• Biomolecules & Therapeutics
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• v.23 no.2
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• pp.189-194
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• 2015

P450 1A2 is responsible for the metabolism of clinically important drugs and the metabolic activation of environmental chemicals. Genetic variations of P450 1A2 can influence its ability to perform these functions, and thus, this study aimed to characterize the functional significance of three P450 1A2 allelic variants containing nonsynonymous single nucleotide polymorphisms (P450 $1A2^*8$, R456H; $^*15$, P42R; $^*16$, R377Q). Variants containing these SNPs were constructed and the recombinant enzymes were expressed and purified in Escherichia coli. Only the P42R variant displayed the typical CO-binding spectrum indicating a P450 holoenzyme with an expression level of ~ 170 nmol per liter culture, but no P450 spectra were observed for the two other variants. Western blot analysis revealed that the level of expression for the P42R variant was lower than that of the wild type, however the expression of variants R456H and R377Q was not detected. Enzyme kinetic analyses indicated that the P42R mutation in P450 1A2 resulted in significant changes in catalytic activities. The P42R variant displayed an increased catalytic turnover numbers ($k_{cat}$) in both of methoxyresorufin O-demethylation and phenacetin O-deethylation. In the case of phenacetin O-deethylation analysis, the overall catalytic efficiency ($k_{cat}/K_m$) increased up to 2.5 fold with a slight increase of its $K_m$ value. This study indicated that the substitution P42R in the N-terminal proline-rich region of P450 contributed to the improvement of catalytic activity albeit the reduction of P450 structural stability or the decrease of substrate affinity. Characterization of these polymorphisms should be carefully examined in terms of the metabolism of many clinical drugs and environmental chemicals.

• Korean Journal of Microbiology
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• v.32 no.3
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• pp.215-221
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• 1994

The complete nucleotide sequence of the cloned pga gene encoding the penicillin G acylase of Bacillus megaterium ATCC 14945 and its 5'- and 3'-flanking regions was determined. The sequence revealed only one large open reading frame (2,406 hp) of the penicillin G acylase (pga) gene. Upstream from ATG of the pga gene, there was a putative ribosome binding site, Shine-Dalgarno sequence. The promoter-like structure, - 10 and - 35 sequences, was also found. Following the stop codon, TAG, a structure reminiscent of the E. coli rho-independent transcription terminator was present. The amino acid sequence was deduced from the nucleotide sequence. The molecular mass of the polypeptide was 91,983 Da. There was a potential signal sequence in its amino-terminal region. A comparison of its deduced amino acid sequence with other characterized penicillin G acylases and the result of SDS-polyacrylamide gel electrophoresis of the purified enzyme showed that a precursor polypeptide of 92 kDa was processed into two dissimilar ${\alpha}$ and ${\beta}$-subunits of 25 and 61 kDa.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.68-72
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• 2010

The ${\beta}$-1,4-glucanase gene of Bacillus licheniformis B1 was expressed in Esherichia coli BL21, and a protein with a mass of 50 kDa that was soluble was overproduced. A protein with a mass of 37 kDa was secreted from B. licheniformis. It seems that the ${\beta}$-1,4-glucanase produced in E. coli contained the leader peptide and unprocessed carboxy-terminal region, but its processing occurred in the carboxyterminal in Bacillus. The optimal temperature of ${\beta}$-1,4-glucanase was $40^{\circ}C$. The enzyme still had 76% maximal activity at $60^{\circ}C$. The optimal pH of the enzyme was 7. The enzyme retained considerable activities over the weak-acidic, neutral, and weak-basic pH range. Acidic fungal cellulases are used in food, detergent, pulp, paper, textile industries. However, studies about neutral and alkaline cellulase are not enough. The cellulase developed in this study may be useful for industrial applications in the fields of biofuel development.

• Journal of Life Science
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• v.21 no.5
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• pp.761-765
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• 2011

Salt stability of enzymes is a crucial practical factor in the food industry. Previously, leucine aminopeptidase (LAP) was purified from Bacillus sp. N2. Here, we present the salt effect of LAP using synthetic substrates. LAP had a hydrolytic activity for L-leucine-${\rho}$-nitroanilide in high concentrations of NaCl (up to 4 M), but not for other neutral salts (LiBr, LiCl, NaBr, KBr, and KCl). It hydrolyzed various synthetic di-peptide substrates with hydrophobic and hydrophilic amino acids at the C-terminal Xaa region, in the presence of 0-4 M NaCl. The result indicated that the hydrolytic action of LAP is not dependent on the hydrophobicity of the amino acid side chain at the scissile bond of the substrate. Remarkably, the hydrolytic activity of LAP was 1-3 folds higher than those of other LAPs and aminopeptidases in 4.5 M NaCl, suggesting that NaCl-tolerant LAP might be used in the food industry as cheese and anchovy sauce.

