• Title/Summary/Keyword: 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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    • v.18 no.3
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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.

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

  • Kim, Soo-Hyun;Choi, Hyun-Jin;Oh, Hyun-Taek;Chung, Mi-Ja;Cui, Cheng-Bi;Ham, Seung-Shi
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.696-701
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    • 2008
  • The objective of this study was to determine whether Codonopsis lanceolata or Platycodon grandiflorum ethyl acetate fraction (CLEA or PGEA) protect cells against sodium nitroprusside (SNP)-induced oxidative stress via the expression of various antioxidant systems. The HepG2 cells exposed for 24 hr to 0.5 mM SNP showed a reduction in the cell viability by an MTT assay. Pretreatment with CLEA and PGEA resulted in an inhibition of SNP-induced cell death. In addition, the effects of CLEA and PGEA on the expression of antioxidant systems via RT-PCR analyses was assessed. The levels of catalase (CAT), glucose-6-phosphate dehydrogenase (G6PD) and metallothionein (MT)-1A mRNA were increased after 24 hr of CLEA exposure. The levels of Mn superoxide dismutase CAT, G6PD, MT-1A, and MT-2A mRNA were increased after PGEA treatment. In conclusion, CLEA and PGEA exert indirect antioxidant effects, perhaps via the induction of a variety of antioxidant systems which, may protect cells against oxidative stress.

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

  • Lee, Jin-Sung;Moon, Ju Yeon;Nam, Ki-Taeg;Park, Seong-Min;Park, Seung-Yong;Jung, Mun Yhung;Son, Yong-Suk
    • Journal of Dairy Science and Biotechnology
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    • v.34 no.4
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    • pp.245-254
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    • 2016
  • This investigation was carried out to study effects of different milking systems on the characteristics of Gouda cheese produced at farmstead milk-processing plants. In the first study, raw milk was collected from automatic milking system and conventional milking system farms, and Gouda cheeses were prepared under the same conditions. After 6 months of aging, the chemical composition was analyzed and free fatty acid (FFA) analysis and sensory evaluation were carried out on the products. In the second study, Gouda cheese samples were collected from seven farmstead milk-processing plants and the chemical composition and texture profile analysis were evaluated. No significant differences were found in the chemical composition of Gouda cheese according to different milking systems or different farms except crude ash. FFA analysis showed significant differences among farms, but sensory evaluation showed no significant differences. No significant differences were observed in FFA content and sensory assessment of different milking systems. Texture profile analysis revealed that there were significant differences in each cheese made at different farm plants, but there were no differences when different milking systems were used.

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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    • v.21 no.4
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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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    • v.14 no.4
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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.

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

  • Ku, Youngwoo;Rho, Younghee;Jeong, Gyujin;Min, Wankee
    • Journal of Korea Technology Innovation Society
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    • v.16 no.3
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    • pp.861-882
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    • 2013
  • This study analyses interorganizational innovation networks empirically in the Korean biotechnology industry, based on systems of innovation approach. Empirical findings of this study are as follows. Innovation networks have not been well developed in the Korean biotechnology industry. Main agents who participate in innovation networks are workers at venture firms, university professors, researchers at government-sponsored research institutions. They all recognize the positive effects of innovation networks. The enhancement of participation willingness and competency of agents, the enrichment of trust and control among agents, and the effective acquisition of the useful technology and resources in innovation networks will largely contribute to development of innovation networks in the Korean biotechnology industry.

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

  • Kim, Seong-Hun;Kwon, Oh-Suk;Oh, Doo-Byoung
    • Journal of Plant Biotechnology
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    • v.37 no.3
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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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    • v.53 no.7
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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.