Earticle

Microalgae is remarkable biomass for producing biofuel. Microalgae can have a rapid growth rate and it consume carbon dioxide for photosynthesis, therefore it can contribute to relieve global warming crisis. Moreover, some microalgae contain a lot of lipid content that can be converted into biodiesel. Maximum lipid content of microlgae have been known as 80% of their dry weight. Lipid content which green algae contain is mainly and fatty acids with a high degree of unsaturation. In this study, we used Neochloris oleoabundans and identified its total lipid, as the pre-step for producing biodiesel from green algae. we used various methods of lipid extraction, and we tried to find a suitable method for microalgae. By this study, we could confirm lipid composition of Neochloris and conclude that Neochloris is proper source for production of biodiesel.

In this study, an increased surface area crystallization process was evaluated to achieve the effective purification of vancomycin, a glycopeptide antibiotic. In addition, the crystallization behaviors (purity, yield, crystallization time, and particle size) were investigated to establish the optimal crystallization strategies. As surface area-increasing materials, diverse types of ion exchange resin were used to increase the surface area per working volume (S/V). When the S/V was increased, in cation exchange resin Amberlite 200, Amberlite IR 120 (Na), and Amberlite IRC 50 and in anion exchange resin Amberlite IRA 400 (Cl) and Amberlite IRA 910 (Cl), vancomycin crystals were successfully generated. The yield of vancomycin increased (> 97%), and the time necessary for crystallization was reduced dramatically (reduced from 24 hr to 12 hr). On the contrary, increase in the S/V hardly affected the purity of vancomycin (> 95%). In addition, when crystallizing after adding ion exchange resin, a surface area-increasing material, it was possible to obtain even smaller vancomycin crystals than when the resin had not been added.

In this study, we evaluated the efficiency and the behavior as well as shapes and sizes of paclitaxel precipitate as the surface area per working volume (S/V) of the reacting solution is increased in an acetone/pentane precipitation process for the purification of paclitaxel. The purity of paclitaxel after 24 hr of precipitation was 54.0% when there was no surface area increase, while it was 54.2% and 77.7% when the surface area was increased by the use of glass beads and anion exchange resin (Amberlite IRA-400OH). The purity was significantly improved when Amberlite IRA-400OH is used as an agent to increase surface area. The yield of paclitaxel improved when glass beads were used but decreased when Amberlite IRA-400OH was used. Compared with the case where no surface area increasing agent was employed, the addition of glass beads or Amberlite IRA-400OH as a surface area increasing agent resulted in a considerable decrease in the size of the paclitaxel precipitate. When Amberlite IRA-400OH was added, the zeta potential value in the precipitation solution was higher (zeta potential value: -21.36 mV) than when no surface area increasing agent was employed (zeta potential value: -0.52 mV), indicating that the precipitated particles are much more stabilized with the addition of Amberlite IRA-400OH.

In this study, we evaluated the effects of the crude extract purities and pure paclitaxel contents on the efficiency and behavior (purity, yield, fractional precipitation time, precipitate shape and size) of fractional precipitation for the purification of paclitaxel from plant cell cultures. As the purity of crude extract and pure paclitaxel content were increased, the yield and purity of paclitaxel were increased and the precipitation time and precipitate size were decreased. Furthermore, we found that the maximum amount of pure paclitaxel content for fractional precipitation was 0.45~0.51% (w/v) regardless of purity of crude extract. By increasing the surface area per working volume (S/V) using a variety of ion exchange resins, fractional precipitation time could be reduced significantly as well as improvement of yield and purity of paclitaxel. Use of an ion exchange resin also resulted in the production of smaller paclitaxel precipitates since it inhibited the growth of particles.

Fractional precipitation is the most effective method for pre-purifying paclitaxel, which is the anticancer agent, extracted from plant cell cultures. However, the fractional precipitation process has been fundamentally problematic due to the lengthy precipitation (~3 days) that is required. An improved fractional precipitation process could considerably reduce the precipitation time by increasing the surface area available for precipitation. Three different types (spiral, round, and twisted types) of reactor with an increased surface area were developed and evaluated for the purification of paclitaxel. This improved purification process serves to minimize solvent usage and the size and complexity of the high performance liquid chromatography operation required for paclitaxel purification.

