Earticle

Successful development of an algae-based bioproducts industry offers world-wide benefits related to sustainable bioenergy and bioproducts production, and environmental benefits. Based on what we have learned, major challenges to the implementation of the industry include cultivation, harvesting, and processing of algae biomass at full-scale. The majority of the research and development of algae-based technologies has emphasized suspended growth of single or mixed cultures in open ponds and closed reactors. However scientific and technological investigations of attached algal biofilm cultures has received minimal attention. This presentation will explore algae science and technology within the context of biofilm reactions and bioreactors. The goal is to develop full-scale algae biorefineries that are technologically efficient and cost-effective.

Human skin is unique. Compared to those of other animals, human skin has less hair and has various colors, such as white, yellow and black. Skin, especially that of the face, is a big factor in determining one’s first impression. Although the human skin color is thought to have been genetically developed depending on their ancestor’s first region of residence, humans tend to prefer certain skin colors, especially those of face. Asians have a tendency to prefer white skin for various reasons. This preference is the basis for the development of depigmenting cosmetics. Functional cosmetics or cosmeceuticals are cosmetics possessing the abilities of curing or improving skin health. In this short review, I would like to address the following research topics; pigmentation process, strategy and development of biochip for depigmenting agent screening.

Biomimetic approaches can provide alternative and more efficient ways of preparing functional substrates and scaffolds for stem cell engineering. One of the interesting molecules for this purpose is 3,4-dihydroxy-Lpheylalanine (dopamine), an unusual amino acid repeatedly observed in Mytilus edulis foot protein-5 of mussel adhesive pads. Mussel adhesive protein promotes mussel adhesion to virtually any type of organic or inorganic material, which is attributed to an extensive repeat of dopamine and lysine residues in the mussel adhesive pads. Dopamine containing both catechol and amine groups polymerizes at an alkaline pH, which is typical of marine environments, to form a polydopamine layer. In this presentation, a mussel-inspired, biomimetic approach will be introduced for surface modification for efficient and reliable manipulation of human neural stem cell (NSC) differentiation and proliferation. Our study demonstrates that polydopamine coating facilitates highly efficient, simple immobilization of neurotrophic growth factors and adhesion peptides onto polymer substrates. The growth factor or peptide-immobilized substrates greatly enhance differentiation and proliferation of human NSCs at a level comparable or greater than currently available animal-derived coating materials (Matrigel) with safety issues. Therefore, polydopaminemediated surface modification can provide a versatile platform technology for developing chemically defined, safe, functional substrates and scaffolds for therapeutic applications of human NSCs.

Homogeneous immunoassays are attractive analytical technologies in food and environmental analyses because they are superior to heterogeneous immunoassays in sample and reagent consumption, analysis time, portability and disposability. Herein, we have developed fluorescence resonance energy transfer (FRET) homogeneous immunoassay for the detection of mycotoxins, such as aflatoxin B1 (AFB1), ochratoxin A (OTA), and zearalenone (ZEN). Owing to intrinsic fluorescence properties of protein and mycotoxins, antibody and antigen played roles as the respective donor and acceptor in the FRET immunoassay. The FRET immunoassay, based on intact antibody, displayed high specificities and detection limits of 1 and 0.85 ng mL-1 for OTA and AFB1, respectively. Furthermore, the sensitivity of Fab fragment-based FRET immunoassay was obtained 10-fold enhancement comparing to the intact antibody-based immunoassay. Practical applications of the FRET immunoassay system were demonstrated by their use in the detection of OTA or AFB1 in spiked grain samples. The observations made in these efforts show that the homogeneous FRET immunoassay strategy, which requires a simple sample preparation procedure, serves as a powerful tool for the rapid and sensitive quantitative determination of mycotoxin in food and environmental samples.

