• 한국응용과학기술학회지
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• 제26권3호
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• pp.248-262
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• 2009

This study is on the feasibility of use of glycoprotein in various areas such as cosmetics and food etc. by extracting, isolating and refining glycoprotein from carrots, red ginseng extract residue, sesame and pine needles using protease(pepsin) and by analyzing general characteristics and measuring various bioactivities. The results of analysis of nutritional composition showed protein contents of glycoprotein. In the analysis of constitutive amino acids, the ratio of contents of hydroxy proline and glycine, the characteristics of glycoproteins appeared similar and the contents of glutamic acid and aspartic acid appeared higher. As a result of measurement contents of total polyphenol and flavonoid, it showed that glycoprotein had more contents generally, and the effect of bioactivity of glycoprotein appeared higher although different kinds of glycoprotein showed a little DPPH radical and nitrite scavenging ability, total antioxidant capacity by ABTS, ACE inhibitory.

• 한국식품과학회지
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• 제41권4호
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• pp.405-412
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• 2009

본 연구는 꾸지뽕 나무 열매로부터 75 kDa의 당단백질(꾸지뽕 당단백질)을 추출한 후 꾸지뽕 당단백질의 첨가에 따른 알레르기성 염증 인자인 histamine 유리 억제능력 및 COX-2의 활성 억제 효과를 평가하였다. RBL-2H3세포를 22시간 동안 IgE로 감작시킨 후, HSA를 처리하여 histamine의 유리양을 측정한 결과 꾸지뽕 당단백질을 처리한 농도가 증가함에 따라 histamine의 유리와 COX-2의 활성 억제율은 증가하였다. 또한 꾸지뽕 당단백질의 처리는 HSA에 의해 유도된 세포내 ROS 생성량을 농도에 의존적으로 억제하였다. 한편 꾸지뽕 당단백질을 농도별로 처리하여 세포내 단백질을 추출하여 western blot을 실시한 결과 100 ${\mu}g$/mL 농도의 꾸지뽕 당단백질을 처리한 그룹에서 ERK1/2, AP-1과 COX-2의 활성 수준은 현저히 억제 되었다(p<0.05). 따라서 이러 한 결과에 미루어볼 때, 꾸지뽕 당단백질은 세포내 해독효소의 활성을 증가시킴으로써 ROS 수준을 감소시켰으므로 꾸지뽕 당단백질의 역할이 다른 천연물 유래의 당단백질과 마찬가지로 특이적인 항산화 능력을 지니고 있음을 나타내며 histamine의 유리 억제와 COX-2의 활성이 억제되었을 것으로 생각된다. 이는 꾸지뽕 당단백질이 항 알레르기 효능을 갖는 물질로써 알레르기성 비염, 아토피 등과 같은 알레르기 관련 질환의 예방 및 치료제로 사용될 수 있을 것 사료된다.

• 한국식품과학회지
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• 제31권2호
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• pp.482-487
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• 1999

과요오드산으로 산화시킨 당을 유리아미노산과 반응시켜서 ${\alpha}-NH_2$기와 펩티딜리신의 ${\varepsilon}-NH_2$기와 반응하여 결합한 것을 확인하였다. 그래서, 제조한 과요드산-산화당은 단백질 분자 표면의 리신의 ${\varepsilon}-NH_2$기와 Schiff 염기 반응으로 공유결합하여 당단백질을 만드는 것으로 볼 수 있다. 과요드산 산화당으로 변형한 고구마 ${\beta}$-아밀라아제는 C, H는 증가, N은 감소하였다. N말단의 ${\alpha}-NH_2$기는 70% (pentamer), 73% (monomer), ${\varepsilon}-NH_2$기는 33% (pentamer), 26% (monomer)가 산화당과 반응하였다. 페놀-황산법에 의한 총당 정량과 DNP 법으로 분석한 결과, $IO_4$-산화말토헥사오스는 고구마 ${\beta}$-아밀라아제 1몰당 6몰이 결합된 것으로 나타났고, 고구마 ${\beta}$-아밀라아제에 결합된 산화가용성 전분은 효소단백질 대비 13.2% (monomer), 13.5% (pentamer), 산화말토헥사오스는 9.7% (pentamer), 9.3% (monomer)로 나타났다.

• The Korean Journal of Physiology and Pharmacology
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• 제26권3호
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• pp.145-155
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• 2022

Multidrug resistance of tumors has been a severe obstacle to the success of cancer chemotherapy. The study wants to investigate the reversal effects of imperatorin (IMP) on doxorubicin (DOX) resistance in K562/DOX leukemia cells, A2780/Taxol cells and in NOD/SCID mice, to explore the possible molecular mechanisms. K562/DOX and A2780/Taxol cells were treated with various concentrations of DOX and Taol with or without different concentrations of IMP, respectively. K562/DOX xenograft model was used to assess anti-tumor effect of IMP combined with DOX. MTT assay, Rhodamine 123 efflux assay, RT-PCR, and Western blot analysis were determined in vivo and in vitro. Results showed that IMP significantly enhanced the cytotoxicity of DOX and Taxol toward corresponding resistance cells. In vivo results illustrated both the tumor volume and tumor weight were significantly decreased after 2-week treatment with IMP combined with DOX compared to the DOX alone group. Western blotting and RT-PCR analyses indicated that IMP downregulated the expression of P-gp in K562/DOX xenograft tumors in NOD/SCID mice. We also evaluated glycolysis and glutamine metabolism in K562/DOX cells by measuring glucose consumption and lactate production. The results revealed that IMP could significantly reduce the glucose consumption and lactate production of K562/DOX cells. Furthermore, IMP could also remarkably repress the glutamine consumption, α-KG and ATP production of K562/DOX cells. Thus, IMP may sensitize K562/DOX cells to DOX and enhance the antitumor effect of DOX in K562/DOX xenograft tumors in NOD/SCID mice. IMP may be an adjuvant therapy to mitigate the multidrug resistance in leukemia chemotherapy.

