Earticle

Exopolysaccharides(EPS) from Aureobasidium pullulans are used for health foods and cosmetics. In order to better exploit EPS production from A. pullulans, medium composition was optimized using stastical methods. And, effect of each nutrient of basal medium was investigated by preliminary studies (one-factor-at-a-time). Maximum growth and EPS production were observed at various initial pH 3-5, while growth and EPS production were also very low at pH 2 and pH 7. Among various nitrogen sources (Ashbya gossypii extract, beef extract, corn steep liquor, corn steep, powder, malt extract, soytone peptone and yeast extract) tested, the highest mycelial growth and EPS production were achieved in the medium containing Ashbya gossypii extract. Also, among various carbon sources, EPS production and the growth of A. pullulans was the best on sucrose. The Plackett–-Burman design was used to evaluate the effects of 7 variables selected from preliminary studies, and three statistically significant variables namely, sucrose, NaNO3, and Ashyba gossiipii extract, were selected. Then, a three-level Box-Behnken design was employed to optimize the medium composition for EPS production in shake-flask. Using this methodology, the quadratic regression model of producing EPS was built and the optimal combinations of media constituents and culture conditions for maximum EPS production (about 30g/L) were determined.

In this study, we propose a new screening method based on differences in metabolism between microorganisms. The novel stains obtained in this way provide information about cultivation of uncultured microorganisms. The key to isolating slow-growing microorganisms from a complex community is eliminating the fast-growing microorganisms. We found that the growth of fast-growing microorganisms could be suppressed by adding phosphate transport inhibitors, fosfomycin or fosmidomycin, to the medium. When microorganisms from the Damupo-seawater were incubated decreasing the amount of phosphate concentration of Marine Media (Difco), the viable cell count decreased by about 33%. In the presence of fosfomycin viable cells decreased markedly and the death rate was 90% and 99% at 5 and 10 mM, respectively. In the case of fosmidomycin, the death rate was 88% at 5 mM. We confirmed that 13 strains of fosfomycin-resistant cells and 20 strains of fosmidomycinresistant cells showed 16S rDNA sequence similarity less than 98% to known database entries. The screening system presented in this work was found to be excellent for searching unculturable microorganisms.

Use of cell free bead as a model to develop a method of vitrification which avoids ice crystal formation by amorphous solidification is an emerging approach to cryopreservation. Developing vitrification strategy, effective cryopreservation of alginate beads has been reported in this study. Beads are produced by spraying alginates into gelling agent using air jet with 20 gauge syringe. And cured in a calcium chloride solution for 10 min. The effect of flow rates, concentration of alginates and calcium chloride liquid on the size, shape, morphology of beads were investigated with a constant nozzle diameter. Among the parameter studied, gas flow rate had pronounced effect on the size of beads as compared to the flow rate of the liquid and concentration of the alginate liquid. The size of bead was reduced to a minimum value 0.55 mm² with increase in the gas flow rate 19 L/Min. At the early stages of the gas flow the rate of the size reduction was relatively low, while dripping mode dominated. Investigation on structural and physical characteristics of 1mm² size beads for SEM &TEM analysis, osmotic pressure test, mechanical stability, swelling and resistance of the beads before and after thawing are carried out.

Acetone/pentane precipitation is a simple, efficient method for pre-purifying paclitaxel extracted from plant cell cultures. However, the acetone/pentane precipitation process has been inherently problematic due to the lengthy precipitation time that is required. An improved acetone/pentane precipitation process could significantly reduce the precipitation time by increasing the surface area per working volume (S/V) of the reacting solution through the addition of an anion exchange resin (Amberlite IRA400) or glass beads. Most of the paclitaxel (>95%) could be obtained after about 24 h of acetone/pentane precipitation by increasing the surface area of reactor. Purity increased with increasing acetone/pentane precipitation time up to 24 h to about 80%, after which it showed little change. The size of paclitaxel precipitates was measured with the electronic microscope, it was observed that the use of an exchange resin had an effect on inhibition of precipitate growth compared to prepared particles without exchange resin. In addition, the amorphous forms of paclitaxel were prepared using acetone/pentane precipitation method. This improved pre-purification process serves to minimize solvent usage and the size and complexity of the high performance liquid chromatography operation required for paclitaxel purification.

