Search results for: yeast

Authors: Swapna Guntupalli, Leela Madhuri Chalasani, Kshatri Jyothi, C. V. Rao, Bondili J. S.

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The study hypothesizes that enhanced biodegradation of Polycyclic Aromatic Hydrocarbons (PAHs) is achievable with an assemblage of microorganisms that are capable of producing biofilm and biosurfactants. Accordingly, PAHs degrading microorganism’s (bacteria, fungi, actinomycetes and yeast) were screened and grouped into different consortia based on their capabilities to produce biofilm and biosurfactants. Among these, Consortium BTSN09 consisting of bacterial fungal cocultures showed highest degradation due to the synergistic action between them. Degradation effiencies were evaluated using HPLC and GC-MS. Within 7days, BTSN09 showed 51% and 50.7% degradation of Phenanthrene (PHE) and Pyrene (PYR) with 200mg/L and 100 mg/L concentrations respectively in a liquid medium. In addition, several degradative enzymes like laccases, 1hydroxy-2-naphthoicacid dioxygenase, 2-carboxybenzaldehyde dehydrogenase, catechol1,2 dioxygenase and catechol2,3 dioxygenase activity was observed during degradation. Degradation metabolites were identified using GC-MS analysis and from the results it was confirmed that the metabolism of degradation proceeds via pthalic acid pathway for both PAHs. Besides, Microbial consortia also demonstrated good biosurfactant production capacity, achieving maximum oil displacement area and emulsification activity of 19.62 cm2, 65.5% in presence of PAHs as sole carbon source. Scanning Electron Microscopy analysis revealed exopolysaccharides (EPS) production, micro and macrocolonies formation with different stages of biofim development in presence of PAHs during degradation.

Keywords: PAHs, biosurfactant, biofilm, biodegradation

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Authors: James Sinclair, Katy Soapi, Brad Carte

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The marine environment has continuously demonstrated to be a rich source of secondary metabolites and bioactive compounds that can address the many pharmaceutical problems facing mankind. The emergence of multidrug resistant pathogens has caused scientists to explore contemporary ways of combating these super bugs. A red-pigmented bacterial strain isolated from a marine alga collected in Fiji was identified to be Pseudoalteromonas rubra from 16s rRNA sequencing. This bacterial strain was cultured using a yeast-peptone media and incubated for five days. The ethyl acetate extract of this bacterium was subjected to chromatographic separation techniques such as vacuum liquid chromatography, flash chromatography, size exclusion chromatography and high-pressure liquid chromatography to yield the pure compound and a number of semi-pure fractions. The crude extract and subsequent purified fractions were analyzed by ultraviolet/visible spectroscopy and mass spectroscopy and was found to contain the compounds ivermectin, stenothricin, cyclo-L-pro-L-val, prodigiosin, mycophenolic acid, phenazine-1-carboxylic acid, eplerenone, staurosporine and pseudoalteromone A. The structure of the pure compound, pseudoalteromone A, was elucidated using NMR 1H, 13C, 1H-1H COSY, HSQC and HMBC spectroscopic data.

Keywords: Pseudoalteromonas rubra, Pseudoalteromone A, secondary metabolites, structure elucidation

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Authors: Farnaz Yusuf, Naseem A. Gaur

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The increase in environmental concerns, rapid depletion of fossil fuel reserves and intense interest in achieving energy security has led to a global research effort towards developing renewable sources of fuels. Second generation biofuels have attracted more attention recently as the use of lignocellulosic biomass can reduce fossil fuel dependence and is environment-friendly. Xylose is the main pentose and second most abundant sugar after glucose in lignocelluloses. Saccharomyces cerevisiae does not readily uptake and use pentose sugars. For an economically feasible biofuel production, both hexose and pentose sugars must be fermented to ethanol. Therefore, it is important to develop S. cerevisiae host platforms with more efficient xylose utilization. This work aims to construct a xylose fermenting yeast strains with engineered oxido-reductative pathway for xylose metabolism. Engineered strain was further improved by adaptive evolutionary engineering approach. The engineered strain is able to grow on xylose as sole carbon source with the maximum ethanol yield of 0.39g/g xylose and productivity of 0.139g/l/h at 96 hours. The further improvement in strain development involves over expression of pentose phosphate pathway and protein engineering of xylose reductase/xylitol dehydrogenase to change their cofactor specificity in order to reduce xylitol accumulation.

Keywords: biofuel, lignocellulosic biomass, saccharomyces cerevisiae, xylose

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Authors: Shohei Maruyama, Yasuo Matsuyama, Sachiyo Aburatani

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The development of the method to annotate unknown gene functions is an important task in bioinformatics. One of the approaches for the annotation is The identification of the metabolic pathway that genes are involved in. Gene expression data have been utilized for the identification, since gene expression data reflect various intracellular phenomena. However, it has been difficult to estimate the gene function with high accuracy. It is considered that the low accuracy of the estimation is caused by the difficulty of accurately measuring a gene expression. Even though they are measured under the same condition, the gene expressions will vary usually. In this study, we proposed a feature extraction method focusing on the variability of gene expressions to estimate the genes' metabolic pathway accurately. First, we estimated the distribution of each gene expression from replicate data. Next, we calculated the similarity between all gene pairs by KL divergence, which is a method for calculating the similarity between distributions. Finally, we utilized the similarity vectors as feature vectors and trained the multiclass SVM for identifying the genes' metabolic pathway. To evaluate our developed method, we applied the method to budding yeast and trained the multiclass SVM for identifying the seven metabolic pathways. As a result, the accuracy that calculated by our developed method was higher than the one that calculated from the raw gene expression data. Thus, our developed method combined with KL divergence is useful for identifying the genes' metabolic pathway.

