• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.12
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• pp.1940-1948
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• 2013

This study is conducted to investigate the antioxidative effect of oyster hydrolysates in the serum and liver of SD-rats through the determination of lipid content, production of free radicals and antioxidant enzyme activities. Two different hydrolysates, Protamex-treated and Neutrase-treated hydrolysate with the cross-linking of protein by transglutaminase (TGPN group) and without (PN group), were fed for 6 weeks. TGPN hydrolysate in serum and liver significantly decreased the total cholesterol in the range of 26.1% to 28.9%, and triglyceride in the liver of up to 6.3%. Superoxide radical in the serum and lipid peroxide radical in the liver were significantly decreased in SD-rats fed 200 mg TGPN hydrolysate. Superoxide dismutase activity was significantly decreased in the liver of SD-rats. These results indicate that TGPN hydrolysate could scavenge the superoxide and hydroxyl radicals, and reduce the superoxide dismutase and catalase activities. The TGPN is also protected the oxidation of protein by the free radicals.

• BMB Reports
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• v.40 no.2
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• pp.218-225
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• 2007

In Arabidopsis thaliana, the microtubule-associated protein AtMAP65-1 shows various functions on microtubule dynamics and organizations. However, it is still an open question about whether AtMAP65-1 binds to tubulin dimers and how it regulates microtubule dynamics. In present study, the tubulin-binding activity of AtMAP65-1 was investigated. Pull-down and co-sedimentation exp eriments demonstrated that AtMAP65-1 bound to tubulin dimers,at a molar ratio of 1 : 1. Cross-linking experiments showed that AtMAP65-1 bound to tubulin dimers by interacting with $\alpha$-tubulin of the tubulin heterodimer. Interfering the bundling effect of AtMAP65-1 by addition of salt and monitoring the tubulin assembly, the experiment results indicated that AtMAP65-1 promoted tubulin assembly by interacting with tubulin dimers. In addition, five truncated versions of AtMAP65-1, namely AtMAP65-1 $\Delta$N339 (amino acids 340-587); AtMAP65-1 $\Delta$N494 (amino acids 495-587); AtMAP65-1 340-494 (amino acids 340-494); AtMAP65-1 $\Delta$C495 (amino acids 1-494) and AtMAP65-1 $\Delta$C340 (amino acids 1-339), were tested for their binding activities and roles in tubulin polymerization in vitro. Four (AtMAP65-1 $\Delta$N339, $\Delta$N494, AtMAP65-1 340-494 and $\Delta$C495) from the five truncated proteins were able to co-sediment with microtubules, and three (AtMAP65-1 $\Delta$N339, $\Delta$N494 and AtMAP65-1 340-494) of them could bind to tubulin dimers in vitro. Among the three truncated proteins, AtMAP65-1 $\Delta$N339 showed the greatest activity to promote tubulin polymerization, AtMAP65-1 $\Delta$N494 exhibited almost the same activity as the full length protein in promoting tubulin assembly, and AtMAP65-1 340-494 had minor activity to promote tubulin assembly. On the contrast, AtMAP65-1 $\Delta$C495, which bound to microtubules but not to tubulin dimers, did not affect tubulin assembly. Our study suggested that AtMAP65-1 might promote tubulin assembly by binding to tubulin dimers in vivo.

• Applied Chemistry for Engineering
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• v.10 no.8
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• pp.1104-1108
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• 1999

In this study it was focused that if the chrome could be removed in the shaving dust, the reusable protein resource could be recycled from the shaving dust. As the chrome forms a Cr-collagenate by cross linking of collagen in the shaving dust, the dust was steeped for swelling and plumping by $Ca(OH)_2$ solution and then chromium in dust was resolved out by $H_2SO_4$ solution in the first stage process. In the second stage one, dust was swelled and plumped by NaOH solution and then the chromium in dust was oxygenated to a hexavalent chrome, which has high solubility in $H_2O_2$ solution. And then the chromium was removed byt he steeping of $H_2SO_4$ solution as the last process. The first stage process was consisted of the sequential steeping of 3%-$Ca(OH)_2$ and $0.8%-H_2SO_4$. In the second stage the total chrome was effectively removed by the sequential steeping of 0.1%-NaOH, $3%-H_2O_2$ and $1%-H_2SO_4$. In the completely dechromated dust, the humidity was measured as 10.68% and the crude protein was contained by 79.81%. Steeping solutions were reused 3 times for chrome removal process, the chrome was entirely removed in shaving dust.

