World Academy of Science, Engineering and Technology

Authors: Tawqeer Ali Syed, Prakash Chandra

Abstract:

This study aims to evaluate the antileishmanial activity of Nigella sativa black seed extract. This well-known plant extract was taken from the botanical garden of Kashmir. Materials and Methods: The methanolic extracts of these plants were screened for their antileishmanial activity against Leishmania major using 3‑(4.5‑dimethylthiazol‑2yl)‑2.5‑diphenyltetrazolium bromide assay or MTT assay. Results: The methanolic extract of Nigella sativa showed potential antileishmanial activity at an inhibition% value of 80.29% ± 0.65%. IC 50 was calculated after 48 hours to be 964.3 µg/ml. Conclusion: Considering these results, these medicinal plants from Kashmir could serve as potential drug sources for antileishmanial compounds.

Keywords: MTT assay, antileishmanial, cell viability, Nigella sativa

Procedia PDF Downloads 193

Authors: Anushaa A.

Abstract:

In this work, silver nanoparticles (AgNP) were manufactured directly without harmful chemicals utilising methanol extract (SNLME) Solanum nigrume leaves. We are using nigrum leaf extract from Solanum, which converts silver nitrate to silver ions, for synthesization purposes. An examination of the AgNP produced was performed using ultraviolet (UV-VIS) spectroscopy, infrared spectroscopy (FTIR) transformed from Fourier and scanning electrons (SEM). Biological activity was also tested. UV-VIS has proven that biosynthesized AgNP exists (420-450 nm). The FTIR spectrum has been utilised to confirm the presence of different functional groups within the biomolecules, which are a nanoparticular capping agent and the spectroscopic and crystal nature of AgNP. The viability of the silver nanoparticles was evaluated using zeta potential calculations. Negative zeta potential of -33.4 mV demonstrated the stability of silver-nanoparticles. The morphology of AgNP was examined using a scanning electron microscope. Greenly generated AgNP showed significant anti-Staphylococcus aureus, Candida, and Escherichia coli action. The green AgNP demonstration indicated that the IC50 for the human teratocarcinoma cell line was 29.24 μg/ml during 24 hours of therapy (PA1 Ovarian cell line). The dose-dependent effects were reported in both antibacterial and cytotoxicity assays and as an effective agent. Finally, the findings of this research showed that silver nanoparticles generated might serve as a viable therapeutic agent to combat microorganisms killing and curing cancer.

Keywords: antimicrobial activity, PA1 ovarian cancer cell line, silver nanoparticles, Solanum nigrum

Procedia PDF Downloads 171

Authors: Hana Hanaee-Ahvaz, Monika Cserjan-Puschmann, Christopher Tauer, Gerald Striedner

Abstract:

Incorporation of noncanonical amino acids (ncAA) into proteins has become an interesting topic as proteins featured with ncAAs offer a wide range of different applications. Nowadays, technologies and systems exist that allow for the site-specific introduction of ncAAs in vivo, but the efficient production of proteins modified this way is still a big challenge. This is especially true for 'hard-to-express' proteins where low yields are encountered even with the native sequence. In this study, site-specific incorporation of azido-ethoxy-carbonyl-Lysin (azk) into an anti-tumor-necrosis-factor-α-Fab (FTN2) was investigated. According to well-established parameters, possible site positions for ncAA incorporation were determined, and corresponding FTN2 genes were constructed. Each of the modified FTN2 variants has one amber codon for azk incorporated either in its heavy or light chain. The expression level for all variants produced was determined by ELISA, and all azk variants could be produced with a satisfactory yield in the range of 50-70% of the original FTN2 variant. In terms of expression yield, neither the azk incorporation position nor the subunit modified (heavy or light chain) had a significant effect. We confirmed correct protein processing and azk incorporation by mass spectrometry analysis, and antigen-antibody interaction was determined by surface plasmon resonance analysis. The next step is to characterize the effect of azk incorporation on protein stability and aggregation tendency via differential scanning calorimetry and light scattering, respectively. In summary, the incorporation of ncAA into our Fab candidate FTN2 worked better than expected. The quantities produced allowed a detailed characterization of the variants in terms of their properties, and we can now turn our attention to potential applications. By using click chemistry, we can equip the Fabs with additional functionalities and make them suitable for a wide range of applications. We will now use this option in a first approach and develop an assay that will allow us to follow the degradation of the recombinant target protein in vivo. Special focus will be laid on the proteolytic activity in the periplasm and how it is influenced by cultivation/induction conditions.

Keywords: degradation, FTN2, hard-to-express protein, non-canonical amino acids

Procedia PDF Downloads 214

Authors: Soultana Konstantinidou, Tiziana Schmidt, Elena Landi, Alessandro De Carli, Giovanni Maltinti, Darius Witt, Alicja Dziadosz, Agnieszka Lindstaedt, Michele Lai, Mauro Pistello, Valentina Cappello, Luciana Dente, Chiara Gabellini, Piotr Barski, Vittoria Raffa

Abstract:

CRISPR/Cas9 technology has gained the interest of researchers in the field of biotechnology for genome editing. Since its discovery as a microbial adaptive immune defense, this system has been widely adopted and is acknowledged for having a variety of applications. However, critical barriers related to safety and delivery are persisting. Here, we propose a new concept of genome engineering, which is based on a nano-formulation of Cas9. The Cas9 enzyme was conjugated to a gold nanoparticle (AuNP-Cas9). The AuNP-Cas9 maintained its cleavage efficiency in vitro, to the same extent as the ribonucleoprotein, including non-conjugated Cas9 enzyme, and showed high gene editing efficiency in vivo in zebrafish embryos. Since CRISPR/Cas9 technology is extensively used in cancer research, melanoma was selected as a validation target. Cell studies were performed in A375 human melanoma cells. Particles per se had no impact on cell metabolism and proliferation. Intriguingly, the AuNP-Cas9 internalized spontaneously in cells and localized as a single particle in the cytoplasm and organelles. More importantly, the AuNP-Cas9 showed a high nuclear localization signal. The AuNP-Cas9, overcoming the delivery difficulties of Cas9, could be used in cellular biology and localization studies. Taking advantage of the plasmonic properties of gold nanoparticles, this technology could potentially be a bio-tool for combining gene editing and photothermal therapy in cancer cells. Further work will be focused on intracellular interactions of the nano-formulation and characterization of the optical properties.