Keywords: metabolic pathways, gene expression data, microarray, Kullback–Leibler divergence, KL divergence, support vector machines, SVM, machine learning

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Authors: Thu Thuy Pham, Thi Chau Loan Tran, Van Duy Nguyen

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Marine fungi play an important role in the marine ecosystems. Marine fungi also supply biomass and metabolic products of industrial value. Currently, the biodiversity of marine fungi along the coastal areas of Vietnam has not yet been studied fully. The objective of this study is to assess the spatial and temporal diversity of planktonic fungi from the coastal waters of Nha Trang Bay and Van Phong Bay in Central Vietnam using culture-dependent and independent approach. Using culture-dependent approach, filamentous fungi and yeasts were isolated on selective media and then classified by phenotype and genotype based on the sequencing of ITS (internal transcribed spacers) regions of rDNA with two primer pairs (ITS1F_KYO2 and ITS4; NS1 and NS8). Using culture-independent approach, environmental DNA samples were isolated and amplified using fungal-specific ITS primer pairs. A total of over 160 strains were isolated from 10 seawater sampling stations at 50 cm depth. They were classified into diverse genera and species of both yeast and mold. At least 5 strains could be potentially novel species. Our results also revealed that planktonic fungi were molecularly diverse with hundreds of phylotypes recovered across these two bays. The results of the study provide data about the distribution and taxonomy of mycoplankton in this area, thereby allowing assessment of their positive role in the biogeochemical cycle of coastal ecosystems and the development of new bioactive compounds for industrial applications.

Keywords: biodiversity, ITS, marine fungi, Nha Trang Bay, Van Phong Bay

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Authors: Hayet El Kolli, Hocine Laouer

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Bunium in the world comprises about 50 to 100 species, mostly distributed in: Algeria, Italy, Pakistan, Iran, and South Africa. Bunium species have several uses like: Bunium persicum which is commonly used as antispasmodic, carminative, anti-obesity and lactogage. This plant have been widely used as an additive in food stuff such as in bread cooking, rice and yoghurt for its carminative, anti-dyspepsia and antispasmodic effect. The B. paucifolium oil has a wide spectrum of action against moulds, yeast and bacteria. The chemical compositions of Bunium incrassatum and Bunium alpinum essential oils were carry out by GC and GC/MS. Therefore, antibacterial activity of two oils was investigated by disk diffusion method against Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 1331, Staphylococcus aureus ATCC 25923, Klebsiella pneumoniae ATCC 700603, Bacillus cereus ATCC 10876, Enterococcus faecalis ATCC 49452, Lysteria monocytogenes ATCC 15313, Citrobacter freundii ATCC 8090, Proteus mirabilis ATCC 35659. A moderate antibacterial activity was found. In conclusion, it is found that essential oils of the two species are rich in sesquiterpens and other oxygenated compounds. These compounds have been reported to show bactericidal activity and the presence of phenolic compounds makes them useful antioxidants so that results confirm some ethnopharmacologique applications of these two oils of Bunium.

Keywords: Bunium alpinum, Bunium incrassatum, apiaceae, essential oil, sesquiterpens, phenols, antibacterial, antioxidant activities

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Authors: Kazem Alirezalu, Javad Hesari, Zabihollah Nemati, Boukaga Farmani

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The application of natural plant extracts which are rich in promising antioxidants and antimicrobial ingredients in the production of frankfurter-type sausages addresses consumer demands for healthier, more functional meat products. The effects of olive leaves, green tea and Urtica dioica L. extracts on physicochemical, microbiological and sensory characteristic of frankfurter-type sausage were investigated during 45 days of storage at 4 °C. The results revealed that pH and phenolic compounds decreased significantly (P < 0.05) in all samples during storage. Sausages containing 500 ppm green tea extract (1.78 mg/kg) showed the lowest TBARS values compared to olive leaves (2.01 mg/kg), Urtica dioica L. (2.26 mg/kg) extracts and control (2.74 mg/kg). Plant extracts significantly (P < 0.05) reduced the count of total mesophilic bacteria, yeast and mold by at least 2 log cycles (CFU/g) than those of control samples. Sensory characteristics of texture showed no difference (P > 0.05) between sausage samples, but sausage containing Urtica dioica L. extract had the highest score regarding flavor, freshness odor, and overall acceptability. Based on the results, sausage containing plant extracts could have a significant impact on antimicrobial activity, antioxidant capacity, sensory score, and shelf life stability of frankfurter-type sausage.

Keywords: antimicrobial, antioxidant, frankfurter-type sausage, green tea, olive oil, shelf life, Urtica dioica L.

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Authors: Angela U. Makolo, Temitayo A. Olagunju

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The knowledge of signaling pathways is central to understanding the biological mechanisms of organisms since it has been identified that in eukaryotic organisms, the number of signaling pathways determines the number of ways the organism will react to external stimuli. Signaling pathways are studied using protein interaction networks constructed from protein-protein interaction data obtained using high throughput experimental procedures. However, these high throughput methods are known to produce very high rates of false positive and negative interactions. In order to construct a useful protein interaction network from this noisy data, computational methods are applied to validate the protein-protein interactions. In this study, a computational technique to identify signaling pathways from a protein interaction network constructed using validated protein-protein interaction data was designed. A weighted interaction graph of the Saccharomyces cerevisiae (Baker’s Yeast) organism using the proteins as the nodes and interactions between them as edges was constructed. The weights were obtained using Bayesian probabilistic network to estimate the posterior probability of interaction between two proteins given the gene expression measurement as biological evidence. Only interactions above a threshold were accepted for the network model. A pathway was formalized as a simple path in the interaction network from a starting protein and an ending protein of interest. We were able to identify some pathway segments, one of which is a segment of the pathway that signals the start of the process of meiosis in S. cerevisiae.

