Earticle

Neurotransmitter release at the synapse requires the fusion of synaptic vesicles with the presynaptic plasma membrane. SNARE (soluble N-ethylmaleimide-sensitive fusion protein-attachment protein receptors) proteins mediate this synaptic fusion event, and the assembly of which to form 4-helical bundle is believed to generate the required force. SNARE complex formation is thought to be initiated at the N-terminal region and zips towards the C-terminal membrane-proximal region (a zipper model). Clostridium botulinum neurotoxins (BoNTs) elicit neuron-specific flaccid paralysis by specifically cleaving neuronal SNARE proteins. After realization of the therapeutic potentials of these otherwise fatal toxins, the controlled use of BoNT/A was approved by the FDA for the treatment of several hypersecretion-related neurological diseases such as strabismus, blepharospam, hemificial spasm and cervical dystonia. Recently, the cosmetic use of these toxins for treating glabellar facial lines and axillary hyperhidrosis has gained immense popularity. Here, we show that SNARE-driven membrane fusion, and the consequent neurotransmitter release, can be controlled by wedging the SNARE zipper with polyphenols. Polyphenols also regulated the degranulation of allergy mediators from mast cells through plopping the SNARE zippers in mast cells. Polyphenols were specific to SNARE complexes. In conclusion, some polyphenols behave like “plant-derived small molecule Botox.”

We developed depigmenting agents of inhibition of MITF (Microphthalmiaassociated transcription factor) binding and melanosome transport for promotion of cosmetics industry and technology diffusion. Our research were progressed by whitening materials selection in specific target using HTS (High throughput screening) Biochip and pharmacophore database virtual screening using molecular design methods of protein target structure. First, we built of skin whitening natural products library and database and evaluated materials by cell-based assay and artificial skin animal alternative test methods. DNA chip is to evaluate the interaction of MITF with DNA binding site E-box by SPR (Surface Plasmon resonance). So, we screened 10 MITF-DNA binding natural products through virtual screening in pharmacophore DB using Structure Based Drug Design (SBDD) and then final 3 ingredients (including N7) selection through E-box DNA and MITF binding by EMSA. In addition, we screened 26 Rab27a- Melanophilin binding inhibitors for the target database to fit the pharmacophore in KFDA natural products 270 and 350 million compounds and then, selected final 10 candidates by establishment of cell based assay for melanosome transport inhibitory activity from 280 compounds library. We developed BMD 27, and it’s containing extracts and completion prototype cosmetic products. This study will contribute to the cosmetics industry and global marketing by application of convergence of BT, IT, NT technology.

현재 국내.외 시장에는 다양한 점도의 내용물의 화장품이 출시되고 있다. 그 중 고점도의 내용물의 화장품은 기존 에어리스 펌프 등을 사용하여 토 출이 되지 않아 아직도 크림용기에 담아 손이나 도구를 이용해 떠서 쓰는 방식으로 사용중이다. 이런 방식으로 사용하다 보면 내용물과 공기가 접 촉해 내용물이 산화 및 경화를 하게 된다. 당사는 이를 방지하기 위해 고점도 내용물도 토출이 가능하고 공기 유입을 차단한 “고점도 무균 용기”를 개발하였다. 개발 제품은 고점도 내용물도 끌어 올려줄수 있는 자바라타입을 적 용시켰 고 기존 배출노즐의 토출공 상에 구비되는 노즐개폐부재의 구조를 개선하 여 화장품 내용물의 고형화 및 공기,이물질 유입을 방지하였다. 이 제품의 개발로 고점도의 아이크림과 영양 크림 등의 제품을 펌프용기에 적용 가능하며 국내 뿐만 아니라 해외 시장까지도 진출이 가능하다고 본 다. 또한 “고점도 무균 용기”로 유기농 제품과 제약 화장품에도 사용 가능 하여 화장품 뿐만 아니라 다양한 분야로 확대 가능하다고 본다. 특허 : 화장품용기 (국내, 2011.9.1일 출원) *해외 출원 준비 중이며 국내 등록 진행중임