Carbohydrate content in algae is one of the present feedstock for bioethanol. The doubling time of microalgae could be range from 3.5 to 6 hours which is short time. Very fast growing microalgae with high content of carbohydrate is a potential feedstock which is renewable and environmental friendly. The present study is carried out for saccharification of Laminaria japonica and Spirulina platensis with acid hydrolysis which is essential to get fermentable sugars. Powder of Laminaria japonica was purchased from Jeon-Won Foods Co where as Spiruilina was cultivated in photobioreactor with spirulina medium, temperature 30℃, light 60~70μmol/m²·s and 0.7 vvm air was provided. The biomass was hydrolysis with sulfuric and hydrochloric acid concentration range from 1, 3, and 5% with temperature 110℃ and 12 0℃ for 30 and 60 minutes. The increase in acid concentration during acid hydrolysis did not significantly affect the total sugar and glucose concentration. In case of biomass higher biomass 1% result higher total sugar yield. In this study, pre-experiment is reported for the most efficient conditions of saccharification using acid hydrolysis for bioethanol production.

Streptomycetes, a major family of actinomycetes, has been intensively investigated, but a potentially-valuable non-streptomycetes rareactinomycetes (NSRA) family is largely uncharacterized. A PCR-based streptomycetes-specific screening was performed to isolate NSRA strains from actinomycetes strain collection which is 180 independentlyisolated antifungal actinomycetes strains (kindly provided by KRIBB) [1, 2]. After screened out most of streptomyces species from the strain collection, several remaining NSRA strains were then confirmed and classified based on 16S rRNA sequence information. To identify the stains whose genes might associate with production of valuable secondary metabolite, PCR-based genome screening was performed using established genetic information such as typeⅠ PKS and NRPS. One strain, putative owning typeⅠ PKS and NRPS genes, shows both antifungal activity and antibacterial activity among the selected strains. These results suggest that a PCR screening strategy for not only NSRA strains but also NSRA genetic manipulation might lead to the isolation of potentially-valuable biosynthetic genes and compounds present in various rare-actinomycetes.

Chinese hamster ovary (CHO) cells are the most widely used mammalian host for the commercial production of recombinant proteins. Recently, various strategies have focused on increasing specific productivity in CHO cell culture. One of the most important mechanisms of regulating of gene expression is the modulation of histones via acetylation. These histone hyperacetylation and enhancement of the expressed mRNA transcription level of target protein were regulated by histone deacetylase inhibitor (HDACi). Effects of HDACi on the enhanced production of recombinant proteins expressed in CHO cells have been reported. To determine the effects of HDACi on the production of recombinant albumin-erythropoietin (Alb-EPO) fusion protein, various inhibitors were added to the cell cultures in exponential growth phase. Among the inhibitors tested, sodium propionate and sodium butyrate were found to be efficient. As these compounds induced cell death, it was hypothesized that HDACi actually induced cell death via apoptosis rather than necrosis. Compared to control cultures without sodium butyrate, the culture with 1 mM sodium butyrate resulted in a 1.7-fold increase in Alb-EPO concentration on day 6. In conclusion, it was found that HDACi enhanced the production of Alb-EPO but also the apoptosis of recombinant CHO cells.

Biodiesel production from microalgae is one of the promising solutions for renewable and sustainable future transport biofuels using inexpensive natural resources (e.g. CO2, H2O). It has been reported that the cells grow under nitrogen deficiency condition have higher lipid level of 50~70%, compared to the normal level of 20~30%. The growth rate, however, is greatly reduced for the cells with higher lipid levels, resulting in the overall lipid productivity unchanged. The objective of this study is to evaluate influence of nitrate concentration and to analyze lipid productivities under different cultivation modes (batch and semi-continuous) for Botryococcus braunii. In order to increase lipid productivity using modified BG-11 (MBG-11) as basis medium, B. braunii cultivated under different nitrate concentration (5%, 25%, 50%, and 100% of nitrate in MBG-11). The culture at 25% of initial nitrate concentration (273 mg/L) of BG-11 showed maximum lipid concentration (1.02 g/L) among tested. B. braunii could accumulate higher lipid level under optimal nitrate concentration. Further optimization of productivity could be achieved by finding optimal cultivation modes

