Earticle

Thermoacidophilic archaea such as Thermoplasma acidophilum and Sulfolobus solfataricus metabolize Dglucose via the non‐phosphorylated metabolic pathway (NMP), an ED‐like pathway in which aldose intermediates are not phosphorylated. The first step of the non‐phosphorylated glycolysis pathway is the oxidation of D‐glucose to D‐gluconate, catalyzed by NADP‐dependent D‐glucose dehydrogenase. DGluconate is then dehydrated by D‐gluconate dehydratase to 2‐keto‐3‐deoxy‐D‐gluconate (KDG), which undergoes an aldolate cleavage to pyruvate and D‐glyceraldehyde by KDG aldolase. During the last ten years we investigated the NMPs in T. acidophilum and S. solfataricus and identified the genes involved in archaeal NMPs. Recently we have also found that oligotrophic marine bacteria metabolize D‐glucose, D‐galactose, Dxylose, L‐arabinose and D‐fucose via the novel NMP where the downstream pathway is different from the previously known NMPs. A BLAST search of the GenBank database has revealed that proteins homologous to the NMP enzymes are present in all three phylogenetic domains of life (Archaea, Bacteria, and Eukarya), which may indicate possible advantages of the non‐phosphorylated metabolic system under oligotrophic or starved conditions. This presentation will review our current understanding of the NMP system in natural environments and our recent progress in the application of the NMP system to the production of bioproducts such as biofuels and bioplastics.

KSBB was established in 1985 to support the development of biotechnology and to provide service to members for collaborative research. KSBB is the representative society of biotechnology in Korea. Organized by members (over 2,500 individuals and 72 organizations) from various BT areas, academics, research institutes, and industries, KSBB strives to support biotechnology development through the following activities. 1. Collaborative research and exchange for the development of biotechnology 2. Academy-Industry collaboration for practical research 3. Organizing and supporting conferences, symposiums and training workshops 4. Publication of journals, newsletter, and proceedings 5. Proposing national BT policies 6. Promoting biotechnology exchange among nations

Public drinking water supplies are clearly at risk for widespread distribution of pathogenic organisms and chemical contaminants that negatively impact human health. The most problematic contaminants in the desert southwest are Cryptosporidium, enterovirus, and Trichloroethylene (TCE). Methods of detection are well developed but require extensive laboratory analysis and time. Measurements need to be made as frequently as customers make demand on the water system, that is, frequently and at nearly all hours of the day. Unfortunately, there are no methods available to continually and cost‐effectively monitor water in real‐time for the presence of these contaminants. We have developed a unique testing facility with a diverse array for continual monitoring sensors for detection of contaminants in water systems. This facility has devices to detect changes in water pH, conductivity, TOC, turbidity, chlorine, nitrate, and microbes based on particle size (to discriminate bacteria, spores, and protozoa). Detection of microbes can be performed based on changes especially in water TOC, turbidity, and the microbial particle size. Viability of the rganisms, as determined by plate counts, has a small role in detection capability; however overall water quality greatly impacts detection based on the role of sensor stability. The goal of this work is to develop an online monitoring scheme for detection of viruses in flowing drinking water. We use multiple approaches. One applies an electrodeposition process that is similar to the use of charged hollow fiber membrane cartridges typically employed for collection of viruses from environmental water samples. The second approach employs a cell culture as a sacrificial layer which is infected by only viable viruses. In both a spectroscopic measurement is performed using the evanescent wave that penetrates no more than 1 um from the surface of an infrared optical element in an attenuated total reflectance measurement scheme. The infrared measurement provides quantitative information on the amount and identity of material deposited from the water. Initial studies of this sensing scheme used proteins reversibly electrodeposited onto germanium (Ge) chips. An off line version of this approach indicated a detection limit of approximately 1,500 viral PFU. The results of those studies were applied to design a method for collection of viruses onto an attenuated total reflectance (ATR) crystal. Spectral signatures can be discriminated between three types of protein and two viruses. There is potential to remove deposited material by reversing the voltage polarity. This work demonstrates a novel and practical scheme for detection of viruses in water systems with potential application to near continual, automated monitoring of municipal drinking water.

We have successfully developed optically coded functional microbeads by coencapsulating both bioluminescent reporter bacterial cells and fluorescent microspheres within a common alginate microbead [1]. This smart functional microbead has a specific stress‐specific bacterial strain and, as an its identification optical code, one of five optical codes generated from fluorescence microspheres such as yellow, green, red, yellow + green, or no fluoresce. These smart microbeads could be randomly scattered on any multi‐well chip plate with the result that, since the cell types are identified on the basis of fluorescent color, the microbead arrays were fabricated without pre‐designation of an individual well. As an example of this method, five different stress specific bioluminescent bacterial strains, each with a different optical code, were successfully implemented to make five different types of optically coded functional microbeads, with a speed of about 30 microbeads/min. This final randomly scattered functional microbeads array biochip, with a fast fabrication of each chip at every 2 min, successfully demonstrated its ability in toxicity screening and monitoring for samples with a few examples for five different stress chemicals. This simple and fast, but not tedious and complicated procedure should be widely and practically used in making the cell array chips for the monitoring of environmental toxicity, new‐borne chemicals, pharmaceutical drugs and the cosmic rays in space station or spaceships in future.