Cellular responses to external signals have been mostly investigated by conventional methods that measure average outcomes for a population of cells. That methods are limited in their throughput, resolution (in space, time, and tracking individual cells instead of population average). To address these problem, high resolution live cell images is increasingly used to detect cellular dynamics in response to drugs and chemicals. But monitoring the real-time behavior of arrays of single cells is only achieved with much experimental difficulty due to the small size. It depends on complex and expensive liquid handling devices that have limited its wider adoption. Extracting single-cell information during cellular responses to external signals in a high-throughput manner is an essential step for quantitative single-cell analyses. Here, we have developed a simple microfluidic platform of trapping single cells in spatially well-defined locations. The device exploits hydrodynamics to trap cells flowing near a narrow aperture. Trap Efficiency were tested by imaging and quantitative analysis. we demonstrate the ease of this method for monitoring multiple single cells over a time course. The simplicity of the design, inexpensive materials make it a device for systems biology experiment.

This study was conducted to screen the alginate-degrading microorganism and purification and characterization of alginate-degrading enzyme. The alginate-degrading marine bacteria was isolated from brown algae Sargassum thunbergii. In the result of 16S ribosomal RNA sequence analysis, the strain was identified as Vibrio sp. V140 and named as Vibrio sp. PKA 1003. The optimum culture conditions for the growth of as Vibrio sp. PKA 1003 were pH 7, 3% NaCl, 25℃ and 24 hr incubation time. To obtained the crude enzyme, as Vibrio sp. PKA 1003 was incubated its optimum culture conditions and the culture medium of the bacterium was centrifuged for 30 min at 12,000 ×g, 4℃. The supernatant was utilized as crude enzyme and the optimal conditions for alginate-degrading ability of enzyme was measured by reducing sugar assay and viscometry. The crude enzyme of alginate-degrading bacteria, as Vibrio sp. PKA 1003, showed highest level of alginate-degrading activity when cultured on pH 9, 30℃, 6% alginic acid and 63 hr incubation time. The crude alginate-degrading enzyme produced by Vibrio sp. PKA 1003 was purified by ammonium sulfate salting, dialysis, DEAE-sephadex, sephadex G-100 and Sephacryl S-200 HR column. The effects of pH, NaCl and temperature on purified enzyme and the metal and substrate specificity was measured by reducing sugar assay.

The anti-inflammatory and anti-atopy effects of the Sargassum micracanthum ethanol extracts(SMEE) and water extracts(SMWE) and tuna eyeball oil extracts(TEOE). To evaluate the anti-inflammatory effects of SMEE, SMWE, and TEOE. The secretion of nitric oxide(NO), TNF-α, IL-6 and IL-1β was measured in LPS-induced RAW 264.7 cells by ELISA. The expression of iNOS, COX-2, and NF-κB p65 protein was studied by immunoblotting. The Balb/c mice were used for an in vivo acute toxicity test and ICR mice were purchased to evaluate croton oil-induced ear edema. It was confirmed that the NO and cytokines secretion significantly diminished compared to the positive control. The rate of formation of edema in the mouse ear was reduced at the highest dose tested compared to that in the control. Moreover, in acute toxicity test, no moralities occurred in mice administered 250 mg/kg body weight of SMEE, SMWE, and TEOE over 2 weeks observation period. To evaluate the anti-atopy effects of SMEE, SMWE, and TEOE, all extracts were applied to the dosal skin of 2,4dinitrochlorobenzene (DNCB)-induced Balb/c mice for an in vivo test. The secretion of IFN-γ, IL-4, and IgE was measured in splenocytes and serum by ELISA. In conclusion, these results suggest that SMEE, SMWE, and TEOE can inhibit inflammatory and atopic dermatitis by modulating cytokines, serum IgE and other substances secretion and improving skin symptoms.

Recently there has been considerable interest in trying to translate microtiter plate based assays into microfluidic platforms. Due to their small scale, microfluidic platforms have many practical advantages such as highly efficient reaction, sensitive detection and massive parallelism. Compared with microtiter plate based assays, the microfluidic platforms can continuously perform more complicated operation. Current method to mimic the microtiter plate based assay in microfluidic platforms categorized into droplet array. We present a programmable multiple droplet array systems that include droplet generation, array, storage, fusion, and elimination process. By integrating pneumatic microvalves with microfluidic system, we can achieve to precisely manipulate individual droplets for delivering, merging and mixing of distinct droplets. This method readily allow us to achieve in-situ monitoring and screening of several types of chemical and biochemical reactions. Thus, we expect that the proposed platform will be a powerful tool to study fundamental biological and chemical reactions, high throughput screening (HTS), or combinatorial synthesis or analysis.