One of the most outstanding features of Saccharomyces cerevisiae sake yeast strains is the high fermentation rate in the sake mash. The molecular mechanism underlying this brewing property has been remained unrevealed, however. Our genome-wide comparative analyses of sake yeast and laboratory yeast strains led to identification of mutations responsible for the high fermentation ability of sake yeast.To identify differences between the sake and laboratory yeast strains that contribute to the superior brewing properties of sake yeast, we compared the gene expression profiles of both types of yeast in fermenting sake mash. Sake yeast showed remarkably reduced expression through Msn2/4p and Hsf1p, representative stress-responsive transcription factors in Saccharomyces cerevisiae. In contrast, yeast stress responses and resultant stress tolerance are generally considered to be important characteristics for effective ethanol fermentation. Therefore, we examined stress tolerance of sake yeast cells in detail, and found that sake yeast exhibited lower survival rates than laboratory yeast under heat shock or ethanol stress. These results demonstrate that sake yeast is defective in stress response in spite of the high fermentation ability.To elucidate the genetic variations responsible for the defective stress response of sake yeast, we searched for mutations in the genes related to Msn2/4p and Hsf1p by utilizing the whole genome sequence data of a representative sake yeast strain Kyokai No. 7. Consequently, we found novel mutations in Kyokai No. 7, including (i) a single nucleotide substitution that causes deletion of the zinc-finger motifs located at the carboxyl terminus of Msn4p, which abrogates the functions of Msn4p as a transcriptional activator; (ii) a single nucleotide insertion that leads to a frameshift of the RIM15 gene, encoding a protein kinase acting upstream of Msn2/4p; and (iii) loss of the entire PPT1 gene locus, whose product is related to regulatory dephosphorylation of Hsf1p. It is noteworthy that all these mutations were distributed only in the sake strains genetically close to Kyokai No. 7. Furthermore, we confirmed that each of these mutations significantly improved the fermentation rate of laboratory strains. Altogether, these sake yeast-specific mutations originate both the stress-sensitive phenotype and the high fermentation rate of sake yeast strains.These results provide novel insights into yeast stress responses as major impediments of effective ethanol fermentation, which are widely applicable to the development and improvement of advanced yeast strains in the bakery, brewing, and biofuel industries.

Single-chain Fv antibodies that VH and VL domains of whole antibodies (whole Abs) are genetifally linked via a flexible linker (G4S)3, have been expected as a next generation of ligand molecular recognition agent in clinical diagnosis. ScFvs can be efficiently produced in an insoluble fraction of recombinant E. coli cells with much lower production cost than the conventional whole Ab, while immobilization method of scFv with high density, activity and uniform orientation is requisite to increase antigen-binding activity of scFv on the solid support to great potential extent. Recently, we succesfully identified the affinity peptide tag which possessed strong binding affintiy against the surface of hydrophilic polystyrene (phi-PS) support for immunodiagnostics. By genetic fusion or chemical conjugation of PS-tag with target proteins such as enzyme as well as antibody, they were able to be immobilized with maintaining much higher density as well as remaining activity due to site-specific attachment of PS-tag onto the surface of phi-PS plate. Especially, it was revealed that use of PS-tag-fused scFvs (scFv-PS) as a ligand antibody resulted in enhancement of sentivity 10 times higher than that of conventional whole Ab. Furthermore, a solid-phase refolding method that refolding and immobilization processes of scFv-PS were simultaniously performed, was successfully developed. Consequently, production cost of scFv-immobilized immunodiagnostic support could become one-tenth or further lower than that of whole Ab-immobilized support. This method was applicable to a variety of antibody species. Thus, the immobilization method for PS-tag-fused scFv developed in this stugy will be considerably useful for production of high-performance antibodyimmobilized diagnostic support.

Microengineering techniques such as lab-on-a-chip technologies are emerging as the next revolution in tools for high throughput cell-based analysis. Cell is the basic unit of life of all organisms and exerts its function by pericellular microenvironment, such as extracellular matrix and other cells adjacent to individual cells. Therefore, to understand, control and predict cellular responses, it is important to build the biomimeticevaluation system. With the development of micro-and nano-technology devices, variety of tools is becoming available for the analysis of cells at the cellular and molecular levels. Since single-cell analysis enables to capture the minimal change within a living organism, it is emerging as less-invasive approach for clinical diagnosis and cell identification. In this study, we developed technologies using magnetic nanoparticles for cellular analysis of individual cells, such as magnetic force-based cell patterning and droplet-based gene expression analysis. Using these methods, the behavior of cancer cells was observed a in the three-dimensional cell culture array along with gene expression analysis.Magnetic force-based cell patterning was conducted using a pin-holder device fabricated by processing electromagnetic soft iron in which each pillar is aligned at center-to-center distance of 250 μm for magnetic convergence. Using this pin-holder device with Nd-B magnet underneath, magnetically labeled cells were allocated on the cell culture dish at single and multiple cells levels by varying the cell seeding density. The magnetic cell patterning technique has been applied for evaluation of invasive capacity with tumor cells in a three dimensional cell culture array, and cell susceptibility assay was performed. Also, a droplet manipulation system using superparamagnetic nanoparticles and magnetic force was developed for gene expression analysis at single cell levels. The magnetic force-based-droplet handling system can be used as a powerful tool for various biochemical applications by moving and coalescing sample droplets using magnetic beads immersed in mineral oil. We optimized the conditions for single cell lysis and RT-PCR analysis and applied this system for detection of Wilms tumor gene mRNA, that is useful as an indicator of minimal residual disease in leukemia. The developed system is quite simple and allows for the handling of droplets and preceding reactions using a magnet without external pumps and valves. These systems are greatly advantageous for cell analysis and related diagnostic applications.

