• Title/Summary/Keyword: surface culture

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Constitutive Expression of Lipase on the Cell Surface of Escherichia coli using OmpC Anchoring Motif

  • Lee, Seung Hwan;Lee, Sang Yup
    • Korean Chemical Engineering Research
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    • v.58 no.2
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    • pp.280-285
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    • 2020
  • We have developed a constitutive display system of the Pseudomonas fluorescens SIK W1 TliA lipase on the cell surface of Escherichia coli using E. coli outer membrane protein C (OmpC) as an anchoring motif, which is an economical compared to induced system. For the constitutive expression of truncated OmpC-TliA fusion proteins, gntT104 promoter was employed. Cell growth was not affected by over expression of fusion protein during entire culture time, suggesting cell lysis was not a problem. The localization of truncated OmpC-TliA fusion protein on the cell surface was confirmed by immunofluorescence microscopy and measuring whole cell lipase activity. Constitutively displayed lipase was very stable, retaining activity enantioselectivity throughout the five repeated reactions. These results suggest that OmpC from E. coli be a useful anchoring motif for displaying enzymes on the cell surface without any inducers, and this stable surface display system can be employed for a broad range of biotechnological applications.

The Biocompatibility of HA Film Deposition on Anodized Titanium Alloy

  • Lee, Kang;Choe, Han-Choel;Kim, Byung-Hoon;Ko, Yeong-Mu
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2009.10a
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    • pp.213-214
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    • 2009
  • A thin film hydroxyapatite (HA) films was deposited on anodized titanium by RF sputtering method. The anodized titanium enhanced the biocompatibility of the Ti and the bioactivity was improved further by the HA deposited on the anodized Ti. $TiO_2$ layer with $0.2{\sim}0.5{\mu}$ diameter pore size was formed on the Ti surface by anodization. Anodized $TiO_2$ layer analysis HA film deposited, oxide pore size and number decreased compared with non-HA deposited surface. The corrosion resistance of HA deposited/anodized Ti was higher than that of the non-treatment Ti alloy in Hank's solution, indicating better protective effect. From the results of cell culture using MTT assays, the best cell proliferation showed in HA deposited surface after anodization of Ti surfaces compared with another surface treatment.

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Fabrication and evaluation of hydrophobic metal stent using electron beam equipment (전자빔 처리를 통한 발수성 금속 스텐트 제작 및 평가)

  • Kim, Jisoo;Park, Jongsung
    • Journal of Sensor Science and Technology
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    • v.30 no.3
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    • pp.165-169
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    • 2021
  • The objective of this study was to fabricate a novel hydrophobic stent for reducing restenosis by employing electron beam equipment. The stent was fabricated from a CoCr alloy tube by using a femtosecond laser and was treated with argon plasma. Subsequently, the stent's surface specification changed from hydrophilic to hydrophobic. Application of the electron beam offers several advantages such as a short processing time, whole surface reforming, and enhancement of material properties. As the surface of the stent was rendered hydrophobic, it can provide equivalent or enhanced mechanical properties and greater functionality with a higher radial force at the extended stent in a blood vessel. The obtained results corresponding to the mechanical properties indicate that the contact angle increased to approximately 130°, and the radial force increased to approximately 3 N. Furthermore, cell culture experiments were conducted for verifying whether cells were cultured on the surface-modified CoCr surface. Based on the obtained results, it is believed that an effective reduction in the restenosis of inserted vascular stents is possible.

Callus Induction and Plant Regeneration from Leaf Tissue. Culture of $\emph{Aralia elata}$ S. (두릅의 엽조직배양에 의한 Callus유기 및 식물체 재분화)

  • 장한호;박철호;조동하;신영범
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.38 no.4
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    • pp.366-370
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    • 1993
  • This study was conducted to determine the optimum culture conditions for inducing callus and regenerating plantlets from cultured leaf tissues of Aralia elata. Young leaf tissues(1cm) of A. elata plant were cultured on MS medium supplemented with 2, 4-D and Thidiazuron. Embryogenic callus was induced along the leaf veins, more efficiently on the medium containing 1.0mg /1 Thidiazuron in 4 weeks after culture initiation. Calli were subcultured to proliferate on MS media containing 2, 4-D, Dicamba, Picloram, and Thidiazuron. Callus was better proliferated on the medium containing Dicamba than on the others.. However, callus subcultured on the medium containing Thidiazuron was more embryogenic and light green-colored, of which some showed embryoid-like structure on the surface. Hormone-free medium was more efficient to regenerate plantlets than media supplemented with Kinetin, BA, and Thidiazuron.