Electrospun Poly(ε-carprolactone) (PCL) nanofibers have been widely used for tissue engineering application. However, poor mechanical properties of the PCL nanofibers their use in hard tissue engineering. Here, we fabricate nanofiber mats from PCL and recombinant mussel coating protein (mfp-1) to mimic a protective coating of mussel (Mytilus species) which is hard and flexible and to improve the mechanical properties of the PCL nanofibers. Stiffness of PCL/Rmfp-1 nanofiber mat linearly increased with the concentration of Fe3+-DOPA in mfp-1, a key component of the mussel protective coating. Mimicking the mussel protective coating is a new strategy for enhancing mechanical properties of the biomedical materials.

Neurodegenerative diseases such as Alzheimer’s disease (AD) and Parkinson’s disease (PD) can be attributed to the specific degeneration of neuronal cells in the brain. However, the natures and action modes of toxic species remain unknown. Here, we present a simple and fast method for the preparation of neurotoxic complex with α-synuclein, which is implicated in PD.

Transgenic plant cell cultures have been used as a production system for therapeutic proteins. Rice cells were transformed to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig). With RAmy3D promoter system, expression of the target protein could be strongly induced by sugar starvation. However, sugar-depleted environment can cause cell death and secretion of proteases into the medium due to cell lysis. In addition, during the mass cultivation and production using bioreactors, various physical factors such as shear stress and bubble burst can damage the cells. In this study, effects of an non-ionic surfactant, Pluronic F-68 (PF-68), as a protective agent in transgenic rice cell cultures were examined to enhance the production. Various concentrations of PF-68 at 1, 3, 5, 7 g/L were added in induction phase. The best hCTLA4Ig production could be obtained with 5 g/L PF-68. It was thought that protective effects against shear stress could enhance hCTLA4Ig production in bioreactor.

Lysosomes are known widely as functional organelles in all eukaryotes by the Belgian cytologist Christian de Duve in 1949. These organelles contain various enzymes like lysozyme to degradate the molecules came from outside. On the other hands, the melanin is a ubiquitous pigment that has many different properties. Too much of melanin leads to make some problems so that we need to maintain amount of melanin normally. To use this functions in lysosomal enzymes, in our study, we found that lysosomes are able to decrease the color intensity of melanin. we measured that superficial intensity of lysosomal activities increased after we treated melanin. Besides when we treated lysosomes isolated from mammalian cells to melanocytes, quantities of melanin decreased. Additionally, to prove biologically lysosomes and melanin, we measured that amount of melanin on different passage. Melanin and lysosomes activities showed differently by aging. Therefore, in our results can show a relations between lysosomes and malenin. We assured that lysosomes are as a useful agent for decreasing amount of melanin.

This study discusses the comparison of oxidative stability of extracted oil from red pepper (Capsicum annum) using supercritical carbon dioxide (SCO2) and hexane. The dried red pepper were crushed by a mechanical blender and sieved (500, 700 ㎛) by mesh. Then these samples were stored at −20℃. The SCO2 extraction was performed using a Supercritical Fluid Chromatography/Extraction (SFC/SFE, JASCO, Japan). The SCO2 extractions were carried out at temperatures 40°C, pressures of 15–30 MPa, CO2 flow rates of 2 mL/min. The hexane extraction was performed by magnetic stirring extraction at 4 0℃ for 12 h. A stir was continuously moving at 300 rpm. The extracts were filtered through a CHMLAB No. F1093-125 filter paper and then evaporated using a rotary evaporator. The sample and extracted oil were refrigerated at −20℃ prior to analysis. Capsaicin in red pepper oil was measured by high performance liquid chromatography (HPLC). The fatty acid compositions of red pepper oil were analyzed by gas chromatography (GC). The extracted oil of red pepper will be assayed for total phenolic content (TPC), radical scavening activities (DPPH) and free fatty acid (FFA).

