Scholarly Research Excellence

Digital Open Science Index

Commenced in January 2007 Frequency: Monthly Edition: International Paper Count: 34

All Types of Base Pair Substitutions Induced by γ-Rays in Haploid and Diploid Yeast Cells

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.

Characteristics of the Storage Stability for Different Saccharomyces cerevisiae Strains

Storage stability is the important factor of baker's yeast quality. Effect of the storage period (fifteen days) on storage sugars and cell viability of baker's yeast, produced from three S. cerevisiae strains (FC-620, FH-620, and FAT-12) as comparison with baker's yeast produced by S. cerevisae F-707 (original strain of baker's yeast factory) were investigated. Studied trehalose and glycogen content ranged from 10.19 to 14.79 % and from 10.05 to 10.69 % (d.w.), respectively before storage. The trehalose and glycogen content of all strains was decreased by increasing the storage period with no significant differences between the reduction rates of trehalose. Meanwhile, reduction rates of glycogen had significant differences between different strains, where the FH-620 and FC-620 strains had lowest rates as 18.12 and 20.70 %, respectively. Also, total viable cells and gassing power of all strains were decreased by increasing the storage period. FH-620 and FC-620 strains had the lowest values of reduction rates as an indicator of storage resistant. Where the reduction rates in total viable cells of FH-620 and FC-620 strains were 22.05 and 24.70%, respectively, while the reduction rates of gassing power were 20.90 and 24.30%, in the same order. On other hand, FAT-12 strain was more sensitive to storage as compared to original strain, where the reduction rates were 35.60 and 35.75%, respectively for total viable cells and gassing power.

Screening of Antagonistic/Synergistic Effect between Lactic Acid Bacteria (LAB) and Yeast Strains Isolated from Kefir

Kefir is a traditional fermented refreshing beverage which is known for its valuable and beneficial properties for human health. Mainly yeast species, lactic acid bacteria (LAB) strains and fewer acetic acid bacteria strains live together in a natural matrix named “kefir grain”, which is formed from various proteins and polysaccharides. Different microbial species live together in slimy kefir grain and it has been thought that synergetic effect could take place between microorganisms, which belong to different genera and species. In this research, yeast and LAB were isolated from kefir samples obtained from Uludag University Food Engineering Department. The cell morphology of isolates was screened by microscopic examination. Gram reactions of bacteria isolates were determined by Gram staining method, and as well catalase activity was examined. After observing the microscopic/morphological and physical, enzymatic properties of all isolates, they were divided into the groups as LAB and/or yeast according to their physicochemical responses to the applied examinations. As part of this research, the antagonistic/synergistic efficacy of the identified five LAB and five yeast strains to each other were determined individually by disk diffusion method. The antagonistic or synergistic effect is one of the most important properties in a co-culture system that different microorganisms are living together. The synergistic effect should be promoted, whereas the antagonistic effect is prevented to provide effective culture for fermentation of kefir. The aim of this study was to determine microbial interactions between identified yeast and LAB strains, and whether their effect is antagonistic or synergistic. Thus, if there is a strain which inhibits or retards the growth of other strains found in Kefir microflora, this circumstance shows the presence of antagonistic effect in the medium. Such negative influence should be prevented, whereas the microorganisms which have synergistic effect on each other should be promoted by combining them in kefir grain. Standardisation is the most desired property for industrial production. Each microorganism found in the microbial flora of a kefir grain should be identified individually. The members of the microbial community found in the glue-like kefir grain may be redesigned as a starter culture regarding efficacy of each microorganism to another in kefir processing. The main aim of this research was to shed light on more effective production of kefir grain and to contribute a standardisation of kefir processing in the food industry.