From the mid‐1980’s into the 1990’s, several national forums in the U.S. debated whether a new engineering discipline based on the science of biology was indeed a relevant idea. There were passionate arguments by engineers and educators against this idea who strongly advocated that already the existing engineering disciplines dealing with biological materials (e.g., agricultural engineering and medical engineering) were biological engineering. They did not appreciate that discoveries in biology at “atomic” levels and the advances in computers and computational methods would make more quantitative representations in biology possible, and that these advances would lead to designing new systems based on principles of living systems and properties of biological materials. The possibilities of engineering living systems and engineered systems having living components were generally discarded or even ridiculed. The Institute of Biological Engineering was born in this environment at a two‐day meeting of ten invited individuals in February 1995 in Atlanta who came to discuss ways for advancing biological engineering within American Society of Agricultural Engineers (ASAE). A consensus emerged on the very first day that an autonomous organization was needed that would attract integrators of biology and engineering to lead the emergence of biological engineering. The ten participants conceptualized IBE with the vision to support the community of scientists and engineers for enhancing and promoting biological engineering in the broadest and most liberal manner through research, education and professional development. IBE became a community of ASAE in June 1995 but spun off in 1999 as an independent society to pursue its broader vision of biological engineering. IBE is the first and the only international society of biological engineering devoted to the development of the fundamental science of engineering in the context of biology. IBE is devoted to the promotion of professional development of members focused on biology‐inspired design. IBE also envisions playing a central role in forming a nexus of scientific and professional societies worldwide that together promote the biological engineering discipline. Partnership with the Korean Society of Biotechnology and Bioengineering (KSBB) is the first successful effort beyond our own country’s borders. Forging partnerships with countries in Asia, Europe and South America are essential to the development of biological engineering. The 21st century is the golden age for biology and with it the use of biology‐inspired engineering and design. Indeed, our era presents the promise of innovative engineering approaches and new designs more harmonious with nature and more sustainable.

The two major forms of the pyrimidine dimers are known to be the cyclobutane pyrimidine dimers (CPDs) and pyrimidine-pyrimidone (6-4) photoproduct. CPD the 6‐4 photoproduct of thyminehave been reported to cause structural changes of DNA, such as bending or kicking by 30℃ and 40℃, respectively. The conventional methods using circular dichroism spectroscopy and atomic force microscopy for monitoring the structural deformation of DNA are time consuming and instrument oriented. In recent, we observed red to purple color change of AuNPs only after mixing with UV‐irradiated DNA in a high salted buffer. The color changes of the mixture was dependent on UV irradiated time and base compositions of the UV‐irradiated DNA. The stability of gold nanoparticles in a high ionic strength solution is maintained by straight ssDNA adsorbed physically on the AuNPs. Single‐stranded and striated DNAs are known to adsorb onto AuNP by uncoiling themselves to facilitate electrostatic interactions between the bases and AuNPs. The UV irradiation to DNA accumulated a conformational deformation of the DNA structure by multiple‐dimer formation. However, the covalent bonds formed in the pyrimidine dimers could impose more stiffness on the UV irradiated DNA. As a result, the UV‐irradiated DNA became more rigid and thus could not effectively uncoil the compacted structure for the electrostatic interaction with AuNP and resulted in AuNP aggregation under the high salt condition. This hypothesis was supported and confirmed with no observation of any mass of fragmented DNA or radical oxygen species under the UV irradiation. It has been also known that the pyrimidine dimers are formed as result of triplettriplet energy transfer by exogenous drugs. Because our method does not require any chemical or biochemical treatments or special instruments for purifying and qualifying the DNA photolesions, it should provide a feasible tool for the studies of the UV‐induced mutagenic or carcinogenic DNA dimers and accelerate screening of a large number of drug candidates.

Biosensor technologies have been rapidly developed in the past several decades as multidisciplinary efforts to meet the needs for rapid detection of biological and chemical agents. A biosensor basically consists of a biosensing material and a transducer and can be used for specific detection of one or multiple analytes. Biosensing materials, including enzymes, antibodies, nucleic acid probes, aptamers, phages, cells, tissues, and organelles, are able to selectively recognize target analytes, and transducers, including electrochemical, optical, piezoelectric, thermal, and magnetic devices, can quantitatively monitor biochemical reactions and convert them into electronic signals. In recent years, biosensor technologies have been integrated with nanomaterials/nanostructures, micro/nanoelectromechanical systems and biotechnology for rapid, specific, sensitive, inexpensive, in‐field, on‐line or real‐time detection of pesticides, antibiotics, bacteria, viruses, toxins, proteins, odors, chemicals, and more in plants, animals, foods, soil, air, and water. Enzyme, ntibody and cell‐based biosensors with amperometric, potentiometric, absorption, and fluorescent transducing methods have been developed for detection of insecticides, herbicides, and fungicides used in agriculture and environment. The detection of pesticide and antibiotic residues in food and environmental samples has been intensively investigated using electrochemical enzymatic, SPR, and bio/chemiluminescence biosensors. Antibody DNA/RNA probe, aptamer and phage‐based electrochemical, optical and piezoelectric biosensors have been specifically studied for quantitative detection of pathogenic bacteria and viruses in plants, animals, foods and environment. Biosensor technologies are still facing with great challenges in improving their sensitivity, high throughput capacity, and multiplex measurement. Biosensors also need to be coupled with new techniques of sample preparation or pretreatment, such as magnetic separation, to make uccessful technology transfer in commercialization. With their abilities and unique features for in‐field, low cost and rapid detection of biological and chemical agents in a complex background, biosensor technologies have shown their great potential for road applications in the areas of agriculture, food and environment.