Sasa bamboo (Sasa borealis) is the smallest species of bamboo in Korea, Japan, and China. The heat processing of bamboo leaves enhances their antioxidant activity as well as their taste and flavor. The total antioxidative activity of roasted S. borealis leaves was shown to be about four times higher than raw S. borealis leaves in an in vitro antioxidative activity test using ABTS+ and DPPH. By using online ABTS+-HPLC analysis, we found that antioxidant components were newly produced via the roasting process; very few, if any, antioxidant components exist in the raw materials. From roasted S. borealis leaves, the following four antioxidative components were isolated and identified using spectroscopic methods: (1) p-formylphenol, (2) p-coumaric acid, (3) N-(p-coumaroyl) serotonin, and (4) N-feruloyl serotonin. The antioxidant activities of each isolated compounds were evaluated via the ABTS+ and DPPH+ methods.

A novel type of hydrolase (BL28) from Bacillus licheniformis was identified, expressed in Escherichia coli, and immobilized. As BL28- immobilization methods, cross-linked enzyme aggregates (CLEAs) was prepared. These CLEA-BL28 aggregates exhibited improved catalytic efficiencies and stabilities compared to free BL28 against harsh conditions of thermal or chemical stress as well as high reusability.

A simple microbe-mediated biosynthetic process was adopted for cobalt oxide nanostructures that retained the shape of the bacterial morphology. Four different types of nanostructures were investigated to determine whether they could be used as effective enzyme carriers. Interestingly, most of esterases were found to be adsorbed well on these nanosturctures, with their activities intact. Furthermore, reusuablities and non-cytotoxicities of these nanostructures were clearly established. Therefore, these nanostrutures have its great potential in biomedical industries and other practical applications.

In this study, we developed a lime addition-capacitive deionization (CDI) hybrid process that can efficiently remove acetic acid and sulfuric acid from the model mixture of glucose, xylose, acetic acid, and sulfuric acid, which are the major components from the biomass hydrolyzate by acid hydrolysis. The key parameters of lime addition process (type of lime, amount of lime, stirrer speed, reaction time) and CDI process (voltage, flow rate, feed concentration) were also optimized. In the lime addition process, the optimal lime type, acids(sulfuric acid and acetic acid)/lime molar ratio, stirrer speed, and reaction time for removal of sulfuric acid were CaCO3, 1:1, 200 rpm, and 6 min, respectively. For the CDI process, the optimal voltage and flow rate were 1.2 V and 20 mL/min. The efficiency of acid removal increased as the initial acetic acid concentration decreased. The developed hybrid process was able to remove 98.08% of sulfuric acid and 76.97% of acetic acid from the mixture of glucose, xylose, acetic acid, and sulfuric acid. It was able to recover most of the sugar (>99%) at high purity (97.53%).

In this study, the tar compounds derived from the plant cell cultures of Taxus chinensis were first identified and quantified via GC/MS (gas chromatography/mass spectrometry) and GC (gas chromatography). 2-Picoline, 2,5-Xylenol, Acenaphthene, 1-Methylnaphthalene and o-Xylene were found as major main tar components in biomass. These compounds were identified and confirmed by comparing their retention times with those of authentic compounds. Each compound also spiked with pure standard. The contents of 2-Picoline, 2,5-Xylenol, Acenaphthene, 1-Methylnaphthalene, and o-Xylene in biomass were 0.251, 0.159, 0.124, 0.094 wt% and 0.053 wt%, respectively. In liquid-liquid extraction, adsorbent treatment tar was removed 41.62, 89.10%, respectively. After hexane precipitation, all of tars were successfully removed.

Phthalocyanine derivatives have perhaps received more attention than any other class of dyes as sensitizers for photodynamic therapy and most favoured are the aluminium and zinc derivatives. Photoinduced electron transfer in donor-acceptor molecules has been extensively studied, in particular for covalently linked dyeds in solution as model systems of natural photosynthesis to harvest efficiently solar energy. In this study, the improvements as electron transport from immobilization of R. sphaeroides on two phthalocyanines were evaluated. In order to determine electron reaction of cell bound phthalocyanines, we are confirmed cyclic voltammetry measurement. R. sphaeroides bound ZnPc in water solvent and CuPc in SDS solvent were shown active electron reactions. It was correlated with the amount of cell bound phthalocyanines. In addition, cell bound phthalocyanine was effected on CO2 reduction under light dark condition cultured R. sphaeroides. Phthalocyanines as electron donoron R. sphaeroides can apply for improving photosynthetic efficiency.