Topical transcutaneous immunization (TCI) presents many clinical advantages, but its underlying mechanism for the induction of mucosal immunity remains unknown. TCI induced Ag-specific IgA Ab-secreting cells expressing CCR9 and CCR10 in the small intestine in a retinoic acid-dependent manner. These intestinal IgA Abs were maintained in Peyer’s patch-null mice but abolished in the Peyer’s patch- and lymph node-null mice. The mesenteric lymph node (MLN) was shown to be the site of IgA isotype class switching after TCI. Unexpectedly, langerin+CD8a- dendritic cells (DCs) emerged in the MLN after TCI; they did not migrate from the skin but rather differentiated rapidly from bone marrow precursors. Depletion of langerin+cells impaired intestinal IgA Ab responses after TCI. Antigens in the intestine transition from skin tissue through the lamina propria into the draining lymphatics, but whether or how this process controls mucosal immune responses is unclear. We demonstrate that small intestinal CX3CR1+ DCs of the lamina propria sample and process both circulatory and luminal antigens. Through cross-presentation of antigen CX3CR1+ DCs induce differentiation of IL-10 secreting CD8a/b T regulatory cells. Taken together, these findings suggest that mucosal DCs are indispensable for the induction of intestinal IgA Abs following skin immunization and that crosstalk between the skin and gut immune systems.

Following repeated reverse-zoonoses of the pandemic H1N1 (pH1N1) 2009 virus in swine populations, novel reassortants between pH1N1 and endemic swine viruses have been detected from pigs worldwide. Through an active surveillance program in pig abattoirs between fall 2011 to spring 2012, we identified reassortant H3N2 and H3N1 swine viruses in Chungbuk Province, South Korea. Whole-genome characterization revealed that the reassortant H3N2 viruses are more genetically closely related to novel H3N2 (rH3N2p) viruses recently detected in North America containing the pandemic matrix gene, which also caused sporadic human infections, rather than previous Korean isolates. In contrast, the variant H3N1 reassortant (rH3N1p) viruses represent four novel genotypes in the backbone of pH1N1. Comparison of growth properties in vitro showed that selected rH3N2p and rH3N1p viruses had comparable kinetics in MDCK and A549 cells but the variant rH3N1p viruses exhibited growth advantage in HBE cells; only three of the 4 rH3N1p viruses persisted in the lungs of infected mice up to 7 dpi but none demonstrated altered virulence. All viruses tested replicated well in the upper respiratory tract of ferrets. Furthermore, serology indicated that these H3 reassortant viruses are antigenically divergent from recent human seasonal H3N2 strains suggesting previous exposure or vaccination may provide limited immunity against these viruses. Establishment of these novel viruses in Korean pigs could increase genetic diversity of endemic swine viruses which may pose threat to public health. Data underscores the need for continuous monitoring to evaluate genetic evolution in this ecologically important virus reservoir.