Comparison of Methods for the Detection of Biofilm Formation in Yeast and Lactic Acid Bacteria Species Isolated from Dairy Products

Lactic acid bacteria (LAB) and some yeast species are common microorganisms found in dairy products and most of them are responsible for the fermentation of foods. Such cultures are isolated and used as a starter culture in the food industry because of providing standardisation of the final product during the food processing. Choice of starter culture is the most important step for the production of fermented food. Isolated LAB and yeast cultures which have the ability to create a biofilm layer can be preferred as a starter in the food industry. The biofilm formation could be beneficial to extend the period of usage time of microorganisms as a starter. On the other hand, it is an undesirable property in pathogens, since biofilm structure allows a microorganism become more resistant to stress conditions such as antibiotic presence. It is thought that the resistance mechanism could be turned into an advantage by promoting the effective microorganisms which are used in the food industry as starter culture and also which have potential to stimulate the gastrointestinal system. Development of the biofilm layer is observed in some LAB and yeast strains. The resistance could make LAB and yeast strains dominant microflora in the human gastrointestinal system; thus, competition against pathogen microorganisms can be provided more easily. Based on this circumstance, in the study, 10 LAB and 10 yeast strains were isolated from various dairy products, such as cheese, yoghurt, kefir, and cream. Samples were obtained from farmer markets and bazaars in Bursa, Turkey. As a part of this research, all isolated strains were identified and their ability of biofilm formation was detected with two different methods and compared with each other. The first goal of this research was to determine whether isolates have the potential for biofilm production, and the second was to compare the validity of two different methods, which are known as “Tube method” and “96-well plate-based method”. This study may offer an insight into developing a point of view about biofilm formation and its beneficial properties in LAB and yeast cultures used as a starter in the food industry.

Extracellular Laccase Production by Co-culture between Galactomyces reesii IFO 10823 and Filamentous Fungal Strains Isolated from Fungus Comb Using Natural Inducer

Extracellular laccases are copper-containing microbial enzymes with many industrial biotechnological applications. This study evaluated the ability of nutrients in coconut coir to enhance the yield of extracellular laccase of Galactomyces reesii IFO 10823 and develop a co-culture between this yeast and other filamentous fungi isolated from the fungus comb of Macrotermes sp. The co-culture between G. reesii IFO 10823 and M. indicus FJ-M-5 (G3) gave the highest activity at 580.20 U/mL. When grown in fermentation media prepared from coconut coir and distilled water at 70% of initial moisture without supplement addition, G3 produced extracellular laccase of 113.99 U/mL.

Decolorization and Phenol Removal of Palm Oil Mill Effluent by Termite-Associated Yeast
A huge of dark color palm oil mill effluent (POME) cannot pass the discharge standard. It has been identified as the major contributor to the pollution load into ground water. Here, lignin-degrading yeast isolated from a termite nest was tested to treat the POME. Its lignin-degrading and decolorizing ability was determined. The result illustrated that Galactomyces sp. was successfully grown in POME. The decolorizing test demonstrated that 40% of Galactomyces sp. could reduce the color of POME (50% v/v) about 74-75% in 5 days without nutrient supplement. The result suggested that G. reessii has a potential to apply for decolorizing the dark wastewater like POME and other industrial wastewaters.
Prophylactic Effects of Dairy Kluyveromyces marxianus YAS through Overexpression of BAX, CASP 3, CASP 8 and CASP 9 on Human Colon Cancer Cell Lines
Colorectal cancer (CRC) is one of the most prevalent cancers and intestinal microbial community plays an important role in colorectal tumorigenesis. Probiotics have recently been assessed as effective anti-proliferative agents and thus this study was performed to examine whether CRC undergo apoptosis by treating with isolated Iranian native dairy yeast, Kluyveromyces marxianus YAS, secretion metabolites. The cytotoxicity assessments on cells (HT-29, Caco-2) were accomplished through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay as well as qualitative DAPI (4',6-diamidino-2-phenylindole staining) and quantitative (flow cytometry assessments) evaluations of apoptosis. To evaluate the main mechanism of apoptosis, Real time PCR method was applied. Kluyveromyces marxianus YAS secretions (IC50) showed significant cytotoxicity against HT-29 and Caco-2 cancer cell lines (66.57 % and 66.34 % apoptosis) similar to 5-Fluorouracil (5-FU) while apoptosis only was developed in 27.57 % of KDR normal cells. The prophylactic effects of Kluyveromyces marxianus (PTCC 5195), as a reference yeast, was not similar to Kluyveromyces marxianus YAS indicating strain dependency of bioactivities on CRC disease prevention. Based on real time PCR results, the main cytotoxicity is related to apoptosis phenomenon and the core related mechanism is depended on the overexpression of BAX, CASP 9, CASP 8 and CASP 3 inducing apoptosis genes. However, several investigations should be conducted to precisely determine the effective compounds to be used as anticancer therapeutics in the future.
Influence of Yeast Strains on Microbiological Stability of Wheat Bread
Problem of food preservation is extremely important for mankind. Viscous damage ("illness") of bread results from development of Bacillus spp. bacteria. High temperature resistant spores of this microorganism are steady against 120°C) and remain in bread during pastries, potentially causing spoilage of the final product. Scientists are interested in further characterization of bread spoiling Bacillus spp. species. Our aim was to find weather yeast Saccharomyces cerevisiae strains that are able to produce natural antimicrobial killer factor can preserve bread illness. By diffusion method, we showed yeast antagonistic activity against spore-forming bacteria. Experimental technological parameters were the same as for bakers' yeasts production on the industrial scale. Risograph test during dough fermentation demonstrated gas production. The major finding of the study was a clear indication of the presence of killer yeast strain antagonistic activity against rope in bread causing bacteria. After demonstrating antagonistic effect of S. cerevisiae on bacteria using solid nutrient medium, we tested baked bread under provocative conditions. We also measured formation of carbon dioxide in the dough, dough-making duration and quality of the final products, when using different strains of S. cerevisiae. It is determined that the use of yeast S. cerevisiae RCAM 01730 killer strain inhibits appearance of rope in bread. Thus, natural yeast antimicrobial killer toxin, produced by some S. cerevisiae strains is an anti-rope in bread protector.
Effect of Different Treatments on Heavy Metal Concentration in Sugar Cane Molasses