Keywords: CRISPR/Cas9, gene editing, gold nanoparticles, nanotechnology

Procedia PDF Downloads 85

Authors: Elene Zhuravliova, Tamar Barbakadze, Irina Kalandadze, Elnari Zaalishvili, Lali Shanshiashvili, David Mikeladze

Abstract:

Background: Multiple sclerosis (MS) is an inflammatory, neurodegenerative disease, which is accompanied by demyelination and autoimmune response to myelin proteins. Among post-translational modifications, which mediate the modulation of inflammatory pathways during MS, methylation is the main one. The methylation of DNA, also amino acids lysine and arginine, occurs in the cell. It was found that decreased trans-methylation is associated with neuroinflammatory diseases. Therefore, abnormal regulation of the methyl cycle could induce demyelination through the action on PAD (peptidyl-arginine-deiminase) gene promoter. PAD takes part in protein citrullination and targets myelin basic protein (MBP), which is affected during demyelination. To determine whether MBP charge isomers are changing the methyl cycle, we have estimated the concentrations of methyl cycle metabolites in MBP-activated primary astrocytes and oligodendrocytes. For this purpose, the action of the citrullinated MBP- C8 and the most cationic MBP-C1 isomers on the primary cells were investigated. Methods: Primary oligodendrocyte and astrocyte cell cultures were prepared from whole brains of 2-day-old Wistar rats. The methyl cycle metabolites, including homocysteine, S-adenosylmethionine (SAM), and S-adenosylhomocysteine (SAH), were estimated by HPLC analysis using fluorescence detection and prior derivatization. Results: We found that the action of MBP-C8 and MBP-C1 induces a decrease in the concentration of both methyl cycle metabolites, S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH), in astrocytes compared to the control cells. As for oligodendrocytes, the concentration of SAM was increased by the addition of MBP-C1, while MBP-C8 has no significant effect. As for SAH, its concentration was increased compared to the control cells by the action of both MBP-C1 and MBP-C8. A significant increase in homocysteine concentration was observed by the action of the MBP-C8 isomer in both oligodendrocytes and astrocytes. Conclusion: These data suggest that MBP charge isomers change the concentration of methyl cycle metabolites. MBP-C8 citrullinated isomer causes elevation of homocysteine in astrocytes and oligodendrocytes, which may be the reason for decreased astrocyte proliferation and increased oligodendrocyte cell death which takes place in neurodegenerative processes. Elevated homocysteine levels and subsequent abnormal regulation of methyl cycles in oligodendrocytes possibly change the methylation of DNA that activates PAD gene promoter and induces the synthesis of PAD, which in turn provokes the process of citrullination, which is the accompanying process of demyelination. Acknowledgment: This research was supported by the SRNSF Georgia RF17_534 grant.

Keywords: myelin basic protein, astrocytes, methyl cycle metabolites, homocysteine, oligodendrocytes

Procedia PDF Downloads 139

Authors: Lali Shanshiashvili, Marika Chikviladze, Nino Mamulashvili, Maia Sepashvili, Nana Narmania, David Mikeladze

Abstract:

Purpose: During demyelinating inflammatory diseases and after the damage of the myelin sheet, myelin-derived proteins, including myelin basic protein (MBP), are secreted into the extracellular space. MBP shows extensive post-translational modifications, including the deimination of arginine residues. Deiminated MBP is structurally less ordered, susceptible to proteolytic attack, and more immunogenic than the unmodified one. It is hypothesized that MBP could change the inflammatory response in astrocytes. Methods: MBP was isolated and purified from bovine brain white matter. Primary astrocyte cultures were prepared from whole brains of 2-day-old Wistar rats. For evaluation of glutamate uptake/release in astrocytes following treatment of cells with MBP charge isomers, Glutamate Assay Kit was used. The expression of EAAT-2 (excitatory amino acid transporters), peroxisome proliferator-activated receptor gamma (PPAR- γ), inhibitor of nuclear factor kappa B (IkB), and high mobility group protein B1 (HMGB1) in astrocytes were assayed by Western Blot analysis. Results: This study investigated the action of deiminated isomer (C8) on the cultured primary astrocytes and compared its effects with the effects of unmodified C1 isomers. The study found that C8 and C1 MBP differently act on the uptake and release of glutamate in astrocytes: nonmodified C1 MBP increases the uptake of glutamate and does not change the release, whereas C8 decreases the release of glutamate but does not alter the uptake. Nevertheless, both isomers increased the expression of PPAR-γ and EAAT2 in the same intensity. However, immunostaining and Western Blots of cell lysates showed a decrease of IkB and increased expression of HMGB1 after the treatment of astrocytes by C8. Moreover, in the presence of C8, astrocytes release more nitric oxide than unmodified C1 isomers. Conclusion: These data suggest that the deiminated isomer of MBP evokes an inflammatory response and enhances the ability of astrocytes to release proinflammatory mediators through activation of NF-kB after the breakdown of myelin sheets. Acknowledgment: This research was supported by the SRNSF Georgia RF17_534 grant.

Keywords: myelin basic protein, glutamate, deimination, astrocytes, inflammation

Procedia PDF Downloads 191

Authors: Scott Nielsen, Luca Longanesi, Chris Chuck

Abstract:

Palm oil is obtained from the flesh and kernel of the fruit of oil palms and is the most productive and inexpensive oil crop. The global demand for palm oil is approximately 75 million metric tonnes, a 29% increase in global production of palm oil since 2016. This expansion of oil palm cultivation has resulted in mass deforestation, vast biodiversity destruction and increasing net greenhouse gas emissions. One possible alternative is to produce a saturated oil, similar to palm, from microbes such as oleaginous yeast. The yeasts can be cultured on sugars derived from second-generation sources and do not compete with tropical forests for land. One highly promising oleaginous yeast for this application is Metschnikowia pulcherrima. However, recent techno-economic modeling has shown that cell lysis and standard lipid extraction are major contributors to the cost of the oil. Typical cell disruption techniques to extract either single cell oils or proteins have been based around bead-beating, homogenization and acid lysis. However, these can have a detrimental effect on lipid quality and are energy-intensive. In this study, a vortex separator, which produces high sheer with minimal energy input, was investigated as a potential low energy method of lysing cells. This was compared to four more traditional methods (thermal lysis, acid lysis, alkaline lysis, and osmotic lysis). For each method, the yeast loading was also examined at 1 g/L, 10 g/L and 100 g/L. The quality of the cell disruption was measured by optical cell density, cell counting and the particle size distribution profile comparison over a 2-hour period. This study demonstrates that the vortex separator is highly effective at lysing the cells and could potentially be used as a simple apparatus for lipid recovery in an oleaginous yeast process. The further development of this technology could potentially reduce the overall cost of microbial lipids in the future.

Keywords: palm oil substitute, metschnikowia pulcherrima, cell disruption, cell lysis

Procedia PDF Downloads 183

Authors: Tereza Branyšová, Martina Kračmarová, Kateřina Demnerová, Michal Ďurovič, Hana Stiborová

Abstract:

Microbial deterioration threatens all objects of cultural heritage, including audio-visual materials. Fungi are commonly known to be the main factor in audio-visual material deterioration. However, although being neglected, bacteria also play a significant role. In addition to microbial contamination of materials, it is also essential to analyse air as a possible contamination source. This work aims to identify bacterial species in the archives of the Czech Republic that occur on audio-visual materials as well as in the air in the archives. For sampling purposes, the smears from the materials were taken by sterile polyurethane sponges, and the air was collected using a MAS-100 aeroscope. Metagenomic DNA from all collected samples was immediately isolated and stored at -20 °C. DNA library for the 16S rRNA gene was prepared using two-step PCR and specific primers and the concentration step was included due to meagre yields of the DNA. After that, the samples were sent to the University of Fairbanks, Alaska, for Illumina MiSeq sequencing. Subsequently, the analysis of the sequences was conducted in R software. The obtained sequences were assigned to the corresponding bacterial species using the DADA2 package. The impact of air contamination and the impact of different photosensitive layers that audio-visual materials were made of, such as gelatine, albumen, and collodion, were evaluated. As a next step, we will take a deeper focus on air contamination. We will select an appropriate culture-dependent approach along with a culture-independent approach to observe a metabolically active species in the air. Acknowledgment: This project is supported by grant no. DG18P02OVV062 of the Ministry of Culture of the Czech Republic.

