Search results for: Saccharomyces cerevisia

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

Procedia PDF Downloads 157

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: 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: Abdol-Hassan Doulah

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In this study, antimicrobial activity of essential oil and ethyl acetate and ether extracts of S. nemorosa were examined against some species of bacteria and fungi. The essential oil of the aerial part of S. nemorosa was examined by GC and GC-MS. In the essential oil of S. nemorosa 26 Compounds have been identified. 2-Nonanone (44.09 %), 2-Undecanone (33.79 %), E-Caryophyllene (3.74 %) and 2-Decanone (2.89 %) were the main components of the essential oil. The essential oil analysis showed greatest antimicrobial activity against Staphylococcus epidermidis (5.3 μg/ml) and S. cerevisiae (9.3 μg/ml). The ethyl acetate showed greatest antimicrobial activity against Bacillus subtilis (106.7 μg/ml), Candida albicans (5.3 μg/ml) and ether extract showed greatest antimicrobial activity against Klebseilla pneumoniae (10.7 μg/ml) and Saccharomyces cerevisiae (10.7 μg/ml). In conclusion, we suggest that the antimicrobial activity of S. nemorosa may be due to its content of germacrene and linalool.

Keywords: antibacterial activity, antifungal activity, Salvia nemorosa L., essential oils, biological activity

Procedia PDF Downloads 461

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

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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: H. O. Udeh, T. E. Kgatla, A. I. O. Jideani

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The effect of Zn2+, Mg2+, and Ba2+ on Saccharomyces pastorianus during very high gravity fermentation was evaluated in this study at independent and three variable combinations. Wort gravity of 21oP was prepared from barley malt, hops and water, to which the metal ions were supplemented in their combinations and subsequently pitched. After 96 h of fermentation, high wort fermentability (%F)= 29.53 was obtained in wort medium containing 900:4 ppm Mg2+ + Ba2+. Increased ethanol titre 7.3491 %(v/v) and 7.1313 %(v/v) were obtained in media containing 900:4 ppm Mg2+ + Ba2+ and 12:900 ppm Zn2+ + Mg2+. Decrease %F= 22.54 and ethanol titre 6.1757% (v/v) was recorded in wort medium containing 12:4 ppm Zn2+ + Ba2+. In media containing the individual metal ions, increased %F= 27.94 and %F= 26.03 were obtained in media containing 700 ppm Mg2+ and 2 ppm Ba2+, with increased ethanol yield of 7.8844% (v/v) and 7.6245% (v/v) respectively. Least %F of 11.75 and 10.80, and ethanol titre of 4.99 (%v/v) and 4.80 (%v/v) were obtained for 10 ppm Zn2+ and 4 ppm Ba2+ respectively.

Keywords: ethanol yield, fermentability, mineral ions, yeast stress, very high gravity fermentation

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

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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: Jessy Mattar, Mohamad Turk, Maurice Nonus, Nikolai Lebovka, Henri El Zakhem, Eugene Vorobiev

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The effects of electro-stimulation of S. cerevisiae cells in colloidal suspension by Pulsed Electric Fields ‎‎(PEF) with electric field strength E = 20 – 2000 V.cm-1 and effective PEF treatment time tPEF = 10^−5 – 1 s were ‎investigated. The applied experimental procedure includes variations in the preliminary fermentation time and ‎electro-stimulation by PEF-treatment. Plate counting was performed.‎ At relatively high electric fields (E ≥ 1000 V.cm-1) and moderate PEF treatment time (tPEF > 100 µs), the ‎extraction of ionic components from yeast was observed by conductivity measurements, which can be related to ‎electroporation of cell membranes. Cell counting revealed a dependency of the colonies’ size on the time of ‎preliminary fermentation tf and the power consumption W, however no dependencies were noticeable by varying the initial yeast concentration in the treated suspensions.‎

Keywords: intensification, yeast, fermentation, electroporation, biotechnology

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

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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

Procedia PDF Downloads 546

Authors: Nassima Behidj-Benyounes, Thoraya Dahmene, Nadjiba Chebout

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This work examines the study of the antimicrobrial effect of alkaloids extracted from the seeds of Peganum harmala L (Zygophyllaceae). This natural substance is extracted by using different solvents (aqueous, ethanolic, and hexane). The evaluation of the antimicrobial activity has only dealt with alkaloids. The antimicrobial effect of alkaloids is evaluated on several microorganisms. It has been tested on eight bacterial strains. The extract has been studied by using two yeasts. Finally, three molds have been studied. It should be noted that these agents are characterized by a high frequency of contamination and pathogenicity. Through this study, we note that Staphylococcus aureus, Saccharomyces cerievisae and E. coli are very sensitive in respect of the ethanol extract. Pseudomonas aerogenosa and Penicillium sp. are resistant to this extract. The other microorganisms are moderately sensitive. The study of the antimicrobial activity of different extracts of the Harmel has shown an optimal activity with the ethanol extract.