Keywords: Bayesian networks, protein interaction networks, Saccharomyces cerevisiae, signalling pathways

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Authors: Anna Gramza-Michałowska, Dominik Kmiecik, Justyna Bilon, Joanna Skręty, Joanna Kobus-Cisowska, Józef Korczak, Andrzej Sidor

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Background: Hypertension is one of the civilization diseases with a global scope. Many research showed that every day diet influences significantly our health, helping with the prophylaxis and diseases treatment. The key factor here is the presence of plant origin natural bio active components. Aim: The following research describes snack health-oriented products for hypertension sufferers enriched with selected plant ingredients. Various analytical methods have been applied to determine product’s basic composition and their antioxidant activity. Methods: Snack products was formulated from a composition of different flours, oil, yeast, plant particles and extracts. Basic composition of a product was evaluated as content of protein, lipids, fiber, ash and caloricity. Antioxidant capacity of snacks was evaluated with use radical scavenging methods (DPPH, ABTS) and ORAC value. Proposed snacks as new product was also characterized with sensory analysis. Results and discussion: Results showed that addition of phyto nutrients allowed to improve nutritional and antioxidative value of examined products. Also the anti radical potential was significantly increased, with no loss of sensory value of a snacks. Conclusions: Designed snack is rich in polyphenolics, that express high antioxidant activity, helpful in hypertension and as low calories product obesity prophylaxis.

Keywords: antioxidant, well-being, hypertension, bioactive compounds

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Authors: Sirikwan Ponprateep, Anchalee Tassanakajon, Vichien Rimphanitchayakit

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A Kazal-type serine proteinase inhibitor, SPIPm2, was abundantly expressed in the hemocytes and secreted into shrimp plasma has anti-viral property against white spot syndrome virus (WSSV). To discover the molecular mechanism of antiviral activity, the binding assay showed that SPIPm2 bind to the components of viral particle and shrimp hemocyte. From our previous report, viral target protein of SPIPm2 was identified, namely WSV477 using yeast two-hybrid screening. WSV477 is an early gene product of WSSV and involved in viral propagation. In this study, the co-immunoprecipitation technique and Tandem Mass Spectrometry (LC-MS/MS) was used to identify the target protein of SPIPm2 from shrimp hemocyte. The target protein of SPIPm2 was cyclophilin A. In vertebrate, cyclophilin A or peptidylprolyl isomerase A was reported to be the immune suppressor interacted with cyclosporin A involved in immune defense response. The recombinant cyclophilin A from Penaeus monodon (rPmCypA) was produced in E.coli system and purified using Ni-NTA column to confirm the protein-protein interaction. In vitro pull-down assay showed the interaction between rSPIPm2 and rPmCypA. To study the biological function of these proteins, the expression analysis of immune gene in shrimp defense pathways will be investigated after rPmCypA administration.

Keywords: cyclophilin A, protein-protein interaction, Kazal-type serine proteinase inhibitor, Penaeus monodon

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Authors: Ganatsios Vassilios, Terpou Antonia, Maria Kanellaki, Bekatorou Argyro, Athanasios Koutinas

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In this study, a new fermentation technology is proposed with potential application in home wine-making. Delignified cellulosic material was used to preserve Tubular Cellulose (TC), an effective fermentation support material in high osmotic pressure, low temperature, and alcohol concentration. The psychrotolerant yeast strain Saccharomyces cerevisiae AXAZ-1 was immobilized on TC to preserve a novel home wine making biocatalyst (HWB) and the entrapment was examined by SEM. Various concentrations of HWB was added in high-density grape must and the mixture was dried immediately. The dried mixture was stored for various time intervals and its fermentation examined after addition of potable water. The percentage of added water was also examined to succeed high alcohol and residual sugar concentration. The effect of low temperature (1-10 oC) on fermentation kinetics was studied revealing the ability of HBW on low-temperature sweet wine making. Sweet wines SPME GC-MS analysis revealed the promotion effect of TC on volatile by-products formation in comparison with free cells. Kinetics results and aromatic profile of final product encouraged the efforts of high-quality sweet wine making in domestic refrigerator and potential marketable opportunities are also assessed and discussed.

Keywords: tubular cellulose, sweet wine, Saccharomyces cerevisiae AXAZ-1, residual sugar concentration

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Authors: Jose A. Aznar-Moreno, Xi Jiang, Stefan Burén, Luis M. Rubio

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Integration of prokaryotic nitrogen fixation (nif) genes into the plastid genome for expression of functional nitrogenase components could render plants capable of assimilating atmospheric N2 making their crops less dependent of nitrogen fertilizers. The nitrogenase Fe protein component (NifH) has been used as proxy for expression and targeting of Nif proteins within plant and yeast cells. Here we use tobacco plants with the Azotobacter vinelandii nifH and nifM genes integrated into the plastid genome. NifH and its maturase NifM were constitutively produced in leaves, but not roots, during light and dark periods. Nif protein expression in transplastomic plants was stable throughout development. Chloroplast NifH was soluble, but it only showed in vitro activity when isolated from leaves collected at the end of the dark period. Exposing the plant extracts to elevated temperatures precipitated NifM and apo-NifH protein devoid of [Fe4S4] clusters, dramatically increasing the specific activity of remaining NifH protein. Our data indicate that the chloroplast endogenous [Fe-S] cluster biosynthesis was insufficient for complete NifH maturation, albeit a negative effect on NifH maturation due to excess NifM in the chloroplast cannot be excluded. NifH and NifM constitutive expression in transplastomic plants did not affect any of the following traits: seed size, germination time, germination ratio, seedling growth, emergence of the cotyledon and first leaves, chlorophyll content and plant height throughout development.

Keywords: NifH, chloroplast, nitrogen fixation, crop improvement, transplastomic plants, fertilizer, biotechnology

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Authors: Linda Chularojmontri, Seewaboon Sireeratawong, Suvara Wattanapitayakul

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Terminalia arjuna Roxb. (family Combretaceae) is commonly known as ‘Sa maw thet’ in Thai. The fruit is used in traditional medicine as natural mild laxatives, carminative and expectorant. Aim of the study: This research aims to study the anti-inflammatory, analgesic and antipyretic activities of Terminalia arjuna extract by using animal models in comparison to the reference drugs. Materials and Methods: The anti-inflammatory study was conducted by two experimental animal models namely ethyl phenylpropionate (EPP)-induced ear edema and carrageenan-induced paw edema. The study of analgesic activity used two methods of pain induction including acetic acid and heat-induced pain. In addition, the antipyretic activity study was performed by induced hyperthermia with yeast. Results: The results showed that the oral administration of Terminalia arjuna extract possessed acute anti-inflammatory effect in carrageenan-induced paw edema. Terminalia arjuna extract showed the analgesic activity in acetic acid-induced writhing response and heat-induced pain. This indicates its peripheral effect by inhibiting the biosynthesis and/or release of some pain mediators and some mechanism through Central nervous system. Moreover, Terminalia arjuna extract at the dose of 1000 and 1500 mg/kg body weight showed the antipyretic activity, which might be because of the inhibition of prostaglandins. Conclusion: The findings of this study indicated that the Terminalia arjuna extract possesses the anti-inflammatory, analgesic and antipyretic activities in animals.

