• 제목/요약/키워드: Systems Biotechnology

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The Actinobacterium Corynebacterium glutamicum, an Industrial Workhorse

  • Lee, Joo-Young;Na, Yoon-Ah;Kim, Eungsoo;Lee, Heung-Shick;Kim, Pil
    • Journal of Microbiology and Biotechnology
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    • 제26권5호
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    • pp.807-822
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    • 2016
  • Starting as a glutamate producer, Corynebacterium glutamicum has played a variety of roles in the industrial production of amino acids, one of the most important areas of white biotechnology. From shortly after its genome information became available, C. glutamicum has been applied in various production processes for value-added chemicals, fuels, and polymers, as a key organism in industrial biotechnology alongside the surprising progress in systems biology and metabolic engineering. In addition, recent studies have suggested another potential for C. glutamicum as a synthetic biology platform chassis that could move the new era of industrial microbial biotechnology beyond the classical field. Here, we review the recent progress and perspectives in relation to C. glutamicum, which demonstrate it as one of the most promising and valuable workhorses in the field of industrial biotechnology.

체세포배발생에 의한 IbOr 유전자 형질전환 카사바 개발 (Development of transgenic cassava plants expressing IbOr gene by somatic embryogenesis)

  • 김선하;김명덕;박성철;정재철;이행순;곽상수
    • Journal of Plant Biotechnology
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    • 제42권2호
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    • pp.88-92
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    • 2015
  • 카사바는 열대와 아열대지역 뿌리작물로서 중요한 식량자원일 뿐만 아니라 동물 사료, 전분, 바이오에탄올 등 다양한 산업소재로서 이용이 가능하다. 그러나 카사바의 산업적 중요성에 비해 형질전환기술을 이용한 신품종 개발은 아직까지 제한적이다. 본 연구에서는 인도네시아 IDB사가 개발한 다수확 카사바 품종을 이용하여 영양강화 및 환경스트레스에 저항성을 향상시킨 카사바를 개발하기 위하여 체세포배를 이용한 식물체 재분화시스템을 확립하였다. 카로티노이드 축적에 관련된 IbOr 유전자를 체세포배를 이용한 Agrobacterium 매개방법으로 카사바에 형질전환하였다. gDNA PCR과 RT-PCR을 통해 19개의 형질전환식물체를 성공적으로 확보하였다. 향후 카로티노이드 함량분석, 환경스트레스 내성분석 등을 통하여 IbOr 카사바 식물체의 농업적 유용성을 검정할 예정이다.

CodA 고발현 형질전환 고구마의 산화 및 건조 스트레스 내성 증가 (Enhanced drought and oxidative stress tolerance in transgenic sweetpotato expressing a codA gene)

  • 박성철;김명덕;김선하;김윤희;정재철;이행순;곽상수
    • Journal of Plant Biotechnology
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    • 제42권1호
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    • pp.19-24
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    • 2015
  • 식물은 여러 환경스트레스에 적응하기 위해 스트레스 내성 유전자의 발현 혹은 proline, trehalose, glycine betaine (GB) 등과 같이 삼투압을 조절하는 compatible solute를 생성하면서 진화해 왔다. GB는 고염, 저온 등 환경스트레스 조건에서 식물의 엽록체에서 축적되는 물질 중 하나이다. 토양 박테리아 Arthrobacter globiformis에서 분리한 choline oxidase (codA) 유전자는 choline을 GB로 전환하는 기능을 한다. 본 연구에서는 산화스트레스 유도성 SWPA2 프로모터의 발현조절 하에 codA 유전자를 엽록체에 과발현시킨 형질전환 고구마 식물체(SC식물체)를 제작하여 다양한 환경스트레스 조건에서의 특성을 분석하였다. SC 식물체는 methyl viologen (MV)에 의한 산화스트레스와 건조 처리 조건에서 내성 증가를 보였다. $5{\mu}M$ MV 처리시 형질전환 식물체는 GB의 함량이 증가하였고 낮은 수준의 이온 전도도를 보였다. 건조 스트레스 조건에서 형질전환 식물체는 codA 유전자의 발현이 증가하였으며, 대조구 보다 높은 상대수분함량을 유지하였다. 따라서 본 연구결과의 SC식물체는 고염, 건조토양 등 조건 불리지역에 재배하면 바이오매스를 증가시킬 수 있을 것으로 예상된다.