Keywords: Peganum harmala L., seeds, alkaloids, bacteria, fungi, yeast, antimicrobial activity

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

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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: F. Halladj, H. Amellal, S. Benamara

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Current consumer trends go towards natural products defined as the products obtained by a traditional manufacturing method. Vinegar is one of those products marketed; it may be industrially obtained by a submerged (fast) or traditional (slow) processes. The latter exhibited a high quality because of its complex microbiological transformations (or two-stage fermentation) by the native must flora. Moreover, although that Acetic acid bacteria have traditionally been considered to play the leading role in vinegar production, some studies have recently highlighted that also yeasts metabolism can affect traditional vinegar chemical properties in a remarkable way. Thus, the aim of this study was to monitor a traditional slow process of vinegar as applied in the south of Algeria using date with hard texture (Degla-Beida variety) to isolate and identify the involved yeasts in order to select them as starter culture. Phenotypic and molecular analysis show that the non-Saccharomyces were the main yeasts species isolated throughout the alcoholic spontaneous fermentation and they included Hanseniaspora guilliermondii and Torulaspora delbrueckii.

Keywords: date vinegar, traditional production, yeasts, Phenotypic, Algeria

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Authors: Muhammad Shahzad Hussain

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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 61

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

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Authors: Vishwesh Kulkarni, Nikhil Bellarykar

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Cellular complexity stems from the interactions among thousands of different molecular species. Thanks to the emerging fields of systems and synthetic biology, scientists are beginning to unravel these regulatory, signaling, and metabolic interactions and to understand their coordinated action. Reverse engineering of biological networks has has several benefits but a poor quality of data combined with the difficulty in reproducing it limits the applicability of these methods. A few years back, many of the commonly used predictive algorithms were tested on a network constructed in the yeast Saccharomyces cerevisiae (S. cerevisiae) to resolve this issue. The network was a synthetic network of five genes regulating each other for the so-called in vivo reverse-engineering and modeling assessment (IRMA). The network was constructed in S. cereviase since it is a simple and well characterized organism. The synthetic network included a variety of regulatory interactions, thus capturing the behaviour of larger eukaryotic gene networks on a smaller scale. We derive a new set of algorithms by solving a nonlinear optimization problem and show how these algorithms outperform other algorithms on these datasets.

Keywords: synthetic gene network, network identification, optimization, nonlinear modeling

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Authors: Kamla Malik

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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 362

Authors: B. Santoso, B. Tj. Hariadi, H. Abubakar

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The aim of the experiment was to evaluate the effect of probiotic addition in concentrate on fermentation characteristics and in vitro nutrient digestibility of the grass Pennisetum purpureophoides. Two strains lactic acid bacteria (LAB) i.e Lactobacillus plantarum and Lactobacillus acidhophilus, and one strain yeast of Saccharomyces cerevisiae were used as probiotic. The probiotics was added at 2% and 4% (v/w) in the concentrate. The result showed the concentrate containing between 1.5 × 106 and 3 × 107 CFU/g of lactic acid bacteria and 3 × 103 CFU/g of S. cerevisiae. The DM, OM and NDF digestibility were higher (P<0.01) in grass substrate with concentrate than in grass alone. Addition of probiotic in concentrate increased (P<0.01) DM, OM and NDF compared to concentrate without probiotic. Total VFA and propionic acid concentrations were higher (P<0.01) in grass substrate with concentrate than in grass alone. Concentration of acetic acid decreased (P<0.01) in grass substrate with concentrate than in grass substrate alone. Addition of L. plantarum and L. acidophilus and S. cerevisiae in concentrate increased (P<0.01) propionic acid concentration. It was concluded that addition of probiotic in concentrate increased propionic concentration and in vitro nutrient digestibility.

