• 제목/요약/키워드: Bioprocess engineering

검색결과 760건 처리시간 0.025초

A Dual Selection Marker Transformation System Using Agrobacterium tumefaciens for the Industrial Aspergillus oryzae 3.042

  • Sun, Yunlong;Niu, Yali;He, Bin;Ma, Long;Li, Ganghua;Tran, Van-Tuan;Zeng, Bin;Hu, Zhihong
    • Journal of Microbiology and Biotechnology
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    • 제29권2호
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    • pp.230-234
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    • 2019
  • Currently, the genetic modification of Aspergillus oryzae is mainly dependent on protoplast-mediated transformation (PMT). In this study, we established a dual selection marker system in an industrial A. oryzae 3.042 strain by using Agrobacterium tumefaciens-mediated transformation (ATMT). We first constructed a uridine/uracil auxotrophic A. oryzae 3.042 strain and a pyrithiamine (PT)-resistance binary vector. Then, we established the ATMT system by using uridine/uracil auxotrophy and PT-resistance genes as selection markers. Finally, a dual selection marker ATMT system was developed. This study demonstrates a useful dual selection marker transformation system for genetic manipulations of A. oryzae 3.042.

Polyhydroxyalkanoate Chip for the Specific Immobilization of Recombinant Proteins and Its Applications in Immunodiagnostics

  • Park, Tae-Jung;Park, Jong-Pil;Lee, Seok-Jae;Hong, Hyo-Jeong;Lee, Sang-Yup
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제11권2호
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    • pp.173-177
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    • 2006
  • In this study, a novel strategy was developed for the highly selective immobilization of proteins, using the polyhydroxyalkanoate (PHA) depolymerase substrate binding domain (SBD) as an active binding domain. In order to determine the appropriacy of this method for immunodiagnostic assays, the single-chain antibody (ScFv) against the hepatitis B virus (HBV) preS2 surface protein and the severe acute respiratory syndrome coronavirus (SARS-CoV) envelope protein (SCVe) were fused to the SBD, then directly immobilized on PH A-coated slides via microspotting. The fluorescence-labeled HBV antigen and the antibody against SCVe were then utilized to examine specific interactions on the PHA-coated surfaces. Fluorescence signals were detected only at the spotted positions, thereby indicating a high degree of affinity and selectivity for their corresponding antigens/antibodies. Furthermore, we detected small amounts of ScFv-SBD (2.7 ng/mL) and SCVe-SBD fusion proteins (0.6ng/mL). Therefore, this microarray platform technology, using PHA and SBD, appears generally appropriate for immunodiagnosis, with no special requirements with regard to synthetic or chemical modification of the biomolecules or the solid surface.

Preparation and Characterization of Resveratrol Nanoemulsions Stabilized by Self-assembly and Complex Coacervation Consisting of Sodium Alginate, Chitosan, and β-Cyclodextrin

  • Choi, Ae-Jin;Jo, Younghee;Cho, Yong-Jin;Kim, Tae-Eun;Kim, Chong-Tai
    • 산업식품공학
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    • 제21권3호
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    • pp.215-224
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    • 2017
  • Resveratrol was incorporated into various combinations of single- and double-layer nanoemulsions, prepared by self-assembly emulsification and complex coacervation with chitosan, alginate, and ${\beta}$-cyclodextrin, respectively. Resveratrol nanoemulsions were composed of medium-chain trigacylglycerols (MCTs), $Tween^{(R)}$ 80, water, chitosan, alginate, and ${\beta}$-cyclodextrin. The corresponding mixtures were formulated for the purpose of being used as a nutraceutical delivery system. Resveratrol nanoemulsions were obtained with particle sizes of 10-800 nm, with the size variation dependent on the emulsification parameters including the ratio of aqueous phase and surfactant ratio. Resveratrol nanoemulsions were characterized by evaluating particle size, zeta-potential value, stability, and release rate. There were no significant changes in particle size and zeta-potential value of resveratrol nanoemulsions during storage for 28 days at $25^{\circ}C$. The stability of resveratrol in the double-layer nanoemulsions complexed with chitosan or ${\beta}$-cyclodextrin was higher, compared with the single-layer nanoemulsions.

Experimental determination of liquid entry pressure (LEP) in vacuum membrane distillation for oily wastewaters

  • Racz, Gabor;Kerker, Steffen;Schmitz, Oliver;Schnabel, Benjamin;Kovacs, Zoltan;Vatai, Gyula;Ebrahimi, Mehrdad;Czermak, Peter
    • Membrane and Water Treatment
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    • 제6권3호
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    • pp.237-249
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    • 2015
  • In this study we investigate a laboratory scale vacuum membrane distillation system to produce pure water from model oil in water emulsion. Experimental determination of liquid entry pressure (LEP) of a commercial Durapore$^{TM}$ GVPH flat sheet membrane using model emulsions in various oil concentrations has been carried out. Two different methods of liquid entry pressure determination - a frequently used, so-called static and a novel dynamic method - have been investigated. In case of static method, LEP value was found to be 2.3 bar. No significant effect of oil content on LEP was detected up to 3200 ppm. In contrast, LEP values determined with dynamic method showed strong dependence on the oil concentration of the feed and decreased from 2.0 bar to a spontaneous wetting at 0.2 bar in the range 0-250 ppm, respectively. Vacuum membrane distillation tests were also performed. The separation performance is evaluated in terms of flux behavior, total organic carbon removal and droplet size distribution of the feed and final retentate. No significant effect of oil content on the flux was found ($5.05{\pm}0.31kgm^{-2}h^{-1}$) up to 250 ppm, where a spontaneous wetting occurred. High separation performance was achieved along with the increasing oil concentration between 93.4-97.0%.

Effects of Medium Components on L-Ornithine Production by Brevibacterium ketoglutamicum

  • TaeHo Lee;Yon
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제1권1호
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    • pp.41-45
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    • 1996
  • Effects of yeast extract and ammonium sulfate were investigated on the production of L-or-nithine by an arginine auxotroph, Brevibacterium ketoglutamicum in flask and batch cultures. Yeast extract as an arginine source and ammonium sulfate as an inorganic nitrogen source had significant effects on L-ornithine production and cell growth. L-ornithine production was repressed by the excessive addition of arginine. Reversion of auxotrophic cells to the wild type was observed when the initial yeast extract concenfration was too low. There existed optimum concentrations of yeast extract and ammonium sulfate for L-or-nithine production. The effects of yeast extract and ammonium sulfate concentrations of the Leudeking-Piret model parameters were examined to analyze the relationship between cell growth and L-ornithine production.

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Analysis of lysosomal membrane proteins exposed to melanin in HeLa cells

  • Bang, Seung Hyuck;Park, Dong Jun;Kim, Yang-Hoon;Min, Jiho
    • Environmental Analysis Health and Toxicology
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    • 제31권
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    • pp.9.1-9.5
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    • 2016
  • Objectives There have been developed to use targeting ability for antimicrobial, anticancerous, gene therapy and cosmetics through analysis of various membrane proteins isolated from cell organelles. Methods It was examined about the lysosomal membrane protein extracted from lysosome isolated from HeLa cell treated by 100 ppm melanin for 24 hours in order to find associated with targeting ability to melanin using by 2-dimensional electrophoresis. Results The result showed 14 up-regulated (1.5-fold) and 13 down-regulated (2.0-fold) spots in relation to melanin exposure. Conclusions It has been found that lysosomal membrane proteins are associated with melanin to decolorize and quantity through cellular activation of lysosome.

Effects of various carbon sources on the production of recombinant phospholipase C (PLC) by Pichia pastoris

  • Kim, Sun-Yong;Han, Kyung-Ah;Rhee, Jong-Il
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2005년도 생물공학의 동향(XVI)
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    • pp.171-174
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    • 2005
  • 20 g/L peptone, 20 g/L dextrose, 10 g/L yeast extract에 100 mg/L zeocin을 첨가하여 동일하게 전배양 한 재조합 Pichia pastoris X-33/pBPT44를 각기 다른 탄소원이 든 배지에 배양하면서 12시간 간격으로 샘플을 채취하여 배양시간에 따른 세포성장, pH, 각 탄소원에 따른 PLC 생산량 등을 측정하였다.

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Expression of various fluorescent protein and their production in shake flasks

  • Park, So-Jung;Han, Kyung-Ah;Rhee, Jong-Il
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2005년도 생물공학의 동향(XVI)
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    • pp.408-411
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    • 2005
  • 대장균을 이용하여 형광단백질을 형질전환하고 배양하면서 그 특성을 관찰하고 형광세기 및 형광 단백질의 크기를 조사하였다.

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