Search results for: S. cerevisiae

Authors: S. Khosravi, X. Chelius, A. Unger, D. Rieger, J. Frickel, T. Sachsenheimer, C. Luechtenborg, R. Schieweck, B. Bruegger, B. Westermann, T. Klecker, W. Neupert, M. E. Harner

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The use of Saccharomyces cerevisiae as a model organism to study eukaryotic cell functions has been used successfully for decades. Like virtually all eukaryotic cells, they contain mitochondria as essential organelles performing various functions, including participation in lipid metabolism. They are separated from the cytosol by a double membrane system consisting of the mitochondrial inner membrane (MIM) and the mitochondrial outer membrane (MOM). This physical separation of the mitochondria requires an exchange of metabolites, proteins, and lipids. Proteinaceous contact sites are thought to be important for this communication. Recently, it was found that Cqd1, in cooperation with Cqd2, controls the distribution of Coenzyme Q within the cell. In this study, a contact site is described, formed by the MOM protein complex Por1-Om14 and the UbiB protein kinase-like MIM protein Cqd1. The present results suggest the additional involvement of Cqd1 in the homeostasis of phospholipids. Moreover, we show that overexpression of the UbiB family proteins also causes tethering of the mitochondria to the endoplasmatic reticulum. Due to the conservation of the subunits of this contact site to higher eukaryotes, its identification in S. cerevisiae might provide promising avenues for further research in other organisms.

Keywords: contact sites, mitochondrial architecture, mitochondrial proteins, yeast mitochondria

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Authors: Natalia Koltovaya, Nadezhda Zhuchkina, Ksenia Lyubimova

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We study the biological effects induced by ionizing radiation in view of therapeutic exposure and the idea of space flights beyond Earth's magnetosphere. In particular, we examine the differences between base pair substitution induction by ionizing radiation in model haploid and diploid yeast Saccharomyces cerevisiae cells. Such mutations are difficult to study in higher eukaryotic systems. In our research, we have used a collection of six isogenic trp5-strains and 14 isogenic haploid and diploid cyc1-strains that are specific markers of all possible base-pair substitutions. These strains differ from each other only in single base substitutions within codon-50 of the trp5 gene or codon-22 of the cyc1 gene. Different mutation spectra for two different haploid genetic trp5- and cyc1-assays and different mutation spectra for the same genetic cyc1-system in cells with different ploidy — haploid and diploid — have been obtained. It was linear function for dose-dependence in haploid and exponential in diploid cells. We suggest that the differences between haploid yeast strains reflect the dependence on the sequence context, while the differences between haploid and diploid strains reflect the different molecular mechanisms of mutations.

Keywords: base pair substitutions, γ-rays, haploid and diploid cells, yeast Saccharomyces cerevisiae

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Authors: Sajjad Ur Rahman, Muhammad Tahir, Mukarram Bashir, Jawad, Aoun Muhammad, Muhammad Zohaib, Hannan Khan, Seemal Javaid, Mariam Azam

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The efficiency of natural metabolites obtained from partially fermented soya hulls and wheat bran using Saccharomyces cerevisiae (DL-22 S/N) ensures a potential impact on the total milk yield and quality of milk production. On attaining a moderate number of Saccharomyces cerevisiae cells around 1×10⁹ CFU/ml, the concentrate was further elevated under in-vivo conditions to study the quality of milk production in lactating buffalo. Ten lactating buffalos of the Nili Ravi breed having the same physical factors were given 12 gm of microbial metabolites daily, along with the palleted feed having 22 % proteins. Another group of 10 lactating animals with the same characteristics was maintained without metabolites. The body score, overall health, incidence of mastitis, milk fat, milk proteins, ash and solid not fat (SNF) were elevated on a weekly basis up to thirty days of trial. It was recorded that the average total increase in quality milk production was 0.9 liter/h/d, whereas SNF in the milk was enhanced to 0.71, and fats were decreased to 0.09 %. Moreover, during all periods of the trial, the overall non-specific immunity of buffalo was increased, as indicated by less than 0.2 % of mastitis incidence compared to 1.8% in the untreated buffalos.

Keywords: natural metabolites, quality milk, milk yield, microorganisms, fermentation, nonspecific immunity, better performing animals

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Authors: Nizar Chaira

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Dates, main products of the oases, due to their therapeutic interests, are considered highly nutritious fruit. Several studies on the valuation biotechnology and technology of dates are made, and several products are already prepared. Isolation of the yeast Saccharomyces cerevisiae, naturally presents in a scrap of date, optimization of growth in the medium based on date syrup and production biomass can potentially expand the range of secondary products of dates. To this end, this paper tries to study the suitability for processing dates technology and biotechnology to use the date pulp as a carbon source for biological transformation. Two strains of Saccharomyces cerevisiae isolated from date syrup (S1, S2) and a commercial strain have used for this study. After optimization of culture conditions, production in a fermenter on two different media (date syrup and beet molasses) was performed. This is followed by studying the kinetics of growth, protein production and consumption of sugars in crops strain 1, 2 and the commercial strain and on both media. The results obtained showed that a concentration of 2% sugar, 2.5 g/l yeast extract, pH 4.5 and a temperature between 25 and 35°C are the optimal conditions for cultivation in a bioreactor. The exponential phase of the specific growth rate of a strain on both media showed that it is about 0.3625 h-1 for the production of a medium based on date syrup and 0.3521 h-1 on beet molasses with a generation time equal to 1.912 h and on the medium based on date syrup, yeast consumes preferentially the reducing sugars. For the production of protein, we showed that this latter presents an exponential phase when the medium starts to run out of reducing sugars. For strain 2, the specific growth rate is about 0.261h-1 for the production on a medium based on date syrup and 0207 h-1 on beet molasses and the base medium syrup date of the yeast consumes preferentially reducing sugars. For the invertase and other metabolits, these increases rapidly after exhaustion of reducing sugars. The comparison of productivity between the three strains on the medium based on date syrup showed that the maximum value is obtained with the second strain: p = 1072 g/l/h as it is about of 0923 g/l/h for strain 1 and 0644 g/l/h for the commercial strain. Thus, isolates of date syrup are more competitive than the commercial strain and can give the same performance in a shorter time with energy gain.

