Search results for: Bovine yeast

Authors: Sunil Dehipawala, Todd Holden, E. Cheung, Robert Regan, P. Schneider, G. Tremberger Jr, D. Lieberman, T. Cheung

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The zinc and iron environments in different growth stages have been studied with EXAFS and XANES with Brookhaven Synchrotron Light Source. Tissue samples included meat, organ, vegetable, leaf, and yeast. The project studied the EXAFS and XANES of tissue samples using Zn and Fe K-edges. Duck embryo samples show that brain and intestine would contain shorter EXFAS determined Zn-N/O bond; as with the cases of fresh yeast versus reconstituted live yeast and green leaf versus yellow leaf. The XANES Fourier transform characteristic-length would be useful as a functionality index for selected types of tissue samples in various physical states. The extension to the development of functional synchrotron imaging for tissue engineering application based on spectroscopic technique is discussed.

Keywords: EXAFS, Fourier Transform, metalloproteins, XANES

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Authors: Dharell B. Siano, Wendy C. Mateo, Victorino T. Taylan, Francisco D. Cuaresma

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Biofuel is one of the renewable energy sources adapted by the Philippine government in order to lessen the dependency on foreign fuel and to reduce carbon dioxide emissions. Rain tree pods were seen to be a promising source of bioethanol since it contains significant amount of fermentable sugars. The study was conducted to establish the complete procedure in processing rain tree pods for village level hydrous bioethanol production. Production processes were done for village level hydrous bioethanol production from collection, drying, storage, shredding, dilution, extraction, fermentation, and distillation. The feedstock was sundried, and moisture content was determined at a range of 20% to 26% prior to storage. Dilution ratio was 1:1.25 (1 kg of pods = 1.25 L of water) and after extraction process yielded a sugar concentration of 22 ⁰Bx to 24 ⁰Bx. The dilution period was three hours. After three hours of diluting the samples, the juice was extracted using extractor with a capacity of 64.10 L/hour. 150 L of rain tree pods juice was extracted and subjected to fermentation process using a village level anaerobic bioreactor. Fermentation with yeast (Saccharomyces cerevisiae) can fasten up the process, thus producing more ethanol at a shorter period of time; however, without yeast fermentation, it also produces ethanol at lower volume with slower fermentation process. Distillation of 150 L of fermented broth was done for six hours at 85 °C to 95 °C temperature (feedstock) and 74 °C to 95 °C temperature of the column head (vapor state of ethanol). The highest volume of ethanol recovered was established at with yeast fermentation at five-day duration with a value of 14.89 L and lowest actual ethanol content was found at without yeast fermentation at three-day duration having a value of 11.63 L. In general, the results suggested that rain tree pods had a very good potential as feedstock for bioethanol production. Fermentation of rain tree pods juice can be done with yeast and without yeast.

Keywords: Fermentation, hydrous bioethanol, rain tree pods, village level.

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Authors: Le Thuy Mai, Vu Nguyen Thanh

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β-Glucosidase is an important enzyme for production of ethanol from lignocellulose. With hydrolytic activity on cellooligosaccharides, especially cellobiose, β-glucosidase removes product inhibitory effect on cellulases and forms fermentable sugars. In this study, β-glucosidase encoding gene (BGL1) from traditional starter yeast Saccharomycosis fibuligera BMQ908 was cloned and expressed in Pichia pastoris. BGL1 of S. fibuligera BMQ 908 shared 98% nucleotide homology with the closest GenBank sequence (M22475) but identity in amino-acid sequences of catalytic domains. Recombinant plasmid pPICZαA/BGL1 containing the sequence encoding BGL1 mature protein and α-factor secretion signal was constructed and transformed into methylotrophic yeast P. pastoris by electroporation. The recombinant strain produced single extracellular protein with molecular weight of 120 kDa and cellobiase activity of 60 IU/ml. The optimum pH of the recombinant β-glucosidase was 5.0 and the optimum temperature was 50°C.