• Toxicological Research
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• v.8 no.2
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• pp.343-357
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• 1992

Tumor necrosis factor-${\alpha}$(TNF), a polypeptide hormone secreted primarily by activated macrophages, was originally identified on the basis of its ability to cause hemorrhagic necrosis and tumor regression in vivo. Subsequently, TNF has been shown to be an important component of the host responses to infection and cancer and may mediate the wasting syndrome known as cachexia. These systemic actions of TNF are reflected in its diverse effects on target cells in vitro. TNF initiates its diverse cellular actions by binding to specific cell surface receptors. Although TNF receptors have been identified on most of animal cells, regulation of these receptors and the mechanisms which transduce TNF receptor binding into cellular responses are not well understood. Therefore, in the present study, the mechanisms how TNF receptors are being regulated and how TNF receptor binding is being transduced into cellular responses were investigated in rat liver plasma membranes (PM) and ME-180 human cervical carcinoma cell lines. $^{125}I$-TNF bound to high ($K_d=1.51{\pm}0.35nM$)affinity receptors in rat liver PM. Solubilization of PM with 1% Triton X-100 increased both high affinity (from $0.33{\pm}0.04\;to\;1.67{\pm}0.05$ pmoles/mg protein) and low affinity (from $1.92{\pm}0.16\;to\;7.57{\pm}0.50$ pmoles/mg protein) TNF binding without affecting the affinities for TNF, suggesting the presence of a large latent pool of TNF receptors. Affinity labeling of receptors whether from PM or solubilized PM resulted in cross-linking of $^{125}I$-TNF into $M_r$ 130 kDa, 90 kDa and 66kDa complexes. Thus, the properties of the latent TNF receptors were similar to those initially accessible to TNF. To determine if exposure of latent receptors is regulated by TNF, $^{125}I$-TNF binding to control and TNF-pretreated membranes were assayed. Specific binding was increased by pretreatment with TNF (P<0.05), demonstrating that hepatic PM contains latent TNF receptors whose exposure is promoted by TNF. Homologous up-regulation of TNF receptors may, in part, be responsible for sustained hepatic responsiveness during chronic exposure to TNF. As a next step, the post-receptor events induced by TNF were examined. Although the signal transduction pathways for TNF have not been delineated clearly, the actions of many other hormones are mediated by the reversible phosphorylation of specific enzymes or target proteins. The present study demonstrated that TNF induces phosphorylation of 28 kDa protein (p28). Two dimensional soidum dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) resolved the 28kDa phosphoprotein into two isoforms having pIs of 6.2 and 6.1. The pIs and relative molecular weight of p28 were consistent with those of a previously characterized mRNA cap binding protein. mRNA cap binding proteins are a class of translation initiation factors that recognize the 7-methylguanosine cap structure found on the 5' end of eukaryotic mRNAs. In vitro, these proteins are defined by their specific elution from affinity columns composed of 7-methylguanosine 5'-triphosphate($m^7$GTP)-Sepharose. Affinity purification of mRNA cap binding proteins from control and TNF treated ME-180 cells proved that TNF rapidly stimulates phosphorylation of an mRNA cap binding protein. Phosphorylation occurred in several cell types that are important in vitro models of TNF action. The mRNA cap binding protein phosphorylated in response to TNF treatment was purifice, sequenced, and identified as the proto-oncogene product eukaryotic initiation factor-4E(eIF-4E). These data show that phosphorylation of a key component of the cellular translational machinery is a common early event in the diverse cellular actions of TNF.