Keywords: cultural heritage, Illumina MiSeq, metagenomics, microbial identification

Procedia PDF Downloads 135

Authors: Pedro Maldonado-Alvarado

Abstract:

An important cassava quality production was detected in Ecuador. However, in this country, few products with low adding-value are produced from the tuber and none from cassava by-products. To our best knowledge, lactic acid was produced from Ecuadorian cassava bagasse starch in a biotechnological way. The objective of this contribution was to study the influence of the fermentation variables (pH and agitation) on the lactic acid production of Ecuadorian cassava varieties from bagasse starch. Enzymatic hydrolysis of cassava bagasse starch for INIAP 650 and INIAP 651 varieties spread in Ecuador was performed using α-amylase and amyloglucosidase. Then, glucose was fermented by Lactobacillus leichmannii strains in different conditions of agitation (0 and 150 rpm) and pH (4.5, 5.0, and 5.5). Significant differences in ash, fibre, protein, lipids, and amylose were found in cassava bagasse starch of INIAP 650 and INIAP 651 with 1.4 and 1.3%, 4.3 and 6%, 1.2 and 2.1%, 1.9 and 1.5%, and 24.3 and 26.5%, respectively. The determination of lactic acid was performed by potentiometric and FTIR analysis. Conversions of cassava bagasse to reduced sugars were 71.7 and 85.1% for INIAP 650 and INIAP 651, respectively. The best lactic acid concentrations were 27.6 and 33.5 g/L, obtained at agitation 150 rpm and pH 5.5 for INIAP 650 and INIAP 651. Qualitative analysis conducted by FTIR spectrophotometry confirmed the presence of lactic acid in the reacted products. This investigation could contribute to the valorisation of residues from promising cassava varieties in Ecuador and hence to increase the development of this country.

Keywords: bagasse starch, cassava, Ecuador, fermentation, lactic acid

Procedia PDF Downloads 179

Authors: Sadia Ilyas, Hyunjung Kim, Rajiv R. Srivastava

Abstract:

Current work has attempted to establish the linkages between circular bio-economy and recycling of copper and gold from urban mine by applying microbial activities instead of the smelter and chemical technologies. Thereafter, based on the potential of microbial approaches and research hypothesis, the structural model has been tested for a significance level of 99%, which is supported by the corresponding standardization co-efficient values. A prediction model applied to determine the recycling impact on circular bio-economy indicates to re-circulate 51,833 tons of copper and 58 tons of gold by 2030 for the production of virgin metals/raw-materials, while recycling rate of the accumulated e-waste remains to be 20%. This restoration volume of copper and gold through the microbial activities corresponds to mitigate 174 million kg CO₂ emissions and 24 million m³ water consumption if compared with the primary production activities. The study potentially opens a new window for environmentally-friendly biotechnological recycling of e-waste urban mine under the umbrella concept of circular bio-economy.

Keywords: urban mining, biobleaching, circular bio-economy, environmental impact

Procedia PDF Downloads 140

Authors: N. Bakradze, T. Dumbadze, N. Gagelidze, L. Amiranashvili, A. D. L. Batako

Abstract:

Cereals are considered as a strategic product in human life and it demand is increasing with the growth of world population. There is always shortage of cereals in various areas of the globe. For example, Georgia own production meets only 15-20% of the demand for grain, despite the fact that the country is considered one of the main centers of wheat origin. In Georgia, there are 14 types of wheat and more than 150 subspecies, and 40 subspecies of common wheat. Increasing wheat production is important for the country. One of the ways to solve the problem is to develop and implement new, environmentally and economically acceptable technologies. Such technologies include pre-sowing treatment of seed with a laser and associative nitrogen-fixing of the Azospirillum brasilensse bacteria. In the region there are Dika and Lomtagora which are among the most common in Georgia. Dika is a frost-resistant wheat, with a high ability to adapt to the environment, resistant to falling and it is sown in highlands. Dicka excellent properties are due to its strong immunity to fungal diseases; Dicka grains are rich in protein and lysine. Lomtagora 126 differs with its winter and drought resistance, and, it has a great ability to germinate. Lomtagora is characterized by a strong root system and a high budding capacity. It is an early variety, fall-resistant, easy to thresh and suitable for mechanized harvesting with large and red grains. The plant is moderately resistant to fungal diseases. This paper presents some preliminary experimental results where, a continuous CO2 laser at a power of 25-40 W/cm2 was used to radiate grains at a flow rate of 10-15 cm/sec. The treatment was carried out on grains of the Triticum aestivum L. var. of Lutescens (local variety name - Lomtagora 126), and Triticum carthlicum Nevski (local variety name - Dika). Here the grains were treated with Azospirillum brasilensse isolate (108-109 CFU / ml), which was isolated from the rhizosphere of wheat. It was observed that the germination of the wheat was not significantly influenced by either laser or bacteria treatment. In the case of the variety Lomtagora 126, when irradiated at an angle of 90°, it slightly improved the growth within 38 days of sawing, and in the case of irradiation at an angle of 90°+1, by 23%. The treatment of seeds with Azospirillum brazilense in both irradiated and non-irradiated variants led to an improvement in the growth of ssprouts. However, in the case of treatment with azospiril alone - by 22%, and with joint treatment of seeds with azospiril and irradiation - by 29%. In the case of the Dika wheat, the irradiation only led to an increase in growth by 8-9%, and the combine treatment of seeds with azospiril and irradiation - by 10-15%, in comparison with the control. Thus, the combine treatment of wheat of different varieties provided the best effect on the growth. Acknowledgment: This work was supported by Shota Rustaveli National Science Foundation of Georgia (SRNSFG) (Grant number CARYS 19-573)

Keywords: laser treatment, Azospirillum brasilensse, seeds, wheat varieties, Lomtagora, Dika

Procedia PDF Downloads 127

Authors: Laura Mejias, Sandra Monteagudo, Oscar Martinez-Avila, Sergio Ponsa

Abstract:

Bioplastics are gaining attention as potential substitutes of conventional fossil-derived plastics and new components of specialized applications in different industries. Besides, these constitute a sustainable alternative since they are biodegradable and can be obtained starting from renewable sources. Thus, agro-industrial wastes appear as potential substrates for bioplastics production using microorganisms, considering they are a suitable source for nutrients, low-cost, and available worldwide. Therefore, this approach contributes to the biorefinery and circular economy paradigm. The present study assesses the solid-state fermentation (SSF) technology for the co-synthesis of exopolysaccharides (EPS) and polyhydroxyalkanoates (PHA), two attractive biodegradable bioplastics, using the leftover of the brewery industry brewer's spent grain (BSG). After an initial screening of diverse PHA-producer bacteria, it was found that Burkholderia cepacia presented the highest EPS and PHA production potential via SSF of BSG. Thus, B. cepacia served to identify the most relevant aspects affecting the EPS+PHA co-synthesis at a lab-scale (100g). Since these are growth-dependent processes, they were monitored online through oxygen consumption using a dynamic respirometric system, but also quantifying the biomass production (gravimetric) and the obtained products (EtOH precipitation for EPS and solid-liquid extraction coupled with GC-FID for PHA). Results showed that B. cepacia has grown up to 81 mg per gram of dry BSG (gDM) at 30°C after 96 h, representing up to 618 times higher than the other tested strains' findings. Hence, the crude EPS production was 53 mg g-1DM (2% carbohydrates), but purity reached 98% after a dialysis purification step. Simultaneously, B. cepacia accumulated up to 36% (dry basis) of the produced biomass as PHA, mainly composed of polyhydroxybutyrate (P3HB). The maximum PHA production was reached after 48 h with 12.1 mg g⁻¹DM, representing threefold the levels previously reported using SSF. Moisture content and aeration strategy resulted in the most significant variables affecting the simultaneous production. Results show the potential of co-synthesis via SSF as an attractive alternative to enhance bioprocess feasibility for obtaining these bioplastics in residue-based systems.

Keywords: bioplastics, brewer’s spent grain, circular economy, solid-state fermentation, waste to product

Procedia PDF Downloads 127

Authors: Bruno Baptista, Andreia S. R. Oliveira, Patricia V. Mendonca, Jorge F. J. Coelho, Fani Sousa

Abstract:

Drug delivery systems are designed to allow adequate protection and controlled delivery of drugs to specific locations. These systems aim to reduce side effects and control the biodistribution profile of drugs, thus improving therapeutic efficacy. This study involved the synthesis of polymeric nanoparticles, based on amphiphilic diblock copolymers, comprising a biocompatible, poly (oligo (ethylene oxide) methyl ether methacrylate (POEOMA) as hydrophilic segment and a pH-sensitive block, the poly (2-diisopropylamino)ethyl methacrylate) (PDPA). The objective of this work was the development of polymeric pH-responsive nanoparticles to encapsulate and carry small RNAs as a model to further develop non-coding RNAs delivery systems with therapeutic value. The responsiveness of PDPA to pH allows the electrostatic interaction of these copolymers with nucleic acids at acidic pH, as a result of the protonation of the tertiary amine groups of this polymer at pH values below its pKa (around 6.2). Initially, the molecular weight parameters and chemical structure of the block copolymers were determined by size exclusion chromatography (SEC) and nuclear magnetic resonance (1H-NMR) spectroscopy, respectively. Then, the complexation with small RNAs was verified, generating polyplexes with sizes ranging from 300 to 600 nm and with encapsulation efficiencies around 80%, depending on the molecular weight of the polymers, their composition, and concentration used. The effect of pH on the morphology of nanoparticles was evaluated by scanning electron microscopy (SEM) being verified that at higher pH values, particles tend to lose their spherical shape. Since this work aims to develop systems for the delivery of non-coding RNAs, studies on RNA protection (contact with RNase, FBS, and Trypsin) and cell viability were also carried out. It was found that they induce some protection against constituents of the cellular environment and have no cellular toxicity. In summary, this research work contributes to the development of pH-sensitive polymers, capable of protecting and encapsulating RNA, in a relatively simple and efficient manner, to further be applied on drug delivery to specific sites where pH may have a critical role, as it can occur in several cancer environments.

Keywords: drug delivery systems, pH-responsive polymers, POEOMA-b-PDPA, small RNAs

Procedia PDF Downloads 244

Authors: Debamitra Chakravorty, Pratap K. Parida

Abstract:

Cold-adapted proteases enhance wash performance in low-temperature laundry resulting in a reduction in energy consumption and wear of textiles and are also used in the dehairing process in leather industries. Unfortunately, the possible drawbacks of using cold-adapted proteases are their instability at higher temperatures. Therefore, proteases with broad temperature stability are required. Unfortunately, wild-type cold-adapted proteases exhibit instability at higher temperatures and thus have low shelf lives. Therefore, attempts to engineer cold-adapted proteases by protein engineering were made previously by directed evolution and random mutagenesis. The lacuna is the time, capital, and labour involved to obtain these variants are very demanding and challenging. Therefore, rational engineering for cold stability without compromising an enzyme's optimum pH and temperature for activity is the current requirement. In this work, mutations were rationally designed with the aid of high throughput computational methodology of network analysis, evolutionary conservation scores, and molecular dynamics simulations for Savinase from Bacillus lentus with the intention of rendering the mutants cold stable without affecting their temperature and pH optimum for activity. Further, an attempt was made to incorporate a mutation in the most stable mutant rationally obtained by this method to introduce oxidative stability in the mutant. Such enzymes are desired in detergents with bleaching agents. In silico analysis by performing 300 ns molecular dynamics simulations at 5 different temperatures revealed that these three mutants were found to be better in cold stability compared to the wild type Savinase from Bacillus lentus. Conclusively, this work shows that cold adaptation without losing optimum temperature and pH stability and additionally stability from oxidative damage can be rationally designed by in silico enzyme engineering. The key findings of this work were first, the in silico data of H5 (cold stable savinase) used as a control in this work, corroborated with its reported wet lab temperature stability data. Secondly, three cold stable mutants of Savinase from Bacillus lentus were rationally identified. Lastly, a mutation which will stabilize savinase against oxidative damage was additionally identified.

Keywords: cold stability, molecular dynamics simulations, protein engineering, rational design

Procedia PDF Downloads 121

Authors: David B. Kisakye, Paul Mugabi

Abstract:

Biomass briquettes have been identified as a plausible and close alternative to commonly used energy fuels such as charcoal and firewood, whose prices are escalating due to the dwindling natural resource base. However, briquettes do not seem to be as popular as would be expected. This study assessed the production, distribution, and acceptability of the briquettes in the Kampala district. A total of 60 respondents, 50 of whom were briquette users and 10 briquette producers, were sampled from five divisions of Kampala district to evaluate consumer acceptability, preference for briquette type and shape. Households and institutions were identified to be the major consumers of briquettes, while community-based organizations were the major distributors of briquettes. The Chi-square test of independence showed a significant association between briquette acceptability and briquette attributes of substitutability and low cost (p < 0,05). The Kruskal Wallis test showed that low-income class people preferred non-carbonized briquettes. Gender, marital status, and income level also cause variation in preference for spherical, stick, and honeycomb briquettes (p < 0,05). The major challenges faced by briquette users in Kampala were; production of a lot of ash, frequent crushing, and limited access to briquettes. The producers of briquettes were mainly challenged by regular machine breakdown, raw material scarcity, and poor carbonizing units. It was concluded that briquettes have a market and are generally accepted in Kampala. However, user preferences need to be taken into account by briquette produces, suitable cookstoves should be availed to users, and there is a need for standards to ensure the quality of briquettes.