Nitrous oxide (N2O) is fourth most green house gas in atmosphere after CO2, CH4, and aqueous vapour. Though its concentration relative low, however N2O gas very difficult ravelled in atmosphere, with impact to global warming up to 310 times compared to CO2. Biofiltration is the latest technology of pollution control for removing N2O with compost as medium filter. This technology has many advantages, such as low installation and operation cost, secure operation, low energy consumption, good stability, and able to remove pollutant with high efficiency. This research was conducted for evaluation influence of pelletized compost to N2O reduction efficiency and microorganisms growth in medium filter. The biofilter was operated at constant flow rates 88 cc/minute using batch flow system for 12 hours. The research indicated that the highest N2O removal efficiency as 62.25% is reached to be obtained at 5x5 mm pelletized containing 40% nutrition. Parameter estimation by adsorption Langmuir equation indicated that maximum biosorption capacities at 5x5 mm pellets size with 40% nutrition supplementation reached to 1.996 g/kg. Parameter which were estimated using Michaelis Menten equation indicated that maximum removal rate (Vm) and Ks (saturation constant) at pellet 5x5 mm with 40% nutrition content respectively reaches to 1215.89 gm‐3h‐1 and 8.51 gm‐3.

Incheon is in an advantageous position of fostering biomedical industries than other areas in Korea as it has many supporting infrastructures such as Korea Biotechnology Commercialization Center, Korea Conformity Laboratories, university clinical centers and research institutes. International airport and an international seaport located in this area is another strength for global business and international trade. Incheon also has some prominent biotechnology companies including Celltrion, Bernabiotech Korea and TS company, etc. Incheon City Government started its plans to nurture a bioindustry in 2004 and has supported to make favorable business environment since then. It has been constructing the Incheon Bioindustry Support Center and Songdo Science Village to provide core R&D facilities, housing and amenities. Incheon Free Economic Zone Authority(IFEZA) has been establishing biomedical clusters in Songdo, Cheongna and Yeongjong area. Its main businesses are to develop the Bio-Medi Park, BIT Port and Midi City. IFEZA decided to develop Songdo area as a high technology industrial complex and has been inducing renowned companies and research institutes from all over the world. Yonsei University Global Academic Complex, Bio Research Complex and Songdo Global Campus, etc. are constructed in this area. As of now, the number of biotechnology companies in Incheon is less than twenty. However, bioindustry in Incheon will encounter rapid growth within a few years, in its size, number, and quality of infrastructures. IFEZA has been working to attract diverse investments and contracts from foreign companies, universities and hospitals. In less than a decade, Incheon is expected to rise as a hub of biomedical industries in the Northeast Asia.

Established by European scientists in 1978, the European Fede‐ration of Biotechnology (EFB) is Europe’ non-profit federation of national Biotechnology Associations, Learned Societies, Univer‐sities, Scientific Institutes, Biotech Companies and individual biotechnologists working to promote biotechnology throughout Europe and beyond. The mission of EFB is to promote the safe, sustainable and beneficial use of the life sciences, to promote research and innovation at the cutting edge of biotechnology, to foster collaboration between academia, research and industry, to provide a forum for interdisciplinary and international cooperation, to improve scientific education and to facilitate an informed dialogue between scientists and the public. The organisation of EFB with its sections, task groups, regional branch offices in 14 different European countries and the EFB Central Office in Barcelona (Spain) will be discussed. An overview of the main EFB‐activities, which include both central and section themes, European and regional events and reach out both from the perspective of scientific and technological disciplines as well as from the geographic perspective. Our global challenges require sustainable value creation for science, industry, human society, and environment. Therefore we look forward to build new bridges of insight, translation and friendship with the Asian Federation of Biotechnology.

Fluorescent pseudomonads are a group of plant growth promoting rhizobacteria (PGPR) which rigorously colonize roots and provide beneficial effects to plant development. The PGPR have been known to directly enhance plant growth by a variety of mechanisms, namely, fixation of atmospheric nitrogen that is transferred to the plant, production of siderophores that chelate iron and make it available to the plant root, solubilization of minerals such as phosphorus and synthesis of phytohormones. PGPR can also indirectly enhance plant growth via suppression of phytopathogens by a variety of mechanisms. One of the mechanisms includes the ability to produce siderophores that chelate iron and make it unavailable to pathogens. Other desirable features for a potent organism are that it should have the ability to synthesize anti-fungal metabolites, such as the antibiotic 2, 4-diacetylphloroglucinol (DAPG), hydrogen cyanide, and cell‐wall lysing enzymes, which suppress the growth of fungal pathogens. The fluorescent pseudomonad strain R81 is a root colonizing rhizobacteria which promotes the growth of many plants. Its broth containing a hydroxamate‐type siderophore and DAPG was used for preparing bioinoculant formulations for agronomical applications. Talc and aluminum silicate powders were used to develop inorganic carrier‐based formulations of the strain. The formulations of the bioinoculant were able to bring the natural contaminants in the non‐sterile carriers to acceptable limits in the presence of siderophore and DAPG, thus obviating the need of costly three‐time sterilization of the carriers. The shelf‐life of talc powder- and aluminium silicate‐based formulations could also be significantly increased. Glycerol was found to be the best carbon source for enhanced biomass production. Splitting of nitrogen source to NH4Cl and urea had a stabilizing effect on pH during batch cultivation. Both batch and fed batch processes were used for production of bacterial biomass and DAPG. Open loop feeding strategy such as exponential feeding of nutrients and closed loop feedback strategies using dissolved oxygen and pH signals were evaluated for fed‐batch cultivation of the pseudomonad for maximal production of DAPG.

