• 한국식품영양과학회지
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• 제14권1호
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• pp.1-12
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• 1985

가열 및 동결저장이 오징어, 굴, 새우 및 명태의 in vitro 소화율과 trypsin indigestible substrate (TIS) 함량 변화에 미치는 영향을 실험한 결과는 다음과 같다. Boiling 했을 때 최고의 운 in vitro 소화율을 나타내는 시간은 오징어 1분, 내장을 제거한 굴 0.5분, 내장을 제거하지 않은 굴은 1분이었고, 명태는 5분 이었으며, 오징어와 굴은 부위에 따라 소화율은 다르게 나타났다. 최고의 in vitro 소화율을 나타내는 steaming 조건은 오징어는 $100^{\circ}C$에서 1분, 굴은 $88^{\circ}C$에서 1분이었으며, 명태는 $100^{\circ}C$에서 $1{\sim}2.5$분이었다. TIS는 진시료에서 in vitro 소화율과 역상관관계를 가지며 변화하였다. 동결건조한 시료가 다른 건조품보다 현저하게 높은 in vitro 소화율을 보였으나, 내장을 제거했거나 지방함량이 낮은 시료를 천일건조했을 때도 동결건조시료에 버금가는 in vitro 소화율을 나타내었다. 동결저장은 지방함량이 낮은 시료의 in vitro 소화율의 저하 및 TIS함량 증가에 효과적이었으나, 지방함량이 높아 산패가 심하게 진행된 시료에는 현저한 효과를 볼 수 없었다. 지방함량은 수산식품단백질을 가열처리했거나 저장했을 때의 소화율 변화에 결정적인 영향을 미침을 알 수 있었으며, 4가지의 효소를 이용한 multi-enzyme digestion technique은 수산식품단백질의 소화율 변화를 측정함에 있어 아주 민감하고 유용한 방법으로 판명되었다.

• 한국수산과학회지
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• 제15권4호
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• pp.263-270
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• 1982

해중의 영양학적인 기초자료를 얻기 위하여 multi-enzyme system을 이용한 체외소화율과 아미노산분석을 기초로 한 단백질수를 측정하였다. 김 (P. tenera)의 체외소화율은 $78.5\sim82.2$로서 산지와 건조조건에 따라 약간의 차이를 보였으며, 잎파래(E. linza)나 다른 갈조류 (미역 U. pinnatifida, 톳 H. fusiforme, 모자반 S. fuuellum)에 비하여 높았고 효소활성 저해물질 (trypsin inhibitor)의 함량은 갈조류에서 높았다. 잎과래는 채외소화율이 김보다 낮음에도 불구하고 효소활성저해물질이 가장 낮은 특이한 결과론 보였다. 전체적으로 해조류의 체의소화을이 다른 연구자들의 생체실험에 의한 소화율 (in vivo digestility)보다 높은 결과를 보인 것은multi-enzyme system을 이용한 체외소화율 측정 방법을 해조류의 정확한 소화율 측정에 적용하기에는 문제성이 있는 것으로 밝혀졌다. 일륜 김에 대한 microwave cooking의 영향은 가열시간이 15분 경과하여도 현저한 소화율 증가는 볼 수 없었으며, 효소활성 저해물질함량은 서서히 감소하는 경향을 보였다. 또한 한국식으로 구운김의 체외소화율은 microwave로 15분 가열한 시료와 비슷하였다. 아미노산 분석 결과를 이용한 단백계수(Factor method)는 김의 경우 6.52로 계산되었고, 잎파래는 6.00, 미역 6.11, 모자반 5.85, 톳은 5.83이었으며, Kjeldahl전소분석결과를 이용한 단백계수(Kjeldahl Method)는 김 6.29, 잎파래 5.83, 미역 5.40, 모자반 5.45, 톳 5.49로 나타나, 종래의 조단백계수(6.25)보다 낮은 결과를 보였다. 해조추의 비단백태질소의 정확한 규명이 없는 상태에서, 해조의 단백질함량 측정에는 아미노산 분석결과를 이용한 새로운 단백계수(Factor Method)를 사용함이 바람직한 것으로 생각되었다.