Levulinic acid (LA) is a versatile chemical building block that can be produced from cellulosic biomass and can be converted into valuable chemicals and fuels. In this study, enzymatic reduction of LA was performed for the synthesis of 4-hydroxyvaleric acid (4HV), which can be used as a monomer of bio-polyester and a precursor of bio-fuels. Because no enzyme that has LA as its native or major substrate has been reported, engineering of the substrate specificity of an enzyme is required to yield the enzymatic reduction of LA. 3-Hydroxybutyrate dehydrogenase (3HBDH) from Alcaligenes faecalis was selected as a target enzyme for the engineering of substrate specificity. A rational design approach with molecular docking simulation was performed for the engineering of the substrate specificity of the enzyme. Through the mutational studies of 3HBDH, the substrate specificity of the enzyme was engineered and a double mutant which has catalytic activity toward LA was selected as a best mutant. Approximately 57% conversion of LA was achieved with this mutant in 24 hr, while no conversion was achieved with the wild-type enzyme.

For developing medical adhesives, many challenging properties are required such as strong and long-term bonding with wet tissues, biocompatibility, and biodegradability. Hydrogel systems based on cross-linked polymeric materials which could provide both adhesion and cohesion in wet environment have been considered as a promising formulation of adhesives. Inspired by marine mussel adhesion, many researchers have tried to use 3,4-dihydroxy-phenylalanine (DOPA) residues as cross-linking mediators of hydrogel which is known to be able to achieve cohesive hardening as well as adhesive bonding with diverse surfaces. Herein, we developed new hydrogel system based on DOPA-containing recombinant mussel coating foot protein type 1 (fp-1). Cross-linking networks were established using oxdiation-induced DOPA-DOPA and iron (Fe3+)-mediated metal-DOPA coordination, which were confirmed by Raman spectroscopy and color change of the gels. Rheological analysis was performed to identify gelation time, frequency effect, and self-healing properties of gels. And also, adhesive strength of gels on porcine skin tissue surfaces in wet condition was measured by lap-shear test. Conclusively, our newly-developed hydrogel based on the DOPA chemistry of mussel adhesive proteins could be potentially used as tissue adhesives and sealants for future applications.

Transplantation of pancreatic islet is one of therapeutic method of Diabetes Mellitus treatment. Islets, however, lose their ECM (extracellular matrix) during isolation process, and these bare islets clusters are vulnerable not only to attack by immune system but also to physical damage. Alginate capsule can solve these problems by making semipermeable physical barrier. Microencapsulation of islet using alginate was performed through air-driven system. However, this method always produces empty capsules, which cause enlargement of transplantation volume. Previous researchers have done the selection of encapsulated islet by hand-picking. In this research, density-gradient method was applied to separate the encapsulated islets and empty capsules. First, density difference of islets and alginate capsules were confirmed. By adjusting concentration of Percoll®, centrifugation speed and time, optimal conditions for gathering encapsulated islets were set up. Final yield of microencapsulated neonatal porcine islet cell clusters (NPCCs) after process was more than 90%.

The use of organic solvents as reaction media for enzymatic reactions has many advantages. Several organic solvents have been proposed as reaction media, especially for transesterifications using Candida antarctica lipase B (CalB). Among organic solvents, tert-butanol is associated with an enhanced conversion rate in biodiesel production. Thus, it is necessary to understand the effect of tert-butanol on CalB to explain the high catalytic efficiency compared with the reaction in other hydrophilic organic solvents. In this study, the effects of tert-butanol on the structure of CalB were investigated by MD simulations. The overall flexibility was increased in the presence of tert-butanol. The substrate entrance and the binding pocket size of CalB in tert-butanol were maintained as in TIP3P water. The distance between the catalytic residues of CalB in tert-butanol indicated a higher likelihood of forming hydrogen bonds. These structural analyses could be useful to understand the effect of tert-butanol effect transesterification by lipases.

The mechanical nature of cation-p interactions in aqueous medium was directly measured by use of a surface forces apparatus in a model system: one surface is coated with poly-L-lysine(PLL) as cations, and the other surface is coated with three kinds of aromatic homo-polymers (poly-L-tryptophan, PTrp; poly-L-tyrosine, PTyr; polystyrene, PS) as p systems. The interaction energy between two surfaces are dependent on types of competing cations (NR4 +, NH4 +, K+, Li+, Na+), contact time, competing cation concentration, and types of aromatic side groups (indole, phenol, benzene).The adhesion to PLL in terms of strength follows the order of PTrp> PS >PTyr and the adhesion to PTrp surface follows the order ofNR4 +> NH3R+≥NH4 +»K+> Li+, Na+, qualitatively consistent with theoretical simulations. The results directly proved the theory of cation-p interactions in aqueous system and verified the hypotheses to explain the physiological phenomena (eg. potassium channel and neuroreceptor).

