• 제목/요약/키워드: Biological systems

검색결과 2,229건 처리시간 0.03초

Heterologous Production of Streptokinase in Secretory Form in Streptomyces lividans and in Nonsecretory Form in Escherichia coli

  • Kim,, Mi-Ran;Choeng, Yong-Hoon;Chi, Won-Jae;Kang, Dae-Kyung;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • 제20권1호
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    • pp.132-137
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    • 2010
  • The skc gene encoding streptokinase (SK) with a molecular mass of approximately 47.4 kDa was cloned from Streptococcus equisimilis ATCC 9542 and heterologously overexpressed in Streptomyces lividans TK24 and E. coli using various strong promoters. When the promoter for sprT [Streptomyces griseus trypsin (SGT)] was used in the host S. lividans TK24, a 47.4-kDa protein was detected along with a smaller hydrolyzed protein (44 kDa), suggesting that posttranslational hydrolysis had occurred as has been reported in other expression systems. The casein/plasminogen plate assay revealed that the plasmid construct containing the SGT signal peptide was superior to that containing the SK signal peptide in terms of SK production. Maximal production of SK was calculated to be about 0.25 unit/ml of culture broth, a value that was five times higher than that obtained with other expression systems using ermE and tipA promoters in the same host. When the skc gene was expressed in E. coli BL21(${\Delta}DE3$)pLys under the control of the T7 promoter, a relatively large amount of SK was expressed in soluble form without hydrolysis. SK activity in E. coli/pET28a-$T7_pSK_m$ was more than 2 units/ml of culture broth, even though about half of the expressed protein formed an inactive inclusion body.

가용화 조성물과 난용성 약물군을 함유하는 고체분산체의 용출양상 (Dissolution Profiles of Solid Dispersions Containing Poorly Water-Soluble Drugs and Solubilizing Compositions)

  • 김태완;최춘영;;권경애;이범진
    • Journal of Pharmaceutical Investigation
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    • 제32권3호
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    • pp.191-197
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    • 2002
  • Polymer based physical mixtures or solid dispersions containing solubilizing compositions[OA, tween80 and SLS] were prepared using a spray-dryer. Lovastatin(LOS), simvastatin(SIMS), aceclofenac(AFC) and cisapride(CSP) were selected as poorly water-soluble drugs. Dextrin, poly(vinylalcohol) (PVA), poly(vinylpyrrolidone)(PVP) and polyethylene glycol(PEG) were chosen as solubilizing carriers for solid dispersions. The solid dispersions containing solubilizing compositions without drug were prepared without using organic solvents or tedious changes of formulation compositions. This system could be used to quickly screen the dissolution profiles of poorly water-soluble drugs by simply mixing with drugs thereafter. In case of solid dispersion containing drug, organic solvent systems could be used to solubilize model drugs. The dissolution rates of the drugs were higher when mixed with drug and solid dispersions containing solubilizing compositions. However, solid dispersions of LOS, AFC, and CSP simultaneously containing drug and solubilizing compositions in organic solvent systems were more useful than physical mixtures of drug and solid dispersions without drug except SIMS. Based on solubilizing capability of polymer based physical mixtures in gelatin hard capsules, optimal solid dispersion system of poorly water-soluble drugs could be formulated. However, it should be noted that dissolution rate of poorly water-soluble drugs were highly dependent on drug properties, solubilizing compositions and polymeric carriers.

Dictyostelium discoideum Ax2 as an Assay System for Screening of Pharmacological Chaperones for Phenylketonuria Mutations

