• 제목/요약/키워드: Systems Biotechnology

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Thermostable ${\beta}$-Glycosidase-CBD Fusion Protein for Biochemical Analysis of Cotton Scouring Efficiency

  • Ha, Jae-Seok;Lee, Young-Mi;Choi, Su-Lim;Song, Jae-Jun;Shin, Chul-Soo;Kim, Ju-Hea;Lee, Seung-Goo
    • Journal of Microbiology and Biotechnology
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    • 제18권3호
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    • pp.443-448
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    • 2008
  • Multidomain proteins for the biochemical analysis of the scouring efficiency of cotton fabrics were constructed by the fusion of a reporter moiety in the N-terminal and the cellulose binding domain (CBD) in the C-terminal. Based on the specific binding of the CBD of Cellulomonas fimi exoglucanase (Cex) to crystalline cellulose (Avicel), the reporter protein is guided to the cellulose fibers that are increasingly exposed as the scouring process proceeds. Among the tested reporter proteins, a thermostable ${\beta}$-glycosidase (BglA) from Thermus caldophilus was found to be most appropriate, showing a higher applicability and stability than GFP, DsRed2, or a tetrameric ${\beta}$-glycosidase (GUS) from Escherichia coli, which were precipitated more seriously during the expression and purification steps. When cotton fabrics with different scouring levels were treated with the BglA-CBD and incubated with X-Gal as the chromogenic substrate, an indigo color became visible within 2 h, and the color depth changed according to the conditions and extent of the scouring.

인간 HepG2 세포에서 더덕 및 도라지 에틸아세테이트 분획물의 항산화 효과에 의한 세포보호 효과 (Cytoprotective Effect by Antioxidant Activity of Codonopsis lanceolata and Platycodon grandiflorum Ethyl Acetate Fraction in Human HepG2 Cells)

  • 김수현;최현진;오현택;정미자;최승필;함승시
    • 한국식품과학회지
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    • 제40권6호
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    • pp.696-701
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    • 2008
  • 이 연구의 목적은 더덕 및 도라지 에틸아세테이트 분획물(CLEA나 PGEA)이 다양한 항산화 시스템을 가동시켜 sodium nitroprusside(SNP)에 의해 유도된 산화적 스트레스에 대항하여 세포를 보호해 줄 수 있을지 연구하는 것이다. 0.5 mM의 SNP를 HepG2 세포주에 24시간 동안 노출시켰을 때 세포 생존율이 감소하였으나, CLEA와 PGEA 천처리에 의해 SNP 노출에 의한 세포 생존율 감소가 저해되었다. 또한 항산화 시스템들 발현에 미치는 CLEA와 PGEA의 영향을 RT-PCR로 알아보았다. Catalase, glucose-6-phosphate dehydrogenase(G6PD) 그리고 metallothionein (MT)-1A mRNA 수준이 CLEA를 세포에 24시간 동안 처리한 후 증가하였고, Mn superoxide dismutase, catalase, G6PD, MT-1A 와 MT-2A mRNA 수준이 PGEA 처리에 의해 증가하였다. 우리는 CLEA와 PGEA가 아마 다양한 항산화 시스템들을 증가시키는 간접적인 항산화 효과를 나타내며, 이들 효과들이 산화적 스트레스로부터 세포를 보호해 줄 것이라 추정하였다.

착유방식이 목장형 유가공으로 제조된 고다치즈의 품질 특성에 미치는 영향 (Effects of Miling System on Gouda Cheese Characteristics made from Farmstead Milk-processing Plant)