Standing of Nepal for traditional medicine is well known. The healers and tribe community are well reputed for their skills in herbal medicine and natural food. High geographical diversity of Nepal made it rich in varieties of flora and fauna which reported as potential sources of drugs. Nepal contains 701 species of medicinal plants (246 are endemic to the country), out of which 75% occupies in high altitude and Himalayan range. At present, large volumes of herbal plants are illegally traded to neighboring countries which enters back to Nepal as herbal medicines in various trade names. Over 70% of drug in Nepal is imported from India, Bangladesh and other abroad countries. Nepal exports USD 8.1 million (approx) worth of crude herbs only to India. Screening of such important medicinal plants of Nepal, finding their importance in drugs, cosmetics or food value and utilization of them in industrial purpose could lead to create a great research potential in Nepal. Besides this, identification of the chemical components present in biologically active extracts of those plants can help to produce better drugs in the country. Many edible plants with drug value can directly be used as a component of food in daily household foods too. Importance of a constant and co-operative model of research industry is essential in Nepal at present which could enhance the area of research and resource of new foods, cosmetics and drugs in this country and the world. The research carried out in three different plants of Himalayan region showed high potency in antimicrobial, cytotoxic and antiproliferative activities which needs to go ahead for further characterization.

【Objectives】Amyloid beta (Aβ) has been implicated in the pathology of Alzheimer's disease (AD). Aβ-peptides are a natively unfolded protein that aggregate into a β-sheet structure of ordered fibrils [1]. This fibril formation proceeds from the assembly of monomers into amorphous aggregates, via proto-fibrils, to produce mature fibrils. Some studies have suggested that these intermediates are the most toxic species. We used cell-sized (>10 μm) model membranes to directly observe spatio-temporal changes in individual vesicles. These biomimetic membranes enable the researcher to manipulate a 'biological' micro-vesicle under a controlled environment. In this study, we examined the interaction of different oligomeric species of Aβ-40 with a lipid vesicle, observing changes in membrane morphology, in real time using a simple phase-contrast microscope.【Methods】Giant liposomes were prepared using the natural swelling method from a dry lipid film. We incubated 80 μM Aβ-40 in 20 mM Tris, pH 7.4 (Tris) at 37 oC for various periods. Immediately after incubation, Aβ-40 (2μM) was added to cell‐sized lipid vesicles composed of dioleoyl-phosphatidylcholine(DOPC), and their interaction was observed in real‐time [2].【Results and Discussion】We found two significantly different membrane‐transformation pathways (slow membrane dynamics and fast membrane dynamics). We characterized the biophysical mechanisms behind these transformations in terms of the change in inner vesicle volume and surface area. Interestingly, mature fibrils, which are often considered inert species, also induced profound membrane changes. The real‐time observation of these morphological transformations, which can be missed in a previous analyses, may help to unlock the mechanisms of AD's Aβ‐induced neuro‐ degeneration [3]. During this symposium, I would like to mention further about our recent resJ. AM. CHEM. SOC. 2010, 132, 10528–10532ults including translocation of Aβ on model membranes [4] and controllable mesoscopic membrane structures [5].

Proliferating cell nuclear antigen (PCNA) is a trimeric ring‐shaped protein that binds to dsDNA and acts as a scaffold for DNA‐related enzymes, such as DNA polymerase and helicase. Although most of PCNAs form homotrimers, three PCNAs found in Sulfolobus solfataricus form a heterotrimer. Fusion proteins between these PCNAs and functional proteins can act as nano‐scale parts, which can self‐assemble to form functional nano‐hybrid complex. Bacterial cytochrome P450 (P450), which is a promising biocatalyst, generally needs to accept electrons from NAD(P)H for its monooxygenase activity, interacting with a ferredoxin that is reduced by a specific ferredoxin reductase with NAD(P)H. Fusing these proteins with PCNAs can generate functional stand‐alone complexes of P450 and electron transfer related proteins. Here we show that three PCNA proteins, fused with either P450 from Pseudomonas putida (P450cam) or one of two electron transfer related proteins (Pdx, PdR), formed a stable heterotrimeric complex (PCNA‐Utilized Protein Complex of P450 and Its Two Electron Transfer Related Proteins, PUPPET). In PUPPET, P450cam, PdX and PdR were in extremely close proximity to each other, enabling efficient electron transfer within the complex (Fig. 1). As a result, PUPPET showed about 60‐fold higher catalytic activities compared with an equimolar mixture of the constituent components. The formation of complexes utilizing PCNA could be a valuable strategy for the construction of complicated multi‐enzymatic reactions, as well as for applications using electron transfer related proteins.