Keywords: β-Glucosidase, Pichia pastoris, Saccharomycopsisfibuligera, recombinant enzyme.

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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: Phakamas Rachamontree, Theerawut Phusantisampan, Natthakorn Woravutthikul, Peerapong Pornwongthong, Malinee Sriariyanun

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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.

Keywords: Single cell protein, response surface methodology, yeast, cassava processing waste.

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Authors: Thidarat Papone, Supaporn Kookkhunthod, Ratanaporn Leesing

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Monoculture and mixed cultures of microalgae and the oleaginous yeast for microbial oil productions were investigated using sugarcane juice as carbon substrate. The monoculture of yeast Torulaspora maleeae Y30, Torulaspora globosa YU5/2 grew faster than that of microalgae Chlorella sp. KKU-S2. In monoculture of T. maleeae Y30, a biomass of 8.267g/L with lipid yield of 0.920g/L were obtained, while 8.333g/L of biomass with lipid yield of 1.141g/L were obtained for monoculture of T. globosa YU5/2. A biomass of 1.933g/L with lipid yield of 0.052g/L was found for monoculture of Chlorella sp. KKU-S2. The biomass concentration in the mixed culture of the oleaginous yeast with microalgae increased faster and was higher compared with that in the monocultures. A biomass of 8.733g/L with lipid yield of 1.564g/L was obtained for a mixed culture of T. maleeae Y30 with Chlorella sp. KKU-S2, while 8.010g/L of biomass with lipid yield of 2.424g/L was found for mixed culture of T. globosa YU5/2 with Chlorella sp. KKU-S2. Maximum cell yield coefficient (YX/S, g/L) was found of 0.323 in monoculture of Chlorella sp. KKU-S2 but low level of both specific yield of lipid (YP/X, g lipid/g cells) of 0.027 and volumetric lipid production rate (QP, g/L/d) of 0.003 were observed. While, maximum YP/X (0.303), QP (0.105) and maximum process product yield (YP/S, 0.061) were obtained in mixed culture of T. globosa YU5/2 with Chlorella sp. KKU-S2. The results obtained from the study shows that mixed culture of yeast with microalgae is a desirable cultivation process for microbial oil production.

Keywords: Microbial oil, Chlorella sp. KKU-S2, Torulaspora maleeae Y30, Torulaspora globosa YU5/2, mixed culture, biodiesel.

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Authors: Elif Gencturk, Ekin Yurdakul, Ahmet Y. Celik, Senol Mutlu, Kutlu O. Ulgen

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Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.  

Keywords: COP, HMF, ribosome biogenesis, thermoplastic microbioreactor, yeast.

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Authors: Mihriban Korukluoglu, Goksen Arik, Cagla Erdogan, Selen Kocakoglu

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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.

Keywords: Antagonistic effect, kefir, lactic acid bacteria (LAB), synergistic, yeast.

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Authors: Yan Ren, Jun Qiao, Xianxia Liu, Pengyan Wang, Qiang Fu, Huijun Shi, Fei Guo, Yuanzhi Wang, Hui Zhang, Jinliang Sheng, Xinli Gu, Xiao-Jun Liu, Chuangfu Chen

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As part of national epidemiological survey on bovine viral diarrhea virus (BVDV), a total of 274 dejecta samples were collected from 14 cattle farms in 8 areas of Xinjiang Uygur Autonomous Region in northwestern China. Total RNA was extracted from each sample, and 5--untranslated region (UTR) of BVDV genome was amplified by using two-step reverse transcriptase-polymerase chain reaction (RT-PCR). The PCR products were subsequently sequenced to study the genetic variations of BVDV in these areas. Among the 274 samples, 33 samples were found virus-positive. According to sequence analysis of the PCR products, the 33 samples could be arranged into 16 groups. All the sequences, however, were highly conserved with BVDV Osloss strains. The virus possessed theses sequences belonged to BVDV-1b subtype by phylogenetic analysis. Based on these data, we established a typing tree for BVDV in these areas. Our results suggested that BVDV-1b was a predominant subgenotype in northwestern China and no correlation between the genetic and geographical distances could be observed above the farm level.

