• Journal of Life Science
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• v.22 no.2
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• pp.266-280
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• 2012

Agar is a cell wall component of macro red algae that can be hydrolyzed by agarase. Agarases are classified into ${\alpha}$-agarase (E.C. 3.2.1.158) and ${\beta}$-agarase (E.C. 3.2.1.81), in accordance with their cleavage pattern, and can be grouped in the glycoside hydrolase (GH)-16, -58, -86, -96, and -118 family according to the amino acid sequences of the proteins. Many agarases and/or their genes have been detected, isolated, and recombinantly expressed from bacteria, and metagenomes have their origins in sea and terrestrial environments. Products of agarases, agarooligosaccharides and neoagarooligosaccharides, represent wide functions such as antitumor, immune stimulation, antioxidation, prebiotic, hepa-protective, antibacterial, whitening, and moisturizing effects; hence, broad applications would be possible in the food industry, cosmetics, and medical fields. In addition, agarases are also used as a tool enzyme for research. This paper reviews the sources, purifications and detection methods, and application fields of agarases. The role of agarases in agar metabolism and the function of their enzymatic products are also surveyed.

• BMB Reports
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• v.32 no.3
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• pp.299-305
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• 1999

Protein carboxyl O-methyltransferase (PCMT) catalyzes the transfer of a methyl group from Sadenosyl-L-methionine to free carboxyl groups of methyl-accepting substrate proteins. Two isozymes were separated by DEAE-Sephacel chromatography from porcine brain cytosol and designated PCMT I and II. Isozymes I and II were further purified by adenosyl homocysteine-Sepharose 4B and Superose HR 12 chromatography. The molecular weights of the purified PCMT I and II were determined by mass spectrometry to be 20,138 Da and 25,574 Da, respectively. The two enzymes displayed different isoelectric points; 7.9 for PCMT I and 5.3 for PCMT II. Isozymes I and II exhibited similar substrate specificities when tested with various methyl-accepting proteins. Myelin basic protein, a component of myelinated neurons, was found to be an excellent methyl-accepting substrate for both PCMT isozymes with different $K_m$ values, $21.1\;{\mu}M$ for PCMT I and $10.6\;{\mu}M$ for PCMT II. The PCMT activity and methyl-accepting capacity displayed similar distribution in the various brain regions with an exception of the lower values in the cerebellum. The overall distribution may relate to a general function of protein repair by PCMT in the brain.

• Water for future
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• v.10 no.2
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• pp.91-100
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• 1977

This is an experimental study to raise higher efficiency of filteration than conve ntional filteration by increasing of filteration capacity for per unit area and by extending of filteration lasting time with biflow filter system which was improved from the conventional rapid sand filteration method in the process of water purification treatment. In order to raise more efficient function of filteration and giving consideration to the filter layer at upper and lower parts of the filter, the fine sand & an thracite were used as a filter medium. Although there is some difficulty than previous fine sand in procurement, it could confirm that such filter medium (fine sand & anthracite) was more effective in the field of load, lasting time and back wash, etc. In consideration of practical effect of filteration. The raw water which was used for this experimental study was not coagulated. As a result of this experiment, the filteration volume could increase more than 2 times than that of conventional method. Besides, much more advantages could be obtained for instance, the requirement of installation area was not much and installation cost could economize. On the other hand, the following results were found. The quality of filtered water became worse as time goes by and the turbidity of filtered water was more influenced by raw water turbidity than by rate of filteration. Lasting time of filteration on change of filteration rate in the filter layer reached 2 times in comparison with previous filter basin, and until loss gead reached to 1.0 meter and 1.5 meter, the following relate formulas between lasting time and rate of filteration were formed. ($T_{1.5}=181.96V^{-0.46},\;T_{1.0}=121.31V^{-0.46}$) Even though the lasting time can be shorten in case of the increase of the filteration rate, but the lasting time was prolonged more than 2 times than of previous method. With taking aim at contribution to the development of water treatment technique, we are planning to study continuously for the future study basing on the results in this papers.