Backgrounds / Objectives: Cholera toxin (CT) is a strong systemic and mucosal adjuvant that greatly enhances IgG, IgA and cellular immune responses. The activity of CT as a mucosal adjuvant has been confirmed with a variety of antigens. The adjuvant properties of CT have been studied in animal models using both in vivo and in vitro experiments. However, CT is a strong enterotoxin. It was known that oral administration of 10ug of CT is sufficient to induce significant diarrhea in humans suggesting that toxicity may limit the clinical use of CT. Experimentals / Methods: CT A1 subunit (CTA1) was genetically fused with protein transduction domain (PTD) which has been known to deliver fused protein highly efficiently into the cells after non-specific binding to the cell surface. To enhance the cellular uptake, we attached PTD at both N-and C- termini of CTA1 (TCTA1T) and analyzed feasibility of TCTA1T as an adjuvant on humoral and cellular immune response. Results: Intranasal delivery of recombinant TCTA1T which was purified from Escherichia coli with ovalbumin (OVA) significantly augmented OVA-specific both systemic and mucosal immune responses which were comparable to that of CT. The adjuvant activity of TCTA1T was enzyme dependent, because the immune response induced by enzymatically inactive TmCTA1T was significantly lower than that of TCTA1T. It also dramatically increased the number of OVA-specific antibody secreting cells and in vivo CTL activity in the lung and spleen. In addition, mucosal administration of TCTA1T resulted in an increase of mucosa-derived dendritic cells in draining lymph node, which is similar to native CT. However, in contrast to CT, TCTAlT was not either reactogenic or toxic in mice and its ADP-ribosyltransferase activity was much lower than that of CT (≈ 5%). Conclusions: Our results demonstrate that non-targeted delivery of enzymatic active TCTA1 is non-toxic and has immune-modulating activity, suggesting a new approach to develop a safe and effective mucosal adjuvant.

The membrane attack complex (MAC) generated after complement activation readily lyses the plasma membranes of most Gram-negative bacteria, but it poorly lyses those of Gram-positive bacteria because of their thick peptidoglycan layers (1). Thus, many reports have suggested that serum-antibody-mediated opsonophagocytotic bacterial killing is necessary to increase the bactericidal efficiency of the MAC against pathogenic Gram-positive bacterial infection (2). Wall teichoic acid (WTA) of Staphylococcus aureus is a major cell envelope-associated glycopolymer that is a key molecule in promoting colonization during S. aureus infection. We recently reported that S. aureus WTA functions as a ligand of human serum mannose-binding lectin (MBL), a recognition molecule of the lectin complement pathway (3). Intriguingly, serum MBL in adults does not bind to WTA because of an inhibitory effect of serum anti-WTA-immunoglobulin (IgG). Here, to examine the biological function of human anti-WTA-IgG, serum, anti-WTA-IgG was recently purified to homogeneity using a purified S. aureus WTA-coupled affinity column. The purified anti-WTA-IgG contained the IgG2 subclass as a major component and specifically induced C4 and C3 deposition on the S. aureus surface in the anti-WTA-IgG-depleted serum, but not in C1q-deficient serum. Furthermore, the anti-WTA-IgG-dependent C3 deposition induced phagocytosis of S. aureus cells by human polymorphonuclear leukocytes. These results demonstrate that serum anti-WTA-IgG is a real trigger for the induction of classical complement-dependent opsonophagocytosis against S. aureus. Our results also support the purified WTA of S. aureus can be used as putative vaccine candidate and the purified human anti-WTA-IgG can be useful for the plausible therapeutic antibodies against methicillin-resistanr S. aureus (MRSA). Furthermore, our results demonstrate that a lack of the lectin complement pathway in MBL-deficient adults is compensated by antigen-specific antibody-mediated adaptive immunity.

GreenPol is the tradename of the SK's environment friendly CO2 polymers synthesized from carbon dioxide and propylene oxide, etc. Microcellular polypropylene carbonate foams produced by a CO2 foaming process and their characterization regarding the influence of the pressure drop rate on the foaming behavior and the property of the final foamed products.GreenPol foams can be adapted to many applications such as construction thermal insulation, package for food and drink, and disposable tray etc.In this reasearch we investigate the GreenPol foams' possibility as an construction thermal insulation.We made thermally stable and high expanded poly(propylene carbonate) foams for construction thermal insulation. Most of the cell structures of this product were closed cells and the foamed ratio of the GreenPol foam were from 10 to 30 times comparing with the mini-pellet which is the base material before foaming process.The estimated demand of the foams for the construction application is more than 4 million tons, and the damand of the foams for the packaging is more than 1.5 million tons.

Development of eco-friendly coating materials from renewable resources with special properties has attracted great attention in recent years. Among the renewable resources, cardanol, a meta-substituted phenol from the cashew nut shell liquid (CNSL), has a variety of advantages due to its unique properties such as improved flexibility and better processability over conventional phenolic resin, heat and electrical resistance, and lower price, etc. Herein, a new eco-friendly coating material was developed from a cardanol without any purification step. It could form transparent, highly glossy film. And surface hardness of the film is higher than that of previously reported poly (cardanol) and cardanol derivatives. Additionally, it showed antibacterial property against E-coli and P. aeruginosa. Conclusively, this coating material is supposed to be a commercially promising candidate in surface coating material market.