• 동아시아식생활학회지
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• 제21권5호
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• pp.731-738
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• 2011

콜레스테롤의 신속하고 정확한 새로운 분석방법을 모색하기 위하여 본 연구에서는 전기적 전도성이 우수한 MWCNT를 이용하여 전극을 제작하였고, 여러 가지 효소고정화 방법을 통해 전기화학적 감응도 분석을 실시하였다. MWCNT의 전도성을 향상시키기 위해 아민기를 도입한 MWCNT-$NH_2$를 제조하였고, MWCNT-$NH_2$/GCE에 PB를 점착하여 작업전극을 제조하였다. 제조한 작업전극은 0.5~500 ${\mu}M$ $H_2O_2$ 농도 범위에서 농도가 증가함에 따라 전류가 비례적으로 증가하였고, 검출한계는 0.1 ${\mu}M$로 나타나 전극이 높은 감도를 가지고 있음을 확인하였다. 또한 콜레스테롤 검출을 위해 적합한 효소 반응기를 제작하기 위해 담체인 aminopropyl glass beads, CNBr-activated sepharose, Na-alginate, toyopearl beads에 cholesterol oxidase를 고정화시켜 바이오센서의 콜레스테롤 표준용액에 대한 감응도를 측정한 결과, aminopropyl glass beads과 CNBr-activated sepharose는 1~100 ${\mu}M$ 범위에서 선형관계를 보였으며, Na-alginate는 5~50 ${\mu}M$의 범위에서, toyopearl beads는 1~50 ${\mu}M$ 범위에서 선형관계를 나타내었다. 검출한계는 제작된 효소반응기 모두 1 ${\mu}M$로 나타나 콜레스테롤에 대한 높은 검출력을 보여주었으나, 특히 CNBr-activated sepharose와 Na-alginate를 이용한 효소반응기가 높은 coupling efficiency와 감응도를 보여 콜레스테롤 검출을 위한 본 바이오 센서 시스템에 적합한 것으로 나타났다.

• Molecules and Cells
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• 제26권6호
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• pp.536-547
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• 2008

Anthocyanidin synthase (ANS, leucoanthocyanidin oxygenase), a 2-oxoglutarate iron-dependent oxygenase, catalyzed the penultimate step in the biosynthesis of the anthocyanin class of flavonoids, from the colorless leucoanthocyanidins to the colored anthocyanidins. The full-length cDNA and genomic DNA sequences of ANS gene (designated as GbANS) were isolated from Ginkgo biloba for the first time. The full-length cDNA of GbANS contained a 1062-bp open reading frame (ORF) encoding a 354-amino-acid protein. The genomic DNA analysis showed that GbANS gene had three exons and two introns. The deduced GbANS protein showed high identities to other plant ANSs. The conserved amino acids (H-X-D) ligating ferrous iron and residues (R-X-S) participating in 2-oxoglutarate binding were found in GbANS at the similar positions like other ANSs. Southern blot analysis indicated that GbANS belonged to a multi-gene family. The expression analysis by real-time PCR showed that GbANS expressed in a tissue-specific manner in G. biloba. GbANS was also found to be up-regulated by all of the six tested abiotic stresses, UV-B, abscisic acid, sucrose, salicylic acid, cold and ethylene, consistent with the promoter region analysis of GbANS. The recombinant protein was successfully expressed in E. coli strain with pET-28a vector. The in vitro enzyme activity assay by HPLC indicated that recombinant GbANS protein could catalyze the formation the cyanidin from leucocyanidin and conversion of dihydroquercetin to quercetin, suggesting GbANS is a bifunctional enzyme within the anthocyanidin and flavonol biosynthetic pathway.

