• Title/Summary/Keyword: Purification system

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Production and Purification of Single Chain Human Insulin Precursors with Various Fusion Peptides

  • Cho, Chung-Woo;Park, Sun-Ho;Nam, Doo-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.2
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    • pp.144-149
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    • 2001
  • For the production and purification of a single chain human insulin precursor, four types of fusion peptides $\beta$-galactosidase (LacZ), maltose binding protein (MBP), glutathione-S-transferase (GST), and (His)(sub)6-tagged sequence (HTS) were investigated. Recombinant E. coli harboring hybrid genes was cultivated at 37$\^{C}$ for 1h, and gene induction occurred when 0.2mM of isopropyl-D-thiogalactoside (IPTG) was added to the culture broth, except for E. coli BL21 (DE3) pLysS harboring a pET-BA cultivation with 1.0mM IPTG, followed by a longer than 4h batch fermentation respectively. DEAE-Sphacel and Sephadex G-200 gel filtration chromatography, amylose affinity chromatography, glutathione-sepharose 4B affinity chromatography, and a nickel chelating affinity chromatography system as a kind of immobilized metal ion affinity chromatography (IMAC) were all employed for the purification of a single chain human insulin precursor. The recovery yields of the HTS-fused, GST-fused, MBP-fused, and LacZ-fused single chain human insulin precursors resulted in 47%, 20%, 20%, and 18% as the total protein amounts respectively. These results show that a higher recovery yield of the finally purified recombinant peptides was achieved when affinity column chromatography was employed and when the fused peptide had a smaller molecular weight. In addition the pET expression system gave the highest productivity of a fused insulin precursor due to a two-step regulation of the gene expression, and the HTS-fused system provided the highest recovery of a fused insulin precursor based on a simple and specific separation using the IMAC technique.

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The development of industrial secure L2 switch and introduction example for management and security improvement of supervisory control network in purification plant (정수장 감시제어망의 관리와 보안개선을 위한 산업용 보안 L2스위치 개발 및 적용사례)

  • Kim, Yunha;Yu, Chool;Oh, Eun;Kim, Chanmoon;Park, Ikdong;Kim, Yongseong;Choi, Hyunju
    • Journal of Korean Society of Water and Wastewater
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    • v.33 no.5
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    • pp.329-339
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    • 2019
  • Recently, the advancement of information and communication technology(ICT) is expanding the connectivity through Internet of Things(IoT), and the media of connection is also expanding from wire/cable transmission to broadband wireless communication, which has significantly improved mobility. This hyperconnectivity has become a key element of the fourth industrial revolution, whereas the supervisory control network of purification plants in korea is operated as a communication network separated from the outside, thereby lagging in terms of connectivity. This is considered the best way to ensure security, and thus there is hardly any consideration of establishing alternatives to operate an efficient and stable communication network. Moreover, security for management of a commercialized communication network and network management solution may be accompanied by immense costs, making it more difficult to make new attempts. Therefore, to improve the conditions for the current supervisory control network of purification plants, this study developed a industrial security L2 switch that supports modbus TCP(Transmission Control Protocol) communication and encryption function of the transmission section. As a result, the communication security performance improved significantly, and the cost for implementing the network management system using Historical Trend and information of HMI(Human Machine Interface) could be reduced by approximately KRW 200 million. The results of this study may be applied to systems for gas, electricity and social safety nets that are infrastructure communication networks that are similar to purification plants.

The Purification Characteristics of the Reactive Soil-Bentonite Landfill Liner (혼합반응차수재의 오염정화특성 연구)

  • 장경준;황의석;안기봉;김학문
    • Proceedings of the Korean Geotechical Society Conference
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    • 2003.03a
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    • pp.585-592
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    • 2003
  • The purpose of this paper is to investigate purification characteristic of soil-bentonite landfill liner and to develop of desirable liner system. In this study, column tests for soil-bentonite, reactive soil-bentonite and existing bentomat and reactive bentomat were carried out under the high pressure with water and leachate.

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Bacterial Overexpression and Denaturing Purification of VPS34-Binding Domain of Beclin 1

  • Baek, Jong-Hyuk;Jung, Juneyoung;Seo, Jeongbin;Kim, Jeong Hee;Kim, Joungmok
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1808-1816
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    • 2016
  • As a scaffolding subunit of the PIK3C3/VPS34 complex, Beclin 1 recruits a variety of proteins to class III phosphatidylinositol-3-kinase (VPS34), resulting in the formation of a distinct PIK3C3/VPS34 complex with a specific function. Therefore, the investigation of a number of Beclin 1 domains required for the protein-protein interactions will provide important clues to understand the PIK3C3/VPS34 complex, of which Beclin1-VPS34 interaction is the core unit. In the present study, we have designed a bacterial overexpression system for the Beclin 1 domain corresponding to VPS34 binding (Vps34-BD) and set up the denaturing purification protocol due to the massive aggregation of Vps34-BD in Escherichia coli. The expression and purification conditions determined in this study successfully provided soluble and functional Vps34-BD.