• Journal of Microbiology and Biotechnology
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• 제8권4호
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• pp.318-324
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• 1998

A ${\beta}$-galactosidase with high transgalactosylic activity was purified from a Bacillus species, registered as KFCC10855. The enzyme preparation showed a single protein band corresponding to a molecular mass of 150 kDa on SDS-PAGE and gave a single peak with the estimated molecular mass of 250 kDa on Sephacryl S-300 gel filtration, suggesting that the enzyme is a homodimeric protein. The amino acid and sugar analyses revealed that the enzyme is a glycoprotein, containing 19.2 weight percent of sugar moieties, and is much more abundant in hydrophilic amino acid residues than in hydrophobic residues, the mole ratio being about 2:1. The pI and optimum pH were determined to be 5.0 and 6.0, respectively. Having a temperature optimum at $70^{\circ}C$ for the hydrolysis of lactose, the enzyme showed good thermal stability. The activity of the enzyme preparation was markedly increased by the presence of exogenous Mg (II) and was decreased by the addition of EDTA. Among the metal ions examined, the most severely inhibitory effect was seen with Ag (I) and Hg (II). Further, results of protein modification by various chemical reagents implied that 1 cysteine, 1 histidine, and 2 methionine residues occur in certain critical sites of the enzyme, most likely including the active site. Enzyme kinetic parameters, measured for both hydrolysis and transgalactosylation of lactose, indicated that the enzyme has an excellent catalytic efficiency for formation of the transgalactosylic products in reaction mixtures containing high concentrations of the substrate.

• Mass Spectrometry Letters
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• 제8권4호
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• pp.69-78
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• 2017

Glycosylation, which is one of the most common post-translation modification (PTMs) of proteins, plays a variety of crucial roles in many cellular events and biotherapeutics. Recent advances have led to the development of various analytical methods employing a mass spectrometry for glycomic and glycoproteomic study. However, site-specific glycosylation analysis is still a relatively new area with high potential for technologies and method development. This review will cover current MS-based workflows and technologies for site-specific mapping of glycosylation ranging from glycopeptide preparation to MS analysis. Bioinformatic tools for comprehensive analysis of glycoprotein with high-throughput manner will be also included.

• Macromolecular Research
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• 제17권4호
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• pp.232-239
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• 2009

This paper describes the simple nanopatterning of proteins on polyelectrolyte surfaces using microcontact printing with a nanopatternable, hydrophilic composite nanomold. The composite nanomold was easily fabricated by blending two UV-curable materials composed of Norland Optical Adhesives(NOA) 63 and poly(ethylene glycol) dimethacrylate(PEG-DMA). NOA 63 provided stable nanostructure formation and PEG-DMA induced high wettability of proteins in the nanomold. Using the composite mold and functionalized surface with polyelectrolytes, the fluorescent, isothiocyanate-tagged, bovine serum albumin(FITC-BSA) was successfully patterned with 8 nm height and 500 nm width. To confirm the feasibility of the protein assay on a nanoscale, a glycoprotein-lectin assay was successfully demonstrated as a model system. As expected, the lectins correctly recognized the nano-patterned glycoproteins such as chicken ovalbumin. The simple preparation of composite nanomold and functionalized surface with a universal platform can be applied to various biomolecules such as DNA, proteins, carbohydrates, and other biomolecules on a nanoscale.

• 한국응용과학기술학회지
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• 제26권4호
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• pp.457-466
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• 2009

This study is manufacturing method and analysis of feasibility about collagen peptide from fish scale. This is processed by enzyme hydrolysis, isolating and refining etc. The results of analysis of nutritional composition showed protein content of collagen peptide. In the analysis of constitutive amino acids, the ratio of contents of hydroxyproline and glycine, the characteristics of collagen peptides appeared similar and the contents of glutamic acid and aspartic acid which are involved in protein metabolism. As a result of measurement of total polyphenol content and total flavonoid, it showed that collagen peptide had more contents generally, and the effect of bioactivity of pig-skin collagen peptide appeared higher although different kinds of scale collagen peptide showed a little DPPH radical scavenging ability, total antioxidant capacity by ABTS, ACE inhibitory.

• 한국잠사곤충학회지
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• 제41권2호
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• pp.75-81
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• 1999

The storage protein-like protein has been purified from the 5th instar larval haemolymph of the Chinese osk silkwom, Antheraea pernyi, and the preparation was shown to be homogeneous by 7.5% native-PAGE. The molecule was consisted of a single subunit with a molecular weight of 80K, but the number of the subunits was not determined. The protein was defied as glycoprotein by Schiff's regent stining. Rabbit antibody prepared against the purified protein crotein crossreacted with the 5th instar larval haemolymph proteins of Antheraea pernyi and antheraea yamamai, but not with those of Bombyx mori and Bombyx mandarina.

• 한국미생물·생명공학회지
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• 제19권2호
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• pp.158-162
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• 1991

Endoinulinase was purified from a commercial inulin preparation produced by Aspergillus ficuum using ion exchange chromatography on CM-Sephadex C-50 and DEAE-Sepharose 6B, HPLC gel filtration on a Protein Pak 125 Colum and HPLC ion exchange chromatography on a TSK DEAE-5pw Column. The endoinulinase had a molecular weight of 72,000${\pm}$1,000 and was glycoprotein with 23 to 25% w/w sugar content. The enzyme was much more active on inulin with random cleavage mode than on sucrose and on palatinose: The ration of activity on inulin and sucrose (I/S ratio) was 10~14.