• Journal of Microbiology and Biotechnology
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• v.14 no.6
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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.

• Transactions of the Korean Society of Mechanical Engineers A
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• v.28 no.8 s.227
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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.

• The Transactions of The Korean Institute of Electrical Engineers
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• v.58 no.1
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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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• v.15 no.5
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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.

• Food Engineering Progress
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• v.14 no.1
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• pp.49-53
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• 2010

Surface plasmon resonance (SPR) which is utilized in thin film refractometry-based sensors has been concerned on measurement of physical, chemical and biological quantities because of its high sensitivity and label-free feature. In this paper, an application of SPR to detection of alcohol content in wine and liquor was investigated. The result showed that SPR sensor had high potential to evaluate alcohol content. Nevertheless, food industry may need SPR sensor with higher sensitivity. Herein, we introduced a nano-technique into fabrication of SPR chip to enhance SPR sensitivity. Using Langmuir-Blodgett (LB) method, gold film with nano-structured surface was devised. In order to make a new SPR chip, firstly, a single layer of nano-scaled silica particles adhered to plain surface of gold film. Thereafter, gold was deposited on the template by an e-beam evaporator. Finally, the nano-structured surface with basin-like shape was obtained after removing the silica particles by sonication. In this study, two types of silica particles, or 130 nm and 300 nm, were used as template beads and sensitivity of the new SPR chip was tested with ethanol solution, respectively. Applying the new developed SPR sensor to a model food of alcoholic beverage, the sensitivity showed improvement of 95% over the conventional one.

• Proceedings of the KWS Conference
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• 2006.05a
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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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• v.28 no.2
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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.

• Speech Sciences
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• v.11 no.4
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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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• v.33 no.6
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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.

• Journal of the Korean Society of Manufacturing Technology Engineers
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• v.19 no.1
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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.