• Clean Technology
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• v.22 no.3
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• pp.175-180
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• 2016

Tyrosinases catalyze the hydroxylation of a monophenol (monophenolase activity) and the conversion of an o-diphenol to o-quinone (diphenolase activity), which are mainly involved in the modification of tyrosine residues into 3,4-dihydroxyphenyl-alanine (DOPA) and DOPA/DOPAquinone-derived intermolecular cross-linking. Previously, we obtained a slightly acidic and cold-active tyrosinase, tyrosinase-CNK, by our recombinant protein approach. The enzyme showed optimal activity at pH 6.0 and 20 ℃ with an abnormally high monophenolase/diphenolase activity ratio and still had approximately 50% activity compared with the highest activity even in ice water. Here, we investigated reaction stability of the recombinant tyrosinase-CNK as a psychrophilic enzyme. The enzyme showed remarkable thermal stability at 0 ℃ and the activity was well conserved in repeated freeze-thaw cycles. Although water-miscible organic solvent as reaction media caused the activity decrease of tyrosinase-CNK as expected, the enzyme activity was not additionally decreased with increased concentration in organic solvents such as ethanol and acetonitrile. Also, the enzyme showed high salt tolerance in chaotropic salts. It was remarkably considered that 2⁺ metal ions might inhibit the incorporation of Cu²⁺ into the active site. We expect that these results could be used to design tyrosinase-mediated enzymatic reaction at low temperature for the production of catechols through minimizing unwanted self-oxidation and enzyme inactivation.

• Journal of Chest Surgery
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• v.30 no.11
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• pp.1092-1096
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• 1997

Thromboelastography(TEG) enables a global assessment of hemostatic function to be made from a single blood sample, documenting the interaction of platelets with protein coagulation cascade from the time of the initial platelet-fibrin interaction, through platelet aggregation, clot strengthening and fibrin cross linking to eventual clot Iysis. Thirty-five patients(mean age 34$\pm$ 12) undergoing open heart surgery from April 1st, 1996 to August 31th, 1996 were investigated at preoperatively and immediate, one hour, and 24 hours after cessation of cardiopulmonary bypass using TEG. Comparisons were made between classic hematological indices and TEG data. There were statistically significant correlation between maximal amplitude(MA) and platelet count before CPB, activating clotting time(ACT) and TEG date(R time, K time and a angle) at 24-hour after CPB. The data on the predictive accuracy for postoperative bleeding at 24-hour after CPB, the TEG was significantly better than ACT(57%) or the coagulation profiles(43%) as a predictor of postoperative bleeding, with an accuracy rate of 100% (P=0.0043). In conclusion, TEG seems to be easy to use, clinically accurate, cost effective and provides data which can effectively manage a patient's hemostasis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.5
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• pp.738-746
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• 2011

We manufactured rice-whole soybean curd by a microbial transglutaminase (MTGase) with a mixture of hydrolyzed rice and micronized whole soybean powder (MWSP) and analyzed its rheological properties, including texture, viscoelasticity, protein cross-linking, and surface structure. A 40% rice suspension digested with a Termamyl enzyme at $85^{\circ}C$ for 20 min showed a 9.0% reducing sugar and a consistency of $1.27\;Pa{\cdot}s^n$, resulting in a great reduction in consistency. A MWSP suspension with 22% solid content was transformed into a typical tofu texture. MWSP curd fortified with 7.5% rice showed enhanced texture properties, with a hardness of 639.6 dyne/$cm^2$, and a springiness of 0.96. In a MWSP suspension (18~22% w/v) treated with 5% MTGase, viscoelasticity increased dependently with MWSP concentration, and a 22% MWSP indicated a G' value of 5.1 Pa and a G'' value of 9.0 Pa. Furthermore, soybean proteins present in the 22% MWSP curd largely disappeared or formed polymers with a high molecular weight by MTGase reaction within 30 min. MWSP (22%) fortified with 7.5% rice showed similar polymerization patterns on SDS-PAGE. The surface structure of the rice-MWSP curds was more dense and homogeneous network due to the addition of hydrolyzed rice. However, the surface structure of all rice-MWSP curds became rough and showed a non-homogeneous network after cold storage.