Keywords: date palm, fermentation, molasses, Saccharomyces, syrup

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Authors: Ganesh Dattatraya Saratale, Min Kyu Oh

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Rice straw is one of the most abundant lignocellulosic waste materials and its annual production is about 731 Mt in the world. This study treats the subject of effective utilization of this waste biomass for biofuels production. We have showed a comparative assessment of numerous pretreatment strategies for rice straw, comprising of major physical, chemical and physicochemical methods. Among the different methods employed for pretreatment alkaline pretreatment in combination with sodium chlorite/acetic acid delignification found efficient pretreatment with significant improvement in the enzymatic digestibility of rice straw. A cellulase dose of 20 filter paper units (FPU) released a maximum 63.21 g/L of reducing sugar with 94.45% hydrolysis yield and 64.64% glucose yield from rice straw, respectively. The effects of different pretreatment methods on biomass structure and complexity were investigated by FTIR, XRD and SEM analytical techniques. Finally the enzymatic hydrolysate of rice straw was used for ethanol production using developed Saccharomyces cerevisiae SR8. The developed yeast strain enabled efficient fermentation of xylose and glucose and produced higher ethanol production. Thus development of bioethanol production from lignocellulosic waste biomass is generic, applicable methodology and have great implication for using ‘green raw materials’ and producing ‘green products’ much needed today.

Keywords: rice straw, pretreatment, enzymatic hydrolysis, FPU, Saccharomyces cerevisiae SR8, ethanol fermentation

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Authors: Mariam Azam, Sajjad Ur Rahman, Mukarram Bashir, Muhammad Tahir, Seemal Javaid, Jawad, Aoun Muhammad, Muhammad Zohaib, Hannan Khan

Abstract:

Products of microbial origin are of great importance as they have proved their value in healthcare and nutrition, use of these microbial metabolites acquired from partially fermented soya hulls and wheat bran along with Saccharomyces cerevisiae (DL-22 S/N) substantiates to be a great source for an increase in the total milk production and quality yield.1×109 CFU/ml cells of Saccharomyces cerevisiae (DL-22 S/N) were further grown under in-vivo conditions for the assessment of quality milk production. Two groups with twelve cows, each having the same physical characteristics (Group A and Group B), were under study, Group A was daily fed with 12gm of biological metabolites and 22% protein-pelleted feed. On the other hand, the animals of Group B were provided with no metabolites in their feed. In thirty days of trial, improvement in the overall health, body score, milk protein, milk fat, yield, incidence rate of mastitis, ash, and solid not fat (SNF) was observed. The collected data showed that the average quality milk production was elevated up to 0.45 liter/h/d. However, a reduction in the milk fats up to 0.45% and uplift in the SNF value up to 0.53% of cow milk was also observed. At the same time, the incidence rate of mastitis recorded for the animals under trial was reduced to half, and improved non specific immunity was reported.

Keywords: microbial metabolites, post-biotics, animal supplements, animal nutrition, proteins, animal production, fermentation

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Authors: Elizabeth Chinyere Amadi

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Yeast (sacchoromyces cerevisiae) was isolated from the fermenting sap of raffia palm (Raphia hookeri) wine. Different concerntrations of the yeast isolate were used to produce bread samples – B, C, D, E, F containing (2, 3, 4, 5, 6) g of yeast isolate respectively, other ingredients were kept constant. Sample A, containing 2g of commercial baker yeast served as control. The proof heights, weights, volumes and specific volume of the dough and bread samples were determined. The bread samples were also subjected to sensory evaluation using a 9–point hedonic scale. Results showed that proof height increased with increased concentration of the yeast isolate; that is direct proportion. Sample B with the least concentration of the yeast isolate had the least loaf height and volume of 2.80c m and 200 cm³ respectively but exhibited the highest loaf weight of 205.50g. However, Sample A, (commercial bakers’ yeast) had the highest loaf height and volume of 5.00 cm and 400 cm³ respectively. The sensory evaluation results showed sample D compared favorably with sample A in all the organoleptic attributes-(appearance, taste, crumb texture, crust colour and overall acceptability) tested for (P< 0.05). It was recommended that 4g compressed yeast isolate per 100g flour could be used to proof dough as a substitute for commercial bakers’ yeast and produce acceptable bread loaves.

Keywords: isolation of yeast, performance evaluation of yeast, Raffia palm wine, used at different concentrations, proofing of bread dough

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Authors: Phillip Minnaar, Neil Jolly, Louisa Beukes, Santiago Benito-Saez

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Dovyalis caffra is a tree found on the African continent. Limited information exists on the effect of acetous fermentation on the phytochemicals of Kei-apple fruit. The phytochemical content of vinegars is derived from compounds present in the fruit the vinegar is made of. Kei-apple fruit juice was co-inoculated with Schizosaccharomyces pombe and Saccharomyces cerevisiae to induce alcoholic fermentation (AF). Acetous fermentation followed AF, using an acetic acid bacteria consortium as an inoculant. Juice had the lowest pH and highest total acidity (TA). The wine had the highest pH and vinegars lowest TA. Total soluble solids and L-malic acid decreased during AF and acetous fermentation. Volatile acidity concentration was not different among vinegars. Gallic, syringic, caffeic, p-coumaric, and chlorogenic acids increased during acetous fermentation, whereas ferulic, sinapic, and protocatechuic acids decreased. Chlorogenic acid was the most abundant phenolic acid in both wines and vinegars. It is evident from this investigation that Kei-apple vinegar is a source of plant-derived phenolics, which evolved through fermentation. However, the AAB selection showed minimal performance with respect to VA production. Acetic acid bacteria selection for acetous fermentation should be reconsidered, and the reasons for the decrease of certain phenolic acids during acetous fermentation needs to be investigated.