Cane molasses is used as a raw material for the production of baker’s yeast (Saccharomyces cerevisiae) in Egypt. The high levels of heavy metals in molasses cause a critical problem during fermentation and cause various kinds of technological difficulties (yield and quality of yeast become lower). The aim of the present study was to determine heavy metal concentrations (cadmium, nickel, lead, and copper) in crude and treated molasses obtained from the storage tanks of the baker’s yeast factory through four seasons. Also, the effect of crude molasses treatment by different methods (at laboratory scale) on heavy metals reduction and its comparison with factory treated molasses were conducted. The molasses samples obtained at autumn season had the highest values of all the studied heavy metals. The molasses treated by cation exchange resin then sulfuric acid had the lowest concentrations of heavy metals compared with other treatments.

Utilization of Whey for the Production of β-Galactosidase Using Yeast and Fungal Culture
Whey is the lactose rich by-product of the dairy industry, having good amount of nutrient reservoir. Most abundant nutrients are lactose, soluble proteins, lipids and mineral salts. Disposing of whey by most of milk plants which do not have proper pre-treatment system is the major issue. As a result of which, there can be significant loss of potential food and energy source. Thus, whey has been explored as the substrate for the synthesis of different value added products such as enzymes. β-galactosidase is one of the important enzymes and has become the major focus of research due to its ability to catalyze both hydrolytic as well as transgalactosylation reaction simultaneously. The enzyme is widely used in dairy industry as it catalyzes the transformation of lactose to glucose and galactose, making it suitable for the lactose intolerant people. The enzyme is intracellular in both bacteria and yeast, whereas for molds, it has an extracellular location. The present work was carried to utilize the whey for the production of β-galactosidase enzyme using both yeast and fungal cultures. The yeast isolate Kluyveromyces marxianus WIG2 and various fungal strains have been used in the present study. Different disruption techniques have also been investigated for the extraction of the enzyme produced intracellularly from yeast cells. Among the different methods tested for the disruption of yeast cells, SDS-chloroform showed the maximum β-galactosidase activity. In case of the tested fungal cultures, Aureobasidium pullulans NCIM 1050 was observed to be the maximum extracellular enzyme producer.
Preservation of Coconut Toddy Sediments as a Leavening Agent for Bakery Products
Toddy sediment (TS) was cultured in a PDA medium to determine initial yeast load, and also it was undergone sun, shade, solar, dehumidified cold air (DCA) and hot air oven (at 400, 500 and 60oC) drying with a view to preserve viability of yeast. Thereafter, this study was conducted according to two factor factorial design in order to determine best preservation method. Therein the dried TS from the best drying method was taken and divided into two portions. One portion was mixed with 3: 7 ratio of TS: rice flour and the mixture was divided in to two again. While one portion was kept under in house condition the other was in a refrigerator. Same procedure was followed to the rest portion of TS too but it was at the same ratio of corn flour. All treatments were vacuum packed in triple laminate pouches and the best preservation method was determined in terms of leavening index (LI). The TS obtained from the best preservation method was used to make foods (bread and hopper) and organoleptic properties of it were evaluated against same of ordinary foods using sensory panel with a five point hedonic scale. Results revealed that yeast load or fresh TS was 58×106 CFU/g. The best drying method in preserving viability of yeast was DCA because LI of this treatment (96%) is higher than that of other three treatments. Organoleptic properties of foods prepared from best preservation method are as same as ordinary foods according to Duo trio test.
