Earticle

Vaccine is a protective clinical measure capable of persuading immune system against infectious agents. Vaccine can be categorized as live attenuated and inactivated. Live attenuated vaccines activate immunity similar to natural infection by replicating living organisms whereas inactivated vaccines are either whole cell vaccines, eliciting immune response by killed organisms,or subunit vaccines, stimulating immunity by non-replicating sub cellular parts. The components of vaccine play a critical role in deciding the immune response mediated by the vaccine. The innate immune responds against the antigen component. Adjuvants represent an importantcomponent of vaccine for enhancing the immunogenicity of the antigens. Subunit vaccines with isolated fractions of killed and recombinant antigens are mostly co-administered with adjuvants. The delivery system of the vaccine is another essential component to ensurethat vaccine is delivered to the right target with right dosage form. Furthermore, vaccine delivery system ensures that the desired immune response is achieved by manipulating the optimal interaction of vaccine and adjuvantwith the immune cell. The aforementioned components along with routes of administration of vaccine are the key elements of a successful vaccination procedure. Vaccines can be administered either orally or by parenteral routes. Many groups had made remarkable efforts for the development of new vaccine and delivery system. The emergence of new vaccine delivery system may lead to pursue the immunization goals with better clinical practices.

In this study, we investigated the ability of luteolin, a plant derived flavonoid on hepatocarcinoma cell growth using HepG2 cell culture system. We found that luteolin increased the Smac/DIABLO releases, a mitochondrial protein that potentiates apoptosis. Luteolin also induced either transcriptional activity or expression of PPAR-gamma, a target of cancer growth that PPAR-gamma agonist sensitizes to apoptosis in certain cancer types. To find the possible upstream target molecules of PPAR-gamma activated by luteolin treatment, we used compound C, a specific inhibitor of AMP-activated protein kinase. Pre-treatment of Compound C significantly restored the activation or expression of PPAR-gamma stimulated by luteolin. This result indicated that AMPK signaling might be involved in the activation or expression of PPAR-gamma signaling pathway stimulated by luteolin. Moreover, we also found that luteolin inhibited the insulin-stimulated Akt phosphorylation as well as AICAR, a specific AMPK activator. These results propose that luteolin significantly induces cancer cell death through modulating survival signal pathways such as PPAR-gamma and Akt. AMPK signaling pathway may be an upstream regulator for survival signal pathways such as PPAR-gamma and Akt stimulated by luteolin.

Marine bacterium producing pigment was isolated from the solar saltern of Mijo-myeon, Namhae, Korea. Based on phenotypic characteristics and 16S rRNA sequence analysis, the strain was identified as Kocuria sp., which produced a yellow pigment. The pigment showed UV absorption maximum at 469nm. The bacterial strain grew well on Marine broth 2216 culture medium. Productivity of the pigment reached the maximum value after 44 hours at 30℃, 2% NaCl and pH 6.0. The pigment was produced best when supplied by 1% lactose as a carbon source and 1% beef extract as a nitrogen source. The result of the color stability study showed that pigment extracted from the strain by ethanol was stable at -20-25℃ and also showed higher stability over 70% for 14 days in light conditions at 25℃. The pigment extract was also stable for all metal ions tested, except for FeCl2.

In this work we have investigated the production of catalase from Bacillus sp. strains, which were screened and identified from soil. These strains were cultivated in shaking flasks with tryptic soy broth (TSB) at 30℃ and 200 rpm. Effects of the temperature and pH on the stability of the native catalase and whole cell viability were studied in the temperature range of 25-60℃ and the pH range of 7-13. Korean natural zeolite was added to culture medium and mixed with microorganisms for 24 hours. The native catalase maintained its activity over 50℃. The enzyme acitiviy of the catalase from Bacillus flexus BKBChE-3 was highest among the Bacillus sp. strains studied. Bacillus flexus BKBChE-3 and immobilized Bacillus cells have survived under extreme conditions of over 50℃ and pH 12. 60 mL of 10.5 mM H2O2 solution were entirely removed within 1 hour with catalase produced from Bacillus sp. on the flask. When Bacillus cells were immobilized on Korean natural zeolite, colony forming unit of Bacillus flexus BKBChE-3 was increased and high efficiency of hydrogen peroxide removal was observed.