Keywords: analgesic activity, anti-inflammatory activity, antipyretic activity, Terminalia arjuna extract

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Authors: Hawbash Jalil

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In this study, the effects of whey protein based films on various properties of kashar cheese were examined. In the study, edible film solutions based on whey protein isolate, whey protein isolate + transglutaminase enzyme and whey protein isolate + chitosan were produced and Kashar cheese samples were coated with these films by dipping method and stored at +4 ºC for 60 days. Chemical, microbiological and textural analyzes were carried out on samples at 0, 30 and 60 days of storage. As a result of the study, the highest dry matter and total nitrogen values were obtained from uncoated control samples This is an indication that the coatings limit water vapor permeability. The highest acidity and pH values obtained from the samples as storage results were 3.33% and 5.86%, respectively, in the control group samples. Both acidity and pH rise in these groups, is a consequence of the buffering of pH changes of hydrolsis products which are as a result of proteolysis occurring in the sample. Nitrogen changes and lipolysis values, which are indicative of the degree of hydrolysis of proteins and triglycerides in kashar cheese, were generally higher in the control group This result is due to limiting the micro organism reproduction by limiting the gas passage of the coatings. Hardness and chewiness values of the textural properties of the samples were significantly reduced in uncoated control samples compared to the coated samples due to maturation. The chitosan film coatings used in the study limited the development of mold yeast until the 30th day but after that did not yield successful results in this respect.

Keywords: chitosan, edible film, transglutaminase, whey protein

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Authors: Chengyu Dong, Shao-Yuan Leu

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Cellulosic ethanol production from lignocellulosic biomass can reduce our reliance on fossil fuel, mitigate climate change, and stimulate rural economic development. The relative low ethanol production (60 g/L) limits the economic viable of lignocellulose-based biorefinery. The ethanol production can be increased up to 80 g/L by removing nearly all the non-cellulosic materials, while the capital of the pretreatment process increased significantly. In this study, a fed-batch prehydrolysis simultaneously saccharification and fermentation process (PSSF) was designed to converse the sulfite pretreated softwood (~30% residual lignin) to high concentrations of ethanol (80 g/L). The liquefaction time of hydrolysis process was shortened down to 24 h by employing the fed-batch strategy. Washing out the spent liquor with water could eliminate the inhibition of the pretreatment spent liquor. However, the ethanol yield of lignocellulose was reduced as the fermentable sugars were also lost during the process. Fed-batch prehydrolyzing the while slurry (i.e. liquid plus solid fraction) pretreated softwood for 24 h followed by simultaneously saccharification and fermentation process at 28 °C can generate 80 g/L ethanol production. Fed-batch strategy is very effectively to eliminate the “solid effect” of the high gravity saccharification, so concentrating the cellulose to nearly 90% by the pretreatment process is not a necessary step to get high ethanol production. Detoxification of the pretreatment spent liquor caused the loss of sugar and reduced the ethanol yield consequently. The tolerance of yeast to inhibitors was better at 28 °C, therefore, reducing the temperature of the following fermentation process is a simple and valid method to produce high ethanol production.

Keywords: cellulosic ethanol, sulfite pretreatment, Fed batch PSSF, temperature

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Authors: Tamar Khardziani, Violeta Berikashvili, Amrosi Chkuaseli, Vladimir Elisashvili

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The main objectives of this proposal are to develop low-cost, innovative, and competitive technologies for the production of mycelial biomass of medicinal mushrooms as a natural food supplement for poultry. To fulfill this task, industrial strains of Lentinus edodes, Ganoderma lucidum, and Pleurotus ostreatus were used in this study. The solid-state fermentation (SSF) of wheat grains, wheat bran, and soy flour was performed in flasks and bags. Among nine mushroom strains, P. ostreatus 2191 appeared to be the most productive in protein biomass accumulation in the SSF of wheat bran. All mushrooms produced exopolysaccharide with the highest yield of 5-8 mg/mL depending on fungal strain and growth substrate. Supplementation of medium with 1% glycerol and 2-4% peptone favored mushroom growth and protein accumulation. Among inorganic nitrogen sources, KNO₃ also provided high biomass and protein production. The SSF of all growth substrates was accompanied by the secretion of cellulase and xylanase activities. The highest CMCase activity (12-13 U/g) was revealed in the cultivation of P. ostreatus 2191 using wheat bran as a growth substrate and ammonium sulfate or yeast extract as a nitrogen source, whereas the highest xylanase activity was detected in the fermentation of soy flour supplemented with peptone. Acknowledgments: This work was supported by the Shota Rustaveli National Science Foundation of Georgia (Grant number STEM-22-2077).

Keywords: mushrooms, plant raw materials, fermentation, biomass protein, cellulase

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Authors: Asma Ansari, Afsheen Aman, Shah Ali Ul Qader

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In the past few years, numerous concerns have been raised against increased bacterial resistance towards effective drugs and become a debated issue all over the world. With the emergence of drug resistant pathogens, the interaction of natural antimicrobial compounds and antibacterial nanoparticles has emerged as a potential candidate for combating infectious diseases. Microbial diversity in the biome provides an opportunity to screen new species which are capable of producing large number of antimicrobial compounds. Among these antimicrobial compounds, bacteriocins are highly specific and efficient antagonists. A combination of bacteriocin along with nanoparticles could prove to be more potent due to broadened antibacterial spectrum with possibly lower doses. In the current study, silver nanoparticles were synthesized through biological reduction using various isolated bacterial, fungal and yeast strains. Spectroscopy and scanning electron microscopy (SEM) was performed for the confirmation of nanoparticles. Bacteriocin was characterized and purified to homogeneity through gel permeation chromatography. The estimated molecular weight of bacteriocin was 10 kDa. Amino acid analysis and N-terminal sequencing revealed the novelty of the protein. Then antibacterial potential of silver nanoparticles and broad inhibitory spectrum bacteriocin was determined through agar well diffusion assay. These synthesized bacteriocin-Nanoparticles exhibit a good potential for clinical applications as compared to bacteriocin alone. This combination of bacteriocin with nanoparticles will be used as a new sort of biocide in the field of nano-proteomics. The advancement of nanoparticles-mediated drug delivery system will open a new age for rapid eradication of pathogens from biological systems.