Selection of Pichia kudriavzevii Strain for the Production of Single-Cell Protein from Cassava Processing Waste
A total of 115 yeast strains isolated from local cassava processing wastes were measured for crude protein content. Among these strains, the strain MSY-2 possessed the highest protein concentration (>3.5 mg protein/mL). By using molecular identification tools, it was identified to be a strain of Pichia kudriavzevii based on similarity of D1/D2 domain of 26S rDNA region. In this study, to optimize the protein production by MSY-2 strain, Response Surface Methodology (RSM) was applied. The tested parameters were the carbon content, nitrogen content, and incubation time. Here, the value of regression coefficient (R2) = 0.7194 could be explained by the model which is high to support the significance of the model. Under the optimal condition, the protein content was produced up to 3.77 g per L of the culture and MSY-2 strain contains 66.8 g protein per 100 g of cell dry weight. These results revealed the plausibility of applying the novel strain of yeast in single-cell protein production.
Fermentation of Xylose and Glucose Mixture in Intensified Reactors by Scheffersomyces stipitis to Produce Ethanol
In this work, two fermentations at different temperatures (25 and 30ºC), with cell recycling, were accomplished to produce ethanol, using a mix of commercial substrates, xylose (70%) and glucose (30%), as organic source for Scheffersomyces stipitis. Five consecutive fermentations of 80 g L-1 (1º, 2º and 3º recycles), 96 g L-1 (4º recycle) and 120 g L-1 (5º recycle)reduced sugars led to a final maximum ethanol concentration of 17.2 and 34.5 g L-1, at 25 and 30ºC, respectively. Glucose was the preferred substrate; moreover xylose startup degradation was initiated after a remaining glucose presence in the medium. Results showed that yeast acid treatment, performed before each cycle, provided improvements on cell viability, accompanied by ethanol productivity of 2.16 g L-1 h- 1 at 30ºC. A maximum 36% of xylose was retained in the fermentation medium and after five-cycle fermentation an ethanol yield of 0.43 g ethanol/g sugars was observed. S. stipitis fermentation capacity and tolerance showed better results at 30ºC with 83.4% of theoretical yield referenced on initial biomass.
Improvement of Monacolin K and Minimizing of Citrinin Content in Korkor 6 (RD 6) Red Yeast Rice

A strain of Monascus purpureus CMU001 was used to prepare red yeast rice from Thai glutinous rice Korkor 6 (RD 6). Adding of different amounts of histidine (156, 312, 625 and 1250 mg in 100 g of rice grains)) under aerobic and air limitation (air-lock) condition were used in solid fermentation. Determination of the yield as well as monacolin K content was done. Citrinin content was also determined in order to confirm the safety use of prepared red yeast rice. It was found that under air-lock condition with 1250 mg of histidine addition gave the highest yield of 37.40 g of dried red yeast rice prepared from 100 g of rice. Highest 5.72 mg content of monacolin K was obtained under air-lock condition with 312 mg histidine addition. In the other hand, citrinin content was found to be less than 24462 ng/g of all dried red yeast rice samples under the experimental methods used in this work.

