• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.1
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• pp.50-56
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• 2002

The $\beta$-galactosidase was purified from the internal organs of edible snail by fractionation with ammonium sulfate, ion exchange chromatography on DEAE-Sephadex, Mono Q HR 5/5 and gel filtration on Sephacryl S-200. Superose 12 HR 10/30 chromatography. The specific activity of the purified $\beta$-galactosidase was 18.8 units/mg protein with 31.3 purification fold from crude extract. The $\beta$-galactosidase had native molecular weight of 144,000 dalton and was composed of two subunits of 72,000 dalton. The isoelectric point of the enzyme was determined 4.1. This enzyme was the most active at pH 3.0 and 6$0^{\circ}C$, and was stable in the pH range 2.0~8.0 and below 5$0^{\circ}C$. The enzyme was inhibited by metal ions and sugars such as fructose, glucose, galactose, maltose and xylose.

• Korean Journal of Microbiology
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• v.31 no.1
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• pp.63-71
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• 1993

A xylanase (xylanase I) was purified 11.9-fold from the culture filtrate of Trichoderma koningii ATCC 26113 by the column chromatography on Sephadex G-75, SP-Sephadex C-50, DEAE-Sephadex A-50 and Sephadex G-50 with an overall yield of 8.2%. The molecular mass determined by gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis was found to be a monomeric polypeptide of ca. 35 kDa. The isoelectric point of the enzyme was estimated to be 9.3. The optimal reaction pH and temperature are 5.8 and 55.deg.C, respectively. The enzyme is stable up to 60.deg.C, while 78% of its activity is lost after the incubation for 10 min at 70.deg.C. The enzyme hydrolyzes sylan with relatively high activity, as well as carboxymethyl cellulose and avicel. The $K_{m}$ values of the enzyme for oat-spelf sylan, larchwood xylan and Avicel were 3.5, 1.6 and 10. 1 mg/ml, respectively. The enzyme hydrolyzed oat-spelt sylan to sylose, sylobiose, sylotriose and arabionoxylobiose, while it degraded larchwood xylan to xylose, xylobiose, xylotriose and arabionoxylobiose, while it degraded larchwood xylan to xylose, xylobiose and xylotriose as the major products. The hydrolysis patterns indicate that xylanase I is endo-enzyme.e.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.3
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• pp.386-392
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• 1998

An objective index for quality estimation of anchovy sauce, our traditional seafood, has been required because inadequate standard causes dispute about quality estimation. Qualities of anchovy sauce made by traditional method and commercial anchor sauce products were compared by investigating their proximate compositions, total nitrogen contents, amino acid contents, development and level of a specific peptide on SDS-polyacrylamde gel electrophoresis (SDS-PAGE) and isoelectric point (pI) of the specific peptide. Also, pIs of IEF were attemped to identify fish species of salt-fermented sauces. The 55,600 and 46,900 dalton of peptide band were identified in all experimental samples. Especially, the area of 55,600 dalton was closely correlated with total nitrogen contents. Also, specific pIs in accordance with fish species were identified. The results suggest that correlation between area of 55,600 dalton and total nitrogen contents are used of index for quality estimation, and specific pIs are used of fish species identification.

• Parasites, Hosts and Diseases
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• v.32 no.4
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• pp.231-242
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• 1994

Pnragonimus westermnni, the lung fluke, is known to migrate to the pulmonary tissue of mammalian hosts and causes pathological changes in the lungs. An acidic thiol-dependent proteinase with a molecular weight of approximately 20,000 daltons was purified to homogeneity using ion-exchange chromatography and gel filtration chromatography. On SDS-PAGE, the molecular weight of the enzyme was 17,500 daltons. Isoelectric point was 6.45. The enzyme was similar to the acidic cysteine proteinase of vertebrates in the properties of pH optimum, substrate specificity and inhibitor sensitivity. Enzymatic activity was stable at pH 5.5 for at least two days when stored at 4℃. The cysteine proteinase was capable of degrading collagen and hemoglobin. Sera of patients with paragonimiasis and mice infected with R westermani reacted in immunoblots with the partially purified proteinase. This result suggested that the cysteine proteinase of P. westermnni may play a role in migration in tissues, and in acquisition of nutrients by parasites from the host. It is also potentially an antigen for the serodiagnosis of paragonimiasis.

