• Korean Chemical Engineering Research
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• v.49 no.3
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• pp.348-351
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• 2011

In this research, we developed a sensor system which can easily detect several chelating agents using polydiacetylene(PDA) vesicles. In comparison to other sensors, PDA based sensor has several advantages. First, detection method is much simpler and faster because it does not require any labeling step in the experiment procedure. Second, significant color-transition from blue to red based upon external stimulus allows us the detection by naked eyes. Finally, it is also possible to perform quantitative analysis of the concentration of the chelating agent by measuring the colorimetric response. In this paper, five types of chelating agents were used, including EDTA, EGTA, NTA, DCTA and DTPA. Among them, EDTA and DCTA triggered especially strong color-transition. In conclusion, this study has led to a successful development of a color transition-based PDA sensor system for easy and rapid detection of chelating agents.

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.105-109
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• 2009

Among signal transduction systems by protein phosphorylation Akt/PKB protein kinase which is one of serine/threonine kinases, is known to regulate the survival and death of the cell and glucose metabolism. Thus, Akt/PKB protein kinase has been used as one of the target proteins to find anti-cancer agents from natural products. In this study, human Akt/PKB protein kinase was expressed in Escherichia coli expression system for the mass production. Human Akt/PKB protein kinase expressed in E. coli formed inclusion body under the general condition. However, most of the expressed protein was solubilized under the culture temperature at $27^{\circ}C$ and 0.01-0.09 mM of IPTG for induction of the protein expression. The expressed protein was purified using $Ni^{2+}$-NTA agarose column and confirmed by using anti-Akt antibody. Subsequently, the purified human Akt/PKB protein kinase was activated by in vitro phosphorylation using cellular extract containing kinases. The activated protein was confirmed to phosphorylate the specific fluorescent peptide specially designed as the artificial substrate for Akt/PKB protein kinase.

• Journal of Dairy Science and Biotechnology
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• v.29 no.1
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• pp.49-54
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• 2011

Bile salt hydrolase (BSH, EC 3.5.1.24) activity, which cleaves amide bond between carboxyl group (bile acid) and amino group (glycine or taurine), is commonly detected in gut-associated species of human and animal. During the screening of BSH active strains from the fecal samples of elderly human volunteers, strain CU30-2 was isolated on the basis of the highly active BSH producing activity. A bsh gene of the isolate was cloned into the pET22b expression vector and overexpressed in Escherichia coli BL21 (DE3) Gold by induction with 1mM IPTG. The overexpressed BSH enzyme with 6x His-tag was purified with apparent homogeneity using a $Ni^+$-NTA agarose column and characterized. The BSH enzyme of E. faecalis CU30-2 exhibited approximately 50 times higher activity against glycol-conjugated bile salts than tauro-conjugated bile salts having the highest activity against glycocholic acid. Considering the prevalence of E. faecalis strains in the human GI tract and glycol-conjugates dominated bile acid composition of human bile, further study is needed to investigate the impact of the BSH activity exerted by E. faecalis strains to the host as well as to the BSH producing strains.

• Journal of Environmental Science International
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• v.1 no.2
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• pp.71-79
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• 1992

Selective extraction procedure has been used to quantify copper and cadmium In association with the various phases of aquatic sediment such as exchangeable/adsorbed, carbonate, manganese oxides, organic matter and iron oxides. Changes of pH influenced on the partitioning of copper in carbonate and exchangeable/ad- sorbed phases and of cadmium in carbonate phase of aquatic sediment. Addition of NTA and EDTA, copper and cadmium associated with carbonate phase were released from sediment to water. Total partitioning coefficient was 8.361 for copper and 0.497 for cadmium. The relative binding strengths of copper and cadmium to each solid phase can be ranked by using the partitioning coefficints. For copper it was observed that carbonate > organic matter > exchangeable/adsorbed > manganese oxides > iron oxides and for cadmiunm it was observed that exchangeable/adsorbed > carbonate > manganese oxides > organic matter > iron oxides.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.241-246
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• 2002

A gene coding for endo-$\beta$-1,3-1,4-glucanase of Bacillus circulans was subcloned into Escherichia coli Ml5 using pQE30 as an expression vector. Endo-$\beta$-1,3-1,4-glucanase produced by the recombinant expression plas-mid pLQ43 was intactly purified to a single protein through a nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography method. The molecular mass of the purified enzyme was estimated to be 28 kDa by SDS-PAGE. The optimum pH and temperature of the enzyme activity were pH 6.8 and $60^{\circ}C$, respectively. This enzyme was fairly stable in the pH ranging 5.5~7.5 and at the temperatures lower than $55^{\circ}C$. The enzyme appeared to be sensitive to most of the metal ions, especially to $Hg^{2＋$, and also to methanol, ethanol, isopropanol or 1-butanol at a concentration of 10%(v/v).