Keywords: consumer acceptability, biomass residues, briquettes, briquette producers, distribution, fuel, marketability, wood fuel

Procedia PDF Downloads 121

Authors: Shuo Mu, Guangzhi Jiang, Jinsa Chen

Abstract:

The analysis of Indel for RNA sequencing of clinical samples is easily affected by sequencing experiment errors and software selection. In order to improve the efficiency and accuracy of analysis, we developed an automatic reporting system for Indel recognition and annotation based on image snapshot of transcriptome reads alignment. This system includes sequence local-assembly and realignment, target point snapshot, and image-based recognition processes. We integrated high-confidence Indel dataset from several known databases as a training set to improve the accuracy of image processing and added a bioinformatical processing module to annotate and filter Indel artifacts. Subsequently, the system will automatically generate data, including data quality levels and images results report. Sanger sequencing verification of the reference Indel mutation of cell line NA12878 showed that the process can achieve 83% sensitivity and 96% specificity. Analysis of the collected clinical samples showed that the interpretation accuracy of the process was equivalent to that of manual inspection, and the processing efficiency showed a significant improvement. This work shows the feasibility of accurate Indel analysis of clinical next-generation sequencing (NGS) transcriptome. This result may be useful for RNA study for clinical samples with microsatellite instability in immunotherapy in the future.

Keywords: automatic reporting, indel, next-generation sequencing, NGS, transcriptome

Procedia PDF Downloads 166

Authors: Abdelghani Tahiri, Youssef Karra, Naima Ait Aabd, Abdelaziz Mimouni

Abstract:

Saffron (Crocus sativus L.), the most expensive spice in the world derived from the stigmas, is an autumn-flowering and sterile triploid (2n=3x=24) geophyte species that belong to the Iridaceae family. This plant species is mainly propagated vegetatively through the formation of daughter corms from the mother one. Low multiplication rates of daughter corms under natural conditions, along with fungal contamination, significantly reduce the productivity and quality of saffron corms. The development of efficient and sustainable strategies for rapid and large-scale production of selected cultivars of saffron will be desired. For this, the main objective of this work is to improve the vegetative propagation of saffron under ex-vitro conditions. Preliminary results of the influence of increasing doses of humic substances (HS) on the growth and multiplication of corms under greenhouse conditions are evaluated. The obtained data shows that the effect of HS depends on the concentration used and the mode of application. Indeed, the application through irrigation has increased the number of shoots and corms, but it has reduced other parameters. On the other hand, the temporary treatment has improved all observed parameters except for the number of shoots and corms. Results obtained in this work suggest that it is possible to improve the propagation of saffron corms under greenhouse conditions.

Keywords: saffron, Crocus sativus L., corm, humic substances

Procedia PDF Downloads 198

Authors: Marie-Elise Beydon, Daniel Sacristan, Isabel Ruppen

Abstract:

MB02 (Alymsys®) is a candidate biosimilar to bevacizumab, which was developed against the reference product (RP) Avastin® sourced from both the European Union (EU) and United States (US). MB02 has been extensively characterized comparatively to Avastin® at a physicochemical and biological level using sensitive orthogonal state-of-the-art analytical methods. MB02 has been demonstrated similar to the RP with regard to its primary and higher-order structure, post- and co-translational profiles such as glycosylation, charge, and size variants. Specific focus has been put on the characterization of Fab-related activities, such as binding to VEGF A 165, which directly reflect the bevacizumab mechanism of action. Fc-related functionality was also investigated, including binding to FcRn, which is indicative of antibodies' half-life. The data generated during the analytical similarity assessment demonstrate the high analytical similarity of MB02 to its RP.

Keywords: analytical similarity, bevacizumab, biosimilar, MB02

Procedia PDF Downloads 257

Authors: S. Najafi, E. Bertini, M. Pezzotti, G.B. Tornielli, S. Zenoni

Abstract:

Grapevine is an important agricultural fruit crop plant consumed worldwide and with a key role in the global economy. Grapevine is strongly affected by both biotic and abiotic stresses, which impact grape growth at different stages, such as during plant and berry development and pre- and post-harvest, consequently causing significant economic losses. Recently global warming has propelled the anticipation of the onset of berry ripening, determining the reduction of a grape color and increased volatilization of aroma compounds. Climate change could negatively alter the physiological characteristics of the grape and affect the berry and wine quality. Modern plant breeding can provide tools such as genome editing for improving grape resilience traits while maintaining intact the viticultural and oenological quality characteristics of the genotype. This study aims at developing a platform for genome editing application in grapevine plants with the final goal to improve berry quality, biotic, and abiotic resilience traits. We chose to directly deliver ribonucleoproteins (RNP, preassembled Cas protein and guide RNA) into plant protoplasts, and, from these cell structures, regenerate grapevine plants edited in specific selected genes controlling traits of interest. Edited plants regenerated by somatic embryogenesis from protoplasts will then be sequenced and molecularly characterized. Embryogenic calli of Sultana and Shiraz cultivars were initiated from unopened leaves of in-vitro shoot tip cultures and from stamens, respectively. Leaves were placed on NB2 medium while stamens on callus initiation medium (PIV) medium and incubated in the dark at 28 °C for three months. Viable protoplasts, tested by FDA staining, isolated from embryogenic calli were cultured by disc method at 1*105 protoplasts/ml. Mature well-shaped somatic embryos developed directly in the protoplast culture medium two months later and were transferred in the light into to shooting medium for further growth. Regenerated plants were then transferred to the greenhouse; no phenotypic alterations were observed when compared to non in-vitro cultured plants. The performed experiments allowed to established an efficient protocol of embryogenic calli production, protoplast isolation, and regeneration of the whole plant through somatic embryogenesis in both Sultana and Shiraz. Regenerated plants, through direct somatic embryogenesis deriving from a single cell, avoid the risk of chimerism during the regeneration process, therefore improving the genome editing process. As pre-requisite of genome editing, an efficient method for transfection of protoplast by yellow fluorescent protein (YFP) marker genes was also established and experiments of direct delivery of CRISPR–Cas9 ribonucleoproteins (RNPs) in protoplasts to achieve efficient DNA-free targeted mutations are in progress.