Effect of Mineral Ion Addition on Yeast Performance during Very High Gravity Wort Fermentation

The effect of Zn2+, Mg2+, and Ba2+ on Saccharomyces pastorianus performance was evaluated in this study at independent and three variable combinations. After 96 h of fermentation, high wort fermentability (%F) = 29.53 was obtained in medium containing 900:4 ppm Mg2+ + Ba2+. Increased ethanol yield 7.35 %(v/v) and 7.13 %(v/v) were obtained in media containing 900:4 ppm Mg2+ + Ba2+ and 12:900 ppm Zn2+ + Mg2+. Decrease %F = 22.54 and ethanol yield 6.18 % (v/v) was obtained in medium containing 12:4 ppm Zn2+ + Ba2+. In media containing the individual ions, increased %F = 27.94 and 26.03 were recorded for media containing 700 ppm Mg2+ and 2 ppm Ba2+ , with ethanol yield of 7.88% (v/v) and 7.62% (v/v) respectively. Reduced %F and ethanol yield was observed for 10 ppm Zn2+ and 4 ppm Ba2+ media. The impact of Ba2+ at 1 and 2 ppm was significant.

Antimicrobial Agents Produced by Yeasts

Natural antimicrobials are used to preserve foods that can be found in plants, animals, and microorganisms. Antimicrobial substances are natural or artificial agents that produced by microorganisms or obtained semi/total chemical synthesis are used at low concentrations to inhibit the growth of other microorganisms. Food borne pathogens and spoilage microorganisms are inactivated by the use of antagonistic microorganisms and their metabolites. Yeasts can produce toxic proteins or glycoproteins (toxins) that cause inhibition of sensitive bacteria and yeast species. Antimicrobial substance producing phenotypes belonging different yeast genus were isolated from different sources. Toxins secreted by many yeast strains inhibiting the growth of other yeast strains. These strains show antimicrobial activity, inhibiting the growth of mold and bacteria. The effect of antimicrobial agents produced by yeasts can be extremely fast, and therefore may be used in various treatment procedures. Rapid inhibition of microorganisms is possibly caused by microbial cell membrane lipopolysaccharide binding and in activation (neutralization) effect. Antimicrobial agents inhibit the target cells via different mechanisms of action.

Statistical Modeling for Permeabilization of a Novel Yeast Isolate for β-Galactosidase Activity Using Organic Solvents

The hydrolysis of lactose using β-galactosidase is one of the most promising biotechnological applications, which has wide range of potential applications in food processing industries. However, due to intracellular location of the yeast enzyme, and expensive extraction methods, the industrial applications of enzymatic hydrolysis processes are being hampered. The use of permeabilization technique can help to overcome the problems associated with enzyme extraction and purification of yeast cells and to develop the economically viable process for the utilization of whole cell biocatalysts in food industries. In the present investigation, standardization of permeabilization process of novel yeast isolate was carried out using a statistical model approach known as Response Surface Methodology (RSM) to achieve maximal b-galactosidase activity. The optimum operating conditions for permeabilization process for optimal β-galactosidase activity obtained by RSM were 1:1 ratio of toluene (25%, v/v) and ethanol (50%, v/v), 25.0 oC temperature and treatment time of 12 min, which displayed enzyme activity of 1.71 IU /mg DW.

Production of 3-Methyl-1-Butanol by Yeast Wild Strain

The biomass-based fuels have become great concern in order to replace the petroleum-based fuels. Biofuels are a wide range of fuels referred to liquid, gas and solid fuels produced from biomass. Recently, higher chain alcohols such as 3-methyl-1-butanol and isobutanol have become a better candidate compared to bioethanol in order to replace gasoline as transportation fuel. Therefore, in this study, 3-methyl-1-butanol was produced through a fermentation process by yeast. Several types of yeast involved in this research including Saccharomyces cerevisiae, Kluyveromyces lactis GG799 and Pichia pastoris (KM71H, GS115 and X33). The result obtained showed that K. lactis GG799 gave the highest concentration of 3-methyl-1-butanol at 274 mg/l followed by S. cerevisiae, P. pastoris GS115, P. pastoris KM71H and P. pastoris X33 at 265 mg/l, 190 mg/l, 182 mg/l and 174 mg/l respectively. Based on the result, it proved that yeast have a potential in producing 3-methyl-1-butanol naturally.