  • Kim, Yu-Min;Yang, Yun Gyeong;Kim, Hye-Lim;Park, Young Shik
    • Journal of Microbiology and Biotechnology
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    • 제25권6호
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    • pp.782-787
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    • 2015
  • In this study, we developed an assay system for missense mutations in human phenylalanine hydroxylases (hPAHs). To demonstrate the reliability of the system, eight mutant proteins (F39L, K42I, L48S, I65T, R252Q, L255V, S349L, and R408W) were expressed in a mutant strain (pah-) of Dictyostelium discoideum Ax2 disrupted in the indigenous gene encoding PAH. The transformed pah - cells grown in FM minimal medium were measured for growth rate and PAH activity to reveal a positive correlation between them. The protein level of hPAH was also determined by western blotting to show the impact of each mutation on protein stability and catalytic activity. The result was highly compatible with the previous ones obtained from other expression systems, suggesting that Dictyostelium is a dependable alternative to other expression systems. Furthermore, we found that both the protein level and activity of S349L and R408W, which were impaired severely in protein stability, were rescued in HL5 nutrient medium. Although the responsible component(s) remains unidentified, this unexpected finding showed an important advantage of our expression system for studying unstable proteins. As an economic and stable cell-based expression system, our development will contribute to mass-screening of pharmacological chaperones for missense PAH mutations as well as to the in-depth characterization of individual mutations.

반도체 공정에서의 APC 기법 및 이상감지 및 분류 시스템 (APC Technique and Fault Detection and Classification System in Semiconductor Manufacturing Process)

  • 하대근;구준모;박담대;한종훈
    • 제어로봇시스템학회논문지
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    • 제21권9호
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    • pp.875-880
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    • 2015
  • Traditional semiconductor process control has been performed through statistical process control techniques in a constant process-recipe conditions. However, the complexity of the interior of the etching apparatus plasma physics, quantitative modeling of process conditions due to the many difficult features constraints apply simple SISO control scheme. The introduction of the Advanced Process Control (APC) as a way to overcome the limits has been using the APC process control methodology run-to-run, wafer-to-wafer, or the yield of the semiconductor manufacturing process to the real-time process control, performance, it is possible to improve production. In addition, it is possible to establish a hierarchical structure of the process control made by the process control unit and associated algorithms and etching apparatus, the process unit, the overall process. In this study, the research focused on the methodology and monitoring improvements in performance needed to consider the process management of future developments in the semiconductor manufacturing process in accordance with the age of the APC analysis in real applications of the semiconductor manufacturing process and process fault diagnosis and control techniques in progress.

Explicating morphophysiological and biochemical responses of wheat grown under acidic medium: Insight into to the antioxidant defense and glyoxalase systems

  • Bhuyan, MHM Borhannuddin;Hasanuzzaman, Mirza;Al Mahmud, Jubayer;Hossain, Md. Shahdat;Alam, Mazhar Ul;Fujita, Masayuki
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.236-236
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    • 2017
  • Low soil pH causes from $H^+$ rhizotoxicity results in nutrients unavailability in the growing media, inhibits plant growth, development and reduces crop yields. The present study was carried out to reveal morpholophysiological and biochemical responses of wheat (Triticum aestivum L.) to acidity stress. Four wheat varieties viz. BARI Wheat-21, BARI Wheat-25, BARI Wheat-26 and BARI Wheat-30 were used in the study. Eight-day-old seedlings were exposed to different pH levels (3.5, 4.5, 5.5 and 6.5) of growing media. Acidity stress at any level reduced biomass, water, and chlorophyll contents in all the varieties; whereas BARI Wheat-26 showed the least damage. $H^+$ rhizotoxicity also caused oxidative stress through excess production of reactive oxygen species and methylglyoxal which increase lipid peroxidation in all the varieties but the lowest oxidative damage was observed in BARI Wheat-26 due to better performance of the antioxidant defense and glyoxalase systems. Considering the growth, physiological and biochemical attributes BARI Wheat-26 may be considered as acidity stress tolerant, among the variety examined.