Keywords: by-products, concentrate, digestibility, probiotics

Procedia PDF Downloads 365

Authors: Afaf Bouchoucha, Karima Si Larbi, Mohamed Amine Bourouaia, Salah.Boulanouar, Safia.Djabbar

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The synthesis, characterization and comparative biological study of manganese (II) and copper (II) complexes with an heterocyclic ligand used in pharmaceutical field (Scheme 1), were reported. Two kinds of complexes were obtained with derivative sulfonamide, [M (L)₂ (H₂O)₂].H₂O and [M (L)₂ (Cl)₂]3H₂O. These complexes have been prepared and characterized by elemental analysis, FAB mass, ESR magnetic measurements, FTIR, UV-Visible spectra and conductivity. Their stability constants have been determined by potentiometric methods in a water-ethanol (90:10 v/v) mixture at a 0.2 mol l-1 ionic strength (NaCl) and at 25.0 ± 0.1 ºC using Sirko program. DFT calculations were done using B3LYP/6-31G(d) and B3LYP/LanL2DZ. The antimicrobial activity of ligand and complexes against the species Escherichia coli, P. aeruginosa, Klebsiella pneumoniae, S. aureus, Bacillus subtilisan, Candida albicans, Candida tropicalis, Saccharomyces, Aspergillus fumigatus and Aspergillus terreus has been carried out and compared using agar-diffusion method. Also, the toxicity study was evaluated on synchesis complexes using Mice of NMRI strain.

Keywords: hetterocyclic ligand, complex, stability constant, antimicrobial activity, DFT, acute and genotoxicity study

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Authors: Lavinia Ruta, Ileana Farcasanu

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The yeast surface display (YSD) technique enables the expression of proteins on yeast cell surfaces, facilitating the identification and isolation of proteins with targeted binding properties, such as nanobodies. Nanobodies, derived from camelid species, are single-domain antibody fragments renowned for their high affinity and specificity towards target proteins, making them valuable in research and potentially in therapeutics. Their advantages include a compact size (~15 kDa), robust stability, and the ability to target challenging epitopes. The project endeavors to establish and validate a platform for producing Green Fluorescent Protein (GFP) and mCherry nanobodies using the yeast surface display method. mCherry, a prevalent red fluorescent protein sourced from coral species, is commonly utilized as a genetic marker in biological studies due to its vibrant red fluorescence. The GFP-nanobody, a single variable domain of heavy-chain antibodies (VHH), exhibits specific binding to GFP, offering a potent means for isolating and engineering fluorescent protein fusions across various biological research domains. Both GFP and mCherry nanobodies find specific utility in cellular imaging and protein analysis applications.

Keywords: YSD, nanobodies, GFP, Saccharomyces cerevisiae

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Authors: Jordi Cuñé, Carlos de Lecea, Laia Marti

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Small molecules known as fermentable oligo-, di-, and monosaccharides and polyols (FODMAPs) are quickly fermented in the large intestine after being poorly absorbed in the small intestine. There is proof that individuals suffering from functional gastrointestinal disorders, like irritable bowel syndrome (IBS), observe an improvement while following a diet low in FODMAPs. Because wheat has a relatively high fructan content, it is a key source of FODMAPs in our diet. A yeast-based method was created in this study to lower the amounts of FODMAP in (whole wheat) bread. In contrast to fermentation by regular baker yeast, the combination of Kluyveromyces marxianus ABB S7 with Saccharomyces cerevisiae allowed a reduction of fructan content by 60% without implying the appearance of other substrates categorized as FODMAP (excess fructose or polyols). The final FODMAP content in the developed whole wheat bread would allow its classification as a safe product for sensitive people, according to international consensus. Cocultures of S. cerevisiae and K. marxianus were established in order to ensure sufficient CO₂ generation; larger quantities of gas were produced due to the strains' synergistic relationship. Thus, this method works well for lowering the levels of FODMAPs in bread.

Keywords: Kluyveromyces marxianus, bakery, bread, FODMAP, IBS, functional gastro intestinal disorders

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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: Daniel Isaac Sanchez Chavez, Salvador Rodríguez Zaragoza, Patricia Velez Aguilar

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In arid and semi-arid regions, plants are essential in the functional activity and productivity, modifying the microclimatic conditions of their environment, which allows many organisms to grow under them. Within these organisms, oribatid mites play a key role in reintegrating nutrients into the soil through the consumption of soil fungi. However, oribatid mites feed on a vast array of fungal species, which is likely to have strong impacts on their population dynamics and their environment. So, in this study, the aim was to determine the gut content of the abundant oribatid mites Zygoribatula sp and Scheloribates sp, under the canopy of the bush P. laevigata in a semi-arid zone through DNA-based analysis. The results showed the presence in the gut of both mites of different fungal taxa. Fungi, such as Aspergillus sp and Mortierella sp, probably served as a food despite the production of deterrent compounds or structures from both fungal species. Saccharomyces sp might serve as well as a food source; however, it might be part of their microbial endosymbionts. On the other hand, the presence of Beauveria sp indicates a probable pathogenicity interaction, instead of fungal consumption, since this fungus is known to be entomopathogenic. Finally, the results might indicate a feeding preference to certain soil fungi according to diverse features from both taxa.

