• Journal of mucopolysaccharidosis and rare diseases
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• v.2 no.1
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• pp.13-16
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• 2016

Mucolipidosis (ML) II/III are autosomal recessive diseases caused by deficiency of post-translational modification of lysosomal enzymes. The mannose-6-phosphate (M6P) residue in lysosomal enzymes synthesized by N-acetylglucosamine 1-phosphotransferase (GlcNAc-phosphotransferase) serves as recognition marker for trafficking in lysosomes. GlcNAc-phosphotransferase is encoded by GNPTAB and GNPTG. Mutations in GNPTAB cause severe ML II alpha/beta and the attenuated ML III alpha/beta. Whereas mutations in GNPTG cause the ML III gamma, the attenuated type of ML III variant. For the diagnostic approaches, increased urinary oligosaccharides excretion could be a screening test in clinically suspicious patients. To confirm the diagnosis, instead of measuring the activity of GlcNAc phosphotransferase, measuring the enzymatic activities of different lysosomal hydrolases are useful for diagnosis. The activities of several lysosomal hydrolases are decreased in fibroblasts but increased in serum of the patients. In addition, the sequence analysis of causative gene is warranted. Therefore, the confirmatory diagnosis requires a combination of clinical evaluation, biochemical and molecular genetic testing. ML II/III show complex disease manifestations with lysosomal storage as the prime cellular defect that initiates consequential organic dysfunctions. As there are no specific therapy for ML to date, understanding the molecular pathogenesis can contribute to develop new therapeutic approaches ultimately.

• Journal of Chest Surgery
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• v.21 no.2
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• pp.246-253
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• 1988

An in vitro model providing with a recirculating perfusion apparatus using an isolated canine heart and its autogenous blood, which was prepared for study of myocardial protection method. This apparatus was easily used by quick connect system and maintained well heart function for about 2 hours. The Langendorff perfusion was initiated for a 10 minute period by introducing perfusate at 37` into the aorta from aortic reservoir located 100 cm above the heart. The isolated perfused working canine heart model was a left heart preparation in which oxygenated perfusion medium [at 37K] entered the cannulated left atrium at a constant flow rate [900ml/ min] under 20 mmHg overflow system and was spontaneously ejected[no electrical pacing] via an cannula against a hydrostatic pressure of 80 cm H2O. During this working period, various indices of cardiac function were measured. The cardiac functions were stable for over 2 hours with perfusion of Krebs-Henseleit solution and autologous blood[1:1] mixture in volume and maintained heart rate ]]3-122/bpm peak systolic pressure 109-113 mmHg, cardiac output 900 ml / min and left atrial mean pressure 8-9 mmHg. In this model, the efficiency of myocardia] protection could be easily measured by means of functional, enzymatic, biochemical and ultrastructural assessment. And also, we believe this model to be a useful assessment screening model of recovery state after long duration of myocardial preservation of donor heart without difficult transplantation procedures.

• Journal of Microbiology and Biotechnology
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• v.15 no.3
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• pp.466-471
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• 2005

An Escherichia coli strain with high phytase activity was screened from pig excreta. The phytase gene, appA, was amplified by PCR technique. To obtain large amounts of appA phytase, the appA gene was subcloned into the baculovirus transfer vector pVL1393 under the control of the Polyhedrin promoter. The recombinant baculovirus harboring the appA gene was obtained after co-transfection and screening. The early $5^{th}$ instar larvae of silkworm were infected with the recombinant virus. Using this system, the appA phytase was overproduced up to 7,710 U per ml hemolymph. SDS-PAGE analysis revealed the baculovirus-derived appA phytase to be approximately 47 kDa in size. The optimal temperature and pH of the expressed phytase were $60^{\circ}C$ and pH 4.5, respectively. The enzymatic activity was increased by the presence of 1 mM $Ca^{2+}$, 1 mM $Mn^{2+}$, or $0.02\%$ Triton X-100.

