• Applied Biological Chemistry
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• v.46 no.2
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• pp.130-133
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• 2003

A method for separating heme-iron from hemoglobin (Hgb) hydrolysate by dialysis was developed. Recovery of heme-iron increased with increasing Hgb concentration, whereas rejection of peptide and separation effciency expressed by HP ratio (heme-iron/peptide) did not show significant differences. HP ratio increased with increases in the degree of hydrolysis of Hgb and $KH_2PO_4$ concentrations of dialysis solution. Recovery of heme-iron decreased with increase in the pH of dialysis solution due to wash-out of heme-iron across the dialysis membrane caused by increase in solubility of heme-iron. Rejections of peptide were 74.5 and 87.5% (2 and 5 kDa of cut off size, respectively), whereas recovery of heme-iron decreased from 86.5 (2 kDa) to 63.1% (25 kDa). Amounts of heme-iron and peptide of dried heme-iron product were 21.7 and 77.0%, and HP ratio and production yield were 28.2 and 6.5%, respectively.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.130.3-131
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• 2003

The biological effects of NADPH-dependent isocitrate dehydrogenase (IDPc) inhibitor. DA-11004, was examined in obese zucker rats or streptozotocin-induced diabetic SD rats. Diabetes was induced by injection of streptozotocin (50mg/kg) dissolved in citrate buffer (pH 4.8) into the tail vein and induction of diabetes was confirmed by the measurement of the tail blood glucose level at 48h. DA-11004 (30mg/kg, po) was injected for successive 7days and significantly reduced the plasma glucose in streptozotocin-induced diabetic rats (P<0.05). (omitted)

• Fisheries and Aquatic Sciences
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• v.21 no.3
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• pp.5.1-5.8
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• 2018

Stimulator of interferon gene (STING) is induced by various inflammatory agents, such as lipopolysaccharide and microbial pathogens, including virus and bacteria. In this study, we obtained a full-length cDNA of a STING homolog from olive flounder using rapid amplification of cDNA ends PCR technique. The full-length cDNA of Paralichthys olivaceus STING (PoSTING) was 1442 bp in length and contained a 1209-bp open reading frame that translated into 402 amino acids. The theoretical molecular mass of the predicted protein sequence was 45.09 kDa. In the PoSTING protein, three transmembrane domains and the STING superfamily domain were identified as characteristic features. Quantitative real-time PCR revealed that PoSTING expressed in all the tissues analyzed, but showed the highest level in the spleen. Temporal expression analysis examined the significantly upregulated expression of PoSTING mRNA after viral hemorrhagic septicemia virus (VHSV) stimulation. In contrast, no significant changes in the PoSTING expression were detected in Edwardsiella tarda-challenged group compared to the un-injected control. The expression of P. olivaceus type I interferon (PoIFN-I) was also highly upregulated upon VHSV challenge. These results suggest that STING might be involved in the essential immune defense against viral infection together with the activation of IFN-I in olive flounder.

• 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.

• Journal of dental hygiene science
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• v.9 no.2
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• pp.189-195
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• 2009

Two diketones, 1,2-phenylpropanedione (PD) and diacetyl (DA) were investigated as new visible light photosensitizers for dental composite resin of bis-GMA in order to improve the photopolymerization effect. The photopolymerization efficiency of bis-GMA composite resin containing PD and DA was studied by IR absorption spectroscopy. And the results were compared with that of camphorquinone (CQ). Relative photopolymerization efficiency of the photosensitizers increased in the order of DA < CQ < PD. Thus. PO is a new visible light photosensitizer for dental composite resin with higher photopolymerization efficiency than that of CQ.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.196.2-197
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• 2003

Parkinson's disease (PO) is a widespread neurodegenerative disorder. Even though PD has been studied in many aspects, it is still unknown the molecular signaling mechanisms linking reactive oxygen species (ROS) and neuronal apoptosis in PD. A better understanding of cellular mechanisms that occur in Parkinson's disease is essential for development of new therapies. In this study we investigated the signaling molecules involved in neuronal apoptosis induced by 6-hydroxydopamine (6-OHDA) in human SK-N-SH neuroblastoma cells as a model cellular system. (omitted)

• Microbiology and Biotechnology Letters
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• v.37 no.1
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• pp.62-68
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• 2009

The culture conditions to maximize the production of laccase (EC 1.10.3.2) from Fomitopsis pinicola mycelia were investigated. Among the tested media for the enzyme production, mushroom complete medium (MCM ; 2% dextrose, 0.2% peptone, 0.2% yeast extract, 0.05% $KH_2PO_4$, and 0.05% $MgSO_4{\cdot}7H_2O$) showed the highest activity of the enzyme. To optimize the culture condition for the laccase activity, influence of various carbon and nitrogen sources was investigated in MCM. Among various carbon and nitrogen sources, 2% glucose and 0.4% peptone showed the highest production of the enzyme, respectively. For the phosphorus and inorganic source, 0.05% $NaH_2PO_4$ and 0.05% $CaCl_2$ were best for the enzyme activity. The enzyme production was reached to highest level after the cultivation for 8 days at $25^{\circ}C$. Native polyacrylamide gel electrophoresis (PAGE) followed by the laccase activity staining using 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as the substrate was performed to identify the laccase under culture conditions studied. Zymogram analysis of the culture supernatant showed a laccase band with molecular mass of 52 kDa. The optimum pH and temperature for the enzyme activity were $80^{\circ}C$ and pH 3.0.

