• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1350-1355
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• 2004

The sprA and sprB genes, encoding the chymotrypsin-like proteases Streptomyces griseus protease A (SGPA) and Streptomyces griseus protease B (SGPB), and the sprT gene that encodes Streptomyces griseus trypsin (SGT) were cloned from S. griseus and were overexpressed in various strains of S. griseus. When the sprT gene was introduced into S. griseus, trypsin activity increased 2-fold in the A-factor deficient mutant strain, S. griseus HH1, and increased 4-fold in the wild strain, S. grise us IFO 13350. However, there was no detectable increase of chymotrypsin activity in the transformants of S. griseus with either sprA or sprB, in contrast to the results obtained from S. lividans as a heterologous host. To solve the negative gene dosage effects in S. griseus, either the sprA or the sprB genes with their own ribosome binding sites were linked to the downstream of the entire sprT gene, and the coexpression efficiency was examined in S. lividans and S. griseus. The transformants of S. lividans with either pWHM3-TA (sprT+sprA) or pWHM3­TB (sprT+sprB) showed 3-fold increase of trypsin activity over that of the control, however, only the transformant of pWHM3-TB demonstrated 7-fold increase in chymotrypsin activity, indicating that the pWHM3-TB has a successful construction for the overexpression of chymotrypsin in Streptomyces. When the coexpression vectors were introduced into S. griseus IFO 13350, the trypsin level sharply increased by more than 4-fold, however, the chymotrypsin level did not increase. These results strongly suggest that the overexpression of the sprA and sprB genes is tightly regulated in S. griseus.

• 대한기계학회논문집A
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• 제28권8호
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• pp.1174-1182
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• 2004

Recently, Self Piercing Rivet(SPR) has been spotlighted in the automotive industry as a substitutive resort of spot welding and has also been watched by the designer as lightening a car body due to their superior assembly processes. Fatigue behavior of SPR joint needs to be investigated experimentally and numerically to predict its structural stiffness and fatigue life. Testing of lap-shear specimens with various material combinations is performed to obtain the joining strength and the fatigue life of SPR connections. The simulation of SPR lap-shear specimens is also conducted to obtain the structural stiffness of SPR connections under different material combinations. A Finite element model of the SPR lap-shear specimen is developed using a FEMFAT SPR pre-processor. The fatigue lift of SPR specimen is predicted using a FEMFAT 4.4e based on the liner finite element analysis.

• 전기학회논문지
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• 제58권1호
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• pp.147-154
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• 2009

This paper presents a grating~integrated SPR (Surface Plasmon Resonance) sensor chip for simple and inexpensive biomolecule detection. The grating-integrated SPR sensor chip has two sensing channels having a nano grating for SPR coupling. An external mirror is used for multi channel SPR sensing. The present sensor chip replaces bulky and expensive optical components, such as fiber-optic switches or special shaped prisms, resulting in a simple and inexpensive wavelength modulated multi-channel SPR sensing system. We fabricate a SPR sensor chip integrated with 835 nm-pitch gratings by a micromolding technique to reduce the fabrication cost. In the experimental characterization, the refractive index sensitivity of each sensing channel is measured as $321.8{\pm}8.1nm$/RI and $514.3{\pm}8.lnm$/RI, respectively. 0.5uM of the target biomolecule (streptavidin) was detected by a $1.13{\pm}0.16nm$ shift of the SPR dip in the 10%-biotinylated sample channel, while the SPR dip in the reference channel for environmental perturbation monitoring remained at the same position. From the experimental results, multi-channel biomolecule detection capability of the present grating-integrated SPR sensor chip has been verified. On the basis of the preliminary experiments, we successfully measured the binding reaction rate for the $2\;nM{\sim}200\;nM$ monoclonal-antibiotin, thus verifying biomolecule concentration detectability of the present SPR sensor chip. The binding reaction rates measured from the present SPR sensor chip agredd well with those from a commercialized SPR sensor.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.1125-1129
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• 2005