‘Development of Environmental Friendly New Constructional Finishing Materials utilizing bio/nano-technology’ can be defined as a development of the most advanced construction materials overcoming limitations in the functions and performances of already existing construction materials by means of environmentally-focused BT/NT convergence technologies resulting in being at the cutting edge of future construction business. PLA, as raw materials of carbon dioxide-reducible biodegradable film with biaxial stretching in that it is used for decorative materials in the field of eco-friendly construction. Flexibility and manufacturing equipment are key technologies of the PLA films. L/D ratio of the PLA enantiomer is able to control the flexibility with additives and PET manufacturing equipment is applid to PLA films with low cost.

Nowadays, as biodegradable carbon-zero materials are inevitably required in all sectors of industry, such as the automobile, construction, electronics industries, lignin may be used as a novel, appropriate, and renewable feedstock. Lignin is an abundant natural material source and waste material. Most industrial lignins are obtained from kraft and sulfite pulping processes. In this work, eco-friendly lignin-based nanocomposites were prepared in a twin-screw extruder at 0-9 wt% of Montmorillonite (MMT) nanoplatelets. The feasibility of using lignin-based polymer as matrix or compatibilizer in composites made from polypropylene (PP) and jute fiber was studied. The properties of the specimens have been studied by the thermogravimetry (TGA), tensile tests and Izod impact tests. The effects of both the lignin based copolyester and the filler particles on tensile properties and izod impact strength were investigated.

The development of eco-friendly coating systems in recent years is a global concern. For this reason, we have developed VOC-free and solvent-free coating system. This coating systems the core technology is epoxy intumescent. This system started as epoxy resin-based corrosion coatings but have had their capabilities extended for fire protection. By their nature they, therefore, provide excellent corrosion resistance properties. This coating system is designed to prevent the calamity from fire, has excellent flame retardant and thermal insulating property. This is film coating that forms an insulating layers, noncombustible thermal barriers preventing flame and heat from contacting substrate, when exposed to fire. This is specially designed for structural steels and used to increase the steels fire endurance time for 3hours.

In 2011, WHO (World Health Organization) reported that electromagnetic waves via cellular phone and other usages can possibly be carcinogenic to human brain [1]. Metallic electromagnetic wave shielding materials have been often used in the past due to their high reflection loss. However, high price of the metals and need of high absorption loss properties required alternative electromagnetic shielding/absorbing materials including nano materials (e.g., carbon nanotube) added epoxy materials. Recently, carbon nanotube added materials having high electromagnetic wave shielding/absorbing characteristics were developed by a number of researchers and their capabilities were evaluated [see e.g., 2,3]. The present paper reviews recent works aimed to develop interior wall board systems with carbon nanotube added materials for shielding/absorbing harmful electromagnetic waves. Details of the summary of the review will be presented.

Although the biological technology and industry needs long-term investment and has high risk, it is the important emerging industry that brings high returns as one of the technology in many government departments supported to develop. In particular, the Ministry of Knowledge and Economy(MKE) has focused on various policies which could foster new industries by supporting companies, universities and research institutes. The Korea Evaluation Institute of Industrial Technology(KEIT) which performs comprehensive planning, evaluation and management of the government(MKE) grants has tried to discover lots of good challenges and create great performance in various industries. In this seminar, I would like to introduce the current trends of bio-industrial development, R&D programs supported by MKE, and the planning, evaluation and management systems of KEIT.

It is reported that the worldwide demands for crops and biomass exceed their supply due to the rapid growth of world population, limitation of cultivation areas, global warming, climate change and subsequent lack of water supply. Crop protection agents have played an important role in promoting productivity of crops yields and biomass. However, for some crop protection agents environmental safety issues and reports of the occurring resistant plant/insect to the introduced agrochemical are increasing. To overcome such problem, recently, researches for the development of new crop protection agents based on greenbio-technology such as searching for new potent molecules from natural resources (plant or microorganism derived organic compounds) and invention of agrochemical molecules with new mode of action as target for resistance control are being carried out actively. In this report the following topics such as development of new herbicides for resistant weeds, development of new insecticides for crop protection, and development of new LOHAS-based biochemical crop protection agents will be discussed.