• Journal of Fisheries and Marine Sciences Education
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• v.26 no.3
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• pp.543-552
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• 2014

Papain family중 하나인 cysteine protease는 근골격계 질환 치료를 위한target molecule로 인식 되어왔으며 Cathepsin B 는 단백질 분해의 초기과정에 관여하는 cysteine proteases 중 하나이다. 본 연구는 넙치의 cathepsin B 유전자의 발현 양상과 넙치 cathepsin B(PoCtB)의 클로닝, 발현 및 효소특성을 분석하였다. cDNA Library Screening을 이용하여 넙치의 cDNA를 클로닝하였다. 넙치의 동정된 cathepsin B 유전자는 993bp의 open reading frame과 330개의 아미노산으로 이루어져있다. Cathepsin B의 propeptide region 내에 GNFD motif와 occluding loop 가 존재함으로써 이것이 명백하게 cathepsin B group이라는 것을 보여주며, 계통 유전학적 분석 결과 다른 종의 cathepsin B에 비해 초창기에 분화되어 나온 것으로 사료된다. mature enzyme인 maPoCtB은 fusion protein인 glutathione S-transferase를 포함하는 pGEX-4T-1 vector에 삽입하여 E.coli 균주인 $DH5{\alpha}$ 내에 발현시켰다. 재조합 단백질인 PoCtB을 과발현 시킨 결과 53kDa의 분자량을 가진다. 넙치 cathepsin B 활성은 Z-Arg-Arg-AMC와 같은 fluorogenic 펩타이드 기질을 이용하여 측정되었고 적정 pH는 pH.7.5 이다.

• KSBB Journal
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• v.27 no.1
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• pp.61-66
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• 2012

In the screening process of organic solvent tolerant bacteria showing good growth in media containing several kinds of organic solvents, one strain was isolated and identified as Bacillus sp. BCNU 5006. The strain was able to tolerate many organic solvents including benzene, toluene, xylene, octane, dodecane, butanol and ethylbenzene. Likewise, it could also utilize these solvents as the sole source of carbon with significant enzyme production. The lipolytic enzyme stability of Bacillus sp. BCNU 5006 was studied in the presence of several kinds of solvents at a 25% (v/v) concentration. The highest enzyme stability was observed in the presence of octane (107%), followed by ethylbenzene (88%), decane (86%), and chloroform (85%). Especially, BCNU 5006 lipase was determined to be more stable than immobilized enzyme (Novozyme 435) in the presence of octane, chloroform and xylene. This organic solvent tolerant Bacillus sp. BCNU 5006 could be expected as a potential bioremediation agent and biocatalyst for biodegradation and provide on organic-solvent-based enzymatic synthetic method in industrial chemical processes.

• Bulletin of the Korean Chemical Society
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• v.32 no.3
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• pp.1000-1006
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• 2011

The discovery of carbohydrate-based bioactive compounds has recently received considerable interest in the drug development. This paper stresses on the application of 1-methoxy-O-glucoside as the central scaffold, whereas salicylic pharmacophores were introduced with diverse spatial orientations probing into the structural preference of an enzymatic target, i.e. protein tyrosine phosphatase 1B (PTP1B). By employing regioselective protection and deprotection strategy, 2,6-, 3,4-, 4,6- and 2,3-di-O-propynyl 1-methoxy-O-glucosides were previously synthesized and then coupled with azido salicylate via click chemistry in forming the desired bidentate salicylic glucosides with high yields. The inhibitory assay of the obtained triazolyl derivatives leads to the identification of the 2,3-disubstituted salicylic 1-methoxy-O-glucoside as the structurally privileged PTP1B inhibitor among this bidentate compound series with micromole-ranged $IC_{50}$ value and reasonable selectivity over other homologous PTPs tested. In addition, docking simulation was conducted to propose a plausible binding mode of this authorized inhibitor with PTP1B. This research might furnish new insight toward the construction of structurally different bioactive compounds based on the monosaccharide scaffold.

• Korean Journal of Food Science and Technology
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• v.27 no.6
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• pp.891-896
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• 1995

In order to screen natural inhibitor of tyrosinase which catalyzes an enzymatic browning of some foods and in vivo synthesis of melanin, inhibitory effect of 129 edible plants and 15 chemical compounds on the in vivo melanin synthesis by mushroom tyrosinase was analyzed. Among leafy vegetables tested, radish bud, red chicory, Shepherd's purse and small green onion were found to have more than 50% tyrosinase inhibition effect in the descending order. Chinese radish and garlic in root vegetables, and nameko, shiitake and oyster mushroom in mushrooms, and teas showed also more than 50% inhibition effect. Among fruit vegetables tested, red pepper, Chinese quince and avocado were found to have more than 50% tyrosinase inhibition effect, while fruits generally showed low inhibitory effect. Medicinal plants which inhibit tyrosinase more than 50% were mume fructus>cinamomi ramulus>rubi fructus>mori cortex>biotae orientalis folium>puerariae radix, and herbs with more than 50% inhibitory effect were allspice>clove>mustard. In some chemical compounds tested, 4-hexylresorcinol, L-cysteine, glutathione, sodium bisulfite and kojic acid showed powerful inhibition effect on mushroom tyrosinase.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.73-79
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• 2010