Batch and Continuous Packed Column Studies Biosorption by Yeast Supported onto Granular Pozzolana

The removal of chromium by living yeast biomass immobilized onto pozzolana was studied. The results obtained in batch experiments indicate that the immobilized yeast on to pozzolana is a excellent biosorbent of Cr(V) with a good removal rates of 85–90%. The initial concentration solution and agitation speed affected Cr(V) removal. The batch studies data were described using the Freundlich and Langmuir models, but the best fit was obtained with Langmuir model. The breakthrough curve from the continuous flow studies shows that immobilized yeast in the fixed-bed column is capable of decreasing Cr(VI) concentration from 15mg/l to a adequate level. 

Microbiological Assessment of Yoghurt Enriched with Flakes from Barley Grain and Malt Extract during Shelf-Life

The effect of flakes from biologically activated hullless barley grain and malt extract on microbiological safety of yoghurt was studied. Pasteurized milk, freeze-dried yoghurt culture YF-L811 (Chr. Hansen, Denmark), flakes from biologically activated hull-less barley grain (Latvia) and malt extract (Ilgezeem, Latvia) were used for experiments. Yoghurt samples with flakes from biologically activated hull-less barley grain and malt extract were analyzed for total plate count of mesophylic aerobic and facultative anaerobic microorganisms, as well yeasts and moulds population during shelflife. Results showed that the changes of pH and titratable acidity affected the concentration of added malt extract. The lowest pH and the highest titratable acidity were determined in samples YFBG5% ME4% and YFBG5% ME6% on the 14th day. The total plate count decreased in all yoghurt samples except sample YFBG5% ME6%, where was determined the increase of microorganisms from 7th till 14th day. The adding of flakes from biologically activated hull-less barley grain in yoghurt samples caused the higher initial content of yeasts and moulds comparing with control. The growth of yeasts and moulds during shelf-life provided the added malt extract in yoghurt samples. Yoghurt enriched with flakes from biologically activated hull-less barley grain and malt extract from a microbiological perspective is safe product.

Culture of Oleaginous Yeasts in Dairy Industry Wastewaters to Obtain Lipids Suitable for the Production of II-Generation Biodiesel
The oleaginous yeasts Lipomyces starkey were grown in the presence of dairy industry wastewaters (DIW). The yeasts were able to degrade the organic components of DIW and to produce a significant fraction of their biomass as triglycerides. When using DIW from the Ricotta cheese production or residual whey as growth medium, the L. starkey could be cultured without dilution nor external organic supplement. On the contrary, the yeasts could only partially degrade the DIW from the Mozzarella cheese production, due to the accumulation of a metabolic product beyond the threshold of toxicity. In this case, a dilution of the DIW was required to obtain a more efficient degradation of the carbon compounds and an higher yield in oleaginous biomass. The fatty acid distribution of the microbial oils obtained showed a prevalence of oleic acid, and is compatible with the production of a II generation biodiesel offering a good resistance to oxidation as well as an excellent cold-performance.
Identification of Binding Proteins That Interact with BVDV E2 Protein in Bovine Trophoblast Cell
Bovine viral diarrhea virus (BVDV) can cause lifelong persistent infection. One reason for the phenomena is attributed to BVDV infection to placenta tissue. However the mechanisms that BVDV invades into placenta tissue remain unclear. To clarify the molecular mechanisms, we investigated the possible means that BVDV entered into bovine trophoblast cells (TPC). Yeast two-hybrid system was used to identify proteins extracted from TPC, which interact with BVDV envelope glycoprotein E2. A PGbkt7-E2 yeast expression vector and TPC cDNA library were constructed. Through two rounds of screening, three positive clones were identified. Sequencing analysis indicated that all the three positive clones encoded the same protein clathrin. Physical interaction between clathrin and BVDV E2 protein was further confirmed by coimmunoprecipitation experiments. This result suggested that the clathrin might play a critical role in the process of BVDV entry into placenta tissue and might be a novel antiviral target for preventing BVDV infection.
Evolutionary Distance in the Yeast Genome
Whole genome duplication (WGD) increased the number of yeast Saccharomyces cerevisiae chromosomes from 8 to 16. In spite of retention the number of chromosomes in the genome of this organism after WGD to date, chromosomal rearrangement events have caused an evolutionary distance between current genome and its ancestor. Studies under evolutionary-based approaches on eukaryotic genomes have shown that the rearrangement distance is an approximable problem. In the case of S. cerevisiae, we describe that rearrangement distance is accessible by using dedoubled adjacency graph drawn for 55 large paired chromosomal regions originated from WGD. Then, we provide a program extracted from a C program database to draw a dedoubled genome adjacency graph for S. cerevisiae. From a bioinformatical perspective, using the duplicated blocks of current genome in S. cerevisiae, we infer that genomic organization of eukaryotes has the potential to provide valuable detailed information about their ancestrygenome.
Hydrolysis of Eicchornia crassipes and Egeria densa for Ethanol Production by Yeasts Isolated from Colombian Lake Fúquene