Keywords: BAC-IB17, multidrug resistance, purification, silver nanoparticles

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Authors: Sumeeta Khurana, Kirti Megha, Amit Gupta, Rakesh Sehgal

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Background: Amoebic keratitis is a painful vision threatening infection caused by a free living pathogenic amoeba Acanthamoeba. It can be misdiagnosed and very difficult to treat if not suspected early. The epidemiology of Acanthamoeba genotypes causing infection in our geographical area is not yet known to the best of our knowledge. Objective: To characterize Acanthamoeba isolates from amoebic keratitis patients. Methods: A total of 19 isolates obtained from patients with amoebic keratitis presenting to the Advanced Eye Centre at Postgraduate Institute of Medical Education and Research, a tertiary care centre of North India over a period of last 10 years were included. Their corneal scrapings, lens solution and lens case (in case of lens wearer) were collected for microscopic examination, culture and molecular diagnosis. All the isolates were maintained in the Non Nutrient agar culture medium overlaid with E.coli and 13 strains were axenised and maintained in modified Peptone Yeast Dextrose Agar. Identification of Acanthamoeba genotypes was based on amplification of diagnostic fragment 3 (DF3) region of the 18srRNA gene followed by sequencing. Nucleotide similarity search was performed by BLAST search of sequenced amplicons in GenBank database (http//www.ncbi.nlm.nih.gov/blast). Multiple Sequence alignments were determined by using CLUSTAL X. Results: Nine out of 19 Acanthamoeba isolates were found to belong to Genotype T4 followed by 6 isolates of genotype T11, 3 T5 and 1 T3 genotype. Conclusion: T4 is the predominant Acanthamoeba genotype in our geographical area. Further studies should focus on differences in pathogenicity of these genotypes and their clinical significance.

Keywords: Acanthamoeba, free living amoeba, keratitis, genotype, ocular

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Authors: Eun-Ji Lee, Hye-Sung Park, Chan-Jung Lee, Won-Sik Kong

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We analyzed the DNA sequence of the ITS (Internal Transcribed Spacer) region of the 18S ribosomal gene and compared it with the gene sequence of T. fuciformis and Hypoxylon sp. in the BLAST database. The sequences of collected T. fuciformis and Hypoxylon sp. have over 99% homology in the T. fuciformis and Hypoxylon sp. sequence BLAST database. In order to select the optimal medium for T. fuciformis, five kinds of a medium such as Potato Dextrose Agar (PDA), Mushroom Complete Medium (MCM), Malt Extract Agar (MEA), Yeast extract (YM), and Compost Extract Dextrose Agar (CDA) were used. T. fuciformis showed the best growth on PDA medium, and Hypoxylon sp. showed the best growth on MCM. So as to investigate the optimum pH and temperature, the pH range was set to pH4 to pH8 and the temperature range was set to 15℃ to 35℃ (5℃ degree intervals). Optimum culture conditions for the T. fuciformis growth were pH5 at 25℃. Hypoxylon sp. were pH6 at 25°C. In order to confirm the most suitable carbon source, we used fructose, galactose, saccharose, soluble starch, inositol, glycerol, xylose, dextrose, lactose, dextrin, Na-CMC, adonitol. Mannitol, mannose, maltose, raffinose, cellobiose, ethanol, salicine, glucose, arabinose. In the optimum carbon source, T. fuciformis is xylose and Hypoxylon sp. is arabinose. Using the column test, we confirmed sawdust a suitable for T. fuciformis, since the composition of sawdust affects the growth of fruiting bodies of T. fuciformis. The sawdust we used is oak tree, pine tree, poplar, birch, cottonseed meal, cottonseed hull. In artificial cultivation of T. fuciformis with sawdust medium, T. fuciformis and Hypoxylon sp. showed fast mycelial growth on mixture of oak tree sawdust, cottonseed hull, and wheat bran.

Keywords: cultivation, optimal condition, tremella fuciformis, nutritional source

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Authors: Yetti Marlida, Harnentis, Yuliaty Shafan Nur

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Background: Tempoyak is a dish derived from fermented durian fruit. Tempoyak is a food consumed as a side dish when eating rice. Besides being eaten with rice, tempoyak can also be eaten directly. But this is rarely done because many cannot stand the sour taste and aroma of the tempoyak itself. In addition, tempoyak can also be used as a seasoning. The taste of tempoyak is acidic, this occurs because of the fermentation process in durian fruit meat which is the raw material. Tempoyak is already very well known in Indonesia, especially in Padang, Bengkulu, Palembang, Lampung, and Kalimantan. Besides that, this food is also famous in Malaysia. The purpose of this research is to improvement production of γ-aminobutyric acid (GABA) by Lactobacillus plantarum isolated from indigenous fermented durian (tempoyak). Selected Lactic Acid Bacteria (LAB) previously isolated from indigenous fermented durian (tempoyak) that have ability to produce γ-aminobutyric acid (GABA). The study was started with identification of selected LAB by 16 S RNA, followed optimation of GABA production by culture condition using different initial pH, temperature, glutamate concentration, incubation time, carbon and nitrogen sources. Results: The result from indentification used polymerase chain reaction of 16S rRNA gene sequences and phylogenetic analysis was Lactobacillus plantarum (coded as Y3) with a sequenced length of 1400bp. The improvement of Gaba production was found highest at pH: 6.0; temperature: 30 °C; glutamate concentration: 0.4%; incubation time: 60 h; glucose and yeast extract as carbon and nitrogen sources. Conclusions: GABA can be produced with the optimum condition fermentation were 66.06 mM.