Keywords: CRISPR-cas9, plant regeneration, protoplast isolation, Vitis vinifera

Procedia PDF Downloads 130

Authors: Techale B., Hongxu Dong, Mihrete Getinet, Aregash Gabizew, Andrew H. Paterson, Kassahun Bantte

Abstract:

The genetic architecture of drought tolerance is expected to involve multiple loci that are unlikely to all segregate for alternative alleles in a single bi-parental population. Therefore, the identification of quantitative trait loci (QTL) that are expressed in diverse genetic backgrounds of multiple bi-parental populations provides evidence about both background-specific and common genetic variants. The purpose of this study was to map QTL related to drought tolerance using three connected mapping populations of different genetic backgrounds to gain insight into the genomic landscape of this important trait in elite Ethiopian germplasm. The three bi-parental populations, each with 207 F₂:₃ lines, were evaluated using an alpha lattice design with two replications under two moisture stress environments. Drought tolerance related traits were analyzed separately for each population using composite interval mapping, finding a total of 105 QTLs. All the QTLs identified from individual populations were projected on a combined consensus map, comprising a total of 25 meta QTLs for seven traits. The consensus map allowed us to deduce locations of a larger number of markers than possible in any individual map, providing a reference for genetic studies in different genetic backgrounds. The mQTL identified in this study could be used for marker-assisted breeding programs in sorghum after validation. Only one trait, reduced leaf senescence, showed a striking bias of allele distribution, indicating substantial standing variation among present varieties that might be employed in improving drought tolerance of Ethiopian and other sorghums.

Keywords: Drought tolerance , Mapping populations, Meta QTL, QTL mapping, Sorghum

Procedia PDF Downloads 163

Authors: Olusegun D. Badewa, E. K. Tsado, A. S. Gana, K. D. Tolorunse, R. U. Okechukwu, P. Iluebbey, S. Ibrahim

Abstract:

Farmers are faced with varying production challenges ranging from unstable weather due to climate change, low yield, malnutrition, cattle invasion, and bush fires that have always affected their livelihood. Research effort must therefore be centered on improving farmers’ livelihood, nutrition, and health by providing early bulking biofortified cassava varieties that could be harvested earlier with reasonable root yield and thereby preventing long stay of the crop on their farmland. This study evaluated cassava genotypes at different harvesting months of 3, 6, 9, and 12 months after planting in order to evaluate their bulking rate at different agroecology of Mokwa and Ubiaja. Data were collected on fresh storage root yield, Harvest index, and Dry matter content. It was shown from the study that traits FSRY, HI, and DM were significant for genotype and months after planting and variable among the genotype while location had no effect on the yield traits. Early bulking genotypes were not high yielding and showed discontinuity at some point across the months. The retrogression in yield performance across months had no effect on the highest yielding. Also, for all the genotypes and across evaluated months, FSRY reduces at 9 MAP due to a reduction in dry matter content during the same month, and the best performing genotype was the genotype IBA90581, followed by IBA120036, IBA130896, and IBA980581 while the least performing was genotype IBA130818.

Keywords: early bulking, dry mater, harvest index, high yielding, root yield

Procedia PDF Downloads 205

Authors: Michael Fundator

Abstract:

Groundbreaking application of biomathematical and biochemical research in neural networks processes to formation of non-canonical bases, islands, and analog bases in DNA and mRNA at or near the transcription that contradicts the long anticipated statistical assumptions for the distribution of bases and analog bases compounds is implemented through statistical and stochastic methods apparatus with addition of quantum principles, where the usual transience of Poisson spike train becomes very instrumental tool for finding even almost periodical type of solutions to Fokker-Plank stochastic differential equation. Present article develops new multidimensional methods of finding solutions to stochastic differential equations based on more rigorous approach to mathematical apparatus through Kolmogorov-Chentsov continuity theorem that allows the stochastic processes with jumps under certain conditions to have γ-Holder continuous modification that is used as basis for finding analogous parallels in dynamics of neutral networks and formation of analog bases and transcription in DNA.

Keywords: Fokker-Plank stochastic differential equation, Kolmogorov-Chentsov continuity theorem, neural networks, translation and transcription

Procedia PDF Downloads 391

Authors: Levylee Bautista, Dawn Grace Santos, Aishwarya Veluchamy, Jesusa Santos, Ghafoor Haque, Jr. I, Rodolfo Rafael

Abstract:

Sesbania grandiflora is widely used in traditional medicine to treat a wide range of ailments. Assessment of its toxic properties is hence crucial when considering public health protection because exposure to plant extracts may pose adverse effects on consumers. This study aimed to investigate the acute intraperitoneal toxicity of S. grandiflora flower methanolic extract (SGFME) in Swiss albino mice. Four different concentrations (11.25, 22.5, 40, and 90 mg/kg) of SGFME were administered intraperitoneally and immediate behavioral and clinical signs were observed. All concentrations of SGFME-treated mice exhibited gasping and faster respiratory rate, writhing, reddening and fanning of the ears, paralysis of the hind leg, and mortality. Such reactions may be attributed to the histamine and saponin content of S. grandiflora. Results of this study suggests that intraperitoneal administration of SGFME produced significant adverse effect in mice, therefore, caution should be exercised in using it as herbal remedy since there is little control over its quality.

Keywords: acute toxicity test, histamine, medicinal plants, Sesbania grandiflora

Procedia PDF Downloads 148

Authors: Levylee Bautista, Dawn Grace Santos, Aishwarya Veluchamy, Jesusa Santos, Ghafoor, Jr. I Haque, Rodolfo Rafael

Abstract:

The booming interest in using natural compounds as an alternative to conventional medications has paved way to focus the attention on plants that provide rich sources of bioactive phytochemicals. For regulatory purposes, evaluation of the genotoxic effects of such alternatives is therefore empirical as part of the plant’s hazard assessment. Sesbania grandiflora is among the plants used as a traditional remedy in folk medicine and a subject of research for its medicinal benefits. This study aimed to evaluate the genotoxic potential induced by S. grandiflora flower methanol extract (SGFME) in terms of the frequency of micronucleus (MN) in polychromatic erythrocyte (PCE) (MNPCE) and PCE ratio employing the micronucleus assay. The frequency of MN was examined in bone marrow cells (BMCs) obtained from male Swiss albino mice exposed in vivo to four different concentrations (11.25, 22.5, 40, and 90 mg/kg) of SGFME and MMC (70 mg/kg; positive control) and sacrificed 24 hours post-intraperitoneal injection. Results showed a significant (p < 0.01) rate of MNPCEs for 11.25 and 22.5 tested concentrations of SGFME and is comparable with the MMC-treated mice. Although PCE ratio values in all doses of SGFME-treated mice were over 0.20, it is worth noting that 40 and 90 tested concentrations of SGFME-treated mice exhibited the lowest value, i.e., 0.22 and 0.28, respectively. The present study has demonstrated that S. grandiflora possesses genotoxic potential for murine BMCs. Such activity could be ascribed from the bioactive compounds present in S. grandiflora that require further isolation and characterization of the active molecules. Likewise, findings of this study warrant a caution of the use of S. grandiflora insomuch as further investigations do not demonstrate their safety.

