• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1810-1818
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• 2015

For the synthesis of various pharmaceuticals, chiral alcohols are useful intermediates. Among them, (R)-ethyl-4-chloro-3-hydroxybutanoate ((R)-ECHB) is an important building block for the synthesis of L-carnitine. (R)-ECHB is produced from ethyl-4-chloro-3-oxobutanoate (ECOB) by a reductase-mediated, enantioselective reduction reaction. The Saccharomyces cerevisiae YOL151W reductase that is expressed in Escherichia coli cells exhibited an enantioselective reduction reaction toward ECOB. By virtue of the C-terminal His-tag, the YOL151W reductase was purified from the cell-free extract using Ni²⁺-NTA column chromatography and immobilized onto Ni²⁺-magnetic microparticles. The physical properties of the immobilized reductase (Imm-Red) were measured using electron microscopy, a magnetic property measurement system, and a zeta potential system; the average size of the particles was approximately 1 μm and the saturated magnetic value was 31.76 emu/g. A neodymium magnet was used to recover the immobilized enzyme within 2 min. The Imm-Red showed an optimum temperature at 45℃ and an optimum pH at 6.0. In addition, Bacillus megaterium glucose dehydrogenase (GDH) was produced in the E. coli cells and was used in the coupling reaction to regenerate the NADPH cofactor. The reduction/oxidation coupling reaction composed of the Imm-Red and GDH converted 20 mM ECOB exclusively into (R)-ECHB with an e.e._p value of 98%.

• 대한치과보철학회지
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• 제47권3호
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• pp.348-355
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• 2009

연구 목적: ShadeEye-Ncc dental chroma meter (Shofu Inc., Kyoto, Japan)와 Shadepilot$^{TM}$ system (Degudent Inc., Hanau, Germany)의 도재색조 재현의 정확성을 비교연구하여 보고자 함이다. 연구 재료 및 방법: ShadeEye-Ncc dental chroma meter와 Shadepilot$^{TM}$ system을 사용하여 Vitapan 3D Master shade guide의 2M2와 3M2 shade tap.을 측색하고, 이 자료를 통하여 shade guide tap. 형태로 도재시편을 제작한다. 시편제작 시 사용하는 porcelain system은 ShadeEye-Ncc dental chroma meter는 Vintage Halo Porcelain system을, Shadepilot$^{TM}$ system은 VitaOmega900 Porcelain system을 사용한다. 모델인 shade tap.과 제작된 도재시편을 spectrophotometer로 측색하고, 그 결과를 토대로하여 ${\Delta}E$를 계산하여, 두 측색기의 정확성을 비교검증하였다. 결과: 1. Shadepilot$^{TM}$ system이 ShadeEye-Ncc dental chroma meter에 비하여 더 적합한 도재색조 재현성을 나타내었다 (P <.05). 2. 모델인 Shade tap.의 shade에 따라서 도재색조 재현성에 차이를 나타내었다. 2M2 shade tap.에 비하여 3M2 shade tap.에서 더 적합한 도재색조 재현성을 나타내었다 (P <.05). 3. 두 측색기 모두에서 제작된 도재시편의 ${\Delta}E$가 평균4.44-6.14로 육안으로 구분 가능한 정도의 색조차이를 보였다. 결론: Shadepilot$^{TM}$ system이 비교적 더 정확한 도재색조 재현성을 나타낸다고 할 수 있으며, ShadeEye-Ncc dental chroma meter를 사용할 때는 치아 전체의 여러부분에서 측색을 하여 정확성을 높이는 것이 필요하리라 생각된다.

• KSBB Journal
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• 제24권4호
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• pp.341-346
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• 2009

본 연구에서는 Candida antarctica로부터 genomic DNA을 추출하여 lipase A(CalA) 유전자를 PCR 증폭하였고, 재조합 pColdIII/CalA, $pPICZ{\alpha}A$/CalA, $pPICZ{\alpha}A$/CalA$his{\times}6$을 구축하였다. 재조합 CalA 유전자의 기능적 발현을 위해 최적화된 시스템을 구축하고자 Escherichia coli와 Pichia pastoris 시스템에서 각각 수행하여 비교, 분석하였다. SDS PAGE gel을 통해 CalA의 발현의 여부 및 발현양을 확인하였고, pNPP를 기질로 한 가수분해 반응을 통해 활성을 측정하였다. E. coli 발현 시스템은 형질전환 방법이 간단하고, 미생물의 성장 속도가 빠르다는 장점을 갖지만 CalA의 활성이 0.02 Unit/ml으로 비교적 낮았으며 세포질 (cytoplasm)에서 발현되므로 비목적 단백질과의 분리 및 정제과정이 필요하다. 재조합 $pPICZ{\alpha}A$/CalA을 P. pastoris 시스템에서 발현한 경우 높은 발현양 뿐만 아니라 분비작용으로 인해 고순도 발현이 용이하였고, 활성 또한 약 0.7 Unit/ml으로 가장 높았다. 결론적으로 CalA의 기능적 발현을 위해 P. pastoris 시스템을 구축하는 것이 가장 적합함을 확인하였다.