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돌말(Epilithic Diatom) 지수를 이용한 국내 주요 하천(금강, 영산강, 섬진강)의 생물학적 수질평가 (Biological Assessment of Water Quality by Using Epilithic Diatoms in Major River Systems (Geum, Youngsan, Seomjin River), Korea)

  • 황순진;김난영;원두희;안광국;이재관;김창수
    • 한국물환경학회지
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    • 제22권5호
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    • pp.784-795
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    • 2006
  • The purpose of this study was to assess biological river water quality by using epilithic diatoms at 40 selected sites in Geum, Youngsan, and Seomjin River systems. The sampling and analyses were performed during three seasons including January, April and June in 2005. Various water quality parameters also were analyzed. We attempted to classify the water quality condition by epilithic diatom indices (DAIpo and TDI) with the results of corresponding analyses of various chemical water quality parameters. A five class system was delivered to describe the water quality condition ranged from "very good" to "very poor." We also proposed a way of classifying water quality condition by combining two diatom indices of DAIpo and TDI. Our results showed that biomass of epilithic diatoms varied not only seasonally but spatially; it was not likely that winter diatoms represent average water quality condition, due to high concentration of nutrients. Water quality status assessed by diatom indices was generally worse than that assessed by BOD, indicating that BOD standard likely underestimates the biological condition of the water body. Importantly, nutrient-based diatom index (TDI) generally overestimated organic matter-based index (DAIpo) at most study sites, indicating that diatoms in studied rivers were likely more affected by nutrients than organic matter. Thus, management strategy to improve river water quality in Korea is suggested to emphasize more on the nutrients than organic matters.

Critical Adjuvant Influences on Preventive Anti-Metastasis Vaccine Using a Structural Epitope Derived from Membrane Type Protease PRSS14

  • Ki Yeon Kim;Eun Hye Cho;Minsang Yoon;Moon Gyo Kim
    • IMMUNE NETWORK
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    • 제20권4호
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    • pp.33.1-33.19
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    • 2020
  • We tested how adjuvants effect in a cancer vaccine model using an epitope derived from an autoactivation loop of membrane-type protease serine protease 14 (PRSS14; loop metavaccine) in mouse mammary tumor virus (MMTV)-polyoma middle tumor-antigen (PyMT) system and in 2 other orthotopic mouse systems. Earlier, we reported that loop metavaccine effectively prevented progression and metastasis regardless of adjuvant types and TH types of hosts in tail-vein injection systems. However, the loop metavaccine with Freund's complete adjuvant (CFA) reduced cancer progression and metastasis while that with alum, to our surprise, were adversely affected in 3 tumor bearing mouse models. The amounts of loop peptide specific antibodies inversely correlated with tumor burden and metastasis, meanwhile both TH1 and TH2 isotypes were present regardless of host type and adjuvant. Tumor infiltrating myeloid cells such as eosinophil, monocyte, and neutrophil were asymmetrically distributed among 2 adjuvant groups with loop metavaccine. Systemic expression profiling using the lymph nodes of the differentially immunized MMTV-PyMT mouse revealed that adjuvant types, as well as loop metavaccine can change the immune signatures. Specifically, loop metavaccine itself induces TH2 and TH17 responses but reduces TH1 and Treg responses regardless of adjuvant type, whereas CFA but not alum increased follicular TH response. Among the myeloid signatures, eosinophil was most distinct between CFA and alum. Survival analysis of breast cancer patients showed that eosinophil chemokines can be useful prognostic factors in PRSS14 positive patients. Based on these observations, we concluded that multiple immune parameters are to be considered when applying a vaccine strategy to cancer patients.

이미다졸계 이온성액체와 염을 포함한 수상이성분계를 이용한 숙신산의 추출 평형 (Extraction Equilibria of Succinic Acid by Using Aqueous Two Phases System Containing Imidazolium Ionic Liquids and Salts)