Keywords: bovine viral diarrhea virus, molecular epidemiology, phylogenetic analysis.

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Authors: A. Daverey, K. Pakshirajan, P. Sangeetha

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Sophorolipids (SLs) production by the yeast Candida bombicola was studied in batch shake flasks using synthetic dairy wastewaters (SDWW) with or without any added external carbon and nitrogen sources. A maximum SLs production of 38.76 g/l was observed with the SDWW supplemented with low cost substrate of sugarcane molasses at 50 g/l and soybean oil at 50 g/l. When the SDWW was supplemented with more costly glucose, yeast extract, urea and soybean oil, the production, however, got lowered to only 29.49 g/l, but with a maximum biomass production of 17.38 g/l together with a complete utilization of the carbon sources.

Keywords: Candida bombicola, dairy wastewater, fat and oil, sophorolipids.

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Authors: N. Cetinkaya, N. H. Ozdemir

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The aim of the study was to investigate the effect of Saccharomyces cerevisiae (SC) live yeast culture on microbial protein supply to small intestine in Kivircik male yearlings when fed with different ratio of forage and concentrate diets. Four Kivircik male yearlings with permanent rumen canula were used in the experiment. The treatments were allocated to a 4x4 Latin square design. Diet I consisted of 70% alfalfa hay and 30% concentrate, Diet II consisted of 30% alfalfa hay and 70% concentrate, Diet I and II were supplemented with a SC. Daily urine was collected and stored at -20°C until analysis. Calorimetric methods were used for the determination of urinary allantoin and creatinine levels. The estimated microbial N supply to small intestine for Diets I, I+SC, II and II+SC were 2.51, 2.64, 2.95 and 3.43 g N/d respectively. Supplementation of Diets I and II with SC significantly affected the allantoin levels in μmol/W0.75 (p<0.05). Mean creatinine values in μmol/W0.75 and allantoin:creatinine ratios were not significantly different among diets. In conclusion, supplementation with SC live yeast culture had a significant effect on urinary allantoin excretion and microbial protein supply to small intestine in Kivircik yearlings fed with high concentrate Diet II (P<0.05). Hence urinary allantoin excretion may be used as a tool for estimating microbial protein supply in Kivircık yearlings. However, further studies are necessary to understand the metabolism of Saccharomyces cerevisiae live yeast culture with different forage:concentrate ratio in Kıvırcık Yearlings.

Keywords: Allantoin, creatinine, Kivircik yearling, microbial nitrogen, Saccharomyces cerevisiae.

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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 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.

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

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Authors: Ramirez D. Edgar I., Angeles H. José J., Ruiz C. Osvaldo, Jacobo A. Victor H., Ortiz P. Armando

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Knowledge of bone mechanical properties is important for bone substitutes design and fabrication, and more efficient prostheses development. The aim of this study is to characterize the viscoelastic behavior of bone specimens, through stress relaxation and fatigue tests performed to trabecular bone samples from bovine femoral heads. Relaxation tests consisted on preloading the samples at five different magnitudes and evaluate them for 1020 seconds, adjusting the results to a KWW mathematical model. Fatigue tests consisted of 700 load cycles and analyze their status at the end of the tests. As a conclusion we have that between relaxation stress and each preload there is linear relation and for samples with initial Young´s modulus greater than 1.5 GPa showed no effects due fatigue test loading cycles.

Keywords: Bone viscoelasticity, fatigue test, stress relaxation test, trabecular bone properties.