As an alternative to the depletion of crude oil reserves, biomass-conversion process to produce fuel and chemicals from renewable resources has been developed worldwide. In 2010, Government announced that biochemistry was selected as a new growth engine to be competitive in chemical engineering and biotechnology industries in the world, and since then this project has been started. 2,3-BDO plays a key role in various biosynthesis of product with low cost. 2,3-BDO can be converted to methyl ethyl ketone by dehydration, which can be used as a liquid fuel additive, and converted to 1,3-butadiene, which is a critical building block for synthetic rubber. The aim of this project is the production of 2,3-BDO and its derivatives by biochemical process. Achievement of the aim can increase competitiveness of energy industry in the world.

최근 차세대 유전체 서열 해독기의 등장은 개인 유전체의 정보를 해독하는데 있어서 시간과 비용적인 측면에서 획기적인 혁명을 가져오고 있으며, 이런 저렴한 비용과 짧 은 해독시간은 개인 간의 유전적인 특성을 반영한 맞춤 의료를 가능하게 하고 있다. 이미 선진국에서는 유전체 정보에 기만한 맞춤 의료 서비스를 미래의 성장 동력으로 만들기 위하여 적극적인 투자를 하고 있으며, 이를 위한 기반 기술 개발을 위한 연구 가 활발히 이루어지고 있다. 맞춤 의료를 실현하기 위하여 현제 큰 문제점은 차세대 유전체 서열 해독기가 만들어 내는 대용량의 데이터를 빠르게 분석하고 해석하기 위한 알고리즘 및 기반 시스템의 구축이다. 차세대 서열 해독기가 만들어내는 정보의 양은 과거 생물학이나 IT분야에 서 다루던 단편적인 정보들과는 비교할 수 없을 만큼 크며, 이 정보가 개인의 유전체 특성을 분석하기 전인 전처리 단계에서 처리되어야 한다는 특징을 가지고 있다. 이를 처리하기 위해서는 이를 전담할 수 있는 대용량의 IT 인프라와 자동화된 시스템이 요 구되고, 이에 따라 이미 선진국에서는 유전체 연구 분야 중, 서열 데이터의 생산과 처 리에 있어 연구소, 대학 등 학문계를 탈피하고, 전문 기업이 해당 분야 기술을 주도하 고 있다. 서열 데이터 처리에서 가장 중요한 병목(bottleneck)을 해결할 생명정보학을 이용한 대용량 유전체 데이터를 분석하는 플랫폼이 필요하며, 미래 바이오산업의 기 초를 다기지 위해서는 기업 주도의 서열 데이터 처리 상용화가 시급할 실정이다. 본 발표에서는 빠른 시간 내에 유전체 정보를 처리하기 위하여 대용량 유전체 정보생 산과, 처리, 자동화, 파이프라인을 통합한 상용화된 소프트웨어 플랫폼 개발과 그 진 행상황에 대하여 소개하고 있다. 또한 이 플랫폼의 성공적인 상용화와 한국의 유전적 특성 반영하기 위한, 한국인 표준유전체의 필요성과 구축과정과 결과에 대한 부분을 포함하고 있다.

In this presentation, a new adaptive water quality management scheme based on real-time and/or near real-time water quality sensors and information technologies is proposed. The sensor-based adaptive management tries to facilitate scientists, engineers and policy makers to perform optimal management of streams, rivers and water infrastructure systems both within a given watershed and nation-wide. The specific elements of this scheme include a real-time sensor-based monitoring network, a predictive modeling framework based on data from sensor network. If a proper conceptual framework is established, multi-scale adaptive management of water and wastewater treatment facilities across a watershed can be possible. However, the approach relies heavily on information collected by sensors. Accurate data collected from properly designed sensors will reduce cost related with uncertainty of the system. Among various sensor technologies, the lab-on-a-chip (LOC)-based sensor technology may be the most suitable for the current water quality management system. Using the technology, small water quality analyzers following the Standard methods can be developed. The LOC-based analyzers can be applied to stream water quality monitoring, and process control for water treatment systems. In this presentation, therefore, the current research regarding the development of the LOC technology-based water quality analyzers and of the water treatment system utilizing the analyzers also will be introduced.