  • 이진성;문주연;남기택;박성민;박승용;정문웅;손용석
    • Journal of Dairy Science and Biotechnology
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    • 제34권4호
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    • pp.245-254
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    • 2016
  • 최근 보급이 증가되고 있는 AMS가 목장형 유가공장에서 생산되는 치즈의 특성에 미치는 영향을 알아보기 위한 연구를 실시하였다. 첫 번째 연구에서는 AMS 및 CMS 농장에서 원유를 채취하여 동일한 조건에서 각각 고다치즈를 제조하여 6개월 숙성시킨 다음 일반 성분 및 유리지방산 분석과 관능평가를 실시하였고, 두 번째 연구에서는 목장형 유가공장 7곳에서 제조한 고다치즈 시료를 채취하여 일반성분과 조직 특성을 비교하였다. 본 연구 결과, 고다치즈의 일반성분은 조회분을 제외하고는 착유 방식이나 목장에 따른 유의적인 차이가 나타나지 않았고, 유리지방산은 각 목장에 따른 차이를 보여주었으나 관능평가에 영향을 미치지 않았으며, 착유 방식에 의한 유리지방산 및 관능평가 결과에서도 유의적인 차이도 발견할 수 없었다. 조직 특성 역시 목장 간에 유의적 차이가 컸던 관계로 착유방식에 따른 차이는 나타나지 않았다. 결론적으로 착유방식은 목장형 유가공장에서 제조한 고다치즈의 특성에는 영향을 미치지 않을 것으로 판단되며, 향후 같은 고다치즈라 하더라도 각 목장별로 차별화하여 전통과 개성이 있는 목장형 유가공 제품으로 발전할 가능성을 보인 것으로 사료된다.

Genome-Wide Response of Deinococcus radiodurans on Cadmium Toxicity

  • Joe, Min-Ho;Jung, Sun-Wook;Im, Seong-Hun;Lim, Sang-Yong;Song, Hyun-Pa;Kwon, Oh-Suk;Kim, Dong-Ho
    • Journal of Microbiology and Biotechnology
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    • 제21권4호
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    • pp.438-447
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    • 2011
  • Deinococcus radiodurans is extremely resistant to various genotoxic conditions and chemicals. In this study, we characterized the effect of a sublethal concentration (100 ${\mu}M$) of cadmium (Cd) on D. radiodurans using a whole-genome DNA microarray. Time-course global gene expression profiling showed that 1,505 genes out of 3,116 total ORFs were differentially expressed more than 2-fold in response to Cd treatment for at least one timepoint. The majority of the upregulated genes are related to iron uptake, cysteine biosynthesis, protein disulfide stress, and various types of DNA repair systems. The enhanced upregulation of genes involved in cysteine biosynthesis and disulfide stress indicate that Cd has a high affinity for sulfur compounds. Provocation of iron deficiency and growth resumption of Cd-treated cells by iron supplementation also indicates that CdS forms in iron-sulfur-containing proteins such as the [Fe-S] cluster. Induction of base excision, mismatch, and recombinational repair systems indicates that various types of DNA damage, especially base excision, were enhanced by Cd. Exposure to sublethal Cd stress reduces the growth rate, and many of the downregulated genes are related to cell growth, including biosynthesis of cell membrane, translation, and transcription. The differential expression of 52 regulatory genes suggests a dynamic operation of complex regulatory networks by Cd-induced stress. These results demonstrate the effect of Cd exposure on D. radiodurans and how the related genes are expressed by this stress.

Production of Recombinant Proteins as Immuno-Analytical Markers of Genetically-Modified Organisms (GMO)

  • Hwang, Ok-Hwa;Park, Hyuk-Gu;Paek, Eui-Hwan;Paek, Se-Hwan;Park, Won-Mok
    • Journal of Microbiology and Biotechnology
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    • 제14권4호
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    • pp.783-788
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    • 2004
  • Marker proteins of genetically-modified organisms (GMO) and their antibodies were prepared and characterized as major components of an analytical system. We selected two GMO markers, neomycin phosphotransferase II and 5- enolpyruvylshikimate-3-phosphate synthase, and produced them from E. coli employing genetic recombination technology. After purification, their structural conformation and binding affinities to the respective antibodies were characterized. The results showed that the recombinant proteins were identical with commercially obtained reference proteins. We further used them as immunogens to raise polyclonal antibodies capable of discriminating GMO containing protein from non-GMO. Well-characterized marker proteins and antibodies will be valuable as immunoreagents in constructing analytical systems such as biosensors and biochips to measure quantities of GMO.