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Authors: T. A. Abegunde, O. B. Oyewole, T. A. Sanni

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A process of conversion of flour from three varieties of cassava, namely Odongbo, ofege and TMS30752 to ethanol using α-amylase locally sourced from germinated unhusked paddy rice and yeast isolated from palm wine was developed. It involves the germination of paddy rice for a period of 15days to produce α-amylase for starch hydrolysis and isolation of yeast from palm wine for fermentation. The results showed that optimum amylase yield of “ofada” rice paddy was at 6^th day germination which was 576.9ml/g. Ethanol yield for TMS30572 (440.3%) was significantly higher than “Odongbo” (160.2%) and “Ofege’’ (115.1%), Sugar conversion efficiency were 311.0%v/v, 268.2%v/v and 186.84%v/v for TMS30572, “Odongbo” and “Ofege” respectively. The ethanol boiling points were 78^oC, 76^oC and 80^oC for TMS30572, “Odongbo” and “Ofege” respectively. This study showed that cassava varieties affects quality of ethanol produced and germination of “ofada” rice for 6 days ensures optimum production of crude amylase enzyme.

Keywords: Cassava, ethanol, fermentation, hydrolysis, α-amylase.

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Authors: P. Chaijak, M. Lertworapreecha, C. Sukkasem

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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.

Keywords: Extracellular laccase, production, yeast, natural inducer.

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Authors: Eva Tvrdá, Marek Halenár, Hana Greifová, Alica Mackovich, Faridullah Hashim, Norbert Lukáč

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Oxidative stress associated with semen cryopreservation may result in lipid peroxidation (LPO), DNA damage and apoptosis, leading to decreased sperm motility and fertilization ability. Curcumin (CUR), a natural phenol isolated from Curcuma longa Linn. has been presented as a possible supplement for a more effective semen cryopreservation because of its antioxidant properties. This study focused to evaluate the effects of CUR on selected oxidative stress parameters in cryopreserved bovine semen. 20 bovine ejaculates were split into two aliquots and diluted with a commercial semen extender containing CUR (50 μmol/L) or no supplement (control), cooled to 4 °C, frozen and kept in liquid nitrogen. Frozen straws were thawed in a water bath for subsequent experiments. Computer assisted semen analysis was used to evaluate spermatozoa motility, and reactive oxygen species (ROS) generation was quantified by using luminometry. Superoxide generation was evaluated with the NBT test, and LPO was assessed via the TBARS assay. CUR supplementation significantly (P<0.001) increased the spermatozoa motility and provided a significantly higher protection against ROS (P<0.001) or superoxide (P<0.01) overgeneration caused by semen freezing and thawing. Furthermore, CUR administration resulted in a significantly (P<0.01) lower LPO of the experimental semen samples. In conclusion, CUR exhibits significant ROS-scavenging activities which may prevent oxidative insults to cryopreserved spermatozoa and thus may enhance the post-thaw functional activity of male gametes.

Keywords: Bulls, cryopreservation, curcumin, lipid peroxidation, reactive oxygen species, spermatozoa.

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Authors: U.K.Ghosh, M.K.Ghosh

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The present experimental investigation brings about a comparative study of lactic acid production by pure strains of Lactobacilli (1) L. delbreuckii (NCIM2025), (2) L. pentosus (NCIM 2912), (3) Lactobacillus sp.(NCIM 2734, (4) Lactobacillus sp. (NCIM2084) and coculture of strain-1 and Stain-2 in solid bed of wheat bran, under the influence of different nitrogen sources such as baker-s yeast, meat extract and proteose peptone. Among the pure cultures, strain-3 attained lowest pH value of 3.44, hence highest acid formation 46.41 g/L, while the coculture attained an overall maximum value 47.56 g/L lactic acid (pH 3.38) at 15 g/L and 20 g/L level of baker-s yeast, respectively.