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Inhibitory Effects of Acinetobacter sp. KTB3 on Infection of Tobacco mosaic virus in Tobacco Plants

  • Kim, Young-Sook;Hwang, Eui-ll;O, Jeong-Hun;Kim, Kab-Sig;Ryu, Myong-Hyun
    • The Plant Pathology Journal
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    • v.20 no.4
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    • pp.293-296
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    • 2004
  • During the screening of antiviral substances having inhibitory effects on Tobacco mosaic virus (TMV) infection on tobacco plants, we found a bacterial isolate KTB3, and identified it as Acinetobacter sp. which strongly inhibited the infection of TMV When the culture filtrate from KTB3 was applied on the upper surface of the Xanthi-nc tobacco leaves at the same time, or 24 hours before TMV inoculation, almost complete inhibition was achieved. Likewise, 86% inhibition was achieved, when the culture filtrate was applied on the underside of the leaves. In field trials, transmission of TMV from diseased seedlings to healthy ones during transplanting work was reduced by 92%, when the culture filtrate was sprayed onto the tobacco seedlings, cv. NC82, 24 hours before transplanting. No toxic effect was observed on the tobacco plants. Antiviral substance from the culture filtrate was purified by ethanol precipitation, dialysis, DEAE-cellulose, and Sephadex G75 gel column chromatography. The partially purified active material which showed positive color reaction to sugar and protein inhibited TMV infection by 60% at 1 ${\mu}$g/ml.

Doxorubicin Productivity Improvement by the Recombinant Streptomyces peucetius with High-Copy Regulatory Genes Cultured in the Optimized Media Composition

  • PARK, HEE-SEOP;KANG, SEUNG-HOON;PARK, HYUN-JOO;KIM, EUNG-SOO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.66-71
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    • 2005
  • Doxorubicin is a clinically important anticancer polyketide compound that is typically produced by Streptomyces peucetius var. caesius. To improve doxorubicin productivity by S. peucetius, a doxorubicin pathway-specific regulatory gene, dnrI, was cloned into a high-copy-number plasmid containing a catechol promoter system. The S. peucetius containing the recombinant plasmid exhibited approximately 9.5-fold higher doxorubicin productivity compared with the wild-type S. peucetius. The doxorubicin productivity by this recombinant S. peucetius strain was further improved through the optimization of culture media composition. Based on the Fractional Factorial Design (FFD), cornstarch, $K_2HPO_4$, and $MgSO_4$ were identified to be the key factors influencing doxorubicin productivity. The Response Surface Method (RSM) results based on 20 independent culture conditions with varying amounts of key factors predicted the highest theoretical doxorubicin productivity of 11.1 mg/l with corn starch of 46.33 g/l, $K_2HPO_4$ of 4.63 g/l, and $MgSO_4$ of 9.26 g/l. The doxorubicin productivity of the recombinant S. peucetius strain with the RSM-based optimized culture condition was experimentally verified to be 11.46 mg/l, which was approximately 30.8-fold higher productivity compared with the wild-type S. peucetius without culture media optimization.

Ethanol Production by the Mixed Culture of Some Aspergilli and Saccharomyces cerevisiae (효모와 고오지 곰팡이의 혼합배양에 의한 주정생산)

  • Choi, Byung-Kwon;Kim, Young-Bae
    • Korean Journal of Food Science and Technology
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    • v.22 no.6
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    • pp.696-699
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    • 1990
  • Some mixed culture systems consisting of koji molds and yeast were tested for the ethanol production by simultaneous saccharification and fermentation using polished rice as the substrate. Aspergillus shirousamii showed the highest ethanol production in the mixed culture with Saccharomyces cerevisiae on steamed rice added with 150 ml water in 250 ml Erlenmeyer flask. The optimum initial pH, temperature and specific surface for the ethanol production in this system were 6.5, $30^{\circ}C$, and 0.1, respectively. Under this condition, 12.9% ethanol was produced with inoculation with $5{\times}10^2$ conidia/ml of A. shirousamii and $5{\times}10^6\;cells/ml$ of S. cerevisiae in 10 days.

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Optimization Studies on Water Treatment Process of Seawater Recirculation Fish Culture Systems 1. Ammonia Removal Kinetics in Seawater Using Rotating Biological Contactor Process (순환여과식 해산 어류 양식장의 수처리 공정 최적화 연구 1. 회전원판법에 의한 해수 중의 암모니아 제거 동력학)

  • CHO Young-Gae;LEE Jae-Kwan;LEE Heon-Mo;YANG Byung-Soo
    • Journal of Aquaculture
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    • v.6 no.4
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    • pp.311-321
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    • 1993
  • Ammonia accumulation is regarded as the limiting factor of the first priority in water qualities of aquatic culture systems. Nitrification efficiency and characteristics in seawater were evaluated using Rotating Biological Contactor (RBC) process as a part of the recycling water treatment facilities for marine fish culture system. Ammonia removal efficiency regarded 99.7 to $83.7\%$ at the ammonia surface loading rates of 48 to $393 mg/m^2$ -day. RBC process was able to withstand to the fluctuation of influent ammonia concentrations and loading and produced the stable effluent. The mathematical model on the fixed-film biological reactor developed by Kornegay seemed to be suitable to RBC process kinetic evaluation for the recycling water treatment of the marine fish culture system. Area capacity constant (P) and half-velocity constant (Ks) in the model were 0.188g/m^2$-day and 1.25mg/l, respectively.

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Aspergillus cumulatus sp. nov., from Rice Straw and Air for Meju Fermentation

  • Kim, Dae-Ho;Kim, Seon-Hwa;Kwon, Soon-Wo;Lee, Jong-Kyu;Hong, Seung-Beom
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.334-336
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    • 2014
  • A new species named Aspergillus cumulatus sp. nov. is described in Aspergillus section Aspergillus (Eurotium state). The type strain (KACC $47316^T$) of this species was isolated from rice straw used in meju fermentations in Korea, and other strains were isolated from the air in a meju fermentation room. The species is characterized by growth at a wide range of water activities and the formation of aerial hyphae on malt extract 60% sucrose agar (ME60S) that resemble a cumulus cloud. Furthermore, A. cumulatus produces yellow ascomata containing small lenticular ascospores (5.1-5.7 ${\mu}m$) with a wide furrow, low equatorial crests, and tuberculate convex surface. The species is phylogenetically distinct from the other reported Aspergillus section Aspergillus species based on multilocus sequence typing using rDNA-ITS, ${\beta}$-tubulin, calmodulin, and RNA polymerase II genes.

HeLa Cell Culture on Nanoimprinted Patterns Using Conducting Polymer (전도성 고분자 나노임프린트 패턴 상의 HeLa 세포 배양)

  • Ahn, Junhyoung;Park, Kyungsook;Lee, Suok;Jung, Sanghee;Lim, Hyungjun;Shin, Yong-Beom;Lee, JaeJong
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.41 no.1
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    • pp.63-67
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    • 2017
  • In bioscience and biotechnology, the research of fundamental life mechanisms and their diseases caused by insufficiency is important. The study of a whole organism is difficult and sometimes impossible because of DNA, RNA, proteins, cellular organelles, various cells, and organs. Cell cultures can provide a simple method for researching cellular mechanisms and conditions, both in terms of physiological performance, and in response to chemical stimulation. According to conventional cell culture methodology, the flat surface is used with surface treatments for cell adhesion on the surface. Micro- and nanoscale patterns have been developed with chemical and biochemical modifications for cell immobilization. In this study, HeLa cell culture on nanostructures patterns was studied, including the 300 nm line and 150 nm pillar structures, using nanoimprint lithography and pyrrole as a biocompatible conducting polymer.