A cryptic plasmid, pMBLR00, was isolated from Leuconostoc mesenteroides subsp. Mesenteroides KCTC 3733. Complete sequencing of pMBLR00 revealed a 4163 bp element with G+C content of 37% harboring three open reading frames (ORFs A, B and C). The ORF A gene product has 98% amino acid identity with the Rep protein of a rolling-circle replication (RCR) plasmid pYS18, isolated from Lactobacillus sakei YSI8. Southern hybridization and nuclease treatment demonstrated that pMBLR00 contains a single-stranded intermediate, confirming that pMBLR00 replicates via RCR mechanism. The ORF B gene product is homologous to mobilization proteins encoded by plasmids isolated from Leuconostoc and Lactobacillus species. An E. coli-Leuconostoc shuttle vector pMBLR01 was constructed based on pMBLR00. The shuttle vector pMBLR01 was stable in Leuconostoc citreum.

The purple nonsulfur photosynthetic bacteria, Rhodobacter sphaeroides, participate in the anoxic cycling of carbon both as the primary producer and as the light-stimulated consumers of the reduced organic compounds. In this study, four different strains, i.e. R. sphaeroides KCTC 1434, MBTLJ-8, MBTLJ-13 and MBTLJ-20 were selected and used to analyze the relationships between the succinic acid and various light conditions. The mutant strain exhibited an enhanced pigments production compared to the R. sphaeroides wild type under all light conditions. Additionally, the growth rate of R. spharoides KCTC1434 and three mutants strains under different light conditions, and the impact on photosynthetic mechanism by various pigment were produced under different light conditions. Therefore, the wave length of light probably affected the growth of all of the strains. Moreover, mutant types occurred the enhanced cell growth phenomenon under the blue colored light emitting diode (LED) conditions and stimulated at low concentrations of succinic acid, which was compensatory for succinic acid. Therefore, R. sphaeroides MBTLJ-8, 13, 20 provieds the strong evidence that enhance cell growth and produce pigments for the succinic acid.

Today a number of nanoparticles (NPs) are applied for medical or medicinal research fields through conjugating with drugs or bio-molecules such as DNA, RNA, and peptides. There are three distinct ways to deliver NPs into animal and human bodies; injection, feeding, and trans-dermal administration. The array of microneedles with length ranging from 600μm to 1.5mm was fabricated using laser-printed PDMS (polydimethylsiloxane) mold and biodegradable polymers such as CMC (Carboxymethyl cellulose), agarose, and etc. The 50 nm fluorescence iron oxide and polystryrene NPs were successfully attached to the surface of microneedles by spraying NPs onto the inner surface of mold. When the array of microneedles was applied to the swine skin, NPs were detected in the hypodermis. Microneedle array conjugated with nanoparticles is expected to be an alternative method to deliver cosmetical or pharmaceutical materials into human skin locally without professional procedures.

Adsorbent treatment is an efficient method to remove waxy and tar compounds from plant cell cultures. In this study, sylopute, active clay (F-1), silica (SiO2), and magnesium oxide (MgO) were applied for the purification of the paclitaxel from plant cell cultures. There was selective adsorption of impurities in the treatment of sylopute and SiO2. Therefore, higher purity paclitaxel could be obtained simply in the following process with a higher yield than with active clay or MgO. The purities of crude paclitaxel after sylopute and SiO2 treatment followed by precipitation were increased up to 57.3% and 59.8%, while the purities for active clay and MgO were 46.9% and 48.4%, respectively. The overall yields of sylopute, SiO2 , active clay, and MgO were 77.3%, 94.2%, 49.1%, and 57.1%, respectively. The best result in removing impurities was obtained from the combination of sylopute or SiO2 treatment followed by precipitation process. Consequently, sylopute and SiO2 can be applied for production scale purification of other natural products as an alternative to synthetic adsorbents other alternative processes.

