drug loaded nanoparticles, the initial burst was reduced which was comparatively higher in drug. Whereas, the formulation with 1mg/ml chitosan in 1.5mg/ml tripolyphosphate solution showed steady release over the entire period of drug release. Then this data was further validated through mathematical modelling to establish the mechanism of drug release kinetics, which showed a typical linear diffusion profile in optimised ETP loaded nanoparticles.

Keywords: ionic gelation, mucoadhesive nanoparticle, semi-permeable dialysis membrane, zeta potential

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Authors: V. Shapaval, N. K. Afseth, D. Tzimorotas, A. Kohler

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Globally there is a dramatic increase in the demand for food, energy, materials and clean water since natural resources are limited. As a result, industries are looking for ways to reduce rest materials and to improve resource efficiency. Microorganisms have a high potential to be used as bio factories for the production of primary and secondary metabolites that represent high-value bio-products (enzymes, polyunsaturated fatty acids, bio-plastics, glucans, etc.). In order to find good microbial producers, to design suitable substrates from food rest materials and to optimize fermentation conditions, rapid analytical techniques for quantifying target bio products in microbial cells are needed. In the EU project FUST (R4SME, Fp7), we have developed a fully automated high-throughput FUST system based on micro-cultivation and FTIR spectroscopy that facilitates the screening of microorganisms, substrates and fermentation conditions for the optimization of the production of different high-value metabolites (single cell oils, bio plastics). The automated system allows the preparation of 100 samples per hour. Currently, The FUST system is in use for screening of filamentous fungi in order to find oleaginous strains with the ability to produce polyunsaturated fatty acids, and the optimization of cheap substrates, derived from food rest materials, and the optimization of fermentation conditions for the high yield of single cell oil.

Keywords: FTIR spectroscopy, FUST system, screening, biotechnology

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Authors: Tania Tzong, Chao-Yi Teng, Tzong-Yuan Wu

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Currently, Chikungunya virus (CHIKV) transmitted to humans by Aedes mosquitoes has distributed from Africa to Southeast Asia, South America, and South Europe. However, little is known about the antigenic targets for immunity, and there are no licensed vaccines or specific antiviral treatments for the disease caused by CHIKV. Baculovirus has been recognized as a novel vaccine vector with attractive characteristic features of an optional vaccine delivery vehicle. This approach provides the safety and efficacy of CHIKV vaccine. In this study, bi-cistronic recombinant baculoviruses vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP were produced. Both recombinant baculovirus can express EGFP reporter gene in insect cells to facilitate the recombinant virus isolation and purification. Examination of vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP showed that this recombinant baculovirus could induce syncytium formation in insect cells. Unexpectedly, the immunofluorescence assay revealed the expression of E1 and E2 of CHIKV structural proteins in insect cells infected by vAc-CMV-CHIKV26S-Rhir-EGFP. This result may imply that the CMV promoter can induce the transcription of CHIKV26S in insect cells. There are also E1 and E2 expression in mammalian cells transduced by vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP. The expression of E1 and E2 proteins of insect and mammalian cells was validated again by Western blot analysis. The vector construction with dual tandem promoters, which is polyhedrin and CMV promoter, has higher expression of the E1 and E2 of CHIKV structural proteins than the vector construction with CMV promoter only. Most of the E1 and E2 proteins expressed in mammalian cells were glycosylated. In the future, the expression of structural proteins of CHIKV in mammalian cells is expected can form virus-like particle, so it could be used as a vaccine for chikungunya virus.

Keywords: chikungunya virus, virus-like particle, vaccines, baculovirus expression vector system

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Authors: Neelofar, Khursheed Alam, Jamal Ahmad

Abstract:

Protein modification in diabetes mellitus may lead to early glycation products (EGPs) or amadori product as well as advanced glycation end products (AGEs). Early glycation involves the reaction of glucose with N-terminal and lysyl side chain amino groups to form Schiff’s base which undergoes rearrangements to form more stable early glycation product known as Amadori product. After Amadori, the reactions become more complicated leading to the formation of advanced glycation end products (AGEs) that interact with various AGE receptors, thereby playing an important role in the long-term complications of diabetes. Millard reaction or nonenzymatic glycation reaction accelerate in diabetes due to hyperglycation and alter serum protein’s structure, their normal functions that lead micro and macro vascular complications in diabetic patients. In this study, Human Serum Albumin (HSA) with a constant concentration was incubated with different concentrations of glucose at 370C for a week. At 4th day, Amadori product was formed that was confirmed by colorimetric method NBT assay and TBA assay which both are authenticate early glycation product. Conformational changes in native as well as all samples of Amadori albumin with different concentrations of glucose were investigated by various biophysical and biochemical techniques. Main biophysical techniques hyperchromacity, quenching of fluorescence intensity, FTIR, CD and SDS-PAGE were used. Further conformational changes were observed by biochemical assays mainly HMF formation, fructoseamine, reduction of fructoseamine with NaBH4, carbonyl content estimation, lysine and arginine residues estimation, ANS binding property and thiol group estimation. This study find structural and biochemical changes in Amadori modified HSA with normal to hyperchronic range of glucose with respect to native HSA. When glucose concentration was increased from normal to chronic range biochemical and structural changes also increased. Highest alteration in secondary and tertiary structure and conformation in glycated HSA was observed at the hyperchronic concentration (75mM) of glucose. Although it has been found that Amadori modified proteins is also involved in secondary complications of diabetes as AGEs but very few studies have been done to analyze the conformational changes in Amadori modified proteins due to early glycation. Most of the studies were found on the structural changes in Amadori protein at a particular glucose concentration but no study was found to compare the biophysical and biochemical changes in HSA due to early glycation with a range of glucose concentration at a constant incubation time. So this study provide the information about the biochemical and biophysical changes occur in Amadori modified albumin at a range of glucose normal to chronic in diabetes. Although many implicates currently in use i.e. glycaemic control, insulin treatment and other chemical therapies that can control many aspects of diabetes. However, even with intensive use of current antidiabetic agents more than 50 % of diabetic patient’s type 2 suffers poor glycaemic control and 18 % develop serious complications within six years of diagnosis. Experimental evidence related to diabetes suggests that preventing the nonenzymatic glycation of relevant proteins or blocking their biological effects might beneficially influence the evolution of vascular complications in diabetic patients or quantization of amadori adduct of HSA by authentic antibodies against HSA-EGPs can be used as marker for early detection of the initiation/progression of secondary complications of diabetes. So this research work may be helpful for the same.

Keywords: diabetes mellitus, glycation, albumin, amadori, biophysical and biochemical techniques

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Authors: M. Cynthya, V. Prabhawathi, D. Mukesh

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Food contamination occurs during post process handling. This leads to spoilage and growth of pathogenic microorganisms in the food, thereby reducing its shelf life or spreading of food borne diseases. Several methods are tried and one of which is use of antimicrobial packaging. Here, papain, a protease enzyme, is covalently immobilized with the help of glutarldehyde on polyurethane and used as a food wrap to protect food from microbial contamination. Covalent immobilization of papain was achieved at a pH of 7.4; temperature of 4°C; glutaraldehyde concentration of 0.5%; incubation time of 24 h; and 50 mg of papain. The formation of -C=N- observed in the Fourier transform infrared spectrum confirmed the immobilization of the enzyme on the polymer. Immobilized enzyme retained higher activity than the native free enzyme. The efficacy of this was studied by wrapping it over S. aureus contaminated cottage cheese (paneer) and cheese and stored at a temperature of 4°C for 7 days. The modified film reduced the bacterial contamination by eight folds when compared to the bare film. FTIR also indicates reduction in lipids, sugars and proteins in the biofilm.

Keywords: cheese, papain, polyurethane, Staphylococcus aureus

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Authors: Ilze Dimanta, Zane Rutkovska, Vizma Nikolajeva, Janis Kleperis, Indrikis Muiznieks

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Majority of word’s steadily increasing energy consumption is provided by non-renewable fossil resources. Need to find an alternative energy resource is essential for further socio-economic development. Hydrogen is renewable, clean energy carrier with high energy density (142 MJ/kg, accordingly – oil has 42 MJ/kg). Biological hydrogen production is an alternative way to produce hydrogen from renewable resources, e.g. using organic waste material resource fermentation that facilitate recycling of sewage and are environmentally benign. Hydrogen gas is produced during the fermentation process of bacteria in anaerobic conditions. Bacteria are producing hydrogen in the liquid phase and when thermodynamic equilibrium is reached, hydrogen is diffusing from liquid to gaseous phase. Because of large quantities of available crude glycerol and the highly reduced nature of carbon in glycerol per se, microbial conversion of it seems to be economically and environmentally viable possibility. Such industrial organic waste product as crude glycerol is perspective for usage in feedstock for hydrogen producing bacteria. The process of biodiesel production results in 41% (w/w) of crude glycerol. The developed lab-scale test system (experimental bioreactor) with hydrogen micro-electrode (Unisense, Denmark) was used to determine hydrogen production yield and rate in the liquid phase. For hydrogen analysis in the gas phase the RGAPro-100 mass-spectrometer connected to the experimental test-system was used. Fermentative bacteria strains were tested for hydrogen gas production rates. The presence of hydrogen in gaseous phase was measured using mass spectrometer but registered concentrations were comparatively small. To decrease the hydrogen partial pressure in liquid phase reactor with a system for continuous bubbling with inert gas was developed. H2 production rate for the best producer in liquid phase reached 0,40 mmol H2/l, in gaseous phase - 1,32 mmol H2/l. Hydrogen production rate is time dependent – higher rate of hydrogen production is at the fermentation process beginning when concentration increases, but after three hours of fermentation, it decreases.