Keywords: acetic acid bacteria, acetous fermentation, liquid chromatography, phenolic acids

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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: Madalina Postaru, Alexandra Tucaliuc, Dan Cascaval, Anca Irina Galaction

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Ergosterol (ergosta-5,7,22-trien-3β-ol) is the precursor of vitamin D2 (ergocalciferol), known as provitamin D2 as it is converted under UV radiation to this vitamin. The natural sources of ergosterol are mainly the yeasts (Saccharomyces sp., Candida sp.), but it can be also found in fungus (Claviceps sp.) or plants (orchids). As ergosterol is mainly accumulated in yeast cell membranes, especially in free form in the plasma-membrane, and the chemical synthesis of ergosterol does not represent an efficient method for its production, this study aimed to analyze the influence of aeration efficiency on ergosterol production by S. cerevisiae in batch and fed-batch fermentations, by considering different levels of mixing intensity, aeration rate, and n-dodecane concentration. Our previous studies on ergosterol production by S. cerevisiae in batch and fed-batch fermentation systems indicated that the addition of n-dodecane led to the increase of almost 50% of this sterol concentration, the highest productivity being reached for the fed-batch process. The experiments were carried out in a laboratory stirred bioreactor, provided with computer-controlled and recorded parameters. In batch fermentation system, the study indicated that the oxygen mass transfer coefficient, kLa, is amplified for about 3 times by increasing the volumetric concentration of n-dodecane from 0 to 15%. Moreover, the increase of dissolved oxygen concentration by adding n-dodecane leads to the diminution for 3.5 times of the produced alcohol amount. In fed-batch fermentation process, the positive influence of hydrocarbon on oxygen transfer rate is amplified mainly at its higher concentration level, as the result of the increased yeasts cells amount. Thus, by varying n-dodecane concentration from 0 to 15% vol., the kLa value increase becomes more important than for the batch fermentation, being of 4 times

Keywords: ergosterol, yeast fermentation, n-dodecane, oxygen-vector

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Authors: Ali Zein Aalabiden Tlais, Alessio Da Ros, Pasquale Filannino, Olimpia Vincentini, Marco Gobbetti, Raffaella Di Cagno

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This study is an example of apple by-products (AP) recycling through a designed fermentation by selected autochthonous Lactobacillus plantarum AFI5 and Lactobacillus fabifermentans ALI6 used singly or as binary cultures with the selected Saccharomyces cerevisiae AYI7. Compared to Raw-, Unstarted- and Chemically Acidified-AP, Fermented-AP promoted the highest levels of total and insoluble dietary fibers, antioxidant activity, and free phenolics. The binary culture of L. plantarum AFI5 and S. cerevisiae AYI7 had the best effect on the bioavailability phenolic compounds as resulted by the Liquid chromatography-mass spectrometry validated method. The accumulation of phenolic acid derivatives highlighted microbial metabolism during AP fermentation. Bio-converted phenolic compounds were likely responsible for the increased antioxidant activity. The potential health-promoting effects of Fermented-AP were highlighted using Caco-2 cells. With variations among single and binary cultures, fermented-AP counteracted the inflammatory processes and the effects of oxidative stress in Caco-2 cells and preserved the integrity of tight junctions. An alternative and suitable model for food by-products recycling to manufacture a dietary supplement fortified with biogenic compounds was proposed. Highlighting the microbial metabolism of several phenolic compounds, undoubted additional value to such downstream wastes was created.

Keywords: apple by-products, antioxidant, fermentation, phenolic compounds

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

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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

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Authors: Nho-Eul Song, Sang-Ho Baik

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Fermentation characteristics of black raspberry vinegar by using static cultures without any additives were has been investigated to establish of vinegar manufacturing conditions and improve the quality of vinegar by optimization the vinegar manufacturing process. The two vinegar manufacturing conditions were prepared; one-step fermentation condition only using mother vinegar that prepared naturally occurring black raspberry vinegar without starter yeast for alcohol fermentation (traditional method) and two-step fermentation condition using commercial wine yeast and mother vinegar for acetic acid fermentation. Approximately 12% ethanol was produced after 35 days fermentation with log 7.6 CFU/mL of yeast population in one-step fermentation, resulting sugar reduction from 14 to 6oBrix whereas in two-step fermentation, ethanol concentration was reached up to 8% after 27 days with continuous increasing yeast until log 7.0 CFU/mL. In addition, yeast and ethanol were decreased after day 60 accompanied with proliferation of acetic acid bacteria (log 5.8 CFU/mL) and titratable acidity; 4.4% in traditional method and 6% in two-step fermentation method. DGGE analysis showed that S. cerevisiae was detected until 77 days of traditional fermentation and gradually changed to AAB, Acetobacter pasteurianus, as dominant species and Komagataeibacter xylinus at the end of the fermentation. However, S. cerevisiae and A. pasteurianus was dominant in two-step fermentation process. The prepared two-step fermentation showed enhanced total polyphenol and flavonoid content significantly resulting in higher radical scavenging activity. Our studies firstly revealed the microbial community change with chemical change and demonstrated a suitable fermentation system for black raspberry vinegar by the static surface method.