The capacity to mediate localized gene expression via substrate-mediated gene delivery would greatly enable numerous applications in gene therapy: i) by reducing systemic spread of vectors at the target delivery site and ii) by reducing immune responses against the vector, potentially preventing side effects in off-target regions. Immobilizing gene delivery vectors onto substrates, which serve for cell adhesion, can function to place the vectors directly inside the cellular microenvironment for subsequent cellular internalization, ultimately inducing localized gene expression adjacent to the substrate and possibly enhancing gene delivery efficiency by sustaining the contact of the vectors with target cells. Importantly, this system has the potential to reduce the vector quantities required for high level of gene expression, such that the use of lower doses can potentially reduce cellular toxicity, which is typically observed by the initial burst of gene vectors (e.g., adenoviral vectors or non-viral vectors) upon direct injection in vivo. Due to these advantages, a variety of substrate-mediated gene delivery systems have been developed, primarily combining non-viral vectors and biomaterials. There have been few attempts to deliver viral vectors from a substrate, presumably due to the lack of moieties on the viral vectors that can specifically interact with substrates. Developing substrate-mediated delivery systems for viral vectors, however, will certainly increase the potential of the system due to substantially enhanced gene delivery capacities of viral vectors compared with non-viral vectors. We have developed a strategy to mediate immobilization of adeno-assoicated viral vectors (AAV) directly onto a substrate to which cells subsequently adhere, thus maintaining high local concentration of AAV vectors with the cell microenvironment as well as increasing the extent of physical contact with the attached cells. The development of systems with the capacity to mediate localized gene expression as well as high efficiency gene delivery will have strong potential for numerous disease therapies and tissue engineering applications.

Recently, various secondary devices have been developed to detect the odorant chemical by using olfactory receptor as a primary transducer. Detection of odorant at very low concentration has become a possibility by these devices. But currently there is no device that shows the odorant binding to the olfactory receptors with a fluorescence imaging. In this research, a cAMP response element reporter gene system was used for odorant screening. When odorant molecules bind to the olfactory receptor, the cAMP level was increased and initiated the phosphorylation of the CREB which binds to CRE promoter to induce the expression of ZsGreen as a reporter. For the high-throughput screening, PEG microwell was fabricated on the glass and reverse transfection technique was also used. Plasmid vector containing olfactory receptor gene and lipofectamin were mixed with gelatin solution and spotted on the PEG microwell. HEK293 cells were seeded on the glass substrate and olfactory receptors were expressed in 48 hours. The odorant binding event was detected by fluorescence of ZsGreen as a reporter.

Lactic acid fermentations were conducted using water hyacinth. It is known that the pretreatment and enzyme hydrolysis process optimize the potential of water hyacinth.1,2 Lactic acid produced by using lactic acid bacteria (Lactobacillus crispatus, L. fermentum, L. casei, L. plantarum, L. helveticus, L. paracasei, L.delbrueckii, and L. reuteri). All cells were grown at 370 C and initial pH 5.5.3 Lactic acid production was measured by HPLC. All lactobacillus strains could produce lactic acid from pretreated Water Hyacinth. The highest lactic acid was achieved when lactic acid fermentation was carried out by L.delbrueckii. The lactic acid concentration was 11.85 g/l by L.delbrueckii. It converted glucose in the medium to lactic acid, perfectly. The other strains also could produce the high level of lactic acid in this medium. These are comparable to the lactic acid produced in complex medium such as MRS medium. This study informed that water hyacinth medium could be alternative medium which can replace the complex and expensive medium for growing Lactobacillus strains in production of lactic acid.

Fatty acid biosynthesis in E. coli is dedicated with conversion of acetyl-CoA to malonyl-CoA with acetyl-CoA carboxylase (ACC) which catalyze the ATP-dependent first step. The fatty acid synthase converts malonyl-CoA using NADPH as a source of reducing equivalence to an acyl carrier protein (ACP). In fatty acid biosynthesis, fabH, fabB and fabF genes constitute an enzyme, β-ketoacyl acyl carrier protein synthase III that catalyze the first committed step in fatty acid biosynthesis(ATP-dependent formation of malonyl-CoA from acetyl-CoA and bicarbonate). fabG is a gene for β-ketoacyl acyl carrier protein reductase and fabI is a gene for NADH-dependent enoyl-acyl carrier protein reductase that is related to the completion of fatty acid elongation. In this study, a vector system was constructed to increase short chain fatty acid (C3-C6) content in E. coli. E. coli W3110 was used for the preparation of genes and E. coli BL21 (DE3) was used as a host cell for the expression of target genes, fabH, fabB, fabF, fabG and fabI.

