• Title/Summary/Keyword: 컬럼

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Extraction of Whitening Agents from Natural Plants and Whitening Effect (천연물에 포함된 미백성분의 추출 및 미백효과)

  • Jin, Yinzhe;Ahn, So Young;Hong, Eun Suk;Li, Guang hua;Kim, Eun-Ki;Row, kyung Ho
    • Applied Chemistry for Engineering
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    • v.16 no.3
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    • pp.348-353
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    • 2005
  • The extracts from natural and fermented products such as Artemisia plants, Rhodiola Salientness, fermented soybeans and soybean paste were used to investigate the whitening effect. 10 g of Artemisia plant were added to 300 mL of ethanol and extracted by sonification at room temperature for 3 h. The extract was further partitioned by the equal volume percent in the order of the n-hexane, chloroform and ethyl acetate. 5 g of Rhodiola salientness was also added to 150 mL of methanol and extracted at the room temperature for 12 h. The effluents from a chromatographic column ($3.9{\times}250mm$, $C_{18}$, $15{\mu}m$) were collected and concentrated in two parts. The extraction of fermented soybeans and soybean paste were done by 60% ethanol. In this work, tyrosinase inhibitory activity and melanin inhibitory effect were measured to confirm the whitening effect. The water layer of Artemisia princeps Pampan showed the good inhibitory of antioxidant, while the hexane layer of Artemisia iwayomogi Kitamura and the chloroform layer of Artemisia princeps Pampan had the excellent melanin inhibitory effect. The Rhodiola salientness had the superior whitening effect to the arbutin in in-vivo melanin production ratio assay. However, the fermented soybeans and soybean paste did not show any whitening effect.

High Throughput Screening and Directed Evolution of Tyrosine Phenol-Lyase (Tyrosine Phenol-Lyase의 고속탐색기술 개발 및 방향성 분자진화)

  • Choi Su-Lim;Rha Eu-Gene;Kim Do-Young;Song Jae-Jun;Hong Seung-Pyo;Sung Moon-Hee;Lee Seung-Goo
    • Microbiology and Biotechnology Letters
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    • v.34 no.1
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    • pp.58-62
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    • 2006
  • Rapid assay of enzyme is a primary requirement for successful application of directed evolution technology. Halo generation on a turbid plate would be a method of choice for high throughput screening of enzymes in this context. Here we report a new approach to prepare turbid plates, by controlling the crystallization of tyrosine to form needle-like particles. In the presence of tyrosine phenol-lyase (TPL), the needle-like tyrosine crystals were converted to soluble phenol rapidly than the usual rectangular tyrosine crystals. When an error-prone PCR library of Citrobacter freundii TPL was spread on the turbid plate, approximately 10% of the colonies displayed recognizable halos after 24 hours of incubation at $37^{\circ}C$. Representative positives from the turbid plates were transferred to LB-medium in 96-wellplates, cultivated overnight, and assayed for the enzyme activity with L-tyrosine as the substrate. The assay results were approximated to be proportional to the halo size on turbid plates, suggesting the screening system is directly applicable to the directed evolution of TPL. Actually, two best mutants on the turbid plates were identified to be $2{\sim}2.5$ and 1.5-fold improved in the activity.

Expression of Recombinant Hybrid Peptide Gaegurin4 and LL37 using Fusion Protein in E. coli (Glutathione S-Transferase에 융합한 재조합 Hybrid Peptide Gaegurin-LL37의 대장균에서의 발현)

  • Bayarbat, Ishvaanjil;Lee, Jae-Hag;Lee, Soon-Youl
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.92-97
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    • 2012
  • Antimicrobial peptides (AMPs) are important components of living organisms acting against Gram-negative and Gram-positive bacterial and fungal pathogens. Cathelicidin human peptides have a variety of biological activities that can be used in clinical applications. AMPs are not produced naturally in large quantities, and chemical synthesis is also economically impractical, especially for long peptides. Therefore, as an alternative, heterologous expression of AMPs by recombinant techniques has been studied as a means to reduce production costs. E. coli is an excellent host for the expression of AMPs, as well as other recombinant proteins, because of the low cost involved and its easy manipulation. However, overexpression of AMPs in E. coli has been shown to cause difficulties resulting from the toxicity of the subsequently produced AMPs. Therefore, fusion expression was theorized to be a solution to this problem. In this study, AMPs were expressed as fused proteins with the glutathione S-transferase (GST) binding protein to protect against the toxicity of AMPs when expressed in E. coli. The LL37, and hybrid gaegurin and LL37 (GGN4(1-16)-LL37(17-32), which we designated as GL32, peptides were expressed as GST-fusion proteins in E. coli and the fusion proteins were then purified by affinity columns. The purified peptides were obtained by removal of GST and were confirmed by western blot analysis. The purified antimicrobial peptides then demonstrated antimicrobial activities against Gram-negative and Gram-positive bacterial strains.

Effects of Activated Carbon Types and Service Life on Adsorption of Tetracycline Antibiotic Compounds in GAC Process (활성탄 재질 및 사용연수에 따른 Tetracycline계 항생물질 흡착특성)

  • Son, Hee-Jong;Jung, Jong-Moon;Hwang, Young-Do;Roh, Jae-Soon;Yu, Pyung-Jong
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.9
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    • pp.925-932
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    • 2008
  • Adsorption performance of tetracycline antibiotic compounds such as tetracycline(TC), oxytetracycline(OTC), chlortetracycline (CTC) and minocycline(MNC) on granular activated carbon(GAC) was evaluated in this study. The coal-based activated carbon was found to be more effective than other carbons in adsorption of tetracycline antibiotic compounds. The wood-based activated carbon was less effective than coconut- and coal-based carbon in adsorption nevertheless having larger pore volume and specific surface area than others carbons. The maximum adsorption capacities(X/M) of coal-based activated carbon for the four tetracycline species was 1.27$\sim$1.36 and 1.69$\sim$1.84 times larger than coconut- and wood-based activated carbon, respectively. Carbon usage rates(CUR) of coal-, coconut- and wood-based activated carbons for tetracycline(TC) were 2.96 g/day, 3.40 g/day and 4.53 g/day, respectively. Similar results were obtained in the adsorption of the rest three tetracycline species. It is concluded that coal-based activated carbon could removed the tetracycline antibiotic compounds better than other material-based activated carbons.

Analysis of gibberellic acid from fruits using HPLC/UV-vis (HPLC/UV-vis을 이용한 과일류 중의 지베렐린 산 분석)