Keywords: genotoxicity, micronucleus, phytochemicals, Sesbania grandiflora

Procedia PDF Downloads 123

Authors: Pauline Ribert, Gaelle Roudaut, Sebastien Dupont, Laurent Beney

Abstract:

Yeasts, anhydrobiotic organisms, can survive extreme water disturbances, thanks to the prolonged and reversible suspension of their cellular activity as well as the establishment of a defense arsenal. This property is exploited by many industrialists. One of the protection systems implemented by yeast is the vitrification of its cytoplasm by trehalose. The thermal resistance of dry yeasts is a crucial parameter for their use. However, studies on the thermal resistance of dry yeasts are often based on yeasts produced in laboratory conditions with non-optimal drying processes. We, therefore, propose a study on the thermal resistance of industrial dry yeasts in relation to their thermophysical properties. Heat stress was applied at three temperatures (50, 75, and 100°C) for 10, 30, or 60-minute treatments. The survival of yeasts to these treatments was estimated, and their thermophysical properties were studied by differential scanning calorimetry. The industrial dry yeasts resisted 60 minutes at 50°C and 75°C and 10 minutes at a temperature close to 100°C. At 100°C, yeast was above their glass transition temperature. Industrial dry yeasts are therefore capable of withstanding high thermal stress if maintained in a specific thermophysical state.

Keywords: dry yeast, glass transition, thermal resistance, vitrification

Procedia PDF Downloads 130

Authors: Ruolan Zhang, Ryo Imai, Naoko Senda, Tomoyuki Sakai

Abstract:

Enzymes have attracted increasing attentions in industrial manufacturing for their applicability in catalyzing complex chemical reactions under mild conditions. Directed evolution has become a powerful approach to optimize enzymes and exploit their full potentials under the circumstance of insufficient structure-function knowledge. With the incorporation of cell-free synthetic biotechnology, rapid enzyme synthesis can be realized because no cloning procedure such as transfection is needed. Its open environment also enables direct enzyme measurement. These properties of cell-free biotechnology lead to excellent throughput of enzymes generation. However, the capabilities of current screening methods have limitations. Fluorescence-based assay needs applicable fluorescent label, and the reliability of acquired enzymatic activity is influenced by fluorescent label’s binding affinity and photostability. To acquire the natural activity of an enzyme, another method is to combine pre-screening step and high-performance liquid chromatography (HPLC) measurement. But its throughput is limited by necessary time investment. Hundreds of variants are selected from libraries, and their enzymatic activities are then identified one by one by HPLC. The turn-around-time is 30 minutes for one sample by HPLC, which limits the acquirable enzyme improvement within reasonable time. To achieve the real high-throughput enzyme screening, i.e., obtain reliable enzyme improvement within reasonable time, a widely applicable high-throughput measurement of enzymatic reactions is highly demanded. Here, a high-throughput screening method using broadband coherent anti-Stokes Raman spectroscopy (CARS) was proposed. CARS is one of coherent Raman spectroscopy, which can identify label-free chemical components specifically from their inherent molecular vibration. These characteristic vibrational signals are generated from different vibrational modes of chemical bonds. With the broadband CARS, chemicals in one sample can be identified from their signals in one broadband CARS spectrum. Moreover, it can magnify the signal levels to several orders of magnitude greater than spontaneous Raman systems, and therefore has the potential to evaluate chemical's concentration rapidly. As a demonstration of screening with CARS, alcohol dehydrogenase, which converts ethanol and nicotinamide adenine dinucleotide oxidized form (NAD+) to acetaldehyde and nicotinamide adenine dinucleotide reduced form (NADH), was used. The signal of NADH at 1660 cm⁻¹, which is generated from nicotinamide in NADH, was utilized to measure the concentration of it. The evaluation time for CARS signal of NADH was determined to be as short as 0.33 seconds while having a system sensitivity of 2.5 mM. The time course of alcohol dehydrogenase reaction was successfully measured from increasing signal intensity of NADH. This measurement result of CARS was consistent with the result of a conventional method, UV-Vis. CARS is expected to have application in high-throughput enzyme screening and realize more reliable enzyme improvement within reasonable time.

Keywords: Coherent Anti-Stokes Raman Spectroscopy, CARS, directed evolution, enzyme screening, Raman spectroscopy

Procedia PDF Downloads 128

Authors: Mariam Dianat Sabet Gilani, Lars M. Blank, Birgitta E. Ebert

Abstract:

Plant-derived lupane-type, pentacyclic triterpenoids are biologically active compounds that are highly interesting for applications in medical, pharmaceutical, and cosmetic industries. Due to the low abundance of these valuable compounds in their natural sources, and the environmentally harmful downstream process, alternative production methods, such as microbial cell factories, are investigated. Engineered Saccharomyces cerevisiae strains, harboring the heterologous genes for betulinic acid synthesis, can produce up to 2 g L-1 triterpenoids, showing high potential for large-scale production of triterpenoids. One limitation of the microbial synthesis is the intracellular product accumulation. It not only makes cell disruption a necessary step in the downstream processing but also limits productivity and product yield per cell. To overcome these restrictions, the aim of this study is to develop an in-situ extraction method, which extracts triterpenoids into a second organic phase. Such a continuous or sequential product removal from the biomass keeps the cells in an active state and enables extended production time or biomass recycling. After screening of twelve different solvents, selected based on product solubility, biocompatibility, as well as environmental and health impact, isopropyl myristate (IPM) was chosen as a suitable solvent for in-situ product removal from S. cerevisiae. Impedance-based single-cell analysis and off-gas measurement of carbon dioxide emission showed that cell viability and physiology were not affected by the presence of IPM. Initial experiments demonstrated that after the addition of 20 vol % IPM to cultures in the stationary phase, 40 % of the total produced triterpenoids were extracted from the cells into the organic phase. In future experiments, the application of IPM in a repeated batch process will be tested, where IPM is added at the end of each batch run to remove triterpenoids from the cells, allowing the same biocatalysts to be used in several sequential batch steps. Due to its high biocompatibility, the amount of IPM added to the culture can also be increased to more than 20 vol % to extract more than 40 % triterpenoids in the organic phase, allowing the cells to produce more triterpenoids. This highlights the potential for the development of a continuous large-scale process, which allows biocatalysts to produce intracellular products continuously without the necessity of cell disruption and without limitation of the cell capacity.