Biological effects of nanomaterials with a focus on toxicity should receive great attention since commercial products as well as medical tools increasingly utilize them. Especially, a fundamental understanding of nanotoxicology is highly desirable both from the material’'s stand point as well as from the biological system’'s point of view. Titanium dioxide (TiO2) is a widely used material in our life: for example, cosmetics of sun-cream to block UV radiation. However, despite its wide array of common applications, its pathogenetic role was always suggested under certain conditions. In this work, the toxicity of TiO2 nanoparicles on human skin- and lung cell was investigated using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfo phenyl)-2H-tetrazolium (MTS) assay- and confocal microscopy for live &dead of cells. The human skin- and lung-cell were purchase from from Korean Cell Line Bank. We observed that TiO2 nanoparticles impedes the cell growth using MTS, and dead cells increase depending on the concentration of TiO2 nanoparticles using confocal microscopy. Based on the results mentioned above, we discuss the toxicity and the safety of TiO2 nanoparicles for the human cells.

Span 80 was selected among the tested detergents for retinal production by metabolically engineered E. coli because the addition of Span 80 increased significantly retinal production. In the flask culture, the optimum concentration of span 80 was determined as 5 g l–1. In the batch culture with 5 g l–1 Span 80, the cell mass, retinal concentration, retinal content, and retinal productivity were 20 g l–1, 61 mg l–1, 3.5 mg g-cells–1, and 3 mg l–1 h–1 after 22 h, respectively. In the fed-batch culture of glycerol with 5 g l–1 span 80, the cell mass, retinal concentration, retinal content, and retinal productivity were 38 g l–1, 335 mg l–1, 8.8 mg g-cells–1, and 16 mg l–1 h–1 after 21 h, respectively. They were approximately 1.1, 3.0-, 2.6- and 3.2-fold higher than those in the fed-batch culture of glycerol without 5 g l–1 Span 80, respectively. This is the first report of retinal production by using the metabolically engineered E. coli. Furthermore, retinal productivity was successfully enhanced by the supplement of Span 80.

Flavonoids are polyphenolic compounds that are ubiquitous in nature and are categorized according to chemical structure, into flavonols, flavones, flavanones, isoflavones, catechins, anthocyanidins and chalcones. Over 4,000 flavonoids have been identified, many of which occur in fruits, vegetables and beverages (tea, coffee, beer, wine and fruit drinks). The flavonoids have aroused considerable interest recently because of their potential beneficial effects on human health-they have been reported to have antiviral, anti-allergic, antiplatelet, anti-inflammatory, antitumor and antioxidant activities. Twelve microorganisms were initially screened for their abilities to catalyze biotransformation of phloretin. Streptomyces avermitilis, two main products were identified in GC/MS analysis. They were interpreted as hydroxylated products of phloretin in A-ring at different position. (mass increase 179 → 267, 192 → 280) This result confirmed hydroxylaion considering BSTFA derivatization of hydroxylated product. Maximum conversion was 6.7 %, which was achieved for 1 hours of reaction, and the substrate (phloretin) and reaction product was completely metabolized after 3 hours of reaction. Three kinds of Cytochrome P450 inhibitor, Coumarin, Erythromycin and Quinidine was added with 0.5mM final concentration to find out the role of P450 enzymes. P450 amplifeid and deleted Streptomycs were also used to confirm the role of P450 enzyme.

2,3-Butanediol(2,3-BDO) is a key starting material for manufacture of bulk chemicals such as methyl ethyl ketone and 1,3-butadien. It also has potential applications in the manufacture of printing ink, perfumes and explosive. Microbial production of 2,3-BDO by using Klebsiella pneumoniae was first investigated in 1906. Klebsiella pneumoniae is a Gram-negative, non-motile, encapsulated, lactose fermenting, facultative anaerobic, rod shaped bacterium found in the normal flora of the mouth, skin, and intestines. In this study, in order to enhanced 2,3-BDO production, budC gene coding for butanediol dehydrogenase was over-expressed and ldhA gene coding for lactate dehydrogenase disrupted by inserting a tetracycline resistance marker in Klebsiella pneumoniae KTCT2242. The fermentation results of both the parent and mutant strain suggested that over-expression of budC and inactivation of ldhA resulted in an improved 2,3-BDO production.

natural antibiotic compound from microbial source are promising for future. Batch and fed-batch culture was carried out to enhance the antibiotics production from B.subtilis produce surfactin-like lipopeptide antibiotics. Dextrose was best carbon source for high production in batch culture. For fed-batch, optimal glucose concentration was 1.5 % around and this concentration showed best antibiotics production rate. Cell growth was also highly enhance by glucose feeding. Ammonium was best nitrogen source for antibiotics production for this strain and it concentration was 4% around. Other salts such as magnesium, manganese, ferrous, and potassium ion was also important for growth as well as production.