Biosensors have recently attracted much attention due to their applications in clinical diagnostics, toxicity analysis, food industries, environmental monitoring and quality control1‐3. Biosensors have the potential to replace or complement the classical analytical methods by simplifying or eliminating sample preparation protocols and making field testing easier and faster with significant decrease in costs per analysis. And nanobiotechnology is rapidly evolving to open new materials useful in solving challenging bioanalytical problems, including specificity, stability and sensitivity. In this context, nanomaterials are being increasingly used for the development of biosensors. Their use has extended into all areas of biosensor research. Among the various nanomaterials, nanostrutured metal oxides are being widely investigated for immobilization of biomolecules like proteins and single stranded DNA oligomers. Among the various nanostructured metal oxides, cerium oxide, zinc oxide, tin oxide have recently attracted much attention for the fabrication of miniaturized biosensing electrodes. I will focus on some of the recent results obtained at our laboratories relating to development of nano‐structured metal oxides based biosensors.

In various fermentation products, cost of the carbon source is a significant contributor to the overall cost of the production. Molasses, a by‐product of the sugar factory, is one of the most common carbon sources. However, glycerol is becoming available and inexpensive as the sequence of the rapid growth of biodiesel industry. Generally, every ton of biodiesel produced, nearly a hundred kilograms of crude glycerol is also produced. In fact, pure glycerol has been used in some fermentation processes and additional cost is added to purify glycerol. Therefore, the application of pure glycerol as a carbon source is limited. Escherichai coli is a common microorganisms frequently used as a host in various industrial applications. Interestingly, E. coli is capable of utilizing a variety of carbon sources that are glucose, xylose and fatty acids including glycerol. L‐phenylalanine is the product from E. coli and it is a raw material of low‐caloric sweetener aspartame production. Due to the increasing demand for soft drinks and low‐caloric food, the commercial value of L‐phenylalanine has increased greatly over the past few years. This work presents the production of L‐phenylalanine from crude glycerol using recombinant E. coli BL21 (DE3). The experiment was divided into two parts. The first part was to optimize the medium formula of E. coli BL21 (DE3) for the biomass and L‐phenylalanine productions using Plackett‐Burman Design (P‐BD) and Central Composite Design (C‐CD). The condition of the first part cultivation was 200 rpm of shaking speed and 7.4 of initial pH at constant cultivation temperature of 37˚C. The results showed that, optimum medium formula for the biomass production were crude glycerol of 46.71 g/l, (NH4)2SO4 of 40.55 g/l, KH2PO4 of 1.742 g/l, K2HPO4 of 1.742 g/l, NaCl of 2.554 g/l, MgCl2 of 0.5800 g/l, CaCl2 of 0.0560 g/l, CoCl2 of 0.0036 g/l, ZnSO4 of 0.0113 g/l, CuSO4 of 0.0064 g/l, Na2MoO4 of 0.0069 g/l and yeast extract of 1.411 g/l. In addition, different medium formula was recommended for the L‐phenylalanine production which were crude glycerol of 25.27 g/l, (NH4)2SO4 of 11.53 g/l, KH2PO4 of 0.585 g/l, K2HPO4 of 0.585 g/l, NaCl of 0.859 g/l, MgCl2 of 0.195 g/l, CaCl2 of 0.0567 g/l, CoCl2 of 0.0036 g/l, ZnSO4 of 0.0115 g/l, CuSO4 of 0.0065 g/l, Na2MoO4 of 0.0069 g/l and yeast extract of 1.4263 g/l. In the second part, the fed‐batch cultivation of E. coli BL21 (DE3) was performed. The optimum medium for the biomass production was applied during 0–16 h of cultivation. After 16th h, the comparison of two different feedings, continuous and pulse feeding operations was carried out. For both feeding the investigation of single substrate feed of crude glycerol and double substrate feeds of crude glycerol and ammonia solution were prepared. The optimum condition of the second part cultivation was 400 rpm of agitation speed, and 6.0 l/min (during 0–16 h and 56–72 h) and 4.0 l/min (during 16–56 h) of aeration rates. The initial volume was 1.3 l, while cultivation temperature and pH were kept constant at 37°C and 7.4. It is obvious that the pulse feeding of crude glycerol and ammonia solution gave the highest biomass and L‐phenylalanine production of 22.71 and 0.781 g/l.

Chirality is one of the most important symbols of nature and living organisms. For decades, chiral compounds have been widely used in pharmaceuticals, flavour, agricultural chemicals and speciality materials. Particularly in pharmaceutical field, the need for new chiral reagents is ever increasing since the human body usually functions via chiral recognition of enzymes. For safety concerns, current regulations of USA Food and Drug Administration (FDA) demand proof that the non-therapeutic isomer should be non-teratogenic. Therefore the target for a commercial process is demanding and an enantiomeric excess (ee) of 98% is a minimum acceptable level. Biocatalysis approaches have remarkable predominance due to many advantages such as mild reaction conditions, eco‐friendliness, few by-products, high enantio-, regio- and chemoselectivity and avoidance of protection/deprotection of functional groups, thereby making it an environmentally benign alternative to conventional chemical process. As reported, the worldwide sales of single-enantiomer drugs have exceeded US $150 billions, among which the contribution rate of the biocatalysis approach is ever growing (up to 15‐20%). In addition to traditional approaches to biocatalyst screening, data mining is becoming increasingly important in the discovery of new biocatalysts due to the exponential growth of protein information in public databases in recent years. This report will discuss both the traditional and modern methods for rapid searching for industrially relavant enzymes for chiral synthesis with typical examples from our laboratory [1‐10].