Keywords: bacteria, black raspberry, vinegar fermentation, yeast

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Authors: Claire B. Salvedia, Enrico P. Supangco, Francisco B. Eligado, Renato Sa Vega, Antonio A. Rayos

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The study was conducted to evaluate the effect of probiotic feeding on weight gain, blood biochemical and hematological indices of crossbred dairy goat kids. Sixteen (16) crossbred Anglo-Nubian x Saanen dairy goat kids, 3 to 4 months old, ranging from 19 to 23kg were randomly assigned into four treatments fed with 5x109 cfu/ml probiotic supplements; Treatment 1 – control; Treatment 2 – lactic acid bacteria (L. plantarum BS and P. acidilactici 3G3); treatment 3 – S. cerevisiae 2030; Treatment 4 – multi-strain probiotics (L. plantarum BS, P. acidilactici 3G3, and S.cerevisiae 2030). Feed ration provided daily for each of the experimental animals were composed of 1kg mixed concentrate feed ((Leucaena leucocephala dried leaves and pollard), and 4 kg fresh Pennisetum purpureum and Gliciridia sepium leaves (50:50). The experimental feeding trial lasted for 9 weeks. Result revealed that treatments fed with probiotics had significantly (P≤0.05) higher weight gain compared to the control. Significant effect on plasma urea nitrogen (PUN) and triglyceride were noted during 30th and 60th day of probiotic feeding. White blood cell counts were significantly affected by probiotic feeding during the 60th day. Concentrations of glucose and cholesterol remained unchanged throughout the experimental period. The findings suggests, under the condition of the experiment, that live probiotic feeding could have a significant role in improving weight gain and metabolism of crossbred dairy goat kids.

Keywords: probiotics, weight gain, blood biochemical indices, crossbred dairy goat kids

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Authors: Sergei Voychuk

Abstract:

Introduction: Adaptive response (AR) is a manifestation of radiation hormesis, which deal with the radiation resistance that may be increased with the pretreatment with small doses of radiation. In the current study, we evaluated the potency of radiofrequency EMF to induce the AR mechanisms and to increase a resistance to UV light. Methods: Saccharomyces cerevisiae yeast strains, which were created to study induction of mutagenesis and recombination, were used in the study. The strains have mutations in rad2 and rad54 genes, responsible for DNA repair: nucleotide excision repair (PG-61), postreplication repair (PG-80) and mitotic (crossover) recombination (T2). An induction of mutation and recombination are revealed due to the formation of red colonies on agar plates. The PG-61 and T2 are UV sensitive strains, while PG-80 is sensitive to ionizing radiation. Extremely high frequency electromagnetic field (EHF-EMF) was used. The irradiation was performed in floating mode and frequency changed during exposure from 57 GHz to 62 GHz. The power of irradiation was 100 mkW, and duration of exposure was 10 and 30 min. Treatment was performed at RT and then cells were stored at 28° C during 1 h without any exposure but after that they were treated with UV light (254nm) for 20 sec (strain T2) and 120 sec (strain PG-61 and PG-80). Cell viability and quantity of red colonies were determined after 5 days of cultivation on agar plates. Results: It was determined that EHF-EMF caused 10-20% decrease of viability of T2 and PG-61 strains, while UV showed twice stronger effect (30-70%). EHF-EMF pretreatment increased T2 resistance to UV, and decreased it in PG-61. The PG-80 strain was insensitive to EHF-EMF and no AR effect was determined for this strain. It was not marked any induction of red colonies formation in T2 and PG-80 strain after EHF or UV exposure. The quantity of red colonies was 2 times more in PG-61 strain after EHF-EMF treatment and at least 300 times more after UV exposure. The pretreatment of PG-61 with EHF-EMF caused at least twice increase of viability and consequent decrease of amount of red colonies. Conclusion: EHF-EMF may induce AR in yeast cells and increase their viability under UV treatment.

Keywords: Saccharomyces cerevisiae, EHF-EMF, UV light, adaptive response

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Authors: Muhammad Zainuddin Arriafdi, Hamudah Hakimah Abdullah, Mohd Helmi Sani, Wan Azlina Ahmad, Muhd Nazrul Hisham Zainal Alam

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The biotechnology process development demands numerous experimental works. In laboratory environment, this is typically carried out using a shake flask platform. This paper presents the design and fabrication of a miniature bioreactor system as an alternative research tool for bioprocessing. The working volume of the reactor is 100 ml, and it is made of plastic. The main features of the reactor included stirring control, temperature control via the electrical heater, aeration strategy through a miniature air compressor, and online optical cell density (OD) sensing. All sensors and actuators integrated into the reactor was controlled using an Arduino microcontroller platform. In order to demonstrate the functionality of such miniature bioreactor concept, series of batch Saccharomyces cerevisiae fermentation experiments were performed under various glucose concentrations. Results attained from the fermentation experiments were utilized to solve the Monod equation constants, namely the saturation constant, Ks, and cells maximum growth rate, μmax as to further highlight the usefulness of the device. The mixing capacity of the reactor was also evaluated. It was found that the results attained from the miniature bioreactor prototype were comparable to results achieved using a shake flask. The unique features of the device as compared to shake flask platform is that the reactor mixing condition is much more comparable to a lab-scale bioreactor setup. The prototype is also integrated with an online OD sensor, and as such, no sampling was needed to monitor the progress of the reaction performed. Operating cost and medium consumption are also low and thus, making it much more economical to be utilized for biotechnology process development compared to lab-scale bioreactors.

Keywords: biotechnology, miniature bioreactor, research tools, Saccharomyces cerevisiae

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Authors: Saeed Ahmed, Tamoor Abbas, M. Amir, M. S. Iqbal, D. Hussain

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This study was conducted to evaluate the effect of supplementation of different levels of yeast culture (Saccharomyces cerevisiae) in Beetal male goats diets on growth performance, digestibility of nutrients and selected blood metabolites. Another objective was to determine the inclusion level of yeast culture for optimal growth performance of Beetal male goats. Eighteen (n=18) Beetal male goats were randomly assigned to three total mixed ration treatments (n=6 goats/treatment): T1, T2 and T3 containing 0gm, 3gm and 6gm/day yeast culture (YC) mixed with total mixed ration (TMR). The diets were iso-nitrogenous and iso-caloric having crude protein 15.2% and ME 2.6Mcal/kg. The total duration of the experiment was 8 weeks. Beetal bucks were fed on TMR diets (T1, T2 and T3) having blend of oat silage, Lucerne hay and concentrate mixed with yeast culture (YC). Bucks were housed individually and feed was offered @ 4% of body weight on dry matter basis. Samples of fresh feed and refusal were collected twice weekly of moisture percentage using hot air oven. Data for daily dry matter intake, body weight gain, nutrient digestibility and selected blood metabolites were analyzed through one-way ANOVA technique under Complete randomised design (SAS Institute Inc, 2002-03). Results were declared significant at P≤0.05. Overall, DMI was not affected (P≥0.05) by dietary treatments. Body weight gain, digestibility of crude protein and crude fibre were improved. Blood glucose concentration was detected higher in the group having supplementation of yeast culture (YC) 6gm/day compared to other two dietary treatments. This study suggested the positive impact of inclusion of yeast culture (YC) up to 6gm/day in the TMR diet for optimal growth performance and digestibility of nutrients in Beetal male goats.