The reaction conditions for the maximal production of 10-hydroxystearic acid from oleic acid by whole cells of Stenotrophomonas nitritireducens were determined as pH 7.5 and 35°C with 0.05% Tween 80. Whole cells of 20 g/L converted all of 30 g/L oleic acid into 10-hydroxystearic acid without detectable byproduct after 4 h under anaerobic conditions, whereas the cells converted 30 g/L oleic acid into 20 g/L 10-hydroxystearic acid with a yield 67% after 3 h under aerobic conditions. These results indicated that anaerobic conditions were more suitable for the production of 10-hydroxystearic acid from oleic acid. Whole cells of S. nitritireducens exhibited the highest productivity and yield of 10-hydroxystearic acid from oleic acid among the previously reported studies

The soil bacterium Bradyrhizobium japonicum induces nodulation on the soybean root, resulting in formation of root nodules where bacteria fix atmosphereic nitrogen into ammonia. Rhizobial lipopolysaccharides (LPS) have been known to be an important factor for initiating and maintaining a symbiotic relationship with their host. In previous studies, a LPS gene region was isolated from B .japonicum strain 61A101C. In this study, we have selected one of the genes whose sequence is highly homologous to lpcC gene, encoding mannosyl transferase. We constructed lpcC knock out mutant to characterize the functions of the lpcC gene in B. japonicum. The lpcC null mutant showed distinct phenotypes from the wild type. The mutant failed to elicit nodules through in vivo plant nodulation test. To examine if LPS deficient mutant becomes sensitive to plant derived defense substance, especially to reactive oxygen species (ROS), wild type and the mutant strains were challenged with hydrogen peroxide at concentrations of 10-15μM. The mutant exhibited a significantly increased sensitivity to hydrogen peroxide. Further studies on the relationship between plant defense mechanism and nodulation are required.

Recently, bio-based fuels and polymers have become important due to the increase of oil price and concerns about environmental pollution. Lactic acid is a very important chemical having various applications in the field of pharmaceutical, cosmetics, textile, leather and food industries. Recently, lactic acid has drawn much attention for monomer of biodegradable polymer, Poly Lactic Acid (PLA). Lactic acid is able to be produced by chemical synthesis or bacterial fermentation, but none of chemical production routes shows technical and economical viability. In this presentation, we will show the production of lactic acid using recombinant lactic acid bacteria.

Recently, bio-based fuels and polymers have become important due to the increase of oil price and concerns about environmental pollution. Gamma aminobutyric acid (GABA) is the non-essential amino acid, which has recently found its possible application for monomer of nylon 4, the biodegradable heat-resistant polymer. Because direct synthesis of GABA from unrelated carbon source such as glucose is hard to achieve, conversion of L-glutamate (MSG) into GABA by enzyme catalysis or whole cell reaction becomes important in an industrial point of view. Several Lactobacillus strains have been reported to naturally convert MSG to GABA through α-decarboxylation of MSG catalyzed by pyridoxal phosphate-dependent glutamate decarboxylase. In this presentation, we report several recombinant bacteria system to achieve high conversion yield of MSG to GABA

Porcine epidemic diarrhea virus (PEDV) is a causative agent for acute enteritis and lethal watery diarrhea in pigs, particularly in neonates, with a high mortality rate. Outbreaks of the disease were reported from numerous European countries as well as Korea, China and Japan. PEDV, a member of the family Coronaviridae, is an enveloped virus possessing a single-stranded positive-sense approximately 28 kb RNA genome. A cell culture system for PEDV propagation was developed and successfully used for virus isolation. However, molecular technologies have been commonly exploited for rapid detection of viruses before detection of virus propagation. Efficient molecular methods for high throughput detection of diverse types of PEDV are not feasible yet. Among several genes of PEDV genome, genes coding for the M protein (a structural membrane glycoprotein, which plays an important role in the assembly process) and the S surface glycoprotein (harbors the specific host cell receptor binding sites) were commonly used for the identification and immunological purpose. In this study, we analyzed diversity of gene sequences of M and S proteins available in public databases and selected gene regions with high variability which could be selected as candidates for design of specific probes.