As a tool for a drug,the preparation of liposomes has been examined for several decades. We investigated to encapsulate lysosomal enzymes extracted from Saccharomyces cerevisiae in liposomes, which are made with L-α-phosphoatidylcholine from egg yolk. We mixed lipid, cholesterol and lysosomal membrane protein isolated from lysosome treated by H2O2 and NH4Cl to modify liposome surface for antimicrobial activity before and after making liposome. The residue after evaporation was mixed using lysosomal enzymes with same ratio and was vortexed to encapsulate them for 1 min. Large lamellar vesicles were sonicated to make a small unilamellar vesicles for 5 sec. The liposomes were confirmed as cell counts against Escherichia coli to evaluate the antimicrobial activity of lysosomal enzymes. The results showed that the antimicrobial activity was not shown without modification of liposome surface. Also, when the liposome was made with among lipid, cholesterol and lysosomal memebrane proteins, we got more good results. Furthermore, to increase the antimicrobial activity, when liposome was used lysosomal membrane proteins treated by H2O2, it was better than others. Therefore, lysosomal membrane proteins for targeting bacteria will play an important a role.

E. coli biosensor strains, carrying a luxCDABE fusion, are sensitive and responsive to various stresses, such as DNA-damaging compounds. With this concept, several E. coli biosensors have been developed in our lab to detect the inhibitory compounds to the fermentative strains which can be derived from the lignin. Also, the limit of the bacterial bioreporters can be overcome with serum complement by making the pores on the cell membrane, resulting in the sensitivity improvement. To test this, E. coli MG1655/recA::luxCDABE strain was exposed to mitomycin C (MMC) with the heat-treated serum. The lowest concentration of detection was lowered by nearly 450-fold, from 31 ppb to 0.07 ppb. Furthermore, real-time qPCR and imaging with a cell memebrane impermeable dye, POPO-3 demonstrated the serum complement activity.

Energy insecurity and global warming are the biggest challenges for the global economy. Overall, transportation sector is the biggest consumer of nonrenewable fossil fuels. Bioethanol is a practical green alternative transportation fuel which has numerous advantages over conventional fossil fuels. However, grains such as corns are used as the feedstock for commercial bioethanol which may cause food insecurity. Therefore, it is vital to produce cellulosic ethanol from non-food biomass sources. However, these materials are quite recalcitrant to be converted into fermentable sugars due to its complex structure. To overcome this barrier, in this research, we searched for new microorganism species by isolating cellulose degraders with high efficiency. Instead of random sampling, several sources were targeted in the nature where cellulosic material was decomposed such as Suncheon bay, silkwarm, termite, and excrement of various grass-eating animals. Total 48 species which showed cellulose degrading ability were isolated both in aerobic and anaerobic conditions by ten-fold serial dilution, heat treatment, and cellulase activity assay with filter paper and Congo red. Several samples from grass-eating animals showed high cellulose-degrading efficiency. In enrichment, five pieces of filter paper were completely degraded and clearing zone around colonies was formed in Congo red plates. The strains of N-b and N-d from Nyala were determined as Pleomorphomonas sp. and Cellulomonas sp. by 16s rDNA sequencing. These strains showed highest growth rate in laboratory-based minimal nutrient broth with glucose, xylose, and CMC as carbon source. It was also observed that both species converted more than 30 percent of the glucose into volatile fatty acid (VFA).

Seaweed is a third-generation biomass that can be used in bioenergy production and has many advantages. The seaweed grows quickly, is lignin-free and is not used as a food crop. The red seaweed, Euchema sponosum, is cultivated in Indonesia. The first step, seaweed slurry was pretreated by thermal acid hydrolysis and enzyme treatment. Optimal pretreatment condition was determined with 11% (w/v) solid content of seaweed and 150 mM sulfuric acid at 121℃ for 40 min. And enzyme saccharification was carried out Celluclast 1.5L and Viscozyme L at pH 4.5, 40℃. However, E. spinosum produced Five-HMF of 6 g/L after thermal acid hydrolysis. To removed 5-HMF, in addition of active charcoal was carried out in the range of 3-6% at room temperature for 2 hour. 5-HMF was reduced from 6 g/L to 1 g/L on 6% active charcoal. The ethanol fermentation was carried out using SHF with Kluyveromyces marxianus KCTC 7150. The ethanol fermentation from 5-HMF removed E. sponosum hydrolysate produced 18 g/L using K. marxianus for 48 h. On the other hand, ethanol fermentation was prolonged when 5-HMF existed.