Keywords: Eco-friendly, lactic acid, lactobacilli, wheat bran

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Authors: Seunghwan Lee

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The lubricating properties of commercially available mucins originating from different animal organs, namely bovine submaxillary mucin (BSM) and porcine gastric mucin (PGM), have been characterized at polymeric surfaces for biomedical applications. Atomic force microscopy (AFM) and pin-on-disk tribometry have been employed for tribological studies at nanoscale and macroscale contacts, respectively. Polystyrene (PS) was employed to represent ‘rigid’ contacts, whereas poly(dimethylsiloxane) (PDMS) was employed to represent ‘soft contacts’. To understand the lubricating properties of mucins in correlation with the coverage on surfaces, adsorption properties of mucins onto the polymeric substrates have been characterized by means of optical waveguide light-mode spectroscopy (OWLS). Both mucins showed facile adsorption onto both polymeric substrates, but the lubricity was highly dependent upon the pH change between 2 and 7.

Keywords: Bovine submaxillary mucin (BSM), Porcine Gastric Mucin (PGM), lubricity, biomedical.

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Authors: Neelima Gupta, Seema Aggarwal

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Most of the biclustering/projected clustering algorithms are based either on the Euclidean distance or correlation coefficient which capture only linear relationships. However, in many applications, like gene expression data and word-document data, non linear relationships may exist between the objects. Mutual Information between two variables provides a more general criterion to investigate dependencies amongst variables. In this paper, we improve upon our previous algorithm that uses mutual information for biclustering in terms of computation time and also the type of clusters identified. The algorithm is able to find biclusters with mixed relationships and is faster than the previous one. To the best of our knowledge, none of the other existing algorithms for biclustering have used mutual information as a similarity measure. We present the experimental results on synthetic data as well as on the yeast expression data. Biclusters on the yeast data were found to be biologically and statistically significant using GO Tool Box and FuncAssociate.

Keywords: Biclustering, mutual information.

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Authors: Karuppaiya Maruthai, Viruthagiri Thangavelu, Manikandan Kanagasabai

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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.

Keywords: cashew apple juice, ethanol, fermentation, yeast, response surface methodology

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Authors: Forough Ataollahi, Belinda Pingguan-Murphy, Wan Abu Bakar Wan Abas, Noor Azuan Abu Osman

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Endothelium proliferation is an important process in cardiovascular homeostasis and can be regulated by extracellular environment, as cells can actively sense mechanical environment. In this study, we evaluated endothelial cell proliferation on PDMS/alumina (Al₂O₃) composites and pure PDMS. The substrates were prepared from pure PDMS and its composites with 5% and 10% Al₂O₃ at curing temperature 50˚C for 4h and then characterized by mechanical, structural and morphological analyses. Higher stiffness was found in the composites compared to the pure PDMS substrate. Cell proliferation of the cultured bovine aortic endothelial cells on substrate materials were evaluated via Resazurin assay and 1, 1’-Dioctadecyl-1, 3, 3, 3’, 3’-Tetramethylindocarbocyanine Perchlorate-Acetylated LDL (Dil-Ac-LDL) cell staining, respectively. The results revealed that stiffer substrates promote more endothelial cells proliferation to the less stiff substrates. Therefore, this study firmly hypothesizes that the stiffness elevates endothelial cells proliferation.

Keywords: Bovine aortic endothelial cells, extra cellular matrix, proliferation, stiffness.

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Authors: P. Ashok, G. M. Kadhar Nawaz

