• Animal cells and systems
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• 제14권1호
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• pp.1-10
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• 2010

Previously we showed that CHIP, a co-chaperone of Hsp70 and E3 ubiquitin ligase, can promote the degradation of mutant SOD1 linked to familial amyotrophic lateral sclerosis (fALS) via a mechanism not involving SOD1 ubiquitylation. Here we present evidence that CHIP functions in the interaction of mutant SOD1 with 26S proteasomes. Bag-1, a coupling factor between molecular chaperones and the proteasomes, formed a complex with SOD1 in an hsp70-dependent manner but had no direct effect on the degradation of mutant SOD1. Instead, Bag-1 stimulated interaction between CHIP and the proteasome-associated protein VCP (p97), which do not associate normally. Over-expressed CHIP interfered with the association between mutant SOD1 and VCP. Conversely, the binding of CHIP to mutant SOD1 was inhibited by VCP, implying that the chaperone complex and proteolytic machinery are competing for the common substrates. Finally we observed that mutant SOD1 strongly associated with the 19S complex of proteasomes and CHIP over-expression specifically reduced the interaction between S6/S6' ATPase subunits and mutant SOD1. These results suggest that CHIP, together with ubiquitin-binding proteins such as Bag-1 and VCP, promotes the degradation of mutant SOD1 by facilitating its translocation from ATPase subunits of 19S complex to the 20S core particle.

• Bulletin of the Korean Chemical Society
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• 제34권9호
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• pp.2725-2730
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• 2013

Single C-reactive protein (CRP) molecules, which are non-specific acute phase markers and products of the innate immune system, were quantitatively detected on a gold-nanopatterned biochip using evanescent field-enhanced fluorescence imaging. The $4{\times}5$ gold-nanopatterned biochip (spot diameter of 500 nm) was fabricated by electron beam nanolithography. Unlabeled CRP molecules in human serum were identified with single-molecule sandwich immunoassay by detecting secondary fluorescence generated by total internal reflection fluorescence (TIRF) microscopy. With decreased standard CRP concentrations, relative fluorescence intensities reduced in the range of 33.3 zM-800 pM. To enhance fluorescence intensities in TIRF images, the distance between biochip surface and CRP molecules was optimally adjusted by considering the quenching effect of gold and the evanescent field intensity. As a result, TIRF only detected one single-CRP molecule on the biochip the first time.

• 한국조리학회지
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• 제16권2호
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• pp.322-329
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• 2010

본 연구의 목적은 된장 식품 구매 시 가장 우선적으로 고려하는 속성을 탐색하고, 그 속성 간 중요도를 도출하는데 그 의의가 있다. 이러한 목적을 달성하기 위해 본 연구에서는 제주 지역 식당업소와 주부들을 대상으로 설문을 통해 된장 식품 구매 시 가장 선호하는 요인을 탐색하고 가장 선호하는 된장들에 대하여 살펴보았다. 분석 결과 국내산 B사 된장을 가장 선호였으며, 국내산 A사 된장, 제주산 재래된장 등의 순으로 선호하는 것으로 나타났으며, 민감도 분석 결과, 제주산 재래된장이 가격적인 측면을 보완하면 시장점유율을 향상시킬 수 있음을 알 수 있었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.550-553
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• 2000

The polymeric matrices made with poly(D,L-lactide-co-glycolide) were prepared using copolymer of poly(D,L-lactide) and poly(ethylene glycol) for application of drug delivery systems. Catalyst made use of stannous actoate. Particle size were differ greatly$(435.3{\pm}11.2{\sim}2284.1{\pm}188.5)$ that nanoparticle made use of according to solvent of various kinds. Polymer could a sharp distinction with copolymerized among LE-1, LE-2 and LE-3 of PLA and PEG of content that to examine $^1H-NMR$ of copolymer make refine and reprecipitation. Drug delivery effect at PLGA nanoparticle : PLA amount more then proved highly drug delivery amount that each LE-1, LE-2, LE-3, drug and solvent was 40mg, 20mg and 10mg. Drug delivery effect proved higher 20mg that change(10mg, 20mg, 40mg) at drug feeding amount with LE-2. The first a lot of drug proved delivery. LE-3 most lactide content proved much delivery since biodegradable on PLGA copolymer result from lactide. Also biodegradable rate was highest at LE-3 much of lactide content, because influence at biodegradable effect of lactide by inclusive of soft PEG.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.89-92
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• 2001

Aspergillus niger has been used as a host system to express many heterologous proteins. It has various advantages over other expression systems in that it is a small eukaryotic GRAS (Generally Recognized aS Safe) organism with a capacity of secreting large amount of foreign proteins. However, it has been known that the presence of an abundant protease is a limiting factor to express a heterologous protein. The proteases deficient mutants of A. niger were obtained using UV -mutagenesis. A total of 1 ${\times}$ $10^5$ spores were irradiated with 10-20% survival dose of UV, 600J/M2 at 280nm, and the resulting spores were screened on the casein -gelatin plates. Ten putative protease deficient mutants were further analyzed on the starch plates to differentiate the pro from the secretory mutant. An endogenous extracellular enzyme, glucose oxidase, was also examined to confirm that the mutant phenotype was due to the proteases deficiency rather than the mutation in the secretory pathway. The reduced proteolytic activity was measured using SDS-fibrin zymography gel, casein degradation assay, and bio-activity of a supplemented hGM -CSF (human Granulocyte-Macrophage Colony Stimulating Factor). Comparing with the wild type strain, less than 30 % of proteolytic activity was observed in the culture filtrate of the protease deficient mutant (pro -20) without any notable changes in cell growth and secretion.