Keywords: lactic acid bacteria, γ-amino butyric acid, indigenous fermented durian, PCR

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Authors: Zainab Bibi, Afsheen Aman, Shah Ali Ul Qader

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Endo-β-1,4-xylanases [EC 3.2.1.8] are one of the major groups of enzymes that are involved in degradation process of xylan and have several applications in food, textile and paper processing industries. Due to broad utility of endo-β-1,4-xylanase, researchers are focusing to increase the productivity of this hydrolase from various microbial species. Harsh industrial condition, faster reaction rate and efficient hydrolysis of xylan with low risk of contamination are critical requirements of industry that can be fulfilled by synthesizing the enzyme with efficient properties. In the current study, a newly isolated thermophile Geobacillus stearothermophilus KIBGE-IB29 was used in order to attain the maximum production of endo-1,4-β-xylanase. Bacterial culture was isolated from soil, collected around the blast furnace site of a steel processing mill, Karachi. Optimization of various nutritional and physical factors resulted the maximum synthesis of endo-1,4-β-xylanase from a thermophile. High production yield was achieved at 60°C and pH-6.0 after 24 hours of incubation period. Various nitrogen sources viz. peptone, yeast extract and meat extract improved the enzyme synthesis with 0.5%, 0.2% and 0.1% optimum concentrations. Dipotassium hydrogen phosphate (0.25%), potassium dihydrogen phosphate (0.05%), ammonium sulfate (0.05%) and calcium chloride (0.01%) were noticed as valuable salts to improve the production of enzyme. The thermophilic nature of isolate, with its broad pH stability profile and reduced fermentation time indicates its importance for effective xylan saccharification and for large scale production of endo-1,4-β-xylanase.

Keywords: geobacillus, optimization, production, xylanase

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Authors: Kubra Dogan, Fatih Tornuk

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Due to increasing consumer demand for the high-quality food products and awareness regarding the health benefits of different nutrients in food minimal processing becomes more popular in modern food preservation. To date, heat treatment is often used for inactivation of spoilage microorganisms in foods. However, it may cause significant changes in the quality and nutritional properties of food. In order to overcome the detrimental effects of heat treatment, several alternatives of non-thermal microbial inactivation processes have been investigated. Ultraviolet (UV) inactivation is a promising and feasible method for better quality and longer shelf life as an alternative to heat treatment, which aims to inhibit spoilage and pathogenic microorganisms and to inactivate the enzymes in vegetable juice production. UV-C is a sub-class of UV treatment which shows the highest microcidal effect between 250-270 nm. The wavelength of 254 nm is used for the surface disinfection of certain liquid food products such as vegetable juice. Effects of UV-C treatment on microbiological load and quality parameter of vegetable juice which is a mix of celery, carrot, lemon and orange was investigated. Our results showed that storing of UV-C applied vegetable juice for three months, reduced the count of TMAB by 3.5 log cfu/g and yeast-mold by 2 log cfu/g compared to control sample. Total phenolic content was found to be 514.3 ± 0.6 mg gallic acid equivalent/L, and there wasn’t a significant difference compared to control. The present work suggests that UV-C treatment is an alternative method for disinfection of vegetable juice since it enables adequate microbial inactivation, longer shelf life and has minimal effect on degradation of quality parameters of vegetable juice.

Keywords: heat treatment, phenolic content, shelf life, ultraviolet (UV-C), vegetable juice

Procedia PDF Downloads 178

Authors: Naser Valipour Motlagh, Majid Aliabadi, Elnaz Rahmani, Samira Ghorbanpour

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Strawberry is one of the most favored fruits all along the world. But due to its vulnerability to microbial contamination and short life storage, there are lots of problems in industrial production and transportation of this fruit. Therefore, lots of ideas have tried to increase the storage life of strawberries especially through proper packaging. This paper works on efficient packaging as well. The primary material used is produced through simple mixing of low-density polyethylene (LDPE) and silver nanoparticles in different weight fractions of 0.5 and 1% in presence of dicumyl peroxide as a cross-linking agent. Final packages were made in a twin-screw extruder. Then, their effect on the quality maintenance of strawberry is evaluated. The SEM images of nano-silver packages show the distribution of silver nanoparticles in the packages. Total bacteria count, mold, yeast and E. coli are measured for microbial evaluation of all samples. Texture, color, appearance, odor, taste and total acceptance of various samples are evaluated by trained panelists and based on 9-point hedonic scale method. The results show a decrease in total bacteria count and mold in nano-silver packages compared to the samples packed in polyethylene packages for the same storage time. The optimum concentration of silver nanoparticles for the lowest bacteria count and mold is predicted to be around 0.5% which has attained the most acceptance from the panelist as well. Moreover, organoleptic properties of strawberry are preserved for a longer period in nano-silver packages. It can be concluded that using nano-silver particles in strawberry packages has improved the storage life and quality maintenance of the fruit.

Keywords: antimicrobial properties, polyethylene, silver nanoparticles, strawberry

Procedia PDF Downloads 119

Authors: Ahmed F. Azmy, Amal E. Saafan, Tamer M. Essam, Magdy A. Amin, Shaban H. Ahmed

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Bacterial strains capable of degradation of malathion from the domestic sewage were isolated by an enrichment culture technique. Three bacterial strains were screened and identified as Acinetobacter baumannii (AFA), Pseudomonas aeruginosae (PS1),andPseudomonas mendocina (PS2) based on morphological, biochemical identification and 16S rRNA sequence analysis. Acinetobacter baumannii AFA was the most efficient malathion degrading bacterium, so used for further biodegradation study. AFA was able to grow in mineral salt medium (MSM) supplemented with malathion (100 mg/l) as a sole carbon source, and within 14 days, 84% of the initial dose was degraded by the isolate measured by high performance liquid chromatography. Strain AFA could also degrade other organophosphorus compounds including diazenon, chlorpyrifos and fenitrothion. The effect of different culture conditions on the degradation of malathion like inoculum density, other carbon or nitrogen sources, temperature and shaking were examined. Degradation of malathion and bacterial cell growth were accelerated when culture media were supplemented with yeast extract, glucose and citrate. The optimum conditions for malathion degradation by strain AFA were; an inoculum density of 1.5x 1012CFU/ml at 30°C with shaking. A specific polymerase chain reaction primers were designed manually using multiple sequence alignment of the corresponding carboxylesterase enzymes of Acinetobacter species. Sequencing result of amplified PCR product and phylogenetic analysis showed low degree of homology with the other carboxylesterase enzymes of Acinetobacter strains, so we suggested that this enzyme is a novel esterase enzyme. Isolated bacterial strains may have potential role for use in bioremediation of malathion contaminated.