Cloning of a 6.6-kb BamHI digested chromosomal DNA from S. griseus IFO13350 revealed the presence of an additional gene encoding a novel trypsin-like enzyme, named SprU. The SprU protein shows a high homology ($79\%$ identity, $88\%$ similarity) with the SGT protease, which has been reported as a bacterial trypsin in the same strain. The amino acid sequence deduced from the nucleotide sequence of the sprU gene suggests that SprU is produced as a precursor consisting of an amino-terminal presequence (29 amino acid residues), prosequence (4 residues), and mature trypsin consisting of 222 amino acids with a molecular weight of 22.94 kDa and a calculated pI of 4.13. The serine, histidine, and aspartic acid residues composing the catalytic triad of typical serine proteases are also well conserved. When the trypsin activity of the SprU was spectrophotometrically measured by the enzymatic hydrolysis of the artificial chromogenic substrate, N-${alpha}$-benzoyl-DL-arginine-p-nitroanilide, the S. lividans transformant with pWHM3-U gave 3 times higher activity than that of control. When the same recombinant plasmid was introduced into S. griseus, however, the gene dosage effect was not so significant, as in the cases of other genes encoding serine proteases, such as sprA, sprB, and sprD. Although two trypsins, SprU and SGT, have a high degree of homology, the pI values, the gene dosage effect in S. griseus, and the gene arrangement adjacent to the two genes are very different, suggesting that the biochemical and biological function of the SprU might be quite different from that of the SGT.

• 산업식품공학
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• 제14권1호
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• pp.49-53
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• 2010

표면 플라즈몬 공명을 이용한 센서는 굴절계 기기의 일종으로서 높은 감도를 가질 뿐만 아니라 비표지 방식이라는 장점을 가지고 있다. 본 연구에서는 재래식 SPR 칩을 이용하여 시판 술 4종의 알코올 함량을 측정하였다. 또한, 재래식 SPR 칩의 감도를 개선하기 위하여 금 박막 위에 금으로 나노형상을 구축하여 나노형상 SPR 칩을 제조하여 모형 술에 대한 감도 개선 효과를 분석하였다. 재래식 SPR 칩을 이용하여 시판 술의 알코올 함량을 측정하기 위한 검량선을 개발하였을 때 시료를 전처리 하지 않고 그대로 측정하였을 때 가장 좋은 검량선을 얻을 수 있었다. 소주, 청주, 이과두주, 탁주 등 시판 술 4종에 대한 1차 회귀식의 검량식에서 결정계수는 각각 0.992, 0.933, 0.918, 그리고 0.984로 나타났다. 한편, 재래식 SPR 칩의 감도를 개선하기 위해 나노형상 SPR 칩을 제조하기 위하여 Langmuir-Blodgett(LB) 방법을 활용하였다. 본 연구에서는 수십 nm 두께의 금 박막을 바닥층으로 하여 그 위에 나노 크기의 실리카 입자를 단분자 층으로 덮어 형틀을 제조하고 다시 그 위에 금을 증착한 후 실리카 입자를 제거하는 방법으로 나노형상을 갖는 SPR 칩을 제조하였다. 나노형상 SPR 칩의 성능을 평가하였을 때 20% 알코올 함량을 가지는 모형 술에 대해서 바닥층의 두께가 50 nm, 나노형상에서 골의 깊이가 20 nm, 나노형상의 배열주기가 300 nm일 때 SPR의 감도가 가장 좋아서 95%의 감도 향상을 얻을 수 있었다. SPR의 감도는 칩과 관련된 인자, 시료의 종류 및 상태에 따라 다르게 나타날 수 있으므로 측정 목적에 알맞은 칩의 설계와 선택이 요구된다.