• Journal of Microbiology
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• v.35 no.2
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• pp.109-116
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• 1997

An extracellular proteinase of Candida albicans was purified by a combination of 0~75% ammonium sulfate precipitation, DEAE Sepharose Fast Flow ion exchange chromatography, and Sephacryl S-200 HR molecular sieve chromatography. Its mlecular weight was approximately 41 kDa on SDS-PAGE and isoelectric point was 4.4. The enzyme was inhibited by pepstain A. Optimum enzyme activity ranged from pH 2.0 to 3.5 with its maximum at pH 2.5 and a temperature of 45$^{\circ}C$. The addition of divalent cations, $Ca^{2+}$, Zn$^{2+}$ and $Mg^{2+}$, resulted in no significant inhibition of enzymatic activity. However, some inhibitory effects were observed by Fe$^{2+}$, Ag$^{2+}$ and Cu$^{2+}$. With BSA as substrate, an apparent $K_m$ was determined to be 7$\times$10$^{-7}$ M and $K_i$, using pepstatin A as an inhibitor, was 8.05$\times$10$^{-8}$ M. N-terminal amino acid sequence was QAVPVTLXNEQ. Degradation of BSA and fibronectin was shown but not collagen, hemoglobin, immunoglobulin G, or lysozyme. The enzyme preferred peptides with Glu and Leu at the P$_1$ position, but the enzyme activity was highly reduced when the P$_2$ position was phe or pro. This enzyme showed antigenicity against sera of patients with candidiasis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.3
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• pp.302-310
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• 2017

Interferon regulatory factor 8 (IRF8) is essential for the development of B and T cells, as well as for the activity of dendritic cells and macrophages. We performed molecular characterization of IRF8 from rock fish, Sebastes schlegelii (Ss), and investigated the spatial and temporal profile of mRNA expression after challenge with lipopolysaccharide (LPS), polyinosinic:polycytidylic acid (poly I:C), or Streptococcus iniae. The full-length cDNA sequence of SsIRF8 was 1,657 bp, containing an ORF of 1,266 bp. The gene had a predicted molecular mass of 47.7 kDa and an isoelectric point of 5.99. The amino acid sequence coded by this gene showed the highest degree of identity (90.8%) and similarity (96.2%) with IRF8 from Oplegnathus fasciatus. The SsIRF8 mRNA was expressed ubiquitously, at varying levels, with the highest level of expression observed in the spleen. To confirm the role of SsIRF8 in mediating the immune response, we measured SsIRF8 mRNA expression in the splenic tissue at different time points after injection with LPS, poly I:C, or S. iniae. The qRT-PCR results showed that SsIRF8 mRNA expression in the poly I:C-injected group was highly upregulated 6 hr after exposure (P<0.05). Expression of SsIRF8 mRNA in the S. iniae-injected group peaked at 24 hr. These results suggest that SsIRF8 might be important in regulating the strength of the rockfish immune response to immunostimulatory agents.

• Journal of Microbiology and Biotechnology
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• v.2 no.3
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• pp.189-196
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• 1992

A $\alpha$-l, 4-D-glucan maltohydrolase $(\beta$-amylase), secreted by the mesophilic aerobic bacterium Bacillus polymyxa No.26, was purified and characterized. The enzyme production was increased after a logarithmic phase of bacterial growth and paralleled with the onset of bacterial sporulation. By applying anion exchange chromatography and gel filtration the enzyme was purified 16.7-fold and had a specific activity of 285.7 units/mg. Two enzyme activities were eluted on a column of DEAE-Sephadex chromatography, and they were designated as E-I for a major enzyme peak and E-II for a minor peak. Of them, E-I enzyme peak was further purified by using gel chromatography. The molecular mass of this enzyme was determined to be 64, 000 daltons and consisted of a single subunit, showing an isoelectric point of 8.9. The enzyme was able to attack specifically the $\alpha$-l, 4-glycosidic linkages in soluble starch and caused its complete hydrolysis to maltose and $\beta$-limited dextrin. This amylolytic enzyme displayed a temperature optimum at $45^\circ{C}$ and a pH optimum at 7.0. The amino acid composition of the purified enzyme was quite similar to the other bacterial $\beta$-amylases reported. Surprisingly, the purified enzyme from this aerobe only exhibited hydrolytic activity on soluble starch, not on starch granules. The degradation of from starch by $\beta$-amylase was greatly stimulated by pullulanase addition. These results differentiated from other $\beta$-amylases reported. Based on a previous result that showed the enzyme system involves in effective degradation of raw starch granules, this result strongly suggested that the purified enzyme (E-I) can be a synergistic part of starch granule-digestion and E-II plays a crucial role in digestion of starch granules.