Biphasic Study to Characterize Agricultural Biogas Plants by High-Throughput 16S rRNA Gene Amplicon Sequencing and Microscopic Analysis

  • Maus, Irena;Kim, Yong Sung;Wibberg, Daniel;Stolze, Yvonne;Off, Sandra;Antonczyk, Sebastian;Puhler, Alfred;Scherer, Paul;Schluter, Andreas
    • Journal of Microbiology and Biotechnology
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    • 제27권2호
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    • pp.321-334
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    • 2017
  • Process surveillance within agricultural biogas plants (BGPs) was concurrently studied by high-throughput 16S rRNA gene amplicon sequencing and an optimized quantitative microscopic fingerprinting (QMF) technique. In contrast to 16S rRNA gene amplicons, digitalized microscopy is a rapid and cost-effective method that facilitates enumeration and morphological differentiation of the most significant groups of methanogens regarding their shape and characteristic autofluorescent factor 420. Moreover, the fluorescence signal mirrors cell vitality. In this study, four different BGPs were investigated. The results indicated stable process performance in the mesophilic BGPs and in the thermophilic reactor. Bacterial subcommunity characterization revealed significant differences between the four BGPs. Most remarkably, the genera Defluviitoga and Halocella dominated the thermophilic bacterial subcommunity, whereas members of another taxon, Syntrophaceticus, were found to be abundant in the mesophilic BGP. The domain Archaea was dominated by the genus Methanoculleus in all four BGPs, followed by Methanosaeta in BGP1 and BGP3. In contrast, Methanothermobacter members were highly abundant in the thermophilic BGP4. Furthermore, a high consistency between the sequencing approach and the QMF method was shown, especially for the thermophilic BGP. The differences elucidated that using this biphasic approach for mesophilic BGPs provided novel insights regarding disaggregated single cells of Methanosarcina and Methanosaeta species. Both dominated the archaeal subcommunity and replaced coccoid Methanoculleus members belonging to the same group of Methanomicrobiales that have been frequently observed in similar BGPs. This work demonstrates that combining QMF and 16S rRNA gene amplicon sequencing is a complementary strategy to describe archaeal community structures within biogas processes.

Salting-out Effects on the Partition of Proteins in Aqueous Two-phase Systems

  • KIM, CHAN-WHA;CHO KYUN RHA
    • Journal of Microbiology and Biotechnology
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    • 제6권5호
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    • pp.352-357
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    • 1996
  • The partition of proteins in the salt-rich phase of polyethylene glycol (pEG)/salt aqueous two-phase systems is limited by the salting-out effects of salt. The logarithm of the concentration of proteins partitioned in the salt-rich phase decreases linearly with increases in the concentration of salt in the salt-rich phase (salting-out). Therefore, the partition of a given protein in the salt-rich phase of aqueous two-phase systems can be estimated from the salting-out constant. The slope of the solubility line (salting-out con-stant) for a given protein is determined by the type of salt in the two-phase systems.