• Journal of Aquaculture
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• v.19 no.1
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• pp.33-39
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• 2006

Anticoagulant activities of a fermented edible brown alga, Laminaria ochotensis was investigated. L. ochotensis was fermented with 15% sugar (w/v) at $25^{\circ}C$ for 10 weeks. Anticoagulant activity was measured from the supernatant of algal mixture at biweekly intervals up to $10^{th}$ week by activated partial thromboplastin (APTT), prothrombin time (PT) and thrombin time (TT) assay using citrated human plasma. Sample having high APTT activity $(6^{th}\;week)$ was filtered, ethanol precipitated and freeze-dried. The polysaccharide compound having anticoagulant activity was purified by DEAE ion exchange chromatography followed by Sepharose-4B gel filtration chromatography. Anticoagulant activity, polysaccharide concentration, and heparin like activity were determined for the collected fractions by APTT, $phenol-H_2SO_4$, and glycosaminoglycan assay, respectively. The anticoagulant activity assay showed that the activity was increased up to $6^{th}$ week, and decreased thereafter. The concentration of our purified compound was $31.0{\mu}g/ml$ and showed higher APTT activity than commercial heparin. At the same concentration of $31.0{\mu}g/ml$, the heparin showed 186.5 sec activity while our purified compound showed an activity of 386 sec. Single spot on agarose gel electrophoresis showed that the compound was purified and polyacrylamide gel electrophoresis (PAGE) results revealed that the molecular mass of the purified polysaccharide compound was between 60 and 500 kDa. Therapeutic interest of the algal polysaccharide as an anticoagulant has recently been in highlighted. This purified anticoagulant compound from fermented L. ochotensis can be used as a model for anticoagulant agent or could be developed as an anticoagulant agent. This study can be extended to identify the structure and chemical composition of the purified polysaccharide, and to establish a relationship between structure and the function of the identified anticoagulant compounds.

• Journal of Ginseng Research
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• v.14 no.2
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• pp.285-290
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• 1990

Ando et al found the root of Ginseng contained an active peptide which had anti-lipolysis funrtion when they studied effective components of Ginseng to cure diabetes mellitus in 1980. In recent years we were making lots of the researches on the Ginseng polypeptide. We obtained a 14 peptide which could decrease levels of blood sugar and hepatic glycogen. However, amino acid composition of the peptide differs from one reported by Ando et al. The results of its purification, structure and functure and function were reported in this paper.

• Journal of Environmental Science International
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• v.23 no.5
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• pp.943-962
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• 2014

Industrial gas drying, dilute gas mixtures purification, air fractionation, hydrogen production from steam reformers and petroleum refinery off-gases, etc are conducted by using adsorptive separation technology. The pressure swing adsorption (PSA) has certain advantages over the other methods, such as absorption and membrane, that are a low energy requirement and cost-effectiveness. A key component of PSA systems is adsorbents that should be highly selective to a gas being separated from its mixture streams and have isotherms suitable for the operation principle. The six standard types of isotherms have been examined in this review, and among them the best behavior in the adsorption of $CO_2$ as a function of pressure was proposed in aspects of maximizing a working capacity upon excursion between adsorption and desorption cycles. Zeolites and molecular sieves are historically typical adsorbents for such PSA applications in gas and related industries, and their physicochemical features, e.g., framework, channel structure, pore size, Si-to-Al ratio (SAR), and specific surface area, are strongly associated with the extent of $CO_2$ adsorption at given conditions and those points have been extensively described with literature data. A great body of data of $CO_2$ adsorption on the nanoporous zeolitic materials have been collected according to pressure ranges adsorbed, and these isotherms have been discussed to get an insight into a better $CO_2$ adsorbent for PSA processes.