• Journal of Life Science
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• v.28 no.6
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• pp.639-647
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• 2018

A new heat shock protein 70 was identified in red-spotted grouper (Epinephelus akaara) based on an expression analysis. The cDNA of red-spotted grouper Hsp70 (designated RgHsp70) was cloned by the rapid amplification of cDNA ends (RACE) techniques. The full-length of RgHsp70 cDNA was 2,152 bp, consisting of a 5'-terminal untranslated region (UTR) of 105 bp, a 3'-terminal UTR of 274 bp, and an open reading frame (ORF) of 1,773 bp that encode a polypeptide of 590 amino acids with a theoretical molecular weight of 64.9 kDa and an estimated isoelectric point of 5.2. Multiple alignment and phylogenetic analyses revealed that the RgHsp70 gene shares a high similarity with other Hsp70 fish genes. RgHsp70 contained all three classical Hsp70 family signatures. The results indicated the RgHsp70 is a member of the heat shock protein 70 family. RgHsp70 mRNA was predominately expressed in the liver, with reduced expression noted in the head-kidney tissues. The expression analysis of different water temperatures (21, 18, 15 and $12^{\circ}C$) for sampled livers revealed that expression gradually increased at $12^{\circ}C$ compared to $21^{\circ}C$. In this study, the effects of water temperature lowering on the physiological conditions were investigated, and the results revealed that novel RgHsp70 may be an important molecule involved in stress responses.

• Bulletin of the Korean Chemical Society
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• v.34 no.9
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• pp.2616-2622
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• 2013

A novel zwitterionic monomer 3-[(2-acrylamido)dimethylammonio]propanesulfonate (DMADAS) was designed and synthesized in this study. Then it was polymerized with acrylamide (AM) by free radical polymerization in 0.5 mol/L NaCl solution with ammonium persulfate ($(NH_4)_2S_2O_8$) and sodium sulfate ($NaHSO_3$) as initiator. The structure and composition of DMADAS and acrylamide-3-[(2-acrylamido)-dimethylammonio]propanesulfonate copolymer (P-AM-DMADAS) were characterized by FT-IR spectroscopy, $^1H$ NMR and elemental analyses. Isoelectric point (IEP) of P-AM-DMADAS was tested by nanoparticle size and potential analyzer. Solution properties of copolymer were studied by reduced viscosity. Antipolyelectrolyte behavior was observed and was found to be enhanced with increasing DMADAS content in copolymer. The results showed that the viscosity of P-AM-DMADAS is 5.472 dl/g in pure water. Electrolyte was added, which weakened the mutual attraction between sulfonic acid group and quaternary ammonium group. The conformation became loose, which led to the increase of reduced viscosity. The ability of monovalent and divalent cation influencing the viscosity of zwitterionic copolymer obeyed the following sequence: $Li^+$ < $Na^+$ < $K^+$, $Mg^{2+}$ < $Ca^{2+}$ < $Ba^{2+}$, and that of anion is in the order: $Cl^-$ < $Br^-$ < $I^-$, $CO{_3}^{2-}$ > $SO{_3}^{2-}{\approx}SO{_4}^{2-}$.

• Korean Journal of Microbiology
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• v.1 no.1
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• pp.10-18
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• 1963

Studies with the Tobacco Mosaic Viruses; W. S Kim, and So, I Y., (Dept. of biology Sung Kyun Kwan Univer. Seoul, Korea.). Using the common strain of tobacco mosaic virus (TMV) which was sent from the Dept. of Plant Pathology, University of Wisconsin, U.S.A. as control virus, a possible new strain of tobacco mosaic virus (SMV) was isolated from tobacco leaves collected from Tobacco Experiment Station farms as well as from various blends of manufactured Korean cigaretts. SMV was isolated by single lesion isolation method and by inoculating the virus through various species of host plants. The two viruses, TMV and SMV were indentified by the difference in symptoms, host range, serological reaction, and electron micrograpy. As the results of the above experiment the author believes the virus isolate SMV is a different strain of TMV. The experimental evidences that SMV belongs to the TMV group are as follows; 1. Both viruses produced local necrotic lesions on Nicotiana glutimosa L. 2. Both showed a dilution end point of $10^8$. 3. Aphid transmission was failed with the viruses. 4. Both had an isoelectric point around pH 3.3. 5. Two viruses were serological reactive. 6. The size of the virus particles was around 270-300mu as they were observed under the electron microscope. The virus SMV, however, is different from the common strain of TMV and the experimental evidences are as follows; 1. SMV produced quite different symptoms from TMV on various host plants like tobacoo(Nicotiana tabacum L., White Burley), Nicotiana rustica L., Chenopodium Koreanse Nakai. Bata vulgaris L., and Datura tatula L., SMV produced distinct local lesions on these host plants whereas TMV incited largely mosaic diseases. 2. The serological titers obtained from the heterologous combinations were lower than those from homologous combinations of antigens and antiser.