• 전기화학회지
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• 제13권1호
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• pp.50-56
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• 2010

Multi-wall carbon nano-tube(MWCNT)를 이용해 screen printed carbon electrodes(SPCEs)을 제작하여 혈당센서의 선택성과 감도가 증가됨을 확인 할 수 있었다. 효소촉매반응을 위한 탄소전극으로의 전자이동의 매개체로 8족 금속 원소인 오스뮴을 중심금속으로 일차 아민을 포함하는 피리딘(pyridine) 리간드를 배위시켜 $[Os(dme-bpy)_2(4-aPy)Cl]^{+/2+}$를 합성하였다. 합성된 오스뮴 착물은 순환 전압전류법을 포함한 다양한 전기화학분석방법을 이용하여 전기적 성질을 조사하였다. 전기적 흡착방법을 이용하여 일차 아민을 갖는 착화합물을 전극위에 고정화 하였다. 오스뮴이 고정화된 MWCNT-SPCEs는 일반적인 carbon electrode보다 약 100배가량의 오스뮴이 흡착됨을 확인 할 수 있었다. (${\tau}_0=2.0\;{\times}\;10^{-9}\;mole/cm^2$) 마지막으로 당(Glucose)과 당 분해효소(Glucose Oxidase, GOx)에 의한 촉매반응의 전류를 확인하였고, 당 농도에 따라 선형 변화하는 전류의 양도 확인하였다.

• Journal of Microbiology
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• 제44권1호
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• pp.106-112
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• 2006

In this study, the genetic diversities of multi-resistant Salmonella typhimurium (ST) isolates were analyzed via the application of both pulsed field gel electrophoresis (PFGE) and Polymerase chain reaction (PCR) analysis methods, using 6 kinds of primers (REP, ERIC, SERE, BOX, P-1254 and OPB-17). And their discriminative abilities (DA) were also compared in order to determine the most effective and reliable analysis method. 118 S. typhimurium isolates, cultured from diverse animals and human patients in Korea beginning in 1993, were analyzed and subjected to a comparison of Simpson's index of diversity (SID), using both PFGE and PCR methods. PFGE by XbaI enzyme digestion allowed for discrimination into 9 pulsotypes, with high SID values (0.991) on the genomic DNA level. This shows that PFGE is a very discriminative genotypic tool, and also that multiple clones of S. typhimurium isolates had existed in domestic animals and humans in Korea since 1993. However, we could ultimately not to trace the definitive sources or animal reservoirs of specific S. typhimurium isolates examined in this study. Depending on the SID values, the combined method (7 kinds of method) was found to be the most discriminative method, followed by (in order) SERE-PCR, REP-PCR, ERIC-PCR, PFGE & OPB-17 (RAPD), P-1254 (RAPD), and BOX-PCR at the $80\%$ clone cut-off value. This finding suggests that the REP-PCR method (which utilizes 4 primer types) may be an alternative tool to PFGE for the genotyping of S. typhimurium isolates, with comparable cost, time, and labor requirement. The establishment of a highly reliable and discriminatory method for epidemiologic analysis is considered necessary in order for researchers to trace the sources of specific pathogens and, consequently, to control and prevent the spread of epidemic S. typhimurium isolates to humans.

• Asian Pacific Journal of Cancer Prevention
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• 제17권1호
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• pp.57-64
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• 2016

Background: Tobacco smoking is considered a risk factor for cervical cancer development due to the presence of tobacco based carcinogenic metabolites in cervical cells of female smokers. In this study, we investigated the role of the T3801C (MspI) polymorphism of CYP1A1, a gene encoding an enzyme necessary for the initiation of tobacco based carcinogen metabolism, on cervical cancer risk. The T to C substitution may alter CYP1A1 activities, potentially elevating cervical cancer risk. Since results of gene-disease association studies vary according to the study population, the multi-ethnic population of Malaysia provides an excellent representative cohort for identifying and comparing the cervical cancer risk among the 3 major ethnics in Southeast Asia in relation to CYP1A1 MspI polymorphism. Materials and Methods: A total of 195 Thin Prep Pap smear samples from HPV negative and cancer free females were randomly selected as controls while 106 formalin fixed paraffin embedded samples from females with invasive cervical cancer were randomly selected for the cases group. The polymorphisms were identified using restriction fragment length polymorphism (RFLP) PCR. Results: We found no significant associations between CYP1A1 MspI polymorphism and cervical cancer in the general Malaysian female population. However, upon ethnic stratification, the variant C/C genotype was significantly associated with a 4.66-fold increase in cervical cancer risk in Malay females (95% CI= 1.21-17.9; p=0.03). No significant association was observed in the Chinese and Indian females. Additionally, there were no significant associations in the dominant model and allele frequency model analysis in both the general and ethnically stratified female population of Malaysia. Conclusions: Our findings suggest that the C/C genotype of CYP1A1 MspI polymorphism is associated with the development of cervical carcinoma in the Malay females of Malaysia.