Keywords: yeast culture, growth performance, digestibility, beetle goat

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Authors: Yee-Seul So, Guiseppe Ianiri, Alex Idnurm, Yong-Sun Bahn

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Tor1 is a serine/threonine protein kinase that is widely conserved across eukaryotic species. Tor1 was first identified in Saccharomyces cerevisiae as a target of rapamycin (TOR). The TOR pathway has been implicated in regulating cellular responses to nutrients, proliferation, translation, transcription, autophagy, and ribosome biogenesis. Here we identified two homologues of S. cerevisiae Tor proteins, CNAG_06642 (Tor1) and CNAG_05220 (Tlk1, TOR-like kinase 1), in Cryptococcus neoformans causing a life-threatening fungal meningoencephalitis. Both Tor1 and Tlk1 have rapamycin-binding (RB) domains but Tlk1 has truncated RB form. To study the TOR-signaling pathway in the fungal pathogen, we attempt to construct the tor1Δ and tlk1Δ mutants and phenotypically analyze them. Although we failed to construct the tor1Δ mutant, we successfully construct the tlk1Δ mutant. The tlk1Δ mutant does not exhibit any discernable phenotypes, suggesting that Tlk1 is dispensable in C. neoformans. The essentiality of TOR1 is independently confirmed by constructing the TOR1 promoter replacement strain by using a copper transporter 4 (CTR4) promoter and the TOR1/tor1 heterozygous mutant in diploid C. neoformans strain background followed by sporulation analysis. To further analyze the function of Tor1, we construct TOR1 overexpression mutant using a constitutively active histone H3 in C. neoformans. We find that the Tor1 overexpression mutant is resistant to rapamycin but the tlk1Δ mutant does not exhibit any altered resistance to rapamycin, further confirming that Tor1, but not Tlk1, is critical for TOR signaling. Furthermore, we found that Tor1 is involved in response to diverse stresses, including genotoxic stress, oxidative stress, thermo-stress, antifungal drug treatment, and production of melanin. To identify any TOR-related transcription factors, we screened C. neoformans transcription factor library that we constructed in our previous study and identified several potential downstream factors of Tor1, including Atf1, Crg1 and Bzp3. In conclusion, the current study provides insight into the role of the TOR signaling pathway in human fungal pathogens as well as C. neoformans.

Keywords: fungal pathogen, serine/threonine kinase, target of rapamycin, transcription factor

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Authors: Alexandra Tucaliuc, Alexandra C. Blaga, Anca I. Galaction, Lenuta Kloetzer, Dan Cascaval

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Ergosterol (ergosta-5,7,22-trien-3β-ol), also known as provitamin D2, is the precursor of vitamin D2 (ergocalciferol), because it is converted under UV radiation to this vitamin. The natural sources of ergosterol are mainly the yeasts (Saccharomyces sp., Candida sp.), but it can be also found in fungus (Claviceps sp.) or plants (orchids). In the yeasts cells, ergosterol is accumulated in membranes, especially in free form in the plasma membrane, but also as esters with fatty acids in membrane lipids. The chemical synthesis of ergosterol does not represent an efficient method for its production, in these circumstances, the most attractive alternative for producing ergosterol at larger-scale remains the aerobic fermentation using S. cerevisiae on glucose or by-products from agriculture of food industry as substrates, in batch or fed-batch operating systems. The aim of this work is to analyze comparatively the influence of aeration efficiency on ergosterol production by S. cerevisiae in batch and fed-batch fermentations, by considering different levels of mixing intensity, aeration rate, and n-dodecane concentration. The effects of the studied factors are quantitatively described by means of the mathematical correlations proposed for each of the two fermentation systems, valid both for the absence and presence of oxygen-vector inside the broth. The experiments were carried out in a laboratory stirred bioreactor, provided with computer-controlled and recorded parameters. n-Dodecane was used as oxygen-vector and the ergosterol content inside the yeasts cells has been considered at the fermentation moment related to the maximum concentration of ergosterol, 9 hrs for batch process and 20 hrs for fed-batch one. Ergosterol biosynthesis is strongly dependent on the dissolved oxygen concentration. The hydrocarbon concentration exhibits a significant influence on ergosterol production mainly by accelerating the oxygen transfer rate. Regardless of n-dodecane addition, by maintaining the glucose concentration at a constant level in the fed-batch process, the amount of ergosterol accumulated into the yeasts cells has been almost tripled. In the presence of hydrocarbon, the ergosterol concentration increased by over 50%. The value of oxygen-vector concentration corresponding to the maximum level of ergosterol depends mainly on biomass concentration, due to its negative influences on broth viscosity and interfacial phenomena of air bubbles blockage through the adsorption of hydrocarbon droplets–yeast cells associations. Therefore, for the batch process, the maximum ergosterol amount was reached for 5% vol. n-dodecane, while for the fed-batch process for 10% vol. hydrocarbon.