• 대한용접접합학회:학술대회논문집
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• 대한용접접합학회 2006년도 춘계 학술대회 개요집
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• pp.310-312
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• 2006

Self-piercing riveting(SPR) is becoming an important joining technique for automotive application of various material sheets and shapes. Fatigue behavior of SPR conections needs to be investigated experimentally and numerically to predict SPR fatigue lives. The simulations of various SPR specimens (Coach-Peel specimen, Cross-Tension specimen, Tensile-Shear specimen, Pure-Shear specimen) are performed to predict the fatigue life of SPR connections under different material combinations. Finite element models of various SPR specimens are developed using a FEMFAT SPOT SPR pre-processor. The fatigue lives of SPR specimens are predicted using a FEMFAT 4.4e based on the liner finite element analysis.

• Journal of Biosystems Engineering
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• 제28권2호
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• pp.167-172
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• 2003

This study was performed to fabricate a batch-type SPR biosensing system using a thiolated E. coli antibody coupling, and to explore the feasibility of real-time detection of E. coii in a stagnant sample solution. In advance. “O” and “K” antigenic serotype E. coli antibodies were thiolated with sulfo-LC-SPDP and dithiothreitol, and immobilized by chemisorption in the gold surface of compact SPR sensors. When the SPR biosensor immobilized with E. coli antibody monitored a E. coli solution, it took 3 to 5 min to stabilize. The SPR biosensing system developed in this study was able to detect E. coli in the range above 10$^4$ CFU/mL at the 0.05 significant level. Also, the SPR biosensor had possibility to significantly detect E. coli in the range of 10$^2$ to 10$^4$ CFU/mL in E. coli solutions. Meanwhile, when the SPR biosensor immobilized with 5. coli antibody was cleaned with NaOH solutions, its ability to detect E. coli largely decreased due to wash-out of the immobilized antibody. In order to reuse the SPR sensor, it should be antibody-immobilized newly.

• 음성과학
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• 제11권4호
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• pp.121-127
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• 2004

This study attempted to provide a spectrum analysis for quantitative evaluation of singing voice quality of singing students rather than the presence or absence of the singer's formant. The regression analysis was used to analyse the relationship between ringing quality, SPR, and SPP of singing voice of college student subjects majoring in music. This study measured singing. power ratio (SPR) in 41 singing students. Digital audio recordings were made in sung vowels for acoustic analyses. Each sample was judged by 1 experienced singing teacher and 4 voice pathologists on one semantic bipolar 7-point scales (ringing-dull). The results showed that the SPR and SPP had significant correlations with ringing quality. The SPR had a significant relationship with ringing quality on singing voice in singing students. The SPR can be an important quantitative measurement for evaluating singing voice quality.

• Journal of Welding and Joining
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• 제33권6호
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• pp.55-59
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• 2015

Self-piercing riveting (SPR) is a sheet-joining method that can be used for materials that are difficult or unsuitable for weld, such as aluminum alloys and other steel sheet metals. The increased application of lightweight materials has initiated many investigations into new SPR conditions for riveting dissimilar materials. However, buckling of the semi-tubular rivet occurs during the riveting of AHSS. In this study, a helical SPR was designed for the riveting of AHSS and Al-alloy. In addition, the reinforced helical SPR which has straight parts was designed. The riveting of AHSS and Al-alloy was simulated. Simulated results were verified by comparison with experimental ones.

• 한국생산제조학회지
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• 제19권1호
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• pp.71-79
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• 2010

One of methods that accomplish fuel-efficient vehicle is to reduce the overall vehicle weight by using aluminum structure typically for cross members, rails and panels in body and chassis. For aluminum structures, the use of Self Piercing Rivet(SPR) is a relatively new joining technique in automotive manufacture. To predict SPR fatigue life, fatigue behavior of SPR connections needs to be investigated experimentally and numerically. Tests and simulations on lap-shear specimen with various material combinations are performed to obtain the joining strength and the fatigue life of SPR connections. A Finite element model of the SPR specimen is developed by using a FEMFAT SPR pre-processor. The fatigue lives of SPR specimens with the curvature are predicted using a FEMFAT 4.4e based on the liner finite element analysis.