• Journal of the Korea Organic Resources Recycling Association
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• v.24 no.3
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• pp.15-21
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• 2016

This experiment was conducted to test desorption and recycling characteristics for silver ion adsorbed into waste coffee grounds by using various desorbing agents such as nitric acid, ethylene diamine triacetic acid (EDTA) and nitrilo triacetic acid (NTA). It is appeared that the highest desorption efficiency for silver ions was obtained as about 97.8 % by 1.0 M of nitric acid solution. Also, in the case of less than 1.0 of the ratio of solid and liquid (S/L) (g/L), silver ions adsorbed onto coffee grounds was desorbed as about 98~100 %, and most of desorption process was completed within 60min. In addition, adsorption capacity of reused waste coffee grounds for silver ions was highly maintained as about 43.9 mg/g until the $2^{nd}$ cycle, as compared with the adsorption capacity with 45.9 mg/g of the adsorption capacity for virgin waste coffee grounds.

• Food Science of Animal Resources
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• v.25 no.2
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• pp.232-237
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• 2005

Lactoferrin is a member of the transferrin family of iron-binding glycoproteins. It is originally found in milk. In addition to its antibacterial and antiviral activities, lactoferrin has many other biological functions include anti-inflammatory properties, antitumor, cell growth-promoting activity as well as antioxidant effect In the present study, we report the production of recombinant bovine lactoferrin and lactoferrin N-lobe in the Rhodococcus erythropolis (R erythropolis) using pTip vector. The expression level was investigated in various range of temperature, and we could successfully expressed the bovine lactoferrin and lactoferrin N-lobe in R erythropolis at low temperature. The recombinant proteins were purified by Nickel-Nitrolotriacetic acid (Ni-NTA). The purified proteins were confirmed by SDS-PAGE and Western blot, which indicating that the recombinant proteins have a molecular weight of 80kDa and 43kDa for bovine lactoferrin and lactoferrin N-lobe, respectively.

• Korean Chemical Engineering Research
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• v.44 no.2
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• pp.166-171
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• 2006

In this study, in using a piezoelectric material of $Pb(Mg_{1/3}Nb_{2/3})O_3-PbTiO_3$ (PMN-PT), which has a high electromechanical coupling coefficient, we have tried to study about this material can be practically available as a new biosensor to detect protein by using surface acoustic wave (SAW). As the results, the filtering of the center frequency of the PMN-PT substrate is a superior result to that of the $LiTaO_3$ (LT) substrate, but the result was not completely satisfactory. Also this study attempts to develop a sensing method to detect mismatched DNA in order to diagnose cancer. We could directly immobilize the MutS to the NTA using the EDC solution. But, we immobilized MutS using nickel and it is judged that is more effective method to detect mismatched DNA.

• The Journal of Natural Sciences
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• v.14 no.1
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• pp.51-61
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• 2004

T7 RNA polymerase is a single subunit RNA polymerase able to accomplish whole transcription process without auxiliary factors. T7 phage lysozyme involcing in destruction of host cell wall repress T7 transcription and affects transcription termination process. Therefore expression vector pT7lys containing T7 phage lysozyme gene was constructed and expressed. T7 phage lysozyme protein was purified to homogeneity by Ni-NTA column chromatography. Also amidase activity of the purified lysozyme was identified. In order to understand the effect of the lysozyme on the sequence specific transcription termination. T7 transcription elongation complexes at the site rrnB T1 transcription termination signal were made in the presence the lysozyme. The results shows that the transcription elongation complexes are unstable in the presence of T7 phage lysozyme.

• KSBB Journal
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• v.19 no.6 s.89
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• pp.462-466
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• 2004

Mycothiol (MSH), a low molecular antioxidant thiol compound, was purified and analyzed from Streptomyns coelicolor A[3]2 by the monobromobimane fluorescence detection method modified by this lab. Through HPLC chromatpgram, MSH fraction was obtained following the elution time of standard MSH (donated by Dr. Robert C. Fahey). That MSH showed the highest concentration among the thiol compounds contained in the cell indicated that MSH was the key thiol compound having antioxidant activity. To understand the role of gene of inositol monophosphatase (I-1-Pase) involved in the MSH biosynthesis, it was isolated from S. coelicolor A(3)2 and cloned and overexpressed in the Escherichia coli. The expressed I-1-Pase was purified through Ni-NTA column. The soluble protein consisted of 281 amino acids, and the molecular weight was 32 kDa. I-1-Pase of S. coelicolor A(3)2 had the sequence homology with those of human and E. coli by 24 and $25\%$, respectively, and had two conserved domains (mofif A and motif B) which were typical of I-1-Pase.