Keywords: bioreactors, ergosterol, fermentation, oxygen-vector

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Authors: Ali Azam Talukder, Jamsheda Ferdous Tuli, Tanzina Islam Reba, Shuvra Kanti Dey, Mamoru Yamada

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Recent global warming created by various pollutants prompted us to find new energy sources instead of fossil fuels. Fossil fuels are one of the key factors to emit various toxic gases in this planet. To solve this problem, along with the scarcity of the worldwide energy crisis, scientists are looking for various alternative options to mitigate the necessity of required future fuels. In this context, bioethanol can be one of the most suitable alternative energy sources. Bioethanol is a renewable, environment-friendly and carbon-neutral sustainable energy. In our previous study, we identified several bioethanol-producing microbes from the natural fermented sources of Bangladesh. Among them, the strain 4C encoded Saccharomyces cerevisiae produced maximum bioethanol when the fermentation temperature was 25˚C. In this study, we have established high-temperature simultaneous saccharification and fermentation process (HTSSF) by co-culturing of thermally adapted thermosensitive 4C as a fermenting agent and Bacillus amyloliquefaciens (C7), as a saccharifying agent under various physiological conditions or treatments. Conventional methods were applied for cell culture, media preparation and other experimental purposes. High-temperature adaptation of strain 4C was made from 30-42ᵒC, using either YPD or YPS media. In brief, for thermal adaptation, the temperature was periodically increased by 2ᵒC, 1ᵒC and 0.5ᵒC when medium growth temperatures were 30-36ᵒC, 36-40ᵒC, and 40-42ᵒC, respectively, where applicable. Amylase activity and bioethanol content were measured by DNS (3, 5-dinitrosalicylic acid) and solvent extraction and dichromate oxidation method, respectively. Among the various growth parameters like temperatures (30˚C, 37˚C and 42˚C), pHs (5.0, 6.0 and 7.0), carbon sources (5.0-10.0%) and ethanol stress tolerance (0.0-12.0%) etc. were tested, maximum Amylase activity (4.0 IU/ml/min) was recorded for Bacillus amyloliquefaciens (C7) at 42˚C, pH 6.0 and 10% starch. On the other hand, 4.10% bioethanol content was recorded when the thermally adapted strain 4C was co-cultured with C7 at 37ᵒC, pH 6.0 and 10.0% starch for 72 hours at HTSSF process. On the other hand, thermally non-adapted strains gave only 0.5-2.0% bioethanol content under the same physiological conditions. The thermally adapted strain 4C and strain C7, both can tolerate ethanol stress up to 12%. Altogether, a comparative study revealed that our established HTSSF process may be suitable for pilot scale and subsequently at industrial level bioethanol production.

Keywords: bioethanol, co-culture, fermentation, saccharification

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Authors: S. Paengkoum, P. Paengkoum, W. Kaewwongsa

Abstract:

Twenty-four male growing goats were randomly assigned to a Randomized Complete Block Design. Dietary treatments were different level of feeding concentrate diet at 1.0, 1.5, 2.0, and 2.5% of body weight (BW). The results showed that average daily gain, microbial N supply, N retention of meat goats in the group of feeding level at 2.0% BW and 2.5% BW were significantly higher (P<0.05) than those goats fed with feeding levels of 1.0% BW and 1.5% BW. Based on this result the conclusion can be made that using 75% fermented cassava pulp by Saccharomyces cerevisiae as the main source of protein to completely replace soybean meal was beneficial to meat goats in terms of feed intake. The feeding concentrate at levels between 2.0-2.5% BW gives highest in the growth of meat goat in this experiment.

Keywords: cassava pulp, yeast, goat, nitrogen retention

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Authors: Angelika-Ioanna Gialleli, Gousi Mantha, Maria Kanellaki, Bekatorou Argyro, Athanasios Koutinas

Abstract:

In this study, a new brewing technology with dry raw materials is proposed with potential application in home brewing. Bio catalysts were prepared by immobilization of the psychrotolerant yeast strain Saccharomyces cerevisiae AXAZ-1 on tubular cellulose. Both the word and the biocatalysts were freeze-dried without any cryoprotectants and used for low temperature brewing. The combination of immobilization and freeze-drying techniques was applied successfully, giving a potential for supplying breweries with preserved and ready-to-use immobilized cells. The effect of wort sugar concentration (7°, 8.5°, 10°Be), temperature (2, 5, 7° C) and carrier concentration (5, 10, 20 g/L) on fermentation kinetics and final product quality (volatiles, colour, polyphenols, bitterness) was assessed. The same procedure was repeated with free cells for comparison of the results. The results for immobilized cells were better compared to free cells regarding fermentation kinetics and organoleptic characteristics.

Keywords: brewing, tubular cellulose, low temperature, biocatalyst

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Authors: Jong H. Kim, Kathleen L. Chan, Luisa W. Cheng

Abstract:

Control of fungal pathogens, such as foodborne mycotoxin producers, is problematic as effective antimycotic agents are often very limited. Mycotoxin contamination significantly interferes with the safe production of foods or crops worldwide. Moreover, expansion of fungal resistance to commercial drugs or fungicides is a global human health concern. Therefore, there is a persistent need to enhance the efficacy of commercial antimycotic agents or to develop new intervention strategies. Disruption of the cellular antioxidant system should be an effective method for pathogen control. Such disruption can be achieved with safe, redox-active compounds. Natural phenolic derivatives are potent redox cyclers that inhibit fungal growth through destabilization of the cellular antioxidant system. The goal of this study is to identify novel, redox-active compounds that disrupt the fungal antioxidant system. The identified compounds could also function as sensitizing agents to conventional antimycotics (i.e., chemosensitization) to improve antifungal efficacy. Various benzo derivatives were tested against fungal pathogens. Gene deletion mutants of the yeast Saccharomyces cerevisiae were used as model systems for identifying molecular targets of benzo analogs. The efficacy of identified compounds as potent antifungal agents or as chemosensitizing agents to commercial drugs or fungicides was examined with methods outlined by the Clinical Laboratory Standards Institute or the European Committee on Antimicrobial Susceptibility Testing. Selected benzo derivatives possessed potent antifungal or antimycotoxigenic activity. Molecular analyses by using S. cerevisiae mutants indicated antifungal activity of benzo derivatives was through disruption of cellular antioxidant or cell wall integrity system. Certain benzo analogs screened overcame tolerance of Aspergillus signaling mutants, namely mitogen-activated protein kinase mutants, to fludioxonil fungicide. Synergistic antifungal chemosensitization greatly lowered minimum inhibitory or fungicidal concentrations of test compounds, including inhibitors of mitochondrial respiration. Of note, salicylaldehyde is a potent antimycotic volatile that has some practical application as a fumigant. Altogether, benzo derivatives targeting cellular antioxidant system of fungi (along with cell wall integrity system) effectively suppress fungal growth. Candidate compounds possess the antifungal, antimycotoxigenic or chemosensitizing capacity to augment the efficacy of commercial antifungals. Therefore, chemogenetic approaches can lead to the development of novel antifungal intervention strategies, which enhance the efficacy of established microbe intervention practices and overcome drug/fungicide resistance. Chemosensitization further reduces costs and alleviates negative side effects associated with current antifungal treatments.