Keywords: Acinetobacter baumannii, biodegradation, malathion, organophosphate pesticides

Procedia PDF Downloads 458

Authors: Astorga-Trejo Rebeca, Fonseca-Peralta Héctor Manuel, Beltrán-Arredondo Laura Ivonne, Castro-Martínez Claudia

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The use of biomass as feedstock for the production of fuels and other chemicals of interest is an ever-growing accepted option in the way to the development of biorefinery complexes; in the Mexican state of Sinaloa, two million tons of residues from corn crops are produced every year, most of which can be converted to bioethanol and other products through biotechnological conversion using yeast and other microorganisms. Therefore, the objective of this work was to take advantage of corn stover and evaluate its potential as a substrate for the production of second-generation bioethanol (2G), enzymes, and xylitol. To produce bioethanol 2G, an acid-alkaline pretreatment was carried out prior to saccharification and fermentation. The microorganisms used for the production of enzymes, as well as for the production of xylitol, were isolated and characterized in our workgroup. Statistical analysis was performed using Design Expert version 11.0. The results showed that it is possible to obtain 2G bioethanol employing corn stover as a carbon source and Saccharomyces cerevisiae ItVer01 and Candida intermedia CBE002 with yields of 0.42 g and 0.31 g, respectively. It was also shown that C. intermedia has the ability to produce xylitol with a good yield (0.46 g/g). On the other hand, qualitative and quantitative studies showed that the native strains of Fusarium equiseti (0.4 IU/mL - xylanase), Bacillus velezensis (1.2 IU/mL – xylanase and 0.4 UI/mL - amylase) and Penicillium funiculosum (1.5 IU / mL - cellulases) have the capacity to produce xylanases, amylases or cellulases using corn stover as raw material. This study allowed us to demonstrate that it is possible to use corn stover as a carbon source, a low-cost raw material with high availability in our country, to obtain bioproducts of industrial interest, using processes that are more environmentally friendly and sustainable. It is necessary to continue the optimization of each bioprocess.

Keywords: biomass, corn stover, biorefinery, bioethanol 2G, enzymes, xylitol

Procedia PDF Downloads 131

Authors: K. Maliki, A. Ajayi, O. M. Makanjuola, O. J. Adebowale

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Cocoyam is herbaceous perennial plant which belongs to the family Araceae and genus xanthosoma or cococasia is mostly cultivated as food crop. It is very rich in Vitamin B6, Magnesium and also in dietary fiber. Matured cocoyam is eaten boiled, Fried or roasted in Nigeria. It can also be dried and used to make flour. Food drying is a method of food preservation in which food is dried, thus inhibit the growth of bacteria yeast and mold through the removal of water. Drying effect on the proximate composition and functional properties of cocoyam flour were investigated. Freshly harvested cocoyam cultivars at matured level were washed with portable water, peeled, sliced into 0.3mm thickness blanch in boiling water at 100°C for 15 minutes and dried using sun drying oven and cabinet dryers. The blanched slices were divided into three lots and were subjected to different drying methods. The dried cocoyam slices were milled into flour using Apex mill and packed into Low Density Polyethylene Film (LDPE) 75 Micron 4 thickness and kept for four months under ambient temperature before analysis. The results showed that the moisture content, ash, crude fiber, fat, protein and carbohydrate ranged from 7.35% to 13.89%, 1.45% to 3.3%, 1.2% to 3.41%, 2.1% to 3.1%, 6.30% to 9.1% and 66% to 82% respectively. The functional properties of the cocoyam flour ranged from 1. 65ml/g to 4.24ml/g water absorption capacity, 0.85ml/g to 2.11ml/g oil absorption capacity 0.56ml/g and 0.78ml/g bulk density and 4.91% to 6.80% swelling capacity. The result showed that there was not significant difference (P ≥ 0.5) across the various drying methods used. Cabinet drying method was found to have the best quality characteristic values than the other drying methods. In conclusion, drying of cocoyam could be used for value addition and provide extension to shelf-life.

Keywords: cocoyam flour, drying, cabinet dryer, oven dryer

Procedia PDF Downloads 211

Authors: Dinesh K. S., D. R. C. V. Jayadevan

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Autism is the most prevalent of a subset of the disorders organized under the umbrella of pervasive developmental disorders. After the publication of Andrew Wakefield's paper in lancet, many critiques deny this connection even without looking in to the matter. The British Medical Journal even put an editorial regarding this issue. BMJ 2010; 340:c1807. But ayurveda has ample of evidences to believe this connectivity. Dysbiosis, yeast growth of the gut, nutritional deficiencies, enzyme deficiencies, essential fatty acid deficiencies, Gastro esophageal reflux disease, indigestion, inflammatory bowel, chronic constipation & its cascade are few of them to note. The purpose of this paper is to present the observed changes in the behavioural symptoms of autistic babies after a gut management protocol which is a usual programme of our autism treatment plan especially after dysbiotic changes after antibiotic administration. Is there any correlation between changes (if significant) in gut symptoms and behavioral problems of autistic babies especially after a dysbiosis induced by antibiotics. Retrospective analysis of the case sheets of autistic patients admitted in Vaidyaratnam P.S.Varier Ayurveda College hospital, kottakkal,kerala, india from September 2010 are taken for the data processing. Autistic patients are used to come to this hospital as a part of their usual course of treatment. We investigated 40 cases diagnosed as autistic by clinical psychologists from different institutions who had dysbiosis induced by antibiotics. Significant change in gut symptoms before and after treatment p<0.05 in most of its components Significant change in behavioral symptoms before and after treatments p<0.05 in most of the components Correlation between gut symptoms change and behavioral symptoms changes after treatment is + 0.86. Conclusion : Selected Polyherbal Ayurveda treatment has significant role to play to make changes abnormal behaviors in autistic babies and has a positive correlation with changes in gut symptoms induced by dysbiosis of antibiotic intake.

