• Title/Summary/Keyword: Membrane bioreactor

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Enhancement of Tissue Type Plasminogen Activator (tPA) Production from Recombinant CHO Cells by Low Electromagnetic Fields

  • Lee, Seo-Ho;Lee, Hyun-Soo;Lee, Mi-Kyoung;Lee, Jin-Ha;Kim, Jong-Dai;Park, Young-Shik;Lee, Shin-Young;Lee, Hyeon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.457-462
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    • 2002
  • Low Electromagnetic Field (EMF) intensity in the range of $1{\mu}T\;to\;10{\mu}T$(Tesla) was found to enhance the growth of CHO cells and the production of tPA in batch and perfusion cultivations. At $1{\mu}T\;intensity,\;1.3{\times}10^7$ viable cells/ml of maximum cell density and 80 mg/l of maximum tPA production were obtained in batch cultivation, compared to $2.8{\times}10^6$ viable cells/ml and 59 mg tPA/1 in unexposed case (control). A similar trend was observed in the perfusion process, where it was possible to obtain $1.2{\times}10^7$ viable cells/ml of maximum cell density and 81 mg tPA/l of maximum tPA production by more than 80 days of cultivation. However, there was not much difference between $1{\mu}T\;and\;10{\mu}T$ in perfusion cultivation, possibly due to better environmental growth conditions being maintained by continuous feeding of fresh medium into the reactor. On the contrary, both cell growth and tPA production were severely inhibited at higher than 1 mT intensity, showing no growth at 10 mT exposure. Specific growth rate was linearly correlated to specific tPA production rate at $1{\mu}T$EMF intensity, which represents a partially growth-related relationship. It was also found that a large amount of $Ca^2+$ was released at low EMF intensity, even though the cell growth was not much affected. Low EMF intensity significantly improved both cell growth and tPA production, and tPA production seemed to be more affected than the cell growth, possibly due to the changes of cell membrane characteristics. It can be concluded that the elaboration of EMF intensity less than $10{\mu}T$ could improve cell growth and tPA production, but mainly tPA secretion through batch or perfusion process in a bioreactor.

Identification and Functional Analysis of Pig β-1,4-N-Acetylglucosaminyltransferase A (MGAT4A) (돼지 유래의 β-1,4-N-acetylglucosaminyltransferase A (MGAT4A) 유전자의 동정 및 기능 분석)

  • Kim, Ji-Youn;Hwang, Hwan-Jin;Chung, Hak-Jae;Park, Mi-Ryung;Byun, Sung June;Kim, Kyung-Woon
    • Journal of Life Science
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    • v.26 no.3
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    • pp.275-281
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    • 2016
  • Glycan modification is important in pharmaceutical industry. Especially, sialic acid affects the bioactivity and stability of medicine. Milk of pig has been used as bioreactor to produce various pharmaceutical proteins. Therefore, it is necessary to modify the glycan chain in pig mammary grand. β-1,4-N-Acetylglucosaminyltransferase A (pMGAT4A) is one of the essential enzymes for increase of sialic acid content, but pig MGAT4A is unclear. In this study, the pMGAT4A was identified and characterized. The pMGAT4A has 1638 nucleotides encoding 535 amino acids and type II membrane topology, which is one of the common features in many glycosyltransferases. The gene was strongly expressed in liver and mammary gland, whereas was weakly expressed in small intestine, stomach and bladder. For functional test, HA-tagged MGAT4A was over-expressed in porcine kidney (PK-15) cell line. Forced expression of pMGAT4A gene was identified by qPCR, and we identified that pMGAT4A is located in Golgi complex by co- staining with HA antibody and BODIPY TR ceramide. In addition, we identified the increase of mannose-β-1,4-N-acetylglucosamine structure by ELISA and immunofluorescence using Datura stramonium agglutinin (DSA), which recognizes mannose-β-1,4-Nacetylglucosamine. Through the specific activity analysis, we showed that pMGAT4A modified bi-antennary to tri-antennary. This event affects sialic acid content. Therefore, we thought that over-expression of pMGAT4A will be necessary in pig mammary grand for improved medicine.

A Study on Management of Seafood Wastewater Treatment Facility using Submerged MBR (침지식 MBR을 이용한 수산물 폐수처리장 운영에 관한 연구)

  • Choi, Yong-Bum;Lee, Hae-Seung;Han, Dong-Joon;Kwon, Jae-Hyouk
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.11
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    • pp.7227-7236
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    • 2015
  • The survey revealed that, due to the discharge characteristics of seafood wastewater, irregular inflow loads were caused, making it difficult to treat the wastewater safely. It is crucial for the operation of pressure and floating tanks for the treatment of high-concentration organic wastewater such as seafood wastewater. The survey of operation factors for the pressure and floating tanks revealed this: A/S ratio 0.05 (design criteria 0.01), the pressurized air pressure 8bar(design criteria 6bar), the pressure tank pressure 6bar (design criteria 4.5bar), and HRT 60sec(design criteria: 10sec). Also, the recirculation rate was changed to over 40%(design criteria: 30%), and the surface load rate was changed to under $13.7m^3/m^2{\cdot}hr$(design criteria: under $17.7m^3/m^2{\cdot}hr$); thus, compared to the initial design criteria, the operation factors were changed according to inflow characteristics, thus enhancing the pressure and floating tank performance. The survey of inflow load revealed BOD 140.7%, $COD_{Mn}$ 120.32%, and SS 106.3%, compared to the inflow design criteria, as well as T-N 135.5% and T-P173.3%, higher than the design criteria. The survey of the treatment facility annual operation cost revealed high portions in sludge treatment cost(27.7%) and chemicals costs(26.0%), and the sludge treatment cost will likely further increase due to the ban on ocean dumping. The unit cost for the treatment of seafood wastewater was found to be KRW 3,858 per ton, more than 27 times higher than the sewage treatment cost(KRW 142.6/ton), presumably because the seafood wastewater contains high-concentration organic substances and nutritive salts.