One-dimensional (1D) nanostructured materials in nanorod form have attracted many attentions due to their unique properties derived from low dimensionality and quantum confinement effect. Many nanorod shaped metal oxide materials have been synthesized using various procedures including a biomimetic method called biomineralization. Among various metal oxides, titanium oxide (TiO2) is stable and harmless to environment, finding its useful applications in many areas including photocatalysts, sensors, memory devices and solar cells. In the present study, we demonstrate the synthesis of nanorod shaped TiO2 using a bacteriophage as a template to guide 1D TiO2 nucleation and subsequent growth along the virus axis. To render this virus-mediated biomineralization of the nanorod shaped TiO2, a genetically engineered phage (f88-STB1) was constructed by genetically fusing TiO2 affinity peptide STB1 (CHKKPSKSC)[1] to a major viral coat protein (pVIII) of the wild-type virus f88. Following confirmation of STB1- fused pVIII expression on the f88-STB1 in approximately 150 copy numbers, it was shown that the f88-STB1 exhibited specific and electrostatic binding affinity to TiO2 at pH 9 where indigenous but nonspecific affinity of the wild-type virus to TiO2 was found abolished almost completely. Since affinity of biomolecules toward TiO2 is necessary for biomineralization, nanorod shaped TiO2 synthesis at pH 9 was enabled only when the reaction was catalyzed by the engineered phages. Furthermore, reaction conditions (e.g. pH, ionic strength, concentrations of viruses and precursors) were found to affect significantly the efficiency of the virus mediated TiO2 biomineralization. Finally, nanorod shaped TiO2 produced in this study was analyzed by SEM, TEM, EDS and XRD to compare its characteristics with the conventional TiO2 in particle form.

High-level expression of recombinant proteins in E. coli often results in accumulation of insoluble aggregates known as inclusion bodies (IBs), requiring further solubilizing and refolding procedures prior to obtaining functionally active products. Although molecular chaperones have been harnessed with some success to refold proteins, the exact function of individual chaperones and the interactions between target protein and chaperones are still unclear. We demonstrated that a refolding cocktail comprising ClpB/DnaKJE, PEG and ATP regeneration system could significantly enhance the refolding efficiency of heat-denatured malate dehyrogenase (MDH).[1][2] To further clarify the individual or synergistic roles of each chaperone, diverse His-tagged molecular chaperones were cloned, expressed and purified, respectively. Heat treatment of MDH at different temperatures resulted in protein aggregates of varying sizes. Following the characterization of the protein aggregates for susceptibility to proteolysis, chaperoning efficiencies on the solubilization and renaturation of the protein aggregates have been studied. Various ratios of chaperones were applied with ATP regeneration system to refold MDH to determine an optimum condition of refolding cocktail.

By using the distillation extractor the kind of large that is 10~15㎝ Phyllostachys bambusoides, Phyllostachys nigra, Phyllostachys pubescens, leaf of Sasa borealis Nakai, stem, root, and the extract material was manufactured. in other words the fresh bamboo was divided into the classificaion and by part and it extracted methanol(100% MeOH, 80℃, 4hr) and the hot water extract(80℃, 4hr) and ethanol extraction (60%EtOH + 40%D·W, 80℃, 4hr) obtained the sample liquid. The content of the inorganic component of this sample liquid contained the nitrogen, phosphorus, calcium, magnesium, sulfur, iron, manganese, and etc. the inorganic material total content average was confirmed as the Sasa borealis Nakai 523.80 mg/L, Phyllostachys bambusoides 409.43 mg/L, Phyllostachys nigra 351.18 mg/L, Phyllostachys pubescens 304.57 mg/L order as the kind. and it w as the bamboo sprout 430.40 mg/L, stem 391.57 mg/L, leaf 343.62 mg/L, root 299.73 mg/L order by part. The inorganic material was extracted method with the ethanol 419.01 mg/L, hydrothermal 348.88 mg/L, methanol 323.49 mg/L order. In the organic acid content analysis, the kind content was shown with the kind of Phyllostachys bambusoides 1,190 mg/L, Phyllostachys nigra 750 mg/L, Phyllostachys pubescens 270 mg/L, and Sasa borealis Nakai 230 mg/L order. The leaf 950 m g/L, root 860 m g/L, stem 590 mg/L, and bamboo sprout 410 mg/L order was displayed by part.