• KSBB Journal
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• 제13권3호
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• pp.244-250
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• 1998

글루코스와 초산을 온라인 모니터링 하기위한 흐름주입분석기 술이 개발되었고 대장균발효공정에 이용되었다. 또한, Epoxy 고 분자 담체에 고정화된 GOD와 SOD 를 이용한 GOD-FIA와 SOD-FlA의 특성을 연구했다. 즉, FIA 의 조작온도, 운반용액 속의 첨 가제 (Triton, EDTA, natrium azid 등), 분석 시 료에 용 해된 신진대사물의 농도, pH, 초산측정시 사코진 농도 등에 따 라 고정화된 GOD와 SOD의 활성도, 즉 검출신호의 높이변화를 고찰했다. 최소배양액 과 복합배양액을 사용한 대장균 발효공정 에서 달루코스와 초산을 동시에 온라인 모니터링 하였으며 그결 과는 오프라인 분석결과와 잘 일치 하였다,

• Genomics & Informatics
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• 제13권1호
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• pp.7-14
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• 2015

During recent years, there has been exponential growth in biological information. With the emergence of large datasets in biology, life scientists are encountering bottlenecks in handling the biological data. This study presents an integrated geographic information system (GIS)-ontology application for handling microbial genome data. The application uses a linear referencing technique as one of the GIS functionalities to represent genes as linear events on the genome layer, where users can define/change the attributes of genes in an event table and interactively see the gene events on a genome layer. Our application adopted ontology to portray and store genomic data in a semantic framework, which facilitates data-sharing among biology domains, applications, and experts. The application was developed in two steps. In the first step, the genome annotated data were prepared and stored in a MySQL database. The second step involved the connection of the database to both ArcGIS and $Prot{\acute{e}}g{\acute{e}}$ as the GIS engine and ontology platform, respectively. We have designed this application specifically to manage the genome-annotated data of rumen microbial populations. Such a GIS-ontology application offers powerful capabilities for visualizing, managing, reusing, sharing, and querying genome-related data.

• Genomics & Informatics
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• 제14권4호
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• pp.241-254
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• 2016

Environmental microbes like Bordetella petrii has been established as a causative agent for various infectious diseases in human. Again, development of drug resistance in B. petrii challenged to combat against the infection. Identification of potential drug target and proposing a novel lead compound against the pathogen has a great aid and value. In this study, bioinformatics tools and technology have been applied to suggest a potential drug target by screening the proteome information of B. petrii DSM 12804 (accession No. PRJNA28135) from genome database of National Centre for Biotechnology information. In this regards, the inhibitory effect of nine natural compounds like ajoene (Allium sativum), allicin (A. sativum), cinnamaldehyde (Cinnamomum cassia), curcumin (Curcuma longa), gallotannin (active component of green tea and red wine), isoorientin (Anthopterus wardii), isovitexin (A. wardii), neral (Melissa officinalis), and vitexin (A. wardii) have been acknowledged with anti-bacterial properties and hence tested against identified drug target of B. petrii by implicating computational approach. The in silico studies revealed the hypothesis that lpxD could be a potential drug target and with recommendation of a strong inhibitory effect of selected natural compounds against infection caused due to B. petrii, would be further validated through in vitro experiments.

• KSBB Journal
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• 제17권3호
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• pp.261-265
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• 2002

생리활성을 가지는 펩타이드에 관한 연구는 각종 질환의 진단, 예방 및 치료약으로서의 응용 가능성이 매우 높아서 현재 많은 연구가 진행 중이다. 펩타이드를 RP-HPLC에 의해 정량, 정성분석 함으로 다양한 연구 분야로의 확대를 모색해 볼 수 있다. 혈액 속에 있는 적혈구내의 펩타이드를 분리하기 위해 0.75% TFA가 포함된 물과 아세토나이트릴을 이동상과 선형적 구배용매 조성을 이용한 실험결과, 펩타이드의 peak 분리도는 상당히 우수하였다. 혈액을 채취한 날짜의 경과에 따라서 새로운 생리활성 물질이 만들어짐을 확인할 수 있었다. 이와 같은 현상은 적혈구내의 세포들이 산소를 계속적으로 소비함으로써 산소 부족현상 때문에 발생하는 것으로 알려져 있고 구조적 변화를 나타내는 생리활성 펩타이드를 정성적으로 확인함으로써 좀더 의학적인 분야로의 연구를 수 행할 수 있게 될 것이다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.554-556
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• 2003

본 연구에서는 고부가가치 의약단백질인 human growth hormone을 고순도로 얻기 위하여 재조합 대장균을 이용하여 ubiquitin이 융합된 형태로 단백질을 발현시키고, 이를 분해하는 ubiquitin-specific pretense를 발현시켜 이를 분리 ${\cdot}$ 정제하고 효소특성을 살펴보았다. UBP1 enzyme을 재조합 대장균을 이용하여 발현하고 분리 ${\cdot}$ 정제한 결과, 분자량은 약 83.5kDa이었으며, $40^{\circ}C$, pH 8.0에서 최대 효소활성을 보였다.