한국 생명공학산업의 혁신네트워크에 관한 실증연구 (An Empirical Study on Innovation Networks in the Korean Biotechnology Industry)

  • 구영우;노영희;정규진;민완기
    • 기술혁신학회지
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    • 제16권3호
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    • pp.861-882
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    • 2013
  • 본 연구는 혁신체제론에 입각해서 국내 생명공학산업의 조직간 혁신네트워크 현황 및 활성화방안을 실증적으로 분석했다. 본 연구의 결과를 요약하면, 다음과 같다. 아직 국내 생명공학산업의 혁신네트워크는 활성화되지 못했지만 혁신네트워크에 참여하고 있는 벤처기업 근무자, 대학교수, 공공연구기관 연구원들은 혁신네트워크 및 혁신네트워크의 성과에 대하여 긍정적으로 평가하고 있다. 이는 국내 생명공학산업의 혁신네트워크가 활성화될 필요성이 있음을 의미한다. 실증분석 결과 혁신네트워크에 참여하는 주체들의 의지와 능력이 제고되고, 혁신네트워크 내 참여주체간 신뢰와 통제가 강화되고, 혁신네트워크내 참여주체들의 기술접근과 자원확보가 잘 이루어진다면, 혁신네트워크는 더욱 활성화될 수 있을 것이다.

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의약용 당단백질에 부가된 당사슬의 중요성 (Impacts of glycans attached to therapeutic glycoproteins)

  • 김성훈;권오석;오두병
    • Journal of Plant Biotechnology
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    • 제37권3호
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    • pp.292-304
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    • 2010
  • High value-added therapeutic proteins have been leading the biologics industry and occupied major portion of the market. More than 60% of the currently available protein therapeutics are glycoproteins attached with glycans which play crucial roles for the protein folding, therapeutic efficacy, in vivo half-life and immunogenecity. This review introduces the process of glycosylation and the impacts of glycans in the aspects of therapeutics. The important glycan structures in therapeutic performances were also summarized focusing on three representative categories of glycoproteins, cytokines, therapeutic antibody and enzyme. Currently, mammalian expression systems such as Chinese hamster ovary cells are preferred for the production of therapeutic glycoproteins due to their ability to synthesize glycans having similar structures with human type glycans. However, recent advances of plant glycoengineering to overcome the limitation originating from different glycan structures will soon allow to develop more efficient and economic plant-based production systems for therapeutic glycoproteins.

Organic matrix-free imaging mass spectrometry

  • Kim, Eunjin;Kim, Jisu;Choi, Inseong;Lee, Jeongwook;Yeo, Woon-Seok
    • BMB Reports
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    • 제53권7호
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    • pp.349-356
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    • 2020
  • Mass spectrometry (MS) is an ideal tool for analyzing multiple types of (bio)molecular information simultaneously in complex biological systems. In addition, MS provides structural information on targets, and can easily discriminate between true analytes and background. Therefore, imaging mass spectrometry (IMS) enables not only visualization of tissues to give positional information on targets but also allows for molecular analysis of targets by affording the molecular weights. Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) MS is particularly effective and is generally used for IMS. However, the requirement for an organic matrix raises several limitations that get in the way of accurate and reliable images and hampers imaging of small molecules such as drugs and their metabolites. To overcome these problems, various organic matrix-free LDI IMS systems have been developed, mostly utilizing nanostructured surfaces and inorganic nanoparticles as an alternative to the organic matrix. This minireview highlights and focuses on the progress in organic matrix-free LDI IMS and briefly discusses the use of other IMS techniques such as desorption electrospray ionization, laser ablation electrospray ionization, and secondary ion mass spectrometry.