• Proceedings of the Korean Association for Survey Research Conference
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• 2003.06a
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• pp.140-160
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• 2003

The purpose of this study is to identify and evaluate the competitiveness of ports in ASEAN(Association of Southeast Asian Nations), which plays a leading role in basing the hub of international logistics strategies as a countermeasure in changes of international logistics environments. This region represents most severe competition among Mega hub ports in the world in terms of container cargo throughput at the onset of the 21st century. The research method in this study accounted for over lapping between attributes, and introduced the HFP method that can perform mathematical operations. The scope of this study was strictly confined to the ports of ASEAN, which cover the top 100 of 350 container ports that were presented in Containerization International Yearbook 2002 wi th reference to container throughput. The results of this study show Singapore in the number one position. Even compared with major ports in Korea (after getting comparative ratings and applying tile same data and evaluation structure), the number one position still goes to Singapore and then Busan(2) and Manila(2), followed by Port Klang(4), Tanjung Priok(5), Tanjung Perak(6), Bangkok(7), Inchon(8), Laem Chabang(9) and Penang(9). In terms of the main contributions of this study, it is the first empirical study to apply the combined at tributes of detailed and representative attributes into the advanced HFP model which was enhanced by the KJ method to evaluate the port competitiveness in ASEAN. Up-to-now, none have comprehensively conducted researches with sophisticated port methodology that has discussed a variety of changes in port development and terminal transfers of major shipping lines. Moreover, through the comparative evaluation among major ports in Korea and ASEAN, the presentation of comparative competitiveness for Korean ports is a great achievement in this study. In order to reinforce this study, it needs further compensative research, including cost factors which could not be applied to modeling the subject ports by lack of consistently qualified data in ASEAN.

• Applied Biological Chemistry
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• v.48 no.4
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• pp.345-351
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• 2005

To access the natural products of uncultured microorganisms, we constructed and screened the metagenomic DNA libraries by using a cosmid vector and DNA inserts isolated directly from soil. Initial screening of the libraries in Escherichia coli resulted in the isolation of several clones that produce a dark brown color when grown in LB medium. One of the positive clones, designed pYS85C, was transposon mutagenized and the DNA surrounding the transposon insertions in cosmids that no longer conferred the production of brown pigment to E. coli was sequenced. Annotation of the pYS85C sequence obtained from the transposon mutagenesis experiment indicated a single 393 amino acid open reading frame (ORF) with a molecular mass of about 44.5 kDa, predicted to be a 4-hydroxyphenylpyruvate dioxygenases (HPPDs), was responsible for the observed brown pigment. In a BLAST search against deposited sequence, the translated protein from this ORF showed moderate-level identity (>60%) to the other known HPPDs and was most conserved in the C-terminal region of the protein. These results show that genes involved in natural product synthesis can be cloned directly from soil DNA and expressed in a heterologous host, supporting the idea that this technology has the potential to provide novel natural products from the wealth of environmental microbial diversity and is a potentially important new tool for drug discovery.

• Food Science and Preservation
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• v.18 no.2
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• pp.184-190
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• 2011

Limonoid UDP-glucosyltransferase (LUGT) is an enzyme that converts limonoids into their corresponding glucosides and ultimately ameliorates limonoid bitterness in Citrus species. In this paper, the LUGT gene was cloned via PCR from 10 Jeju Citrus species. All the deduced glucosyltransferase proteins harbored a highly conserved plant secondary product glucosyltransferase (PSPG) motif within the C terminal region. Phylogenetic analysis based on the amino acid sequence comparison of the LUGT proteins from 10 Citrus species generated three distinct types. The expression patterns of LUGT gene in three representative species from each type were quite different with that of C. unshiu Marc. cv. Miyagawawase(Gungcheon), which his without distinctive juice delayed bitterness. Ourresultssho wth at some Citrus speciessuchas Citrusleiocarpa HORT(Bingul), Citruserythrosa HORT (Dongjunggul), and Citrustachibana TANAKA(Honggul) end emicin Jeju maybe susceptible to intense juice delayed bitterness due to delay inexpression of LUGT.