Oleaginous yeast has attracted great attention as high potentials for the biodiesel production because it grows fast and its final cell density is high. However, the oleaginous yeast utilizes organic substrates for its growth, and it causes the price of biodiesel to rise sharply [1]. This research focuses on optimizing conditions for the cultivation of oleaginous yeast Cryptococcus sp, which was isolated from fermented seafood, using low-cost substrate, Jerusalem artichoke.The major component of Jerusalem artichoke is inulin, which is composed of fructose units terminated by a glucose unit [2]. It has excellent environmental adaptability, is easy to cultivate, and cannot be digested by the human enzyme system. For these reasons, Jerusalem artichoke seems to be a potential feedstock candidate [3]. To utilize Jerusalem artichoke as a microbial substrate, pretreatment process is absolutely needed. The pretreatment factors, such as temperature, reaction time, and the concentration of nitric acid were optimized through response surface methodology (RSM). An optimal pretreatment condition was at 101℃, 0.7% of nitric acid, and for 10 min, which resulted in 53.4% of fructose yield.

In our present investigation, for the isolation of bioactive substances that could potentially inhibit the over expressions of matrix metalloproteinases, we worked on a marine derived fungal species Aspergillus. The fungal specimen was collected at Cheongsapo, Busan, South Korea. The culturing of the fungal strain was done at large scale and bioactive components were fractioned from the broth and the mycelium as well. The resultant extracts were fractioned with the aid of silica gel flash chromatography that resulted in fractions that were further purified by applying Octadecyl functionalized silica gel chromatography. Later the pure compounds from these fractions were obtained by using High-performance liquid chromatography (HPLC). The resultant pure compounds were analyzed for their respective structures and stereoscopic properties on the basis of Electron Ionization Mass Spectrometer spectrum (EI-MS), 1-dimensional and 2-dimensional Nuclear Magnetic resonance comprehensive analyses. The exploitation of marine fungal strain has yielded 2 compounds 2-Benzyl-4H-pyran-4-One (BHPO), and N-(1- (4-methoxy-2-oxo-2H-pyran-6-yl)-2-phenylethyl) acetamide (NOPPA) Pyrophen. Following to that, cell viability of these secondary metabolites were also evaluated to check their potentiality for in vitro tests. Our investigation suggests that there was no considerable cytotoxic effects exhibited by these metabolites and hence could be recommended for in vitro studies for their beneficial biological effects.

With increasing demand for the natural substances with sustainable production as pharmaceutical agents, marine microalgae have been recognized to provide chemical and pharmacological novelty and diversity. In this study marine microalga Chlamydomonas sp. was hydrolyzed with the combination of food-grade microbes, Candida utilis and Bacillus subtilis. The microbial hydrolysate of the Chalydomonas sp. was purified. The successful assay guided purification leads to isolation of bioactive peptide ENLDDLE – 846.35 Da (H-P-3). Activation of gastric enteric glial cells (EGC) was found to incur damages to the intestinal epithelial barriers. The isolated peptide H-P-3 showed a protective effect on bacterial lipopolysaccharide (1 μg/ml) and interferone-γ (10 ng/ml) activated enteric glial cells mediated disruption of the intestinal epithelial barriers. The peptide H-P-3 significantly attenuated the activated EGC mediated damage of tight junction proteins ZO-1 (Zonula occludens-1) and occludin with simultaneous suppression of enteric cell mortality. It was observed that the inflammatory cascade of the activated EGC create an oxidative stress condition in intestinal epithelial cells (IEC-6) which leads to DNA damage and cell cycle arrest. The molecular pathway studies showed that the peptide is protecting the intestinal epithelial cells by down-regulating the inflammatory signaling pathway NF-κB in EGC which suppress the activation of ATM/ATR dependent DNA damage in IEC-6 epithelial cells.