• Journal of the Korean GEO-environmental Society
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• v.24 no.9
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• pp.5-13
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• 2023

Cement-based reinforcement materials, which are representative slope reinforcement materials, can cause contamination of ground and groundwater when ground injection or surface application is applied. Accordingly, slope reinforcement materials using eco-friendly biopolymers are attracting attention as a means of replacing existing materials, but the biopolymers currently used are easily dissolved when exposed to groundwater or rainfall environments, reducing strength. In order to solve this problem, the cross-linking of protein between sodium casein and Transglutaminase (TGase, C₂₀H₁₆N₄O₂S₂) was used to increase the water resistance of biopolymers, and a rainfall slope test was conducted to evaluate their usability and applicability as a slope reinforcing material. In the case of reinforcement with only sodium casein, the precipitation dissolved sodium casein, and the slope was completely destroyed in 1 hour. On the other hand, it was observed that the slope reinforced by adding a small amount of TGase (0.5%) do not collapse even after 80 hours of rainfall duration due to increased water resistance. Strength and water resistance increases due to the addition of a small amount of TGase, and its applicability as an eco-friendly reinforcement is confirmed.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.10-17
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• 2016

Enzyme immunoassay to analyze specific binding activity of antibody to antigen uses horseradish peroxidase (HRP) or alkaline phosphatase (AP). Chemical methods are usually used for coupling of these enzymes to antibody, which is complicated and random cross-linking process. As results, it causes decreases or loss of functional activity of either antibody or enzyme. In addition, most enzyme assays use secondary antibody to detect antigen binding activity of primary antibody. Enzymes coupled to secondary antibody provide a binding signal by substrate-based color development, suggesting secondary antibody is required in enzyme immunoassay. Additional incubation time for binding of secondary antibody should also be necessary. More importantly, non-specific binding activity caused by secondary antibody should also be eliminated. In this study, we cloned AP isolated from Escherichia coli (E. coli) chromosome by PCR and fused to) hAY4 single-chain variable domain fragment (ScFv) specific to death receptor (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand (TRAIL). hAY4 ScFv-AP expressed in E. coli showed 73.8 kDa as a monomer in SDS-PAGE. However, this fusion protein shown in size-exclusion chromatography (SEC) exhibited 147.6 kDa as a dimer confirming that natural dimerization of AP by non-covalent association induced ScFv-AP dimerization. In several immunoassay such as ELISA, Western blot and immunocytochemistry, it showed antigen binding activity by color development of substrates catalyzed by AP directly fused to primary hAY4 ScFv without secondary antibody. In summary, hAY4 ScFv-AP fusion protein was successfully purified as a soluble dimeric form in E. coli and showed antigen binding activity in several immunoassays without addition of secondary antibody which sometimes causes time-consuming, expensive and non-specific false binding.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.6
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• pp.718-724
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• 1999

This study was investigated the changes of the properties of matter such as the gel formation of the combined fish based on the mixed rate between the ocean jumbo squid and Alaska pollock surimi, and compared the relationships between the gel formation and water holding capacity. The changes of the gel formation based on 20 min fish grinding time and $2.5\%$ salt concentration according to the mixed rate was thought as the optimal addition limit. There was no significant function of gel product more than $20\%$ Jumbo squid meat. The more squid meat in the mixed meat could make the lower breaking stress but 7:3 rate of pollock : squid could retain breaking strain. The effect of the moisture content on mixed fish meat was studied and the drastic decrease of the gel formation and water holding capacity was indicated in $78\%$.