Lysosomes (LY) with antimicrobial (AM) and anticancerous (AC) functions, were altered by some stimuli factor. Unlike LY, peroxisomes (PE) have different cellular functions because they contain oxidative stress related enzymes. In this study, the LY activity under the several stresses have characterized to separate function of LY and PE based on subcellular distribution under several stressful conditions. The AM and AC activity of LY was enhanced as a result of subcellular distribution of LY and PE, and also affected the reduction of melanin pigments intensity. Aminopeptidase Y (APE3) was found as the highest expressed protein in response to oxidative stress by 2-DE. Then, the gene encoding for the APE3 gene was used to construct the recombinant Saccharomyces cerevisiae in order to enhance its activity in LY. As a result, recombinant S. cerevisiae have showed more enhanced AM and AC activity comparing with normal cells. Finally we found that a bacterial growth in blood was inhibited by LY, when we injected LY for therapy of bacterimia in rat. Our results indicated the increase in endogenous levels of LY and their activity following genetic modification may have various applications as AM agent and apoptosis-inducing materials for cancer cells. Therefore, it is very promising to use the organelles from genetic modified cells for improving in vitro and in vivo functions and PE extract as useful agent for skin lightening cosmetics and treatment of hyperpigmentation disorders.

It becomes important to develop novel biotechnologies that can subside the global climate change problem, mainly revoked by emission of carbon dioxide (CO2). Since carbonic anhydrase (CA, EC 4.2.1) accelerates the hydration of CO2, CA-based CO2 capture tool is considered as one of the potential systems for solving the problems. Recently, we cloned several types of CAs from marine microalgae or marine bacteria and we re-constructed their sequences optimized for highly-efficient production in E. coli system. Several CAs were obtained successfully in microbial expression system and their properties were characterized in terms of their CO2 sequestration ability. In particular, the acquired CAs could mediate catalysis of the dissolved CO2 to bicarbonate (HCO3 -), which can be precipitated as CaCO3 in the presence of Ca2+. CA-based CO2 mineralization can be used as potent platform technology for CO2 capture and use. Furthermore, HCO3 - mediated by CAs can be attached with PEP (phosphoenolpyruvate) by phosphoenolpyruvate carboxylase (PEPCase, EC 4.1.1.31) to make four-carbon organic acid, oxaloacetate (OAA). These approaches based on marine CA will be platform system for development of efficient CO2 capture/use technologies.

It becomes important to develop novel biotechnologies that can subside the global climate change problem, mainly revoked by emission of carbon dioxide (CO2). Since carbonic anhydrase (CA, EC 4.2.1) accelerates the hydration of CO2, CA-based CO2 capture tool is considered as one of the potential systems for solving the problems. Recently, we cloned several types of CAs from marine microalgae or marine bacteria and we re-constructed their sequences optimized for highly-efficient production in E. coli systemThe purpose of this study was to develop a general method for the facile detection of food-borne pathogens. As a proof-of-concept, the fusion proteins containing P1 and P2 domain of Norwalk virus capsid proteins were constructed, expressed and characterized. Recombinant E. coli BL21(DE3) strains harboring pET22-6His-P1 and pET22-6His-P2 were cultivated for the production of fusion proteins. Then, soluble or insoluble fusion proteins were purified in HPLC and measured its binding ability and their secondary structure. The majority of the 6His-P2 fusion proteins were expressed in soluble proteins, while 6His-P1 fusion proteins were expressed in inclusion bodies. After solubilization with urea, 6His-P1 recombinant proteins were further purified by affinity chromatography. A total of 1 mg purified 6His-P2 fusion proteins and 0.7 mg of purified 6His-P1 fusion proteins were obtained with the overall yield from 1 L of bacterial culture. When tested the binding affinity of two fusion proteins in ELISA assay, we observed the 6His-P2 fusion proteins could be used as recognition element for the use of biosensor of food-borne pathogens. Much detailed results will be presented. [This study was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2011-0010312)]

Over the past few decades, toxic red tides have had a negative effect on human health, the fisheries industry, and coastal environments, resulting in significant economic losses. We demonstrate a method for the control and suppression of harmful algal blooms (HABs) through selective algal lysis. The method involves the algicidal effect of HABs with aminoclay and has negligible effects on non-harmful phytoplankton, zooplankton, and fish farmed. The use of positively charged colloidals of aminoclays produced cell lysis in HABs within several minutes due to the ammonium characteristics of the aminoclay and the electrostatic attraction between the clay nanoparticles and the algal cells. We show that selective algal lysis by water-dispersed aminoclays in batch mode produces 100% cell death. In contrast, treatment with yellow loess caused algal flocs with little cell lysis. The amount of aminoclay loading needed for the mitigation of HABs is at least ten times lower than that of the conventional yellow loess used as a mechanical treatment.