• 대한기계학회논문집
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• 제16권4호
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• pp.649-657
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• 1992

본 연구에서는 스테핑 모터의 토크특성과 전동시스템의 미스얼라인 먼트에 의 한 플리편심을 고려하여 스페핑 모터 구동 타이밍벨트 전동시스템의 전동오차에 관한 이론 및 실험적 연구를 수행하고, 전동오차 감소를 위한 설계 개선 기초자료를 얻고자 한다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권2호
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• pp.144-149
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• 2001

For the production and purification of a single chain human insulin precursor, four types of fusion peptides $\beta$-galactosidase (LacZ), maltose binding protein (MBP), glutathione-S-transferase (GST), and (His)(sub)6-tagged sequence (HTS) were investigated. Recombinant E. coli harboring hybrid genes was cultivated at 37$\^{C}$ for 1h, and gene induction occurred when 0.2mM of isopropyl-D-thiogalactoside (IPTG) was added to the culture broth, except for E. coli BL21 (DE3) pLysS harboring a pET-BA cultivation with 1.0mM IPTG, followed by a longer than 4h batch fermentation respectively. DEAE-Sphacel and Sephadex G-200 gel filtration chromatography, amylose affinity chromatography, glutathione-sepharose 4B affinity chromatography, and a nickel chelating affinity chromatography system as a kind of immobilized metal ion affinity chromatography (IMAC) were all employed for the purification of a single chain human insulin precursor. The recovery yields of the HTS-fused, GST-fused, MBP-fused, and LacZ-fused single chain human insulin precursors resulted in 47%, 20%, 20%, and 18% as the total protein amounts respectively. These results show that a higher recovery yield of the finally purified recombinant peptides was achieved when affinity column chromatography was employed and when the fused peptide had a smaller molecular weight. In addition the pET expression system gave the highest productivity of a fused insulin precursor due to a two-step regulation of the gene expression, and the HTS-fused system provided the highest recovery of a fused insulin precursor based on a simple and specific separation using the IMAC technique.

• 대한한방소아과학회지
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• 제34권4호
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• pp.22-30
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• 2020

Objectives This study was conducted to confirm the inhibitory effect of β-glucan on epithelial inflammation induced by atopic dermatitis through Endocannabinoid system (ECS) activity. Methods Six-week-old NC/Nga mice were divided into a control group (Ctrl), atopic dermatitis elicitation group (ADE), and a β-glucan-treated group (β-glucan treatment after atopy dermatitis elicitation, β-GT). After 3 weeks, CB1, CB2, and GPR55 were observed to confirm the regulation of ECS activity, and filaggrin in the stratum corneum and Kallikrein-related peptidase (KLK) 7 in the stratum corneum and protease activated receptor (PAR)-2 were observed to confirm the inhibition of the inflammation, Phosphorylated extracellular signal-related kinase (p-ERK), Phosphorylated mammalian target of rapamycin (p-mTOR), and E-Cadherin were observed to confirm microenvironmental regulation. Results β-GT was significantly increased in CB1, CB2, and GPR55 positive reactions compared to that of the ADE. In positive reaction of the filaggrin in the stratum corneum, β-GT was significantly increased than that of the ADE. For KLK7 positive and PAR2 positive, β-GT was significantly reduced compared to the ADE. The p-ERK-positive and p-mTOR-positive reactions were significantly reduced in β-GT than in ADE. E-cadherin positive reaction was significantly increased in β-GT than in ADE (All p < 0.01). Conclusions It was confirmed that β-glucan has the effect of inhibiting the epithelium induced by atopic dermatitis through the ECS activity.

• 한국동물생명공학회지
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• 제35권2호
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• pp.155-162
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• 2020