In Sarawak, East Malaysia, agro-residues from sago starch processing industries are abundant and readily available. It has been estimated approximately 7 tonnes of sago pith waste was produced daily from a single sago starch processing mill. Currently these residues were washed off into nearby streams together with wastewater and deposited in the factory’s compound, which can lead to serious environmental problems. Bioconversion of the agro‐residue offers the possibility of creating marketable value‐added products. Utilization of sago residue not only reduce the polluting effects from the sago processing industries, but will also provide an economic solution for waste management system at sago processing mills. This study emphasized on utilizing the trapped starch in sago hampas as a source for glucose production via enzymatic hydrolysis which will later use as substrate for bioethanol fermentation. Initially, three different loading amount (w/v: 5%, 7% and 9%) of sago hampas were examined for glucose production using commercial enzyme. As a result, almost all of trapped starch in 5% and 7% of sago hampas were converted into glucose after 60 min of enzymatic hydrolysis. However for 9% sago hampas, some starch still left behind even though the same procedure of hydrolysis was carried out. Alternative method which is recycles the solution of hydrolyzed sago hampas (HSH) was introduced. Greater improvement of glucose concentration (175.85 g/L) was achieved successfully. Ethanol concentration of 40.3 g/L was obtained from 84.7 g/L of initial glucose by batch fermentation using commercial baker yeast. This study shows the ability of waste starch from sago hampas to be utilized as a feedstock for bioethanol production.

The objective of this study was to design nanoscale biocatalyst system by utilizing the membrane as nanoporous media. The nanostructure was modified by two step methods: simple adsorption and continue with pressure driven filtration. Two types of polymeric membranes mixed cellulose ester (MCE) and polyethersulfone (PES) were used as matrices for immobilization of lipase from Pseudomonas fluorescens. The lipase solution was allowed to permeate through the membrane and lipase molecule was then adsorbed onto inner wall of pores. The porosity and membrane matrices influenced enzyme loading. The best result for enzyme loading in membrane matric was 3,75 g/m2 using PES membrane with incubation time of 18 hours. PES membrane was selected for further continuous transesterification studies. We evaluated the transesterification activity by converting triolein and methanol to methyl oleate and glycerol. The reaction was carried out in situ within the pores of membrane matric, so that its pores behaved as nanoreactor during formation of product. The triolein conversion using this kind of nanoreactor was about 80% with 30 minute of residence time. The productivity of immobilized lipase within the pores were 45 fold higher than that of native free lipase.

cell‐specific immunosuppressive compound with novel mode of pharmacological action both in vivo and in vitro, whose chemical structure was shown to be identical to a previously‐reported antifungal compound named tautomycetin (TMC). Inhibition of T cell proliferation with TMC was observed at concentrations 100‐fold lower than those needed to achieve maximal inhibition with cyclosporin A (CsA). TMC is classified as a type I polyketide derived metabolite based on its chemical structure, specifically the presence of a linear branched‐chain fatty acid‐like moiety. The unique chemical structure of TMC that includes an ester bond linkage between a cyclic C8 dialkylmaleic anhydride at one terminus, and a linear polyketide chain bearing a terminal alkene at the other, indicated that the corresponding biosynthetic pathway features a number of unique biochemical steps with significant potential for generating novel TMC derivatives. Sequence information revealed two multi‐modular type I polyketide synthases (PKSs) and several additional gene products presumably involved in TMC biosynthesis. The deduced roles for most of the TMC PKS catalytic domains were consistent with the expected functions necessary for TMC chain elongation and processing. In addition, a tmcN with a deduced product of 1,029 amino acid residues, located on the 3’‐terminus of an approximately 70‐kb contiguous TMC biosynthetic gene cluster, was found to have amino acid sequence homology with a bacteria Large ATP‐binding regulators of the LuxR (LAL) protein family. Gene disruption of tmcN from the Streptomyces sp. CK4412 chromosome resulted in significantly reduced antifungal activity as well as the absence of TMC. In addition, overexpression of tmcN stimulated TMC biosynthesis, strongly suggesting that TmcN is a pathway‐specific positive‐regulator that activates transcription of the TMC biosynthetic pathway genes in Streptomyces sp. CK4412.