The aquatic plants are a promising renewable energy resource. Lake Fúquene polluting macrophytes, water hyacinth (Eichhornia crassipes C. Mart.) and Brazilian elodea (Egeria densa Planch.), were saccharifiedby different treatments and fermented to ethanol by native yeasts. Among the tested chemical and biological methods for the saccharification, Pleurotus ostreatus at 10% (m/v) was chosen as the best pre-treatment in both macrophytes (P<0.01). Subsequently 49 yeasts were isolated from Lake Fúquene and nine strains were selected, which presented the highest precipitates characteristic of ethanol in the iodoform test. The fermentations from water hyacinth and Brazilian elodea hydrolysates using these yeasts produced ethanol at a rate between 0.38 to 0.80gL-1h-1 and 0.15 to 0.27gL-1h-1 respectively. The ethanol presence was confirmed by gas chromatography–mass spectrometry. The nine yeasts chosen were preliminarily identified as belonging to the genera Candida spp., Brettanomyces sp. and Hansenula spp.

Statistical Screening of Medium Components on Ethanol Production from Cashew Apple Juice using Saccharomyces diasticus

In the present study, effect of critical medium components (a total of fifteen components) on ethanol production from waste cashew apple juice (CAJ) using yeast Saccharomyces diasticus was studied. A statistical response surface methodology (RSM) based Plackett-Burman Design (PBD) was used for the design of experiments. The design contains a total of 32 experimental trails. The effect of medium components on ethanol was studied at two different levels such as low concentration level (-) and high concentration levels (+). The dependent variables selected in this study were ethanol concentration (g/L) and cellmass concentration (g/L). Data obtained from RSM on ethanol production were subjected to analysis of variance (ANOVA). In general, initial substrate concentration significantly influenced the microbial growth and product formation. Of the medium components evaluated, CAJ concentration, yeast extract, (NH4)2SO4, and malt extract showed significant effect on ethanol fermentation. A second-order polynomial model was used to predict the experimental data and the model fitted the data with a high correlation coefficient (R2 > 0.98). Maximum ethanol (15.3 g/L) and biomass (6.4 g/L) concentrations were obtained at the optimum medium composition and at optimum condition (temperature-30°C; initial pH-6.8) after 72 h fermentation using S.diasticus.

Enzymatic Esterification of Carboxylic Acids and Higher Alcohols in Organic Medium
The studying of enzymatic esterification of carboxylic acids and higher alcohols was performed by esterase Saccharomyces cerevisiae in water-organic medium. Investigation of the enzyme specificity to acetic substrates showed the best result with acetic acid in esterification reactions with ethanol whereas within other carboxylic acids the esterification decreased with acids: hexanoic > pentanoic > butyric > decanoic. In relation to higher alcohols C3-C5, esterification increased with alcohols propanol < butanol < amylol. Also it was determined that esterase was more specific to alcohols with branched chain such as isobutyl alcohol and isoamyl alcohol. Data obtained may have important practical implications, for example, for application of yeast esterase in producing various volatile esters as well as in enzymatic transformation of volatile acids and toxic fusel alcohols into volatile esters by providing the production of the high quality alcoholic beverages with redused content of higher alcohols as well as with improved degustational and hygienic properties.
Integrated Cultivation Technique for Microbial Lipid Production by Photosynthetic Microalgae and Locally Oleaginous Yeast