A thiazolidinedione derivative, TD49 with the highly selective algicide to red tide was newly synthesized and its acute toxicity was examined in order to evaluate the effect on aquatic ecosystems of coast. Major three species having important role in the food chain of marine ecosystem, such as Skeletonema costatum of microalgae, Daphnia magna of crustacea, Paralichthys olivaceus of flatfish fingerling were employed for the acute toxicity assessment. EC50 or LC50 as the assessment criterion was investigated to each specie, and NOEC (No Observed Effect Concentration) and PNEC (Predicted No Effect Concentration) from most sensitive specie to toxicity of TD49 were further calculated. EC50 of S. costatum in 96-hour, EC50 of D. magna in 48-hour, and LC50 of P. olivaceus in 72-hour for TD49 were 0.34 μM, 0.68 μM, and 0.58 μM, respectively. NOEC from the results of S. costatum was estimated to be 0.20 μM and PNEC was estimated as 3.40 nM by applying factor value of 100 to EC50 0.34 μM of S. costatum. In addition, it was revealed that Solutol used as the dispersing agent of TD49 had very little toxic influence under the concentration range of 0~0.4 μM used in TD49 toxicity experiment. Although the estimated concentration of TD49 that will be sprayed onto the coastal field for the algicide is higher than NOEC value, it is considered that the spraying concentration would not be a considerable problem due to a dilution effect by tide at the opened coast.

We investigated the effect of inhibitory compounds derived lignocellulosic hydrolysates on cell growth, sugar consumption and ethanol productivity, and also we intended to identify the potential for ethanol production based on lignocellulosic hydrolysates. Cell growth and ethanol production in the presence of acetate were initiated after 12 hr. Furans showed a longer lag time and phenolics showed a significant effect on strain and ethanol production in comparison to other model compounds. In the case of lignocellulosic hydrolysates, the acetate strongly affected cell growth and ethanol production.

In this work, mass production of Bacillus licheniformis SCD121067 through medium optimization by statistical experimental method was studied. First, galactose, yeast extract and potassium phosphate dibasic were selected as carbon, nitrogen and phosphate sources for mass production of B. licheniformis SCD121067 by using one factor at a time method. Second, according to the result of Plackett-Burman experimental design, key factors was yeast extract and K2HPO. Finally, the response surface methodology was performed to obtain the optimum concentrations of two selected variables. The optimized medium composition consisted of 20 g/L galactose, 36 g/L yeast extract, 0.41 g/L K2HPO4, 0.25 g/L Na2CO3, 0.4g/L MgSO4 and 0.01g/L CaCl2. Dry cell weight (15.4 g/L) by optimum production medium were increased 10 times, as compared to that determined with basic production medium (1.5 g/L). Fermentation conditions were examined for the mass production of B. licheniformis. The effect of temperature, agitation speed, pH and aeration rate on the mass production of B. licheniformis were also studied in a batch fermenter which was carried out in a 2.5 L bioreactor with a working volume of 1.5 L containing optimized production medium. As a result, dry cell weight of batch culture was 30.7 g/L at 42℃, 300 rpm, pH 8.0 and 2 vvm.

The recent bloom of a very large jellyfish Nemopilema nomurai has caused a danger to sea fishery and sea bathers. Presently, Nemopilema nomurai is thrown away through a separator system in the sea. The objective of this work was to produce bio-gas from Nemopilema nomurai by using anaerobic digestion. The bio-gas includes the hydrogen or the methane gases. It relates that Nemopilema nomurai is effectually changed into the renewable energy. When the jellyfish biomass was used as an organic carbon source the bio-gases were evolved. The aim of this study was to determine the optimal conditions for hydrogen and methane gases production according to the substrate concentrations of Nemopilema nomurai, optimal culture condition and the sludge-pretreatment without pH control. The optimal culture condition was found to be 35℃ and the heat-treatments of jellyfish was done at 120℃ for 30 min. The production rate of hydrogen and methane gas were found to be 8.8 mL/L/h, 37.2 mL/L/h from 1.5 g of dry Nemopilema nomurai.