Malaysia has abundant of agricultural wastes which can be utilized for production of fine chemicals of commercial value from microbial sources such as enzymes, antibiotics, flavouring compounds and also microbial biomass which was used as animal feeds through fermentation and enzymatic processes. Lignocellulosic agricultural wastes such as palm kernel cake, sugar cane baggase, paddy straw, rice husks, pineapple wastes, and cynobogon (serai) leaves contribute to more than 5 million tones of wastes per year. Lignocellullose consists of three types of polymers, cellulose, hemicelluloses and lignin that are strongly intermeshed and chemically bonded by non‐covalent forces and by covalent cross‐linkages. These components can be degraded by lignocellulytic fungi or bacteria for production of biofuel (ethanol and biobutanol), organic acid, bioflavour (biovanillin), biopesticide, enzymes and polyoses. Beside of lignocellulosic materials, Malaysia also produce more than 20 million tones of shellfish waste such as shrimps, crabs and crawfish which can be utilized for production of value added products. Chitin is a major component in shellfish waste such as shrimps, crabs and crawfish. Chitin from seafood waste have been developed and used in various field such as environmental, agriculture, medical and food industries. The end product of chitin decompose have been used for chitinase, chitooligochitin, N‐acetylglucosamine production. Instead of chitinolytic enzymes, shellfish waste can be used as a substrate for induction of fibrinolytic enzymes which widely used in pharmaceutical industry, food industry and medical field. The used of shellfish waste in medium formulation wil give a great impact in formulation of cost effective medium for fibrinolytic enzymes production.

Production cost of cellulases is critical for the effective cellulosic ethanol production processes. A fast-growing cellulolytic fungus Penicillium decumbens has been isolated in our laboratory in 1979. It has a well balanced lignocellulose degrading enzyme system. The higher productive and catabolite-derepression mutants (JU-A10 and JU-A10-T) were isolated by mutagenesis and screening. The mutants have been successfully used to produce cellulase preparations and cellulosic ethanol in industrial or pilot scale in China. Higher cellulase activity was achieved by optimizing culture conditions of the strain and composition of production medium with corncob residue (CCR) from xylitol industry as the substrate. The cellulase activity reached 18.9 IU/ml and the cost of cellulase fermentation was reduced. To learn how cellulase production was regulated, genome sequencing of wild type strain 114-2, resequencing of JU-A10 and JU-A10-T genomes were performed to identify the difference in their genomes. The genome size is about 30 Mb, containing 9984 putative genes. 23 genes were found having CBM1 (carbohydrate binding module 1), which is highest number comparing with other known cellulase producing fungi. The digital gene expression tag profiling and Fluorescent Difference 2D Gel Electrophoresis were used to analyze the gene expression patterns and transcriptomics of P.decumbens strains grown in medium containing cellulose-wheat bran or glucose. More than 7 million tags were sequenced for each RNA sample. When P.decumbens was grown in medium containing cellulose as carbon source, transcripts of many cellulases and hemicellulases were expressed at higher levels relative to glucose-grown cultures. Up-regulated genes in the presence of wheat bran included: genes encoding various biomass-degrading enzymes, such as cellobiohydrolases, endoglucanases, β-glucosidase, swollenin, xylanases, arabinosidase, esterases, α-amylase, chitinase and cutinase. It was found that a single nucleotide deletion occurred in the gene of CreA, a repressor of cellulase and xylanase expression, in the mutants, and it was confirmed to be the reason of derepression. Our analysis will provide genome-wide insights into the changes in P. decumbens mutants. The results provided valuable information for the study on the mechanism of cellulase synthesis in P.decumbens. The cellulase and cellulosic ethanol production processes were studied to etermine the optimal techniques for cellulase fermentation and related parameters for alcohol fermentation, in order to provide reference for industrialization of cellulosic ethanol in China.

Recombinant protein therapeutics have opened new therapy options particularly in disease areas where previously no, or only insufficient, therapies were available. Some 165 biopharmaceutical products have gained approval. In 2008, total recombinant protein-based drug sales were $94 billion, and are expected to increase to $145 billion in 2014. The rising pressure of cost-containment in all major markets is driving the uptake of generics and also creates a demand for biosimilars. And the patent and regulatory data protection periods for the first and second waves of biopharmaceuticals based on recombinant proteins have started to expire, opening the way for other manufacturers to place follow-on products. However, the cost and duration of development for biosimilars are much greater than for small-molecule generics, and presents a significant barrier to entry and a resistor of biosimilars market growth. On the other side, the innovator companies have been focusing on differentiated next-generation products which bring about convincing medical progress and make the first-generation, off-patent products obsolete, such as pegylated interferons, long-acting epoetins and fast-acting/slow-acting insulins. In this context, it has even been considered that competition in future indeed might not primarily be between innovators and price-cutting copiers, but rather with second-generation biopharmaceuticals (so-called 'biobetters') based on improved formulation or delivery systems, or derivatized biologics with improved performance. HanAll BioPharma had acquired a French company having proprietary technology for the design and engineering of therapeutic proteins that allowed the identification of key entry sites for proteolysis (or protease-mediated degradation). Through systematic point mutations at selected positions for proteolysis, Hanall was able to create novel versions of therapeutic proteins with decreased susceptibility to protease-mediated degradation in vitro and in vivo, while the biological activity was not affected. These protease-resistant molecules showed advantageous pharmacokinetic properties when administered to animals by different routes (subcutaneous, intravenous or even oral). The presentation concerns a novel IFN-α-2b (HanferonTM) derived from this technology.