Keywords: microenvironment, endosymbionts, Oribatida, fungi

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Authors: F. A. Ogundolie, F. Okogue, D. V. Adegunloye

Abstract:

Greywater samples were obtained from three restaurants in the Federal University of Technology; Akure coded SSR, MGR and GGR. Fungi isolates obtained include Rhizopus stolonifer, Aspergillus niger, Mucor mucedo, Aspergillus flavus, Saccharomyces cerevisiae. Of these fungi isolates obtained, R. stolonifer, A. niger and A. flavus showed significant degradation ability on grey water and was used for this research. A simple bioreactor was constructed using biodegradation process in purification of waste water samples. Waste water undergoes primary treatment; secondary treatment involves the introduction of the isolated organisms into the waste water sample and the tertiary treatment which involved the use of filter candle and the sand bed filtration process to achieve the end product without the use of chemicals. A. niger brought about significant reduction in both the bacterial load and the fungi load of the greywater samples of the three respective restaurants with a reduction of (1.29 × 108 to 1.57 × 102 cfu/ml; 1.04 × 108 to 1.12 × 102 cfu/ml and 1.72 × 108 to 1.60 × 102 cfu/ml) for bacterial load in SSR, MGR and GGR respectively. Reduction of 2.01 × 104 to 1.2 × 101; 1.72 × 104 to 1.1 × 101, and 2.50 × 104 to 1.5 × 101 in fungi load from SSR, MGR and GGR respectively. Result of degradation of these selected waste water by the fungi showed that A. niger was probably more potent in the degradation of organic matter and hence, A. niger could be used in the treatment of wastewater.

Keywords: Aspergillus niger, greywater, bacterial, fungi, microbial load, bioreactor, biodegradation, purification, organic matter and filtration

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Authors: Byeong-Uk Lim, Young-Ran Song, Sang-Ho Baik

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This study aimed to develop yeast starters for scientific alcohol production and systematic quality control of whisky. A total of 389 yeast strains were isolated from traditional Korean fermentation starter (nuruk) and rice wine (makgeolli), and ten strains were finally selected for their high alcohol productivities, in which their alcohol productions were above 17.3% (v/v) during 10 days under two steps of glucose feeding condition (30% and then 15%, w/v). By 18s rDNA sequence analysis, all strains were identified as Saccharomyces cerevisiae (SC), and they can grow well under 50% (w/v) glucose and 10% (v/v) ethanol conditions. Furthermore, the capability of ten different SC strains to ferment rice wine for whisky was studied. Rice wine was fermented with only steamed rice, water, and two types of enzymes (glucoamylase and α-amylase) during 14 days at 25 °C, and then their oenological properties have been determined. As the results, the fermented rice wines indicated the final pH range of 4.24-4.38 and acidity range of 0.12-0.18. The highest ethanol production of 20.2% (v/v) was found in the fermentation with a SC-156 strain, whereas SC-92 (16.8%) and SC-119 (16.4%) showed significantly lowest ethanol productions. In addition, the residual sugar contents showed negative correlation with alcohol contents. Moreover, this study focused on nucleotide polymorphisms in the MSN2 and MSN4 genes to investigate the cause of the defective stress responses in yeast. Consequently, it was also confirmed that the deletion of the N termini of Msn4p from identified point mutations in SC-63, SC-95, SC-156, SC-158, and SC-160 strains.