  • 이용화;강정원;홍연기;김기섭
    • 청정기술
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    • 제20권4호
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    • pp.349-353
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    • 2014
  • 숙신산은 의약품, 식품 첨가물, 청정용매와 같은 종래의 응용 이외에도 생분해성 고분자의 중요한 전구물질로 주목 받고 있다. 그러나 최근 화석 연료의 높은 가격과 화석 원료 유래 제품의 전주기에 걸친 온실가스 배출로 인해 재생 가능한 바이오매스에 의한 숙신산의 생물학적 생산에 대한 관심이 높아지고 있다. 본 연구에서는 숙신산의 선택적인 추출과 농축을 동시에 수행할 수 있는 공정으로서 이미다졸계 이온성 액체/$K_2HPO_4$에 의한 수상이성분계를 적용하였다. 실험 결과 숙신산에 존재하는 $K_2HPO_4$ 수용액에 적정량의 이미다졸계 이온성 액체가 첨가됨에 따라 안정한 수상이성분계가 형성됨을 확인할 수 있었으며 수상이성분계 형성 능력은 [HMIm][Br]${\fallingdotseq}$[OMIm][Br]>[BMIm][Br]>[EMImBr]의 순서로 나타났다. 숙신산의 최대 추출 효율은 약 90%였으며 이온성 액체에서 양이온 사슬길이가 증가할수록 상부상으로 동반 추출되는 물의 양이 줄어들어 숙신산의 선택적 농축효과가 크게 나타났다. 따라서 본 연구에서 사용된 이미다졸계 이온성 액체와 $K_2HPO_4$으로 구성된 수상이성분계는 숙신산의 선택적 추출과 농축에 효과적임을 알 수 있었다.

친수성 이온성 액체를 이용한 수상이성분계에서의 아크릴산 추출 평형 (Extraction Equilibrium of Acrylic Acid by Aqueous Two-Phase Systems Using Hydrophilic Ionic Liquids)

  • 이용화;이우윤;김기섭;홍연기
    • Korean Chemical Engineering Research
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    • 제52권5호
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    • pp.627-631
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    • 2014
  • 본 연구에서는 아크릴산 추출을 위한 효과적인 분리공정으로서 모폴린계 이온성 액체/$K_2HPO_4$에 의한 수상이성분계를 적용하여 수상이성분계 형성 특성 및 아크릴산 추출 효율에 대한 모폴린계 이온성 액체의 양이온 알킬사슬 길이의 영향을 고찰하였다. 실험 결과 $K_2HPO_4$ 수용액에 적정량의 모폴린계 이온성 액체를 첨가함에 따라 안정한 수상이성분계가 형성됨을 확인할 수 있었으며 이성분계 형성능력은 [HMMor][Br]>[OMMor][Br]>[BMMor][Br]> [EMMor][Br]의 순서로 나타났다. $K_2HPO_4$ 수용액을 기준으로 모폴린계 이온성 액체의 수상이성분계 형성 능력은 기존의 이미다졸계와 큰 차이가 없었으며 친수성알콜/$K_2HPO_4$와 비교했을 경우에는 적은양으로도 수상이성분계 형성이 가능하였다. 모폴린계 이온성 액체를 이용한 수상이성분계 추출에 있어 아크릴산 추출 효율은 50~90%의 범위를 가지며 이 값들은 이미다졸계와 큰 차이가 없었다. 모폴린계 이온성 액체가 이미다졸계 이온성 액체에 비해 경제적으로 생산할 수 있으므로 모폴린계 이온성 액체와 $K_2HPO_4$로 구성된 수상이성분계는 아크릴산을 포함한 각종 생물공정에 의한 제품 분리에 효과적임을 알 수 있었다.

Taxonomic Position and Affinities of Isopyrum mandshuricum within Korean Isopyroideae (Ranunculaceae) Based on Molecular Data

  • Lee, Nam-Sook;Yeau, Sung-Hee;Kim, Ji-Hyun;Kim, Min-Ju
    • Animal cells and systems
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    • 제3권2호
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    • pp.133-141
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    • 1999
  • To examine the taxonomic position and affinities of Isopyrum mandshuricum (Ranunculaceae) and related taxa, genetic analysis were carried out on the basis of isozyme patterns and ITS sequences. Molecular data, both isozyme patterns and ITS sequences suggest that I. mandshuricum is closely related to Enemion raddeanum than to Semiaquilegia adoxoides. The estimation of genetic identities by isozyme analysis reveals that I. manshuricum is genetically distant from E. raddeanum. The phylogenetic tree based on molecular data is rather congruent with the phenogram based on quantitative morphological characteristics, but not consistent with one based on qualitative morphological characteristics. Incongruencies between molecular and qualitative morphological data provide clues to re-evaluate several morphological features.

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