Keywords: antifungals, antioxidant system, benzo derivatives, chemosensitization

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Authors: Sara Heidari, Jafar Mohammadzadeh Milani, Elmira Pouladi Borj

Abstract:

In this research, stability of Ochratoxin A (OTA) during bread making process including fermentation with yeasts (Saccharomyces cerevisiae) and Sourdough (Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus and Lactobacillus fermentum) and baking at 200°C were examined. Bread was prepared on a pilot-plant scale by using wheat flour spiked with standard solution of OTA. During this process, mycotoxin levels were determined after fermentation of the dough with sourdough and three types of yeast including active dry yeast, instant dry yeast and compressed yeast after further baking 200°C by high performance liquid chromatography (HPLC) with fluorescence detector after extraction and clean-up on an immunoaffinity column. According to the results, the highest stability of was observed in the first fermentation (first proof), while the lowest stability was observed in the baking stage in comparison to contaminated flour. In addition, compressed yeast showed the maximum impact on stability of OTA during bread making process.

Keywords: Ochratoxin A, bread, dough, yeast, sourdough

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Authors: Boukhiar Aissa, Iguergaziz Nadia, Halladj Fatima, Lamrani Yasmina, Benamara Salem

Abstract:

Determining the alcohol content in alcoholic fermentation bioprocess has a great importance. In fact, it is a key indicator for monitoring this fermentation bioprocess. Several methodologies (chemical, spectrophotometric, chromatographic...) are used to the determination of this parameter. However, these techniques are very long and require: rigorous preparations, sometimes dangerous chemical reagents, and/or expensive equipment. In the present study, the date juice is used as a substrate of alcoholic fermentation. The extracted juice undergoes an alcoholic fermentation by Saccharomyces cerevisiae. The study of the possible use of refractometry as a sole means for the in situ control of this process revealed a good correlation (R2 = 0.98) between initial and final ° Brix: ° Brix f = 0.377× ° Brixi. In addition, we verified the relationship between the variation in final and initial ° Brix (Δ ° Brix) and alcoholic rate produced (A exp): CΔ° Brix / A exp = 1.1. This allows the tracing of abacus isoresponses that permit to determine the alcoholic and residual sugar rates with a mean relative error (MRE) of 5.35%.

Keywords: refractometry, alcohol, residual sugar, fermentation, brix, date, juice

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Authors: Preshanthan Moodley, E. B. Gueguim Kana

Abstract:

This study examines the kinetics of S. cerevisiae BY4743 growth and bioethanol production from sugarcane leaf waste (SLW), utilizing two different optimized pretreatment regimes; under two fermentation modes: steam salt-alkali filtered enzymatic hydrolysate (SSA-F), steam salt-alkali unfiltered (SSA-U), microwave salt-alkali filtered (MSA-F) and microwave salt-alkali unfiltered (MSA-U). The kinetic coefficients were determined by fitting the Monod, modified Gompertz, and logistic models to the experimental data with high coefficients of determination R² > 0.97. A maximum specific growth rate (µₘₐₓ) of 0.153 h⁻¹ was obtained under SSA-F and SSA-U whereas, 0.150 h⁻¹ was observed with MSA-F and MSA-U. SSA-U gave a potential maximum bioethanol concentration (Pₘ) of 31.06 g/L compared to 30.49, 23.26 and 21.79g/L for SSA-F, MSA-F and MSA-U respectively. An insignificant difference was observed in the μmax and Pm for the filtered and unfiltered enzymatic hydrolysate for both SSA and MSA pretreatments, thus potentially reducing a unit operation. These findings provide significant insights for process scale up.

Keywords: lignocellulosic bioethanol, microwave pretreatment, sugarcane leaves, kinetics

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Authors: Deokyeong Choe, Keonwook Nam, Young Hoon Roh

Abstract:

Wood waste is a potentially significant resource for economic and environment-friendly recycling. Wood waste represents a key sustainable source of biomass for transformation into bioethanol. Unfortunately, wood waste is highly recalcitrant for biotransformation, which limits its use and prevents economically viable conversion into bioethanol. As a result, an effective pretreatment is necessary to degrade cellulose of the wood waste, which improves the accessibility of cellulase. In this work, a H₃PO₄-acetone pretreatment was selected among the various pretreatment methods and used to dissolve cellulose and lignin. When the H₃PO₄ and acetone were used, 5–6% of the wood waste was found to be very appropriate for saccharification. Also, when the enzymatic saccharification was conducted in the mixture of the wood waste and 0.05 M citrate buffer solution, glucose and xylose were measured to be 80.2 g/L and 9.2 g/L respectively. Furthermore, ethanol obtained after 70 h of fermentation by S. cerevisiae was 30.4 g/L. As a result, the conversion yield from wood waste to bioethanol was calculated to be 57.4%. These results show that the pretreated wood waste can be used as good feedstocks for bioethanol production and that the H₃PO₄-acetone pretreatment can effectively increase the yield of ethanol production.