Since the market approval of recombinant human insulin as a drug for diabetes treatment in 1982, many recombinant DNA pharmaceutical products including TNF‐alpha blockers have been developed. In general, these products have relatively lower incident of adverse events and high efficacy rates with their targeting properties. As approaching the expiration of patents for the first group of the originator bioproducts, developing “biosimilar” products to be a first provider in the market has been an issue amongst pharmaceutical companies. However, due to large and complex molecular structure of biologics, manufacturing processes including types of cell lines and media are highly influencing on the product quality. Even when a minuscule modification is applied in the manufacturing process of a biological product, it is hardly assure that the product is the same as the one made through previous process. Therefore, the comparability to original products is an essential component in biosimilar product development. Comparability is not only an issue of quality but issues carefully treated in safety and efficacy evaluation. In this presentation, we will share the experience gained through the biosimilar development and discuss about potential problems in depth.

Regulatory environment encourage pharmaceutical company to develop biosimilar product and issued relevant guideline as a general principles to be applied and addressed for development of Biosimilar and also issued product specific guideline. Celltrion is major biosimilar developing player in monoclonal antibody area and already achieved outstanding progress. Celltrion took systematic strategy with global consulting group for IND submission via scientific advisory meeting with EMA and local regulatory agencies. The presentation includes trends of biosimilar development, regulatory environment, Celltrion's approach and case study of our leading biosimilar project.

Vascular endothelial growth factor (VEGF-A) is a key cytokine in the development of normal blood vessels as well as the development of vessels in tumors and other tissues undergoing abnormal angiogenesis. There are two market‐launched humanized antibodies derived from a common mouse anti‐VEGF antibody - bevacizumab, a full‐length IgG1 approved for the treatment of specified cancer indications, and ranibizumab, an affinity‐matured antibody Fab domain approved for use in age‐related macular degeneration (AMD). In clinical trial, these two anti-VEGF antibodies, bevacizumab (Avastin_ anti-VEGF antibody) and ranibizumab (Lucentis anti‐VEGF antibody), have demonstrated therapeutic utility in blocking VEGF‐induced angiogenesis. Avastin, however, has showed its limitation in expanding its clinical application because of efficacy and safety issues such as hypertension, bleeding, bowel/nasal perforation, thrombotic angiopathy, encephalopathy, coronary/perifheral artery diseases. With this view, next generation anti‐VEGF antibodies might have improved VEGF affinity or might more efficiently block VEGF activity through binding to a novel epitope. An additional desirable quality for new anti‐VEGF antibodies is an ability to bind both mouse and human VEGF. This allows testing of the antibodies in a greater variety of model systems before progression to primate studies and addresses the contribution of host VEGF to xenograft growth. As an effort to develop a next generation anti‐VEGF antibody to overcome the efficacy and safety limitation of Avastin, fully human antibody F6 has been developed. The detailed profile of F6 will be introduced. Also the efficacy of F6 antibody was tested in vivo side by side with Avastin, and showed more potent activity to inhibit esophageal cancer growth in regression xenograft model.

The global market of Bio drugs has increased substantially during the last decade since the introduction of recombinant insulin in 1982. The intellectual properties of these bio drugs are beginning to expire, so that the attention of biosimilar products has increased since the European government published the regulatory guideline for biosimilar products in 2005. Due to the heavy financial burden of health care budget by use of expensive innovative bio drugs, governments of industrialized countries are very active on the approval of biosimilar products. Biosimilar products, however, are facing significant challenge of the successful marketing. Intense competition among major pharmaceutical companies, heavy development cost of the products globally and significant challenge by original drug makers cast doubts on the success of Biosimilars in the future. The advance of new drug delivery technology such as longer acting bio drugs enables the companies to develop “Bio‐better” products. At present, the global market of bio drugs is moving toward the bio‐better products. The long acting biobetters are adding values to the existing bio drugs and thus increasing the market size of total bio drugs. Development of Bio‐better is the main focus of the drug development among major harmaceutical companies and new long acting technology is much anticipated. Alteogen Inc developed a new proprietary long acting technology using a carrier molecule called NexPTM. Protein drugs such as hGH, GCSF or interferons are genetically fused to NexPTM, and these fusion bio drugs showed substantially improved PK profiles which are expected to be suitable for weekly or bi‐weekly injection cycle rather than daily or bi‐daily injections of the original bio drugs. NexPTM fused bio drugs also demonstrated impressive in vivo biological efficacies shown by PD experiments in animal models. NexP fused bio‐betters, therefore, can be a uccessful model for bio drug development targeting global market and eventually replacing very crowded and low profit biosimilars with technical edge.

With the LightCycler® family of PCR systems, Roche Applied Science has set a standard in real-time PCR. LightCycler® Instruments are well known for their speed, accuracy, and flexibility, which are due to the symmetrical "one-well–like"design, air-based heating and cooling, and the use of capillaries as reaction vessels. Following along the same lines, the new LightCycler® 480 System efficiently facilitates the delivery of higher sample throughputs when performing gene expression or mutation analysis in life-science research. For the first time, a real-time PCR platform now offers the LightCycler® Systems’ unique combination of accuracy and speed for multiwell-plate based assays with comparable performance with 96- or 384 sample throughputs, and for all relevant qPCR applications. In addition, the highly modular concept of the LightCycler® 480 System hardware and software allows scientists to customize the system to best suit their laboratory’s specific research needs.