Keywords: yeast, high-alcohol, whisky, rice wine

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Authors: P. Rahnemoon, M. Sarabi Jamab, M. Javanmard Dakheli, A. Bostan

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In recent years, tendency to use of natural antimicrobial agents in food industry has increased. Pomegranate peels containing phenolic compounds and anti-microbial agents, are counted as valuable source for extraction of these compounds. In this study, the extraction of pomegranate peel extract was carried out at different ethanol/water ratios (40:60, 60:40, and 80:20), temperatures (25, 40, and 55 ˚C), and time durations (20, 24, and 28 h). The extraction yield, phenolic compounds, flavonoids, and anthocyanins were measured. ‎Antimicrobial activity of pomegranate peel extracts were determined against some food-borne ‎microorganisms such as Salmonella enteritidis, Escherichia coli, Listeria monocytogenes, ‎‎Staphylococcus aureus, Aspergillus niger, and Saccharomyces cerevisiae by agar diffusion and MIC methods. Results showed that at ethanol/water ratio 60:40, 25 ˚C and 24 h maximum amount of phenolic compounds ‎‏(‎‏‎349.518‎‏ ‏mg gallic acid‏/‏g dried extract), ‎flavonoids (250.124 mg rutin‏/‏g dried extract), anthocyanins (252.047 ‎‏‏mg ‎cyanidin‏‎3‎‏glucoside‏/‏‎100 g dried extract), and the strongest antimicrobial activity were obtained. ‎All extracts’ antimicrobial activities were demonstrated against every tested ‎‎microorganisms‏.‎‏ Staphylococcus aureus showed the highest sensitivity among the tested ‎‎‎microorganisms.

Keywords: antimicrobial agents, phenolic compounds, pomegranate peel, solvent extraction‎

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Authors: Jae-Hyung Jin, Kyung-Tae Lee, Yee-Seul So, Eunji Jeong, Yeonseon Lee, Dongpil Lee, Dong-Gi Lee, Yong-Sun Bahn

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Cryptococcus neoformans causes cryptococcal meningoencephalitis mainly in immunocompromised patients as well as immunocompetent people. But therapeutic options are limited to treat cryptococcosis. Some signaling pathways including cyclic AMP pathway, MAPK pathway, and calcineurin pathway play a central role in the regulation of the growth, differentiation, and virulence of C. neoformans. To understand signaling networks regulating the virulence of C. neoformans, we selected the 114 putative phosphatase genes, one of the major components of signaling networks, in the genome of C. neoformans. We identified putative phosphatases based on annotation in C. neoformans var. grubii genome database provided by the Broad Institute and National Center for Biotechnology Information (NCBI) and performed a BLAST search of phosphatases of Saccharomyces cerevisiae, Aspergillus nidulans, Candida albicans and Fusarium graminearum to Cryptococcus neoformans. We classified putative phosphatases into 14 groups based on InterPro phosphatase domain annotation. Here, we constructed 170 signature-tagged gene-deletion strains through homologous recombination methods for 91 putative phosphatases. We examined their phenotypic traits under 30 different in vitro conditions, including growth, differentiation, stress response, antifungal resistance and virulence-factor production.

Keywords: human fungal pathogen, phosphatase, deletion library, functional genomics

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Authors: Aida Kalantari, Boyang Ji, Tao Chen, Ivan Mijakovic

Abstract:

3-hydroxypropanoic acid (3-HP) is one of the most important biomass-derivable platform chemicals that can be converted into a number of industrially important compounds. There have been several attempts at production of 3-HP from renewable sources in cell factories, focusing mainly on Escherichia coli, Klebsiella pneumoniae, and Saccharomyces cerevisiae. Despite the significant progress made in this field, commercially exploitable large-scale production of 3-HP in microbial strains has still not been achieved. In this study, we investigated the potential of Bacillus subtilis to be used as a microbial platform for bioconversion of glycerol into 3-HP. Our recombinant B. subtilis strains overexpress the two-step heterologous pathway containing glycerol dehydratase and aldehyde dehydrogenase from various backgrounds. The recombinant strains harboring the codon-optimized synthetic pathway from K. pneumoniae produced low levels of 3-HP. Since the enzymes in the heterologous pathway are sensitive to oxygen, we had to perform our experiments in micro-aerobic conditions. Under these conditions, the cell produces lactate in order to regenerate NAD+, and we found the lactate production to be in competition with the production of 3-HP. Therefore, based on the in silico predictions, we knocked out the glycerol kinase (glpk), which in combination with growth on glucose, resulted in improving the 3-HP titer to 1 g/L and the removal of lactate. Cultivation of the same strain in an enriched medium improved the 3-HP titer up to 7.6 g/L. Our findings provide the first report of successful introduction of the biosynthetic pathway for conversion of glycerol into 3-HP in B. subtilis.

Keywords: bacillus subtilis, glycerol, 3-hydroxypropanoic acid, metabolic engineering

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Authors: Elizabeth Chinyere Amadi

Abstract:

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

Abstract:

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

Procedia PDF Downloads 90