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Microcontact Printing of Biotin for Selective Immobilization of Streptavidin-fused Proteins and SPR Analysis

  • Lee, Sang-Yup;Park, Jong-Pil;Lee, Seok-Jae;Park, Tae-Jung;Lee, Kyung-Bok;Park, Insung S.;Kim, Min-Gon;Chung, Bong-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권2호
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    • pp.137-142
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    • 2004
  • In this study, a simple procedure is described for patterning biotin on a glass substrate and then selectively immobilizing proteins of interest onto the biotin-patterned surface. Microcontact printing (CP) was used to generate the micropattern of biotin and to demonstrate the selective immobilization of proteins by using enhanced green fluorescent protein (EGFP) as a model protein, of which the C-terminus was fused to a core streptavidin (cSA) gene of Streptomyces avidinii. Confocal fluorescence microscopy was used to visualize the pattern of the immobilized protein (EGFP-cSA), and surface plasmon resonance was used to characterize biological activity of the immobilized EGFP-cSA. The results suggest that this strategy, which consists of a combination of $\mu$CP and cSA-fused proteins. is an effective way for fabricating biologically active substrates that are suitable for a wide variety of applications. one such being the use in protein-protein assays.

Report on 31 unrecorded bacterial species in Korea that belong to the phylum Actinobacteria

  • Choi, Jung-Hye;Cha, Ju-Hee;Bae, Jin-Woo;Cho, Jang-Cheon;Chun, Jongsik;Im, Wan-Taek;Jahng, Kwang Yeop;Jeon, Che Ok;Joh, Kiseong;Kim, Seung Bum;Seong, Chi Nam;Yoon, Jung-Hoon;Cha, Chang-Jun
    • Journal of Species Research
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    • 제5권1호
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    • pp.1-13
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    • 2016
  • To discover and characterize indigenous species in Korea, a total of 31 bacterial strains that belong to the phylum Actinobacteria were isolated from various niches in Korea. Each strain showed the high sequence similarity (>99.1%) with the closest bacterial species, forming a robust phylogenetic clade. These strains have not been previously recorded in Korea. According to the recently updated taxonomy of the phylum Actinobacteria based upon 16S rRNA trees, we report 25 genera of 13 families within 5 orders of the class Actinobacteria as actinobacterial species found in Korea. Cellular morphology, Gram staining, basic biochemical characteristics are described in the species description.

Production and characterization of lentivirus vector-based SARS-CoV-2 pseudoviruses with dual reporters: Evaluation of anti-SARS-CoV-2 viral effect of Korean Red Ginseng

  • Jeonghui Moon;Younghun Jung;Seokoh Moon;Jaehyeon Hwang;Soomin Kim;Mi Soo Kim;Jeong Hyeon Yoon;Kyeongwon Kim;Youngseo Park;Jae Youl Cho;Dae-Hyuk Kweon
    • Journal of Ginseng Research
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    • 제47권1호
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    • pp.123-132
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    • 2023
  • Background: Pseudotyped virus systems that incorporate viral proteins have been widely employed for the rapid determination of the effectiveness and neutralizing activity of drug and vaccine candidates in biosafety level 2 facilities. We report an efficient method for producing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pseudovirus with dual luciferase and fluorescent protein reporters. Moreover, using the established method, we also aimed to investigate whether Korean Red Ginseng (KRG), a valuable Korean herbal medicine, can attenuate infectivity of the pseudotyped virus. Methods: A pseudovirus of SARS-CoV-2 (SARS-2pv) was constructed and efficiently produced using lentivirus vector systems available in the public domain by the introduction of critical mutations in the cytoplasmic tail of the spike protein. KRG extract was dose-dependently treated to Calu-3 cells during SARS2-pv treatment to evaluate the protective activity against SARS-CoV-2. Results: The use of Calu-3 cells or the expression of angiotensin-converting enzyme 2 (ACE2) in HEK293T cells enabled SARS-2pv infection of host cells. Coexpression of transmembrane protease serine subtype 2 (TMPRSS2), which is the activator of spike protein, with ACE2 dramatically elevated luciferase activity, confirming the importance of the TMPRSS2-mediated pathway during SARS-CoV-2 entry. Our pseudovirus assay also revealed that KRG elicited resistance to SARS-CoV-2 infection in lung cells, suggesting its beneficial health effect. Conclusion: The method demonstrated the production of SARS-2pv for the analysis of vaccine or drug candidates. When KRG was assessed by the method, it protected host cells from coronavirus infection. Further studies will be followed for demonstrating this potential benefit.