Keywords: bio-hydrogen, fermentation, experimental bioreactor, crude glycerol

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Authors: Mohammad Alfaifi

Abstract:

Organophosphates are among the most widely used synthetic insect pesticides. The widespread use of organophosphates has stimulated research into the possible existence of effects related with their reproductive toxic activity. The present study aimed to assess the effects of diazinon (DIZ) on male reproductive system. DIZ at the dose levels of 1.5, 3.0 and 9.0 mg/kg b. wt./day was administered orally to male rats of Wistar strain for 30 days to evaluate the toxic alterations in testicular histology, biochemistry, sperm dynamics, and testosterone levels. The body weight of animals did not show any significant changes, however, a significant reduction was observed in testes weight. DIZ also brought about marked reduction in epididymal and testicular sperm counts in exposed males and a decrease in serum testosterone concentration. Histopathological examination of testes showed mild to severe degenerative changes in seminiferous tubules at various dose levels. Fertility test showed 79% negative results. All these toxic effects are moderate at low doses and become severe at higher dose levels. From the results of the present study it is concluded that DIZ induces severe testicular damage and results in reduction in sperm count and thus affect fertility. Small changes in sperm counts are known to have adverse affects on human fertility. Therefore, application of such insecticide should be limited to a designed programme.

Keywords: reproductive toxicity, fertility, diazinon, sperm count

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Authors: Mona Heydari, Ehsan Motamedian, Seyed Abbas Shojaosadati

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Prediction of perturbations after genetic manipulation (especially gene knockout) is one of the important challenges in systems biology. In this paper, a new algorithm is introduced that integrates microarray data into the metabolic model. The algorithm was used to study the change in the cell phenotype after knockout of Gss gene in Escherichia coli BW25113. Algorithm implementation indicated that gene deletion resulted in more activation of the metabolic network. Growth yield was more and less regulating gene were identified for mutant in comparison with the wild-type strain.

Keywords: metabolic network, gene knockout, flux balance analysis, microarray data, integration

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Authors: S. V. Ncube, A. I. O. Jideani, E. T. Gwata

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The plight of food insecurity in developing countries has led to renewed interest in underutilized legumes. Their nutritional versatility, desirable functionality, pharmaceutical value and inherent bioactive compounds have drawn the attention of researchers. This has provoked the development of value added products with the aim of commercially exploiting their full potential. However processing of these legumes leads to changes in nutritional composition as affected by processing variables like pH, temperature and pressure. There is therefore a need for process control and quality assurance during production of the value added products. However, conventional methods for microbiological and biochemical identification are labour intensive and time-consuming. Biosensors offer rapid and affordable methods to assure the quality of the products. They may be used to quantify nutrients and anti-nutrients in the products while manipulating and monitoring variables such as pH, temperature, pressure and oxygen that affect the quality of the final product. This review gives an overview of the types of biosensors used in the food industry, their advantages and disadvantages and their possible application in processing of legumes.

Keywords: legume processing, biosensors, quality control, nutritional versatility

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Authors: Ehsan Motamedian

Abstract:

Finding all optimal flux distributions of a metabolic model is an important challenge in systems biology. In this paper, a new algorithm is introduced to identify all alternate optimal solutions of a large scale metabolic network. The algorithm reduces the model to decrease computations for finding optimal solutions. The algorithm was implemented on the Escherichia coli metabolic model to find all optimal solutions for lactate and acetate production. There were more optimal flux distributions when acetate production was optimized. The model was reduced from 1076 to 80 variable fluxes for lactate while it was reduced to 91 variable fluxes for acetate. These 11 more variable fluxes resulted in about three times more optimal flux distributions. Variable fluxes were from 12 various metabolic pathways and most of them belonged to nucleotide salvage and extra cellular transport pathways.

Keywords: flux variability, metabolic network, mixed-integer linear programming, multiple optimal solutions

Procedia PDF Downloads 426