The effects of Sonication on Pervaporation for recovery of n-butanol from aqueous solutions using a silicone rubber tubing was investigated. Temperature (37 °C, 50 °C and 60 °C) and n-butanol feed concentration (10-50 g/L) were varied for the investigation. Permeate n-butanol concentration as well as flux increase with increase in temperature and n-butanol feed concentration. As for the effect of sonication, the permeate n-butanol concentration was enhanced by as much as 20% as compared to pervaporation with no sonication.

Coffee is one of the most popular beverages and traded agricultural commodities in the world. Coffee has been shown the positive or negative impact in humans. Lactobacillus species are used industrially for the production of foods, probiotics and biotherapeutics. Various Lactobacillus sp. were grown on coffee medium and the effect of coffee on the growth was observed. It was found that Lactobacillus sp. were able to grow in coffee media. L. helveticus, L. delbrueckii, L. fermentum, L. plantarum, L. reuteri, L. casei and L. paracasei were used in this study. We cultured the strains in coffee medium and MRS medium. Lactobacillus helveticus , Lactobacillus delbrueckii and other lactobacillus showed different growth patterns when they were grown on coffee medium.

Bdellovibrio bacteriovorus(BALOs) is predatory bacteria, small rod shape, and very motile. BALOs grow to predate other bacteria, especially gram negative bacteria. Many people tried to figure out an easy way to study of BALOs predation. But they are very small , so it was very hard to show optically. So, this study focused on the prey bacteria which were various based upon the optical properties. For this study, pUCDK and pHKT3 were transformed into E.coli MG1655, and they show bioluminescence and red fluorescence, respectively. First, test was MOI test. This test result show bioluminescence decreased depends on BALOs number. In second test, it shows bioluminescence and optical density depend on predation time and BALOs number. For making different BALOs, BALOs were treated different physical conditions. Bioluminescence was decreased depend on predation time, and BALOs numbers. For visible effect we tested not only biolumenescence but also fluorescence. Fluorescence is RFP, but RFP was not decreased like bioluminescence, so we determined biuminescence is better than fluorescence for visible predation test. For different BALOs number, BALOs treated different method, filteration or centrifugation or both, and centrifugation is most effective. So, determine centrifugation affection, we change the centrifuge speed, and predation test was done. The result was centrifuge speed is effected collect BALOs number. References 1. Carey Lambert, Karen A Morehouse, Chien-Yi Chang and R Elizabeth Sockett, Bdellovibrio: growth and development during the predatory cycle, Bdellovibrio: growth and development during the predatory cycle(2006), ScienceDirect, 9:639-6442. Amy C. Vollmer, Shimshon Belkin, Dana R, Smulski, Tina K, Van Dyk, and Robert A. Larossa, Detection of DNA Damage by Use of Escherichia coli Carrying recA'::lux, uvrA'::lux, or alkA'::lux Reporter Plasmids(1997), Applied and Environmenal Microbiology, 2566-2571

Gene expression involves the modification and mobilization of nucleosomes, the fundamental unit of chromatin. Nucleosomes are mobilized by chromatin remodeling complexes to alter DNA-histone contact and reveal DNA elements recognized by transcriptional factors. RSC is a nucleosome-remodeling complex essential for growth that can alter histone-DNA interaction by using the energy of ATP hydrolysis. The yeast RSC complexes contain RSC8. To confirm the existence of the complex, we tagged Rsc8 with FIAG tag and purified them in A. nidulans. Also, the rsc8 conditional mutant strain was generated to test the intracellular localization pattern and its function. This mutant has only one full-length rsc8 gene under the control of inducible promoter, alcA(p), and a truncated 5' rsc8 sequence. When mCherry-RSC8 fusion was induced on media containing glycerol as the sole carbon source, RSC8 produced a smaller colony size compared to the control R153 strain. To test whether rsc8 mutation would suppress the loss-of-function sidB mutation, we generated the sidB-rsc8 double mutant. On rich media containing glucose, the rsc8 mutation showed lethal phenotype indicating that the rsc8 mutation didn’t suppress the sidB mutation. On minimal media containing glycerol as the sole carbon source, the rsc8 mutation partially suppress the sidB mutation. Also, we checked its protein localization pattern. As a result, rsc8 localized at nucleus. In this observation, we confirmed the RSC8 mutation did not affect activity of SIDB.