• Microbiology and Biotechnology Letters
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• v.41 no.2
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• pp.145-152
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• 2013

The culture conditions to maximize the production of endoglucanase (EC 3.2.1.4) from the brown rot fungus Fomitopsis pinicola MKACC 54347 mycelia were investigated. Among the tested media for endoglucanase production, Mandel's mineral salts medium (MSM; 1% cellulose, 0.1% peptone, 0.14% $(NH_4)_2SO_4$, 0.03% urea, 0.2% $KH_2PO_4$, 0.03% $MgSO_4{\cdot}7H_2O$, 0.03% $CaCl_2$, and 0.1% trace metal solution (19.8 mM $FeSO_4$, 13.0 mM $MnSO_4$, 12.2 mM $ZnSO_4$, and 15.4 mM $CoCl_2$)) produced the highest activity of the enzyme. To optimize the medium composition for enzyme activity, the effects of various carbon, nitrogen, phosphorus, and inorganic sources were investigated in MSM. Maximal enzyme production was accomplished using a medium containing 2% carboxymethyl cellulose (CMC), 2% yeast extract, 0.2% $KH_2PO_4$, 0.03% $MnSO_4$, and 0.3% trace metal solution. Different physiological conditions, like incubation period and temperature, were also examined to assess their influence on enzyme production. Enzyme production from F. pinicola reached its highest level after cultivation for 8 days at $25^{\circ}C$. Nondenaturing polyacrylamide gel electrophoresis (PAGE), followed by the endoglucanase activity staining using CMC as the substrate, was performed to identify the endoglucanase under the culture conditions studied. Zymogram analysis of the culture supernatant revealed an endoglucanase band with a molecular mass of 52 kDa. The optimum pH and temperature for enzyme activity were $55^{\circ}C$ and pH 5.0, respectively.

• Journal of Life Science
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• v.25 no.6
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• pp.673-679
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• 2015

The culture conditions needed to maximize the production of laccase from Pholiota highlandensis mycelia were investigated. Among the tested media for laccase production, Coriolus versicolor medium (CVM; 2% dextrose, 0.4% peptone, 0.6% yeast extract, 0.046% KH₂PO₄, 0.1% K₂HPO₄, 0.05% MgSO₄·7H₂O) showed the highest activity for the enzyme. Then, to optimize culture conditions for laccase activity, the influences of various carbon, nitrogen, phosphorus, and inorganic salt sources in CVM were investigated. The optimum culture medium was 2% fructose, 0.4% peptone with 0.6% yeast extract, 0.05% NaH₂PO₄, and 0.05% MgSO₄·7H₂O as carbon, nitrogen, phosphorus, and inorganic salt sources, respectively. Several aromatic compounds in the medium enhanced laccase activity to varying degrees. Guaiacol induced maximum laccase production, yielding 114.1 U/ml laccase activity after cultivation for 11 days at 25℃. The optimum pH and temperature for laccase production were 8.0 and 35℃, respectively. Native polyacrylamide-gel electrophoresis (PAGE) followed by laccase-activity staining with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as the substrate was performed to identify the presence of laccase under the optimum conditions studied. Zymogram analysis of the supernatant culture showed an enzymatic band with a molecular mass of about 90 kDa.

• Journal of Microbiology and Biotechnology
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• v.17 no.8
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• pp.1271-1283
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• 2007

A fibrinolytic protease (PoFE) was purified from the cultured mycelia of the edible oyster mushroom Pleurotus ostreatus, using a combination of various chromatographies. The purification protocol resulted in an 876-fold purification of the enzyme, with a final yield of 6.5%. The apparent molecular mass of the purified enzyme was estimated to be 32 kDa by SDS-PAGE, fibrin-zymography, and size exclusion using FPLC. The optimal reaction pH value and temperature were pH 6.5 and $35^{\circ}C$, respectively. PoFE effectively hydrolyzed fibrinogen, preferentially digesting the $A{\alpha}$-chain and the $B{\beta}$-chain over the ${\gamma}$-chain. Enzyme activity was enhanced by the addition of $Ca^{2+},\;Zn^{2+},\;and\;Mg^{2+}$ ions. Furthermore, PoFE activity was potently inhibited by EDTA, and it was found to exhibit a higher specificity for the chromogenic substrate S-2586 for chymotrypsin, indicating that the enzyme is a chymotrypsin-like metalloprotease. The first 19 amino acid residues of the N-terminal sequence were ALRKGGAAALNIYSVGFTS, which is extremely similar to the metalloprotease purified from the fruiting body of P. ostreatus. In addition, we cloned the PoFE protein, encoding gene, and its nucleotide sequence was determined. The cDNA of cloned PoFE is 867 nucleotides long and consists of an open reading frame encoding 288 amino acid residues. Its cDNA showed a high degree of homology with PoMEP from P. ostreatus fruiting body. The mycelia of P. ostreatus may thus represent a potential source of new therapeutic agents to treat thrombosis.