Equine follicle stimulating hormone receptor (eFSHR) has a large extracellular domain and an intracellular domain containing approximately 10 phosphorylation sites within the G protein-coupled receptor. This study was conducted to analyze the function of phosphorylation sties at the eFSHR C-terminal region. We constructed a mutant of eFSHR, in which the C-terminal cytoplasmic tail was truncated at residue 641 (eFSHR-t641). This removed 10 potential phosphorylation sites from the C-terminal region of the intracellular loop. The eFSHR-wild type (eFSHR-wt) and eFSHR-t641 cDNAs were subcloned into the pCMV-ARMS1-PK2 expression vector. These plasmids were transfected into PathHunter CHO-K1 Parental cells expressing β-arrestin 2 enzyme acceptor fusion protein and analyzed for agonist-induced cAMP response. The cAMP response in cells expressing eFSHR-t641 was lower than the response in cells expressing eFSHR-wt. EC₅₀ values of eFSHR-wt and eFSHR-t641 were 1079 ng/mL and 1834 ng/mL, respectively. eFSHR-t641 was approximately 0.58-fold compared with that of eFSHR-wt. The maximal response in eFSHR-wt and eFSHR-t641 was 24.7 nM and 16.7 nM, respectively. The Rmax value of phosphorylation sites in eFSHR-t641 was also decreased to approximately 68.4% of that in eFSHR-wt. The collective data implicate that the phosphorylation sites in the eFSHR C-terminal region have a pivotal role in signal transduction in PathHunter CHO-K1 cells, and indicate that β-arrestin is involved in coupling the activated receptors to the internalization system.

• 한국정보통신학회논문지
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• 제21권12호
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• pp.2261-2269
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• 2017

기존의 상용 LTE 통신망은 도시 단위 EPC(Evolved Packet Core)에 기지국(eNodeB)을 지역 단위로 고정 배치하여 서비스를 제공한다. 하지만, 개별 노드 단위의 서비스 제공이 필요하고 이동을 전제로 하는 재난망, 군 전술망에는 적합하지 않는 구조이다. 본 논문에서는 분산된 각 노드에서 서비스가 가능하도록 기지국과 사용자 단말 그리고 EPC를 하나의 장비로 통합한 LTE 시스템을 제안하고, 노드 간 통신을 위해 M2M(Machine to Machine) 타입의 사용자 단만을 노드 단위로 탑재하여 무선 In-band backhaul 링크를 구성하였다. 제안된 시스템에서는 LTE의 모든 구성 항목들이 하나로 통합되어 있어 일반 사용자 단말 접속 후 이동하여 접속된 노드가 바뀌면 IP 앵커가 되는 P-GW의 변경이 필요하다. 노드 간 이동성 지원을 위해 UPFE(User Packet Forwarding Extension)방식을 정의하고 P-GW가 변경되어도 이동성을 지원하는 EPC의 핸드오버 절차를 구현하였다. 또한, 통합 LTE 시스템에서 구현된 핸드오버 절차 대하여 통신 노드 추가에 따른 Cell Range를 분석하고 기존 시스템과의 핸드오버 지연시간을 비교한다.

• The Korean Journal of Physiology and Pharmacology
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• 제5권6호
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• pp.511-519
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• 2001

Nimopidine, one of dihydropyridine derivatives, has been widely used to pharmacologically identify L-type Ca currents. In this study, it was tested if nimodipine is a selective blocker for L-type Ca currents in sensory neurons and heterologous system. In mouse dorsal root ganglion neurons (DRG), low concentrations of nimodipine $(<10\;{\mu}M),$ mainly targeting L-type Ca currents, blocked high-voltage-activated calcium channel currents by ${\sim}38%.$ Interestingly, high concentrations of nimodipine $(>10\;{\mu}M)$ further reduced the 'residual' currents in DRG neurons from ${\alpha}_{1E}$ knock-out mice, after blocking L-, N- and P/Q-type Ca currents with $10\;{\mu}M$ nimodipine, $1\;{\mu}M\;{\omega}-conotoxin$ GVIA and 200 nM ${\omega-agatoxin$ IVA, indicating inhibitory effects of nimodipine on R-type Ca currents. Nimodipine $(>10\;{\mu}M)$ also produced the inhibition of both low-voltage-activated calcium channel currents in DRG neurons and ${\alpha}_{1B}\;and\;{\alpha}_{1E}$ subunit based Ca channel currents in heterologous system. These results suggest that higher nimodipine $(>10\;{\mu}M)$ is not necessarily selective for L-type Ca currents. While care should be taken in using nimodipine for pharmacologically defining L-type Ca currents from native macroscopic Ca currents, nimodipine $(>10\;{\mu}M)$ could be a useful pharmacological tool for characterizing R-type Ca currents when combined with toxins blocking other types of Ca channels.

• 한국광학회지
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• 제4권1호
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• pp.101-107
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• 1993

3준위 레이저 rate equation 및 overlap integral로부터 파장 1.48 ${\mu}m$에서 여기된 E$r^{3+}$ 첨가 광섬유 광증폭기를 위한 광섬유 매개 변수의 최적 조건을 계산하였다. 이 계산으로부터 Er3+ 첨가 광섬유 광섬유 광증폭기의 소신호 이득(small signal gain) 특성을 개구수 (N.A.), V값, 광섬유 길이, 차단 파장(cutoff wavelength) 등의 함수로 알아 보았으며 또한, 최대 소신호 이득을 갖기 위한 광섬유 매개 변수를 결정하였다.