Increasing demands on fossil fuel have led to the unprecedented attraction to microalgal biofuel as an alternative energy. In this study, we investigated growth and lipid productions of microalga Nannochloris oculata under various carbon dioxide or nitrogen source concentrations and irradiance conditions. Biomass production of N. oculata was highest under 2% CO2 with 0.3 flow rate (vvm). In addition, biomass productivities were proportional to the concentration of nitrogen source, whereas lipid biosynthesis was suppressed under higher nitrogen concentration (up to 50 mg/L). High irradiation (160~180 μmol/m2·s) enhanced growth rate and lipid production of N. oculata.

A new hydrogen-oxidizing bacterium, Aeromonas sp. strain JS-1, that can fix CO2 via the reductive pentose phosphate cycle (Calvin-Benson cycle) under chemoautotrophic conditions but not photoautotrophic conditions was isolated from fresh water. Strain JS-1 showed considerable CO2 fixation ability during continuous cultivation even at high CO2 concentration. Strain JS-1 used H2 and CO2 fixation as energy and carbon sources, respectively. Carbon dioxide fixation is carried out through the Calvin-Benson cycle, in which ribulose-1,5-bisphosphate carboxylase/ oxygenase (RubisCO) is the key enzyme. Hydrogen-oxidizing chemoautotrophic Aeromonas sp. strain JS-1 exhibited remarkedly strong RubisCO [EC 4.1.1.39] activity. RubisCO was purified as an L8S8-type hexadecamer with molecular mass of 560 kDa by gel filtration. The enzyme consisted of two different subunits eight large (56 kDa) and eight small (15 kDa), as demonstrated by SDS-PAGE. The specific activity of the purified enzyme was about 3.31 unit/mg and stable up to 45℃. The Km values for RuBP, CO2, and Mg2+ were estimated to be 0.25 mM, 5.2 mM and 0.91 mM, respectively.

To develop thermostable ethanol fermentative yeast strain for lignocellulosic simultaneous saccharification and fermentation, high ethanol producing yeast, Saccharomyces cerevisiae CHY1012 and thermostable yeast, Kluyveromyces marxianus CHY1703 were fused by protoplast fusion. The thermostable fusant, CHY1612 was identified as a Kluyveromyces marxianus by phenotypic and physiological characteristics, as well as molecular analysis based on the D1/D2 domains of the large subunit (26S) rDNA gene and the internally transcribed spacer (ITS) 1 + 2 regions. For lignocellulosic ethanol production, AFEX pretreated barley straw at 150℃ for 90 min was used in a simultaneous saccharification and fermentation (SSF) process using thermotolerant CHY1612. The SSF from 16% pretreated barley straw at 43℃ gave a saccharification ratio of 90.5%, a final ethanol concentration of 38.5 g/L, and a theoretical yield of 91.2%. These results show that K. marxianus CHY1612 has potential for lignocellulosic ethanol production through simultaneous saccharification and fermentation with further development of process.

Biological products, such as live varicella vaccine, are composed of biological substances derived from biological organisms. It is very difficult to identify these biologics’ characteristics by analysis of simple physical and chemical methods alone. So the reference material is essential in order to evaluate the quality of bilogics. The 1’st national standard for varicella live vaccine was manufactured, established in 2002 and 2003, and have been used for the manufacturer’s quality control and national lot release since then. As the lack of its availability and the decrease of its stability, this study was initiated by National Institute of Food and Drug Safety Evaluation (NiFDS) in 2008 to manufacture and establish the 2nd national standard for varicella live vaccine. The candidate material was manufactured from one of domestic manufacterers and the joint research of the NiFDS and manufacturers of varicella live vaccine was conducted to estimate of the reliable virus content. In the collaborative study, 3 laboratories including NiFDS performed the virus content test more than 7 times and all assay results were statistically analyzed. The mean coefficient of variation (CV) was 1.24%, and the geometric mean titre (GMT) variation range of each laboratory was low. On the basis of the results of this study, the candidate material of 2nd national standard for varicella live vaccine was assigned a potency of 4.26 log10 pfu/0.5 mL, when reconstituted in 0.7 mL.