Mass spectrometry is a promising tool for both qualitative and quantitative analysis of glycans owing to mass accuracy, efficiency, and reproducibility, but it has been of limited success in quantitative analysis for both neutral and sialylated glycans in a high-throughput manner. Here, we present an improved high-throughput N-glycan preparation method that enables N-glycan liberation to permethylation using a stack of three 96-well plates (i.e., PVDF, PGC, and sodium hydroxide plates). N-glycans from chicken ovalbumin glycoprotein and porcine thyroglobulin glycoprotein have been identified by using this high-throughput system. Moreover, the absolute quantity of total N-glycans including neutral and sialylated oligosaccharides was determined using internal standards. Finally, we applied this technique to confirm the N-glycan from human serum prostate specific antigen (PSA). To the best of our knowledge, this study presents a first attempt to develop an in situ N-glycan preparation method with a high-throughput platform.

Our previous study demonstrated that some polyphenols could stop neuronal SNARE zippering [1]. Membrane fusion intermediates such as hemifusion and partially zipped SNARE complexes, which are otherwise transient, could be captured and dissected by the identified polyphenols [1]. The polyphenols did indeed impair neuroexocytosis by inhibiting SNARE complex formation in neuron and paralyzed muscles [2]. Based on these results, we hypothesized that some polyphenols interfere with SNARE complex formation in cells other than neuronal cells, though the functional SNARE complex-comprising proteins might be different. Among the various cells in which SNARE proteins are involved in trafficking pathways and exocytosis, mast cell was selected to demonstrate our hypothesis, as the secretory behaviors of mast cells have been relatively well-characterized. Here, it was demonstrated that polyphenols inhibited SNARE complex formation in mast cells in a SNARE-specific manner, which might provide a clue to how polyphenols exhibit anti-allergic activity. Polyphenols regulated SNARE complex formation in mast cells, resulting in degranulation inhibition. As a result of selective or common binding of polyphenols to two different SNARE complexes, membrane fusion of the corresponding granules with plasma membrane was selectively or commonly inhibited, leading to polyphenols’ differential degranulation inhibition of distinct subsets of granules. Because all intracellular membrane fusion events are mediated by SNARE-family members and SNARE proteins are distributed on nearly every membrane of all eukaryotic cells, our results implicate the possibility that formation of a variety of SNARE complexes in numerous cell types is controlled by polyphenols, which, in turn, might regulate corresponding membrane trafficking.

Aptamers are powerful capturing probes against various targets such as proteins, small organic compounds, metal ions, and even cells. In this study, we isolated and characterized the ssDNA aptamers against E. coli. Total twenty eight ssDNAs were isolated after 10 rounds selection via bacterial cell-SELEX process. Other bacterial cells (Klebsiella pneumoniae, Citrobacter freundii, Enterobacter aerogenes, and Staphylococcus epidermidis) were used for counter selection to enhance the specificity of ssDNA aptamers against E. coli. With priority based-on 2D-structural prediction, ten ssDNA sequences were experimented for their affinity and specificity.. Finally, four ssDNA aptamers showed high affinity and specificity to E. coli The dissociation constant (Kd) of these four ssDNA aptamers to E. coli were estimated in a range of 12.4 to 25.2 nM, and they didn't bound to other bacterial cells including four counter cells. For other E. coli strains, they showed a similar or slightly weaker affinity compared with target strain. Based on the competitive binding assay, we assumed that two among four aptamers may bind to similar moiety on the surface of E. coli. In addition, the aptamer cocktails composed with selected aptamer pools showed a higher binding signal to E. coli compared with using only single aptamer.

Commonly used techniques for analyzing gene expression, such as real-time PCR, Northern blot, micro-arrays, and in situ hybridization, have proven invaluable in understanding RNA processing and regulation. However, these techniques rely on the use of lysed and/or fixed cells and are therefore limited in their ability to provide important spatialtemporal information. This has led to the development of numerous techniques for imaging RNA in living cells, some of which have already provided important insight into the dynamic role RNA plays in dictating cell behavior. Here I present the nano-sized oligonucleotide fluorescent probes, molecular beacons, that have allowed for RNA imaging in living cells. Molecular beacons have the potential to provide a powerful tool for rapid RNA detection in living cells. I also discuss their design, structure, uses, and issues, as well as applications such as disease marker detections and specific stem cell isolations. It is expected that continued advancements in live cell imaging of RNA using molecular beacons will open new and exciting opportunities in a wide range of biological and medical applications.