Keywords: betulinic acid, biocompatible solvent, in-situ extraction, isopropyl myristate, process development, secondary metabolites, triterpenoids, yeast

Procedia PDF Downloads 129

Authors: Sylvana A. Vega, Cesar E. Huilinir, Carlos J. Gonzalez

Abstract:

Co-digestion in anaerobic biodigesters is a technology improving hydrolysis by increasing methane generation. In the present study, the dimensional computational fluid dynamics (CFD) is numerically analyzed using Ansys Fluent software for agitation in a full-scale Continuous Stirred Tank Reactor (CSTR) biodigester during the co-digestion process. For this, a rheological study of the substrate is carried out, establishing rotation speeds of the stirrers depending on the microbial activity and energy ranges. The substrate is organic waste from industrial sources of sanitary water, butcher, fishmonger, and dairy. Once the rheological behavior curves have been obtained, it is obtained that it is a non-Newtonian fluid of the pseudoplastic type, with a solids rate of 12%. In the simulation, the rheological results of the fluid are considered, and the full-scale CSTR biodigester is modeled. It was coupling the second-order continuity differential equations, the three-dimensional Navier Stokes, the power-law model for non-Newtonian fluids, and three turbulence models: k-ε RNG, k-ε Realizable, and RMS (Reynolds Stress Model), for a 45° tilt vane impeller. It is simulated for three minutes since it is desired to study an intermittent mixture with a saving benefit of energy consumed. The results show that the absolute errors of the power number associated with the k-ε RNG, k-ε Realizable, and RMS models were 7.62%, 1.85%, and 5.05%, respectively, the numbers of power obtained from the analytical-experimental equation of Nagata. The results of the generalized Reynolds number show that the fluid dynamics have a transition-turbulent flow regime. Concerning the Froude number, the result indicates there is no need to implement baffles in the biodigester design, and the power number provides a steady trend close to 1.5. It is observed that the levels of design speeds within the biodigester are approximately 0.1 m/s, which are speeds suitable for the microbial community, where they can coexist and feed on the substrate in co-digestion. It is concluded that the model that more accurately predicts the behavior of fluid dynamics within the reactor is the k-ε Realizable model. The flow paths obtained are consistent with what is stated in the referenced literature, where the 45° inclination PBT impeller is the right type of agitator to keep particles in suspension and, in turn, increase the dispersion of gas in the liquid phase. If a 24/7 complete mix is considered under stirred agitation, with a plant factor of 80%, 51,840 kWh/year are estimated. On the contrary, if intermittent agitations of 3 min every 15 min are used under the same design conditions, reduce almost 80% of energy costs. It is a feasible solution to predict the energy expenditure of an anaerobic biodigester CSTR. It is recommended to use high mixing intensities, at the beginning and end of the joint phase acetogenesis/methanogenesis. This high intensity of mixing, in the beginning, produces the activation of the bacteria, and once reaching the end of the Hydraulic Retention Time period, it produces another increase in the mixing agitations, favoring the final dispersion of the biogas that may be trapped in the biodigester bottom.

Keywords: anaerobic co-digestion, computational fluid dynamics, CFD, net power, organic waste

Procedia PDF Downloads 99

Authors: Chinumani Choudhury

Abstract:

Heavy metal contamination is an alarming problem, which poses a serious risk to human health and the surrounding geology. Soils get contaminated with heavy metals due to the un-regularized industrial discharge of the toxic metal-rich effluents. Under such a condition, the remediation of the contaminated sites becomes imperative for a sustainable, safe, and healthy environment. Phytoextraction, which involves the removal of heavy metals from the soil through root absorption and uptake, is a viable remediation technique, which ensures extraction of the toxic inorganic compound available in the soil even at low concentrations. The soil present in the Silghat Region of Assam, India, is mostly contaminated with Zinc (Zn) and Lead (Pb), having concentrations as high as to cause a serious environmental problem if proper measures are not taken. In the present study, an extensive experimental study was carried out to understand the effectiveness of two commonly planted trees in Assam, namely, i) Indian Pennywort and ii) Fenugreek, in the removal of heavy metals from the contaminated soil. The basic characterization of the soil in the contaminated site of the Silghat region was performed and the field concentration of Zn and Pb was recorded. Various long-term laboratory pot tests were carried out by sowing the seeds of Indian Pennywort and Fenugreek in a soil, which was spiked, with a very high dosage of Zn and Pb. The tests were carried out for different concentration of a particular heavy metal and the individual effectiveness in the absorption of the heavy metal by the plants were studied. The concentration of the soil was monitored regularly to assess the rate of depletion and the simultaneous uptake of the heavy metal from the soil to the plant. The amount of heavy metal uptake by the plant was also quantified by analyzing the plant sample at the end of the testing period. Finally, the study throws light on the applicability of the studied plants in the field for effective remediation of the contaminated sites of Assam.

Keywords: phytoextraction, heavy-metals, Indian pennywort, fenugreek

Procedia PDF Downloads 105

Authors: Carlos A. Garcia-Mogollon, Juan C. Quintero-Diaz, Claudio Avignone-Rossa

Abstract:

Clostridium saccharoperbutylacetonicum 1-4N (Csb 1-4N) is an industrial reference strain for Acetone-Butanol-Ethanol (ABE) fermentation. Csb 1-4N is a solventogenic clostridium and H₂ producer with a metabolic profile that makes it a good candidate for Bioelectrosynthesis System (BES). The aim of this study was to evaluate the electroactivity of Csb 1-4N by cyclic voltammetry technique (CV). The Bioelectrosynthesis fermentation (BES) started in a Triptone-Yeast extract (TY) medium with trace elements and vitamins, Complex Nitrogen Source (CNS), and bicarbonate (NaHCO₃, 4g/L) as a carbon source, run at -600mVAg/AgCl and adding 200uM NADH. The six BES batches were performed with different media composition with and without NADH, CNS, HCO₃⁻ , and applied potential. The CV was performed as three-electrode system: platinum slice working electrode (WE), nickel contra electrode (CE) and reference electrode Ag/AgCl (ER). CVs were run in a potential range of -0.7V to 0.7V vs. VAg/AgCl at a scan rate 10mV/s. A CV recorded using different NaHCO₃ concentrations (0.25; 0.5; 1.0; 4g/L) were obtained. BES fermentation samples were centrifuged (3000 rpm, 5min, 4C), and supernatant (7mL) was used. CVs were obtained for Csb1-4N BES culture cell-free supernatant at 0h, 24h, and 48h. The electrochemical analysis was carried out with a PalmSens 4.0 potentiostat/galvanostat controlled with the PStrace 5.7 software, and CVs curves were characterized by reduction and oxidation currents and reduction and oxidation peaks. The CVs obtained for NaHCO₃ solutions showed that the reduction current and oxidation current decreased as the NaHCO₃ concentration was decreased. All reduction and oxidation currents decreased until exponential growth stop (24h), independence of initial cathodic current, except in medium with trace elements, vitamins, and NaHCO3, in which reduction current was around half at 24h and followed decreasing at 48. In this medium, Csb1-4N did not grow, but pH was increased, indicating that NaHCO₃ was reduced as the reduction current decreased. In general, at 48h reduction currents did not present important changes between different mediums in BES cultures. In terms of intensities in the peaks (Ip) did not present important variations; except with Ipa and Ipc in BES culture with NaHCO₃ and NADH added are higher than peaks in other cultures. Based on results, cathodic and anodic currents changes were induced by NaHCO₃ reduction reactions during Csb1-4N metabolic activity in different BES experiments.

Keywords: clostridium saccharoperbutylacetonicum 1-4N, bioelectrosynthesis, carbon dioxide fixation, cyclic voltammetry

Procedia PDF Downloads 120