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Rough set theory is used to handle uncertainty and incomplete information by applying two accurate sets, Lower approximation and Upper approximation. In this paper, the rough clustering algorithms are improved by adopting the Similarity, Dissimilarity–Similarity and Entropy based initial centroids selection method on three different clustering algorithms namely Entropy based Rough K-Means (ERKM), Similarity based Rough K-Means (SRKM) and Dissimilarity-Similarity based Rough K-Means (DSRKM) were developed and executed by yeast dataset. The rough clustering algorithms are validated by cluster validity indexes namely Rand and Adjusted Rand indexes. An experimental result shows that the ERKM clustering algorithm perform effectively and delivers better results than other clustering methods. Outlier detection is an important task in data mining and very much different from the rest of the objects in the clusters. Entropy based Rough Outlier Factor (EROF) method is seemly to detect outlier effectively for yeast dataset. In rough K-Means method, by tuning the epsilon (ᶓ) value from 0.8 to 1.08 can detect outliers on boundary region and the RKM algorithm delivers better results, when choosing the value of epsilon (ᶓ) in the specified range. An experimental result shows that the EROF method on clustering algorithm performed very well and suitable for detecting outlier effectively for all datasets. Further, experimental readings show that the ERKM clustering method outperformed the other methods.

Keywords: Clustering, Entropy, Outlier, Rough K-Means, validity index.

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Authors: Anjali Haloi, Debabrata Das

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Silver nanoparticles inhibit a wide variety of microorganisms. The mechanism of inhibition is not entirely known although it is recognized to be concentration dependent and associated with the disruption of membrane permeability. Data on differential gene expression as a response to nanoparticles could provide insights into the mechanism of this inhibitory effect. Silver nanoparticles were synthesized in yeast growth media using a modification of the Creighton method and characterized with UV-Vis spectrophotometry, transmission electron microscopy (TEM), and X-ray diffraction (XRD). In yeasts grown in the presence of silver nanoparticles, we observed that at concentrations below the minimum inhibitory concentration (MIC) of 48.51 µg/ml, the total RNA content was steady while the cellular protein content declined rapidly. The analysis of the expression levels of KRR1 and PWP2, two important genes involved in rRNA maturation in yeasts, showed up to 258 and 42-fold decreases, respectively, compared to that of control samples. Whether silver nanoparticles have an adverse effect on ribosome assembly and function could be an area of further investigation.

Keywords: Ag NP, yeast, qRT-PCR, KRR1, PWP2.

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Authors: A. Manivannan, R. T. Narendhirakannan

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The objective of this research was to investigate biodegradation of water hyacinth (Eichhornia crassipes) to produce bioethanol using dilute-acid pretreatment (1% sulfuric acid) results in high hemicellulose decomposition and using yeast (Pachysolen tannophilus) as bioethanol producing strain. A maximum ethanol yield of 1.14g/L with coefficient, 0.24g g^-1; productivity, 0.015g l^-1h^-1 was comparable to predicted value 32.05g/L obtained by Central Composite Design (CCD). Maximum ethanol yield coefficient was comparable to those obtained through enzymatic saccharification and fermentation of acid hydrolysate using fully equipped fermentor. Although maximum ethanol concentration was low in lab scale, the improvement of lignocellulosic ethanol yield is necessary for large scale production.

Keywords: Acid hydrolysis, Biodegradation, Hemicellulose, Pachysolen tannophilus, Water hyacinth.

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Authors: Eva Tvrdá, Boris Botman, Marek Halenár, Tomáš Slanina, Norbert Lukáč

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In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P<0.05; Time 24 h). At the same time, a long-term exposure of spermatozoa to concentrations ranging between 0.05 µg/mL and 2 µg/mL had a negative impact on the mitochondrial metabolism (P<0.05; Time 24 h). The viability staining revealed that 0.001-1 µg/mL Salvia extract had no effects on bovine male gametes, however 2 µg/mL Salvia had a persisting negative effect on spermatozoa (P<0.05). Furthermore 0.05-2 µg/mL Salvia exhibited an immediate ROS-promoting effect on the sperm culture (P>0.05; Time 0 h and 2 h), which remained significant throughout the entire in vitro culture (P<0.05; Time 24 h). Our results point out to the necessity to examine specific effects the biomolecules present in Salvia officinalis may have individually or collectively on the in vitro sperm vitality and oxidative profile.

Keywords: Bulls, CASA, MTT test, reactive oxygen species, sage, Salvia officinalis, spermatozoa.