• Journal of Microbiology and Biotechnology
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• 제19권1호
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• pp.65-71
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• 2009

Microbial oxidoreductive systems have been widely used in asymmetric syntheses of optically active alcohols. However, when reused in multi-batch reaction, the catalytic efficiency and sustainability of non-growing cells usually decreased because of continuous consumption of required cofactors during the reaction process. A novel method for NADPH regeneration in cells was proposed by using pentose metabolism in microorganisms. Addition of D-xylose, L-arabinose, or D-ribose to the reaction significantly improved the conversion efficiency of deracemization of racemic 1-phenyl-1,2-ethanediol to (S)-isomer by Candida parapsilosis cells already used once, which afforded the product with high optical purity over 97%e.e. in high yield over 85% under an increased substrate concentration of 15 g/l. Compared with reactions without xylose, xylose added to multi-batch reactions had no influence on the activity of the enzyme catalyzing the key step in deracemization, but performed a promoting effect on the recovery of the metabolic activity of the non-growing cells with its consumption in each batch. The detection of activities of xylose reductase and xylitol dehydrogenase from cell-free extract of C. parapsilosis made xylose metabolism feasible in cells, and the depression of the pentose phosphate pathway inhibitor to this reaction further indicated that xylose facilitated the NADPH-required deracemization through the pentose phosphate pathway in C. parapsilosis. moreover, by investigating the cofactor pool, the xylose addition in reaction batches giving more NADPH, compared with those without xylose, suggested that the higher catalytic efficiency and sustainability of C. parapsilosis non-growing cells had resulted from xylose metabolism recycling NADPH for the deracemization.

• BMB Reports
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• 제48권1호
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• pp.25-29
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• 2015

Ubiquitination is a post translational modification which mostly links with proteasome dependent protein degradation. This process has been known to play pivotal roles in the number of biological events including apoptosis, cell signaling, transcription and translation. Although the process of ubiquitination has been studied extensively, the mechanism of polyubiquitination by multi protein E3 ubiquitin ligase, SCF complex remains elusive. In the present study, we identified UbcH5a as a novel stimulating factor for poly-ubiquitination catalyzed by $SCF^{hFBH1}$ using biochemical fractionations and MALDI-TOF. Moreover, we showed that recombinant UbcH5a and Cdc34 synergistically stimulate $SCF^{hFBH1}$ catalyzed polyubiquitination in vitro. These data may provide an important cue to understand the mechanism how the SCF complex efficiently polyubiquitinates target substrates.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권1호
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• pp.7-12
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• 2000

The expression of the mouse $\alpha-amylase$ gene in the methylotrophic yeast, P pastoris was investigated. The mouse $\alpha-amylase$ gene was inserted into the multi-cloning site of a Pichi a expression vector, pPIC9, yielding a new expression vector pME624. The plasmid pME624 was digested with SalI or BglII, and was introduced into P. pastoris strain GSl15 by the PEG1000 method. Fifty-three transformants were obtained by the transplacement of pME624 digested with SaiII or BglII into the HIS4locus $(38\;of\;Mut^+\;clone)$ or into the AOX1 locus $(15\;of\;Mut^s\;clone)$. Southern blot was carried out in 11 transformants, which showed that the mouse $\alpha-amylase$ gene was integrated into the Pichia chromosome. When the second screening was performed in shaker culture, transformant G2 showed the highest $\alpha-amylase$ activity, 290 units/ml after 3-day culture, among 53 transformants. When this expression level of the mouse $\alpha-amylase$ gene is compared with that in recombinant Saccharomyces cerevisiae harboring a plasmid encoding the same mouse $\alpha-amylase$ gene, the specific enzyme activity is eight fold higher than that of the recombinant S. cerevisiae.