Keywords: ayurveda, autism, dysbiosis, antibiotic

Procedia PDF Downloads 597

Authors: Seyed Reza Aghili, Tahereh Shokohi, Lotfollah Davoodi, Zahra Kashi, Azam Moslemi, Mahdi Abastabar, Iman Haghani, Sabah Mayahi, Asoudeh A.

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Introduction: In diabetic foot ulcers, if fungal agents such as Candida species penetrate into the cutaneous or depth of ulcer, can increase the degree of the wound and cause Candia infection and make it more difficult to heal. Material & Methods: A cross-sectional study was performed on 100 diabetic foot ulcer patients in 2020 in Sari, Iran. patient's data and wound grade were recorded in a questionnaire. Candida infection was diagnosed with direct microscopic examination and culture of samples. Colony-PCR molecular method was used for ITS region of DNA and then PCR-RFLP with Msp1 enzyme and using HWP1 specific gene to determine species of Candida agent. Results: Of 100 patients, the mean age 62.1 ± 10.8 years, 95% type 2 diabetes, 83%>10 years duration diabetes, 59% male, 66%> poor education level, 99% married, 52% rural, 95% neuropathic symptoms, 88% using antibiotics, 69%HbA1C >9%, and mean ulcer degree 2.6±1.05 were. Candida infection was seen in 13% of the deep tissue of the wound and 7% cutaneous around the wound. The predominant Candida isolated was C. parapsilosis (71.5%), C .albicans (14.3%). Fungal infections caused by mold fungi were not detected. There was a statistically significant relationship between yeast infection and gender, rural, HbA1C and ulcer degree. Conclusion: Mycological evaluations often are ignored. Candida parapsilosis is the most common infectious agent in these patients and may require specific treatment. Therefore, more attention of physicians to Candida infections particularly, early diagnosis and effective treatment can help faster recovery and prevent amputation.

Keywords: diabetic foot ulcer, candida infection, risk factors, c. parapsilosis

Procedia PDF Downloads 159

Authors: Adebola Ajayi, Francis B. Aiyeleye, Olakunke M. Makanjuola, Olalekan J. Adebowale

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Cocoyam, a generic term for both xanthoma and colocasia, is a traditional staple root crop in many developing countries in Africa, Asia and the Pacific. It is mostly cultivated as food crop which is very rich in vitamin B6, magnesium and also in dietary fiber. The cocoyam starch is easily digested and often used for baby food. Drying food is a method of food preservation that removes enough moisture from the food so bacteria, yeast and molds cannot grow. It is a one of the oldest methods of preserving food. The effect of drying methods on the proximate composition and functional properties of extracted cocoyam starch flour were studied. Freshly harvested cocoyam cultivars at matured level were washed with portable water, peeled, washed and grated. The starch in the grated cocoyam was extracted, dried using sun drying, oven and cabinet dryers. The extracted starch flour was milled into flour using Apex mill and packed and sealed in low-density polyethylene film (LDPE) 75 micron thickness with Nylon sealing machine QN5-3200HI and kept for three months under ambient temperature before analysis. The result showed that the moisture content, ash, crude fiber, fat, protein and carbohydrate ranged from 6.28% to 12.8% 2.32% to 3.2%, 0.89% to 2.24%%, 1.89% to 2.91%, 7.30% to 10.2% and 69% to 83% respectively. The functional properties of the cocoyam starch flour ranged from 2.65ml/g to 4.84ml/g water absorption capacity, 1.95ml/g to 3.12ml/g oil absorption capacity, 0.66ml/g to 7.82ml/g bulk density and 3.82% to 5.30ml/g swelling capacity. Significant difference (P≥0.5) was not obtained across the various drying methods used. The drying methods provide extension to the shelf-life of the extracted cocoyam starch flour.

Keywords: cocoyam, extraction, oven dryer, cabinet dryer

Procedia PDF Downloads 255

Authors: Juliana Lukša, Bazilė Ravoitytė, Elena Servienė, Saulius Serva

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Totiviridae L-A virus is a persistent Saccharomyces cerevisiae dsRNA virus. It encodes the major structural capsid protein Gag and Gag-Pol fusion protein, responsible for virus replication and encapsulation. These features also enable the copying of satellite dsRNAs (called M dsRNAs) encoding a secreted toxin and immunity to it (known as killer toxin). Viral capsid pore presumably functions in nucleotide uptake and viral mRNA release. During cell division, sporogenesis, and cell fusion, the virions remain intracellular and are transferred to daughter cells. By employing high throughput RNA sequencing data analysis, we describe the influence of solely L-A virus on the expression of genes in three different S. cerevisiae hosts. We provide a new perception into Totiviridae L-A virus-related transcriptional regulation, encompassing multiple bioinformatics analyses. Transcriptional responses to L-A infection were similar to those induced upon stress or availability of nutrients. It also delves into the connection between the cell metabolism and L-A virus-conferred demands to the host transcriptome by uncovering host proteins that may be associated with intact virions. To better understand the virus-host interaction, we applied differential proteomic analysis of virus particle-enriched fractions of yeast strains that harboreither complete killer system (L-A-lus and M-2 virus), M-2 depleted orvirus-free. Our analysis resulted in the identification of host proteins, associated with structural proteins of the virus (Gag and Gag-Pol). This research was funded by the European Social Fund under the No.09.3.3-LMT-K-712-19-0157“Development of Competences of Scientists, other Researchers, and Students through Practical Research Activities” measure.

Keywords: totiviridae, killer virus, proteomics, transcriptomics

Procedia PDF Downloads 102