• Journal of Korea Port Economic Association
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• v.26 no.3
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• pp.143-164
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• 2010

Japanese government has instituted and carried out Super-core Ports Policy to improve their operating efficiencies of international container ports from 2004. Despite of diverse institutional and administrative supports within this policy, this paper has assessed that they couldn't accomplished goals at the end of 2009 when the policy ended. In short, Japanese Super-core ports have explicitly lost the competitiveness for functioning as a hub-port in the East Asia region. We could find some defaults in the policy i.e. some limits of the private terminal operating units, inefficiencies in teaming between ports within Super-core port, lacking with system for collecting cargoes. In 2010, Japan is going to reinforce International Strategic Container Port Policy following Super-core Ports Policy. With this policy, they ought to prepare for the last leap on the basis of selection and concentration rules in international port. This new policy is particularly focused on recapturing their T/S cargo via Busan port. Regarding these changes, Busan port needs to prepare counter measurements for preserving Japanese T/S cargo firmly.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.9
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• pp.1226-1234
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• 2012

3,4,3',5'-Tetrahydroxy-trans-stilbene (piceatannol) is a derivative of resveratrol with a variety of biological activities, including anti-inflammatory, anti-proliferative, and anti-cancer activities. We assessed the mechanisms by which piceatannol inhibits inflammatory responses using lipopolysaccharide (LPS)-treated Raw264.7 murine macrophages. Piceatannol (0~10 ${\mu}mol/L$) decreased LPS-induced release of nitric oxide, tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, IL-$1{\beta}$, and inhibited LPS-induced protein expression of inducible nitric oxide synthase (iNOS). Activation of nuclear factor-kappaB (NF-${\kappa}B$), activator protein (AP)-1, and signal transducer and activator of transcription 3 (STAT3) are crucial steps during an inflammatory response. Piceatannol prevented LPS-induced degradation of inhibitor of ${\kappa}B$ ($I{\kappa}B$), translocation of p65 to the nucleus, and phosphorylation of stress-activated protein kinase/c-Jun NH2-terminal kinase (SAPK/JNK). Additionally, piceatannol inhibited LPS-induced phosphorylation of STAT3 and IL-6-induced translocation of STAT3 to the nucleus. Furthermore, piceatannol increased the protein and mRNA levels of hemeoxygenase (HO)-1, the rate-limiting enzyme of heme catabolism that plays a critical role in mediating antioxidant and anti-inflammatory effects. Piceatannol further induced antioxidant response elements (ARE)-driven luciferase activity in Raw264.7 cells transfected with an ARE-luciferase reporter construct containing the enhancer 2 and minimal promoter region of HO-1. These results suggest that piceatannol exerts anti-inflammatory effects via the down-regulation of iNOS expression and up-regulation of HO-1 expression.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.17 no.4
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• pp.447-455
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• 1995

Cavernous sinus thrombosis is one of the major complications of abscesses of the maxillofacial region. The initial symptoms of CST are usually pain in the eye and tenderness to pressure. this is associated with high fluctuating fever, chills, rapid pulse, and sweating. Venous obstruction subsequently causes edema of the eyelids, lacrimation, proptosis, chemosis and retinal hemorrhages. Blindness is sometimes an accompaniment of cavernous sinus thrombosis when the infection also involves the orbit. There is also cranial nerve involvement (oculomotor, troclear, abducence) and ophthalmoplegia, diminished or absent corneal reflex, ptosis, and dilation of the pupil occur. The terminal stages bring signs of advanced toxemia and meningitis. Infections of the face can cause a septic thrombosis of the cavernous sinus. Furunculosis and infected hair follicles in the nose are frequent causes. Extractions of maxillary anterior teeth in the presence of acute infection and especially curettage of the sockets under such circumstances can cause this condition. The infection is usually staphylococcal. The inflection may spread directly through the pterygoid plexus of veins and the pterygomaxillary space and then ascend into the sinus or it may spread directly from the pterygopalatine space to the orbit. This is possible because of the absence of valves in the angular, facial, and ophthalmic veins. The treatment is empirical antibiotic therapy followed by specific anbibiotic therapy based on blood or pus culture. The inflection usually involves one side, however, it may easily spread to the opposite side through the circulus sinus. Unless it is treated early, the prognosis is poor even in this doses. Occasionally the antibiotics will not adequately resolve the septic thrombus, and death ensues. the use of anticoagulants to prevent venous thrombosis has been recommended, but the efficacy of such therapy has not been substantiated. Surgical access through eye enucleation has been suggested. We report a case which demonstrates cavernous sinus thrombosis by the infection after the functional neck dissection and the intraoral reconstruction with auriculomastoid fascio-cutaneous island flap.