The Korea Polar Research Institute (KOPRI) has assembled a culture collection of cold-adapted bacterial strains from both the Arctic and Antarctic. To identify excellent protease-producers among the proteolytic bacterial collection (874 strains), 78 strains were selected in advance according to their relative activities and were subsequently re-examined for their extracellular protease activity on $0.1{\times}$ ZoBell plates supplemented with 1% skim milk at various temperatures. This rapid and direct screening method permitted the selection of a small group of 15 cold-adapted bacterial strains, belonging to either the genus Pseudoalteromonas (13 strains) or Flavobacterium (2 strains), that showed proteolytic activities at temperatures ranging between $5-15^{\circ}C$. The cold-active proteases from these strains were classified into four categories (serine protease, aspartic protease, cysteine protease, and metalloprotease) according to the extent of enzymatic inhibition by a class-specific protease inhibitor. Since highly active and/or cold-adapted proteases have the potential for industrial or commercial enzyme development, the protease-producing bacteria selected in this work will be studied as a valuable natural source of new proteases. Our results also highlight the relevance of the Antarctic for the isolation of protease-producing bacteria active at low temperatures.

• Applied Biological Chemistry
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• v.37 no.6
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• pp.451-455
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• 1994

To detect naturally occurring bioactive compounds from unused marine resources, the screening for the 5-lipoxygenase(potato lipoxygenase-II) inhibitors in Asterina pectinifera, Halocynthia roretzi skin, Nototodarus sloani ink, Anthocidaris crassispina skin, Sargassum horneri, Agarum cribrosum, Odonthalia corymbifera and Desmarestia ligulata was carried out. Water, ether, acetone and methanol fractions extracted from Sargassum horneri had strong inhibitory effect on enzymatic lipid oxidation by potato lipoxygenase-II, and their $IC_{50}$ were 320, 18, 9.5 and $100\;{\mu}g/mL$, respectively. The $IC_{50}$ of ether fraction extracted from Asterina pectinifera and acetone fraction extracted from Nototodarus sloani ink were 29.5 and $34.3\;{\mu}g/mL$, and these extracts showed relatively excellent inhibitory activity. Nonpolar solvent (ether, acetone) extracts of tested marine organisms had more inhibitory effect against 5-lipoxygenase than the polar solvent(water) extracts.

• Mycobiology
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• v.37 no.1
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• pp.53-61
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• 2009

The process of biodegradation in lingo-cellulosic materials is critically relevant to biospheric carbon. The study of this natural process has largely involved laboratory investigations, focused primarily on the biodegradation and recycling of agricultural by-products, generally using basidiomycetes species. In order to collect super white rot fungi and evaluate its ability to degrade lingo-cellulosic material, 35 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye. In the laccase enzymatic analysis chemical test, 33 white rot fungi and 2 brown rot fungi were identified. The degradation ability of polycyclic aromatic hydrocarbons (PAHs) according to the utilized environmental conditions was higher in the mushrooms grown in dead trees and fallen leaves than in the mushrooms grown in humus soil and livestock manure. Using Poly-R 478 dye to assess the PAH-degradation activity of the identified strains, four strains, including Agrocybe pediades, were selected. The activities of laccase, MnP, and Lip of the four strains with PAH-degrading ability were highest in Pleurotus incarnates. 87 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye on solid media. Using Poly-R 478 dye to assess the PAHdegrading activity of the identified strains, it was determined that MKACC 51632 and 52492 strains evidenced superior activity in static and shaken liquid cultures. Subsequent screening on plates containing the polymeric dye poly R-478, the decolorization of which is correlated with lignin degradation, resulted in the selection of a strain of Coriolus versicolor, MKACC52492, for further study, primarily due to its rapid growth rate and profound ability to decolorize poly R-478 on solid media. Considering our findings using Poly-R 478 dye to evaluate the PAH-degrading activity of the identified strains, Coriolus versicolor, MKACC 52492 was selected as a favorable strain. Coriolus versicolor, which was collected from Mt. Yeogi in Suwon, was studied for the production of the lignin-modifying enzymes laccase, manganese-dependent peroxidase (MnP), and lignin peroxidase (LiP).