The objective of this research is to study of microbial lipid production by locally photosynthetic microalgae and oleaginous yeast via integrated cultivation technique using CO2 emissions from yeast fermentation. A maximum specific growth rate of Chlorella sp. KKU-S2 of 0.284 (1/d) was obtained under an integrated cultivation and a maximum lipid yield of 1.339g/L was found after cultivation for 5 days, while 0.969g/L of lipid yield was obtained after day 6 of cultivation time by using CO2 from air. A high value of volumetric lipid production rate (QP, 0.223 g/L/d), specific product yield (YP/X, 0.194), volumetric cell mass production rate (QX, 1.153 g/L/d) were found by using ambient air CO2 coupled with CO2 emissions from yeast fermentation. Overall lipid yield of 8.33 g/L was obtained (1.339 g/L of Chlorella sp. KKU-S2 and 7.06g/L of T. maleeae Y30) while low lipid yield of 0.969g/L was found using non-integrated cultivation technique. To our knowledge this is the unique report about the lipid production from locally microalgae Chlorella sp. KKU-S2 and yeast T. maleeae Y30 in an integrated technique to improve the biomass and lipid yield by using CO2 emissions from yeast fermentation.

Effect of Commercial or Bovine Yeasts on the Performance and Blood Variables of Broiler Chickens Intoxicated with Aflatoxins
The effects of commercial or bovine yeasts on the performance and blood variables of broiler chickens intoxicated with aflatoxin were investigated in broilers. Four hundred eighty broilers (Arbor Acres; 3-wk-old) were randomly assigned to 4 groups. Each group (120 broiler chickens) was further randomly divided into 6 replicates of 20 chickens. The treatments were control diet without additives (treatment 1), 250 ppb AFB1 (treatment 2), commercial yeast, Saccharomyces cerevisiae, (CY 2.5 x 107 CFU/g) + 250 ppb AFB1 (treatment 3) and bovine yeast, Saccharomyces cerevisiae, (BY 2.5 x 107 CFU/g + 250 ppb AFB1 (treatment 4). Complete randomized design (CRD) was used in the experiment. Feed consumption and body weight were recorded at every five-day period. On day 42, carcass compositions were determined from 30 birds per treatment. While chicks were sacrificed, 3-4 ml blood sample was taken and stored frozen at (-20°C) for serum chemical analysis to determine effects of consumption of diets on blood chemistry (total protein, albumin, glucose, urea, cholesterol and triglycerides). There were no significant differences in ADFI among the treatments(P>0.05). However, BWG, FCR and mortality were highly significantly different (P
Microbial Oil Production by Isolated Oleaginous Yeast Torulaspora globosa YU5/2
Microbial oil was produced by soil isolated oleaginous yeast YU5/2 in flask-batch fermentation. The yeast was identified by molecular genetics technique based on sequence analysis of the variable D1/D2 domain of the large subunit (26S) ribosomal DNA and it was identified as Torulaspora globosa. T. globosa YU5/2 supported maximum values of 0.520 g/L/d, 0.472 g lipid/g cells, 4.16 g/L, and 0.156 g/L/d for volumetric lipid production rate, and specific yield of lipid, lipid concentration, and specific rate of lipid production respectively, when culture was performed in nitrogen-limiting medium supplemented with 80g/L glucose. Among the carbon sources tested, maximum cell yield coefficient (YX/S, g/L), maximum specific yield of lipid (YP/X, g lipid/g cells) and volumetric lipid production rate (QP, g/L/d) were found of 0.728, 0.237, and 0.619, respectively, using sweet potato tubers hydrolysates as carbon source.
Microbiological Contamination of Outdoor Air in Marine Durres's Harbour, Albania

Microbial air contamination of the outdoor air in Marine Durres-s Harbour (Durres, Albania) was estimated by sedimentation technique in August-October 2008. The sampling areas were: Ferry Terminal (FT), Fishery Harbor (FH), East Zone (EZ), Fuel Quay (FQ) and Apollonian Beach (AB). The aim of this study was to measure the number of aerobic plate count (mesophilic aerobic bacteria) and fungi (yeasts and molds) in the outdoor air in these areas. The number of colonies that were formed determines the number of cells at the moment in the outdoor air; respectively the number of mesophilic aerobic bacteria and yeasts and molds. The measure of bacteria and fungi used is CFU (Colony Forming Units) per Petri dish. It is said that marine harbours are very polluted areas. The aim of study was the definition of mesophilic aerobic bacteria and yeasts and molds number, and the comparison of microorganisms number in air sampling areas.

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