• Journal of the Korean Magnetic Resonance Society
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• v.19 no.2
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• pp.67-73
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• 2015

The microcystin is a cyclic heptapeptide from metabolites of cyanobacteria in the genera mycrocystis, anabaeba as a result of eutrophication. It has been known that microcystin-LR is a potent inhibitor of the catalytic subunits of protein phosphatase-1 (PP-1) as well as powerful tumor promoter. The active site of microcystin actually has two metal ions $Fe^{2+}/Zn^{2+}$ close to the nucleophilic portion of PP-1-microcystin complex. We report the isolation and purification of this microcystin-LR from cyanobacteria (blue-green algae) obtained from Daechung Dam in Chung-cheong Do, Korea. Microcystin-LR was extracted from solid-phase extraction (SPE) sample preparation using a CN cartridge. The cyanobacteria extract was purified to obtain microcystin-LR by HPLC method and identified by LC/MS. The detail structural studies that can elucidate the possible role of monovalent and divalent metal ions in PP-1-microcystin complexation were carried out by utilizing molecular dynamics. Conformational changes in metal binding for ligands were monitored by molecular dynamic computation and potential of mean force (PMF) using the method of the free energy perturbation. The microcystin-metal binding PMF simulation results exhibit that microcystin can have very stable binding free energy of -10.95 kcal/mol by adopting the $Mg^{2+}$ ion at broad geometrical distribution of $0.5{\sim}4.5{\AA}$, and show that the $K^+$ ion can form a stable metal complex rather than other monovalent alkali metal ions.

• The Korean Journal of Food And Nutrition
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• v.9 no.4
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• pp.404-408
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• 1996

One of the major objectives of the food industry is the enrichment of the functional properties and nutritional value of soybean protein. To attain this goal, an expression system of cDNA encoding native and protein-engineered soybean proteins in a microorganism must be developed and the function then ability of self-assembly and the functionalities of the expressed proteins should be evaluated before the modified genes are transfered to soybean plants. The pro-$\beta$-conglycinin synthesized in E. coli BL21(DE3) comprised approximately 20% of the total bacterial proteins and the expressed protein are formed soluble and trimer such as native protein in E. coli cells. The highly expressed protein was purified to homogeneity by salt precipitation with 20~40$ Ammonium sulfate ion-exchange chromatography with Q-Sepharose and hydrophobic column chromatography with Butyltoyopearl. Therefore, we concluded that the high-level expression system of $\beta$-conglycinin cDNA was established and a relatively simple and rapid method for purifying pro-$\beta$-conglicinin was also developed.

• The Korean Journal of Food And Nutrition
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• v.11 no.3
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• pp.347-353
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• 1998

An antibiotic for methicillin resistant Staphylococcus aureus(MRSA) produced by Streptomyces lydicus YSK-681 was extracted by chloroform, and then purified by the C18 reversed-phase HPLC and silica gel column HPLC. The molecular weight of the purified antibiotic was determined from the FAB analysis MS an m/z 1022.4 and 1036.4(M+H)+, indicating that the isolated antibiotic consisted of two similar compounds with the molecular weight difference of 14 m/z value. With the aid of the various nuclear magnetic resonance(NMR) spectroscopic techniques such as 1H NMR, 13C NMR, DEPT and HMQC spectroscopy, the characteristics of function al groups were deduced as the hydroxyl group and leucine. The MIC values of the purified antibiotic were observed at 1∼32 $\mu\textrm{g}$/$m\ell$ against Gram-positive bacteria compared to > 125 $\mu\textrm{g}$/$m\ell$ against Gram-negative bacteria or fungi. The antibiotic was active at 8 $\mu\textrm{g}$/$m\ell$ of MIC90 and S180 at the concentration of 500 $\mu\textrm{g}$/$m\ell$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.4
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• pp.370-375
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• 2014

BACE2 is a membrane-bound aspartic protease that is highly homologous with BACE1. While BACE1 processes the amyloid precursor protein (APP) at a key step in generating ${\beta}$-amyloid peptide and presumably causes Alzheimer's disease (AD), BACE2 has not been demonstrated to be involved in APP processing directly, and its physiological functions are unknown. To determine its function and to develop inhibitors from marine sources, we constructed an overexpression vector for producing BACE2. The gene encoding human BACE2 protease was amplified using the polymerase chain reaction and cloned into the pET11a expression vector, resulting in pET11a/BACE2. Recombinant BACE2 protease was overexpressed successfully in E. coli as inclusion bodies, refolded using the rapid-dilution method, and purified via two-step fast protein liquid chromatography using Sephacryl S-300 gel filtration and Resource-Q column chromatography. The BACE2 protease produced was an active form. This study provides an efficient method not only for studying the basic properties of BACE2, but also for developing inhibitors from natural marine sources.