A novel fractional precipitation process, both simple and efficient, was developed for producing (+)-dihydromyricetin in high purity and high yield from crude extracts. The optimal acetone composition in water, initial (+)-dihydromyricetin concentration in crude extract, storage temperature, storage time, and pH were 1/5 (v/v), 0.1 g/mL, 4 °C, 32 h, and 9.0, respectively. Crude extracts were efficiently pre-purified using fractional precipitation of (+)-dihydromyricetin by differences of solubility in an acetone solution, increasing purity from 55.0 to over 84.9% with an overall yield of 97.5%. The use of fractional precipitation for pre-purification allowed for rapid and efficient separation of (+)- dihydromyricetin from interfering compounds and dramatically increased the purity of crude (+)-dihydromyricetin for subsequent purification step.

Domestic bamboo classified into Phyllostachys nigra var. Henonis, hyllostachys bambusoides Siebold et Zucc., Phyllostachys pubescens, and Sasa borealis (Hack.) Makino analyzed in order to inquire into the antibacterial activity of the bamboo and pest control effect. The antimicrobial activity experiment, insect pest control experiment, and insect pest control packing experiment was done after extracting the bamboo with the hydrothermal and ethanol and methanol, this extract. The pest control effect about 4 species was the best when extracting the stem of Phyllostachysnigra var. henonis and Sasa borealis (Hack.) Makino and leaf with the ethanol. The pathogenic bacteria blocking capability could search for the effect that it excels in Rosemary. And in the Phyllostachys nigra var. Henonis extract and Bambusae Calulis in Liquamen, the little bit of the blocking capability could be confirmed. The effect was in the Fusarium oxysporum and Bacillus Anthracis and the stem of the Phyllostachys nigra var. Henonis and leaf extract bought with the Phyllostachys nigra var. Henonis leaf extract and Sasa borealis (Hack.) Makino, stem extract was exposed to have the effect about the two-spotted spider mite.

It is well established that cancer is death-causing factor in the world. H. fusiforme has potential as a new functional food component that evidences immunomodulatory activities and anticancer activity This study was conducted to investigate the effects of enzyme extracts Hizikia fusiforme (Viscozyme, Celluclast, Alcalase, Lysing enzyme, flavozyme, pectinex, Neutrase, AMG, Termamyl) on Human cancer cell (HCT-116(colorectal carcinoma:KCLB 10247), HeLa(cervix, uterine ademocarinoma: KCLB 10002), MKN-45(stomach adenocarcinoma: KCLB 30177). Enzyme extracts had on apoptosis of the cancer cell. we confirm the cell activity 0, 0.25, 0.5, 1 ㎍/ml extract of the H.fusiforme. Enzyme extract was treated at 50℃, 24 hr. We determined the cytotoxic effect of these fractions against human cancer cell lines using MTT assay. A s a result, the apoptosis of the human cancer cell lines (HCT-116, MKN-45, HeLa) were efficiency in the sample treated Viscozyme, Flavourzyme, Neutrase respectively.

IgY (Immunoglobulin Yolk) was separated from a hen's egg yolk. IgY in egg yolk corresponds to IgG in animal serum and plays an important role as immunological proteins in intestines. Carragenan and arabic gum were used as pretreatment agents to purify IgY from fresh egg yolk. DEAE sepharose column in FPLC (Fast Protein Liquid chromatography) was an ion exchange tool to remove contaminants as well as to elute IgY from the column. GF HPLC (Gel Filtraion High Performance Liquid Chromatograpgy) enables to measure the molecular weights of IgY and to identify the purified IgY by comparing the molecular weight of standard IgY with the purified one. IgY is a heterogeneous group of different molecular weight and ionic properties, which was investigated with various IE HPLC (Ion Exchange High Performance Liquid Chromatograpgy) columns such as AX, CX and SCX. Three peaks of IgY were separated in the AX column under the conditions of 0.5 M NaCl and pH=8. The SCX column also gave the three peaks of IgY at pH=5 and NaCl = 0.5 M.

Capsosiphon fulvescens, a green alga, grows in copious amounts along the south-western coast of Korea and also widely throughout the world, including European and North american coasts. Although the C. fulvescens has been traditionally used in Korea as a functional food for centuries, the uses and potential values of bioactive materials from this species have not been well studied. The enzyme hydrolysates prepared with Alcalase (Novozyme, Denmark) was tested for ACE(Angiotensin converting enzyme) inhibitory effects. The maximum inhibitory activity was observed after Alcalase hydrolysis for 6hrs(72.9%). We separated of Alcalase hydrolysate by UF membrane system(Hydro-2K, Sartorius). The total carbohydrate of the below MWCO 2000 fraction was determined by the phenol-sulfuric acid method at 490 nm. The amount of sulfate residues was determined by Loui's method using Na2SO4 as a standard. Uronic acid content was determined by the carbazole reaction using D-glucuronic acid as a standard. Protein was quantified by Bradford method. The monosaccharide composition of the below MWCO 3000 was analyzed by HPLC after 2 M TFA hydrolysis and labeling of the hydrolysate with 1-phenyl-3-methyl-5- pyrazolone (PMP). Alcalase hydrolysate contained 67.1% total carbohydrate, 29.8% uronic acid, 0% sulfate and 3.1% protein. Below MWCO 2K contained 2.8% protein, 42.9% uronic acid, 0% sulfate and 54.3% total carbohydrate, which is mainly composed of glucose(95%).

The purpose of our study is discovery of biomarker with elucidation of relationships between aberrant glycosylation and cancer. Glycosylation changes in the serum of cancer patients have been identified, and many of these can be derived from the induced cancer. In order to elucidate relationships between glycosylation and gastric cancer, we analyzed different glycosylation patterns of serum proteins of patients with gastric cancer compare to normal subjects. As results of blotting and LC/MS, we have been known that β-haptoglobin (β-Hp) is target glycoprotein which has 4 N-, 1O- glycosylation site for gastric cancer diagnosis. We analyzed glycosylation status of β-haptoglobin (β-Hp) purified from 10 cancer patients and 10 normal subjects. Lectin blotting index of serum b-haptoglobin with AAL was clearly higher for cancer patients than for healthy subjects. The results indicated that fucosylated glycan were increased in b-haptoglobin of gastric cancer patients than that of normal by and large. Increase in blotting index of b-haptoglobin of gastric cancer with PHAL which recognized tetra-structure due to high β1-6 GlcNAc blanching was also detected compare with that of healthy controls. Lectin blotting of b-haptoglobin with PHA-E, MAL, SNA was performed with some cases of cancer and healthy subjects, but the degree of this association was less than that for AAL and PHAL. We are now in the process of confirming these results with lager groups of cancer patients and other recent technology

Agarose, one of algal galactan, was treated by acid catalyst such as HCl or H2SO4 to produce galactose, HMF (hydroxymethyl furfural), and levulinic acid. Since HMF and levulinic acid are inhibitors to microorganisms, they should be removed before the subsequent step of fermentation to produce bioethanol or other useful compounds. In this study, we investigated recovery of galactose and removal of impurities from algal galactan hydrolysate by nanofiltration. For a model solution containing 2 g/L galactose, 2 g/L HMF and 2 g/L levulinic acid, at an operating pressure of 200 psig, the rejections of galactose, HMF and levulinic acid were 99.3 %, 8.7 % and 37.5 %, respectively. This results clearly suggests that HMF and levulinic acid can be effectively removed into the permeate while galactose is retained in the feed solution. For the optimization of operating conditions, effects of the operating pressures (50 ~ 400 psig), the pH (4.0 ~ 8.5), temperature (20 ~ 35℃) and the concentration of each component (0 ~ 20 g/L) on the solution flux and rejection were investigated.. The pH affected the rejection of levulinic acid only. The rejection increased dramatically with pH. As the operating temperature increase, the solution flux increased rapidly, while the rejections of all the components decreased. The increase in concentration caused a decrease in the solution flux and rejection for all of the components.