Keywords: wood waste, H₃PO₄-acetone, bioethanol, fermentation

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Authors: Muhammad Shahzad Hussain

Abstract:

Two major problems are facing generally by conventional poultry farming that is disease outbreaks and poor performance, which results due to improper management. To enhance the growth performance and efficiency of feed and reduce disease outbreaks, antibiotic growth promoters (AGPs) which are antibiotics at sub-therapeutic levels, are extensively used in the poultry industry. European Union has banned the use of antibiotics due to their presence in poultry products, development of antibiotic-resistant pathogens, and disturbance of normal gut microbial ecology. These residues cause serious health concerns and produce antibiotic resistance in pathogenic microbes in human beings. These issues strengthen the need for the withdrawal of AGPs from poultry feed. Nowadays, global warming is a major issue, and it is more critical in tropical areas like Pakistan, where heat stress is already a major problem. Heat stress leads to poor production performance, high mortality, immuno-suppression, and concomitant diseases outbreak. The poultry feed industry in Pakistan, like other countries of the world, has been facing shortages and high prices of local as well as imported feed ingredients. Prebiotics are potential replacer for AGP as prebiotics has properties to enhance the production potential and reduce the growth of harmful bacteria as well as stimulate the growth/activity of beneficial bacteria. The most commonly used prebiotics in poultry includes mannan oligosaccharide (MOS). MOS is an essential component of the yeast cell wall (YCW) (Saccharomyces cerevisiae); therefore, the YCW wall possesses prebiotic properties. The use of distillery yeast wall (YCW) has the potential to replace conventional AGPs and to reduce mortality due to heat stress as well as to bind toxins in the feed. The dietary addition of YCW has not only positive effects on production performance in poultry during normal conditions but during stressful conditions. A total of 168-day-old broilers were divided into 6 groups, each of which has 28 birds with 4 replicates (n=7).Yeast cell wall (YCW) supplementation @ 0%, 1%, 1.5%, 2%, 2.5%, 3% from day 0 to 35. Heat stress was exposed from day 21 to 35 at 30±1.1ᵒC with relative humidity 65±5%. Zootechnical parameters like body weight, FCR, Organ development, and histomorphometric parameters were studied. A significant weight gain was observed at group C supplemented @ 1.5% YCW during the fifth week. Significant organ weight gain of Gizzard, spleen, small intestine, and cecum was observed at group C supplemented @ 1.5% YCW. According to morphometric indices Duodenum, Jejunum, and Ileum has significant villus height, while Jejunum and Ileum have also significant villus surface area in the group supplemented with 1.5% YCW. IEL count was only decreased in 1.5% YCW-fed group in jejunum and ileum, not in duodenum, that was less in 2% YCW-supplemented group. Dietary yeast cell wall of saccharomyces cerevisiae partially reduced the effects of high ambient temperature in terms of better growth and modified gut histology and components of mucosal immune response to better withstand heat stress in broilers.

Keywords: antibiotics, AGPs, broilers, MOS, prebiotics, YCW

Procedia PDF Downloads 68

Authors: Boukhiar Aissa, Halladj Fatima, Iguergaziz Nadia, Lamrani yasmina, Benamara Salem

Abstract:

Determining the alcohol content in alcoholic fermentation bioprocess is of great importance. In fact, it is a key indicator for monitoring this bioprocess. Several methodologies (chemical, spectrophotometric, chromatographic) are used to the determination of this parameter. However, these techniques are very long and they require: rigorous preparations, sometimes dangerous chemical reagents and/or expensive equipment. In the present study, the date juice is used as the substrate of alcoholic fermentation. The extracted juice undergoes an alcoholic fermentation by Saccharomyces cerevisiae. The study of the possible use of refractometry as a sole means for the in situ control of alcoholic fermentation revealed a good correlation (R2=0.98) between initial and final °Brix: °Brixf=0.377×°Brixi. In addition, the relationship between Δ°Brix and alcoholic content of the final product (A,%) has been determined: Δ°Brix/A=1.1. The obtained results allowed us to establish iso-responses abacus, which can be used for the determination of alcohol and residual sugar content, with a mean relative error (MRE) of 5.35%.

Keywords: alcoholic fermentation, date juice, refractometry, residual sugar

Procedia PDF Downloads 316

Authors: Kamla Malik

Abstract:

For ethanol production from paddy straw firstly pretreatment was done by using sodium hydroxide solution (2.0%) at 15 psi for 1 hr. The maximum lignin removal was achieved with 0.5 mm mesh size of paddy straw. It contained 72.4 % cellulose, 15.9% hemicelluloses and 2.0 % lignin after pretreatment. Paddy straw hydrolysate (PSH) with fruits wastes (5%), such as sweet lime, apple, sapota, grapes, kinnow, banana, papaya, mango, and watermelon were subjected to simultaneous saccharification and co-fermentation (SSCF) for 72 hrs by co-culture of Saccharomyces cerevisiae HAU-1 and Candida sp. with 0.3 % urea as a cheap nitrogen source. Fermentation was carried out at 35°C and determined ethanol yield at 24 hours interval. The maximum production of ethanol was produced within 72 hrs of fermentation in PSH + sapota peels (3.9% v/v) followed by PSH + kinnow peels (3.6%) and PSH+ papaya peels extract (3.1 %). In case of PSH+ banana peels and mango peel extract the ethanol produced were 2.8 % and 2.2 % (v/v). The results of this study suggest that wastes from fruits that contain fermentable sugar should not be discarded into our environment, but should be supplemented in paddy straw which converted to useful products like bio-ethanol that can serve as an alternative energy source.

Keywords: ethanol, fermentation, fruit wastes, paddy straw

Procedia PDF Downloads 371