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Authors: Amir Saber Gharamaleki, Beitollah Alipour, Zeinab Faghfoori, Ahmad YariKhosroushahi

Abstract:

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.

Keywords: Anticancer, anti-proliferative, apoptosis, cytotoxicity, yeast.

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Authors: D.T. Mirzarakhmetova

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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.

Keywords: enzymes in non-conventional media, esterification, higher alcohols, volatile esters, yeast esterase

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Authors: M. T. Asadollahzadeh, A. Ghasemian, A. R. Saraeian, H. Resalati, P. R. Lennartsson, M. J. Taherzadeh

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Since filamentous fungi are capable of assimilating several types of sugars (hexoses and pentoses), they are potential candidates for bioconversion of spent sulfite liquor (SSL). Three filamentous fungi such as Aspergillus oryzae, Mucor indicus, and Rhizopus oryzae were investigated in this work. The SSL was diluted in order to obtain concentrations of 50, 60, 70, 80, and 90% and supplemented with two types of nutrients. The results from cultivations in shake flask showed that A. oryzae and M. indicus were not able to grow in pure SSL and SSL90% while R. oryzae could grow only in SSL50% and SSL60%. Cultivation with A. oryzae resulted in the highest yield of produced fungal biomass, while R. oryzae cultivation resulted in the lowest fungal biomass yield. Although, the mediums containing yeast extract, (NH₄)₂SO₄, KH₂PO₄, CaCl₂∙2H₂O, and MgSO₄∙7H₂O as nutrients supplementations produced higher fungal biomass compared to the mediums containing NH₄H₂PO₄ and ammonia, but there was no significant difference between two types of nutrients in terms of sugars and acetic acid consumption rate. The sugars consumption in M. indicus cultivation was faster than A. oryzae and R. oryzae cultivation. Acetic acid present in SSL was completely consumed during cultivation of all fungi. M. indicus was the best and fastest ethanol producer from SSL among the fungi examined, when yeast extract and salts were used as nutrients supplementations. Furthermore, no further improvement in ethanol concentration and rate of sugars consumption was obtained in medium supplemented with NH₄H₂PO₄ and ammonia compared to medium containing yeast extract, (NH₄)₂SO₄, KH₂PO₄, CaCl₂∙2H₂O, and MgSO₄∙7H₂O. On the other hand, the higher dilution of SSL resulted in a better fermentability, and better consumption of sugars and acetic acid.

Keywords: Ethanol, filamentous fungi, fungal biomass, spent sulfite liquor.

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Authors: S. C. Santos, S. R. Dionísio, A. L. D. De Andrade, L. R. Roque, A. C. Da Costa, J. L. Ienczak

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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.

Keywords: 5-carbon sugar, cell recycling fermenter, mixed sugars, xylose-fermenting yeast.

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Authors: A. Ghaffari, A. S. Mostafavi

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Complex assemblies of interacting proteins carry out most of the interesting jobs in a cell, such as metabolism, DNA synthesis, mitosis and cell division. These physiological properties play out as a subtle molecular dance, choreographed by underlying regulatory networks that control the activities of cyclin-dependent kinases (CDK). The network can be modeled by a set of nonlinear differential equations and its behavior predicted by numerical simulation. In this paper, an innovative approach has been proposed that uses genetic algorithms to mine a set of behavior data output by a biological system in order to determine the kinetic parameters of the system. In our approach, the machine learning method is integrated with the framework of existent biological information in a wiring diagram so that its findings are expressed in a form of system dynamic behavior. By numerical simulations it has been illustrated that the model is consistent with experiments and successfully shown that such application of genetic algorithms will highly improve the performance of mathematical model of the cell division cycle to simulate such a complicated bio-system.

Keywords: Cell cycle, Cyclin-dependent kinase, Fission yeast, Genetic algorithms, Mathematical modeling, Wiring diagram

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