Numerical Assessment for Coastal Water Purification Utilizing a Tidal Jet System (조석분류를 이용한 연안해역의 수질정화에 관한 수치적 평가)

  • Park, Jong-Chun
    • Journal of Ocean Engineering and Technology
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    • v.20 no.4 s.71
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    • pp.58-63
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    • 2006
  • When the costal zone surrounded by a breakwater has a narrow vertical opening, currents in the vicinity of a narrow entrance can result in a jet flow, coinciding with the tide. Such a Tidal-Jet Generator(TJG) can change the water mass distribution and transport processes in the domain of influence of the jet. Also, it can decrease the residual time of them. In the present study, the water purification utilizing tidal jets in the coastal zone over constant bathymetry are estimated numerically, using a finite-difference numerical scheme, named the NS-MAC-TIDE method, which isbased on the fully 3D Navier-stokes(NS) equations. The shear velocity near the inlet of the TJG are predicted from the flow field simulation, and are assessed qualitatively with the development of scouring or sediment that is caused by the change of diffusion or sweeping flowup from the seabed of sediment particles. Finally, through solving a transport equation of concentration, the residual time related on mass transport processes and the flushing mechanism for water purification are investigated.

Recombinant Expression, Isotope Labeling, and Purification of Cold shock Protein from Colwellia psychrerythraea for NMR Study

  • Moon, Chang-Hun;Jeong, Ki-Woong;Kim, Hak-Jun;Heo, Yong-Seok;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.30 no.11
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    • pp.2647-2650
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    • 2009
  • Cold shock proteins (Csps) are a subgroup of the cold-induced proteins on reduction of the growth temperature below the physiological temperature. They preferentially bind to single-stranded nucleic acids to translational regulation via RNA chaperoning. Csp plays important role in cold adaptations for the psychrophilic microorganism. Recently, Cold shock protein from psychrophilic bacteria, Colwellia psychrerythraea (CpCsp) has been identified. Three dimensional structures of a number of Csps from various microorganisms have been solved by NMR spectroscopy or X-ray crystallography, but structures of psychrophilic Csps were not studied yet. Therefore, cloning and purification protocols for further structural study of psychrophilic Csp have been optimized in this study. CpCsp was expressed in E. coli with pET-11a vector system and purified by ion exchange, size exclusion, and reverse phase chromatography. Expression and purification of CpCsp in M9 minimal media was carried out and $^{15}N$-labeled proteins with high purity over 90% was obtained. Further study will be carried out to investigate the tertiary structure and dynamics of CpCsp.

A Rapid and Simple Homogenizing Method for the Purification of Single-walled Carbon Nanotubes

  • Choi, Sang-Kyu;Jung, Seung-Il;Lee, Seung-Beck
    • Transactions on Electrical and Electronic Materials
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    • v.9 no.5
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    • pp.209-212
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    • 2008
  • We developed a simple and effective purification method to obtain high-purity single-walled carbon nanotubes (SWCNTs) with low surface damage. The purification process consists of oxidization at $430^{\circ}C$ for 1 h in a furnace system of air atmosphere and homogenization in dilute hydrochloric acid solution for extremely short time. The role of homogenizer was examined during purification process in terms of purity and quality of purified SWCNTs. High-purity and low surface damage of SWCNT products was obtained using homogenizer which was operated at 8500 rpm for 10 min in the environment of 7 % HCI solution. From XRD spectra, we observed that metal catalysts were thoroughly removed. Raman spectra showed that the intensity values of crystallization ($I_{G}/I_{D}$) of purified SWCNTs were very similar with that of pristine SWCNTs. Moreover, the structure damage of purified SWCNTs was hard to find from electron microscopy. Consequently, homogenizing, which is a quick and simple manner, can be promising method for obtaining final SWCNTs with clearly high purity and crystallinity.

Application of water control by high fiux MF membrane (고 플럭스 MF막의 정수처리 실용화에 관한 연구)

  • Yong, Hwang-Sang
    • Journal of environmental and Sanitary engineering
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    • v.23 no.2
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    • pp.55-63
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    • 2008
  • All over the world, the existing process of water purification needs more flocculants and chlorine due to a gradual decline in the quality of source water. Therefore, the problem of the remaining aluminium and DBPs in purified water is on the rise. To solve this problem, the process of membrane filter has recently come into the spotight. This study reaches the following conclusions concerning TMP variation in order to solve the dropping of flux throgh a membrane filter when operating a membrane filter system in the process of water purification. 1. In case that a cohesion-precipitation process was introduced to pre-treatment of a membrane filter, initial TMP was very satisfactory(0.27kg/cm) in producing the constantly safe quality of water, $0.04{\sim}0.1$(mean 0.05) NTU by pouring 2mg/l of PACI(10% $Al_2O_3$) used for the existing process of water purification in high-density turbidity at a dry or flood season and at occurrence of high algae. 2. As flux increased at 0.5m/day.m, TMP increased 0.05 kgf/cm. 3. As filtering, operation mode of PVDF MF membrane filtering was 48 minutes and 1 cycle of back washing was 42 minutes, flux was increased 1.5m/day.m and TMP increased $0.25{\sim}0.27kgf/cm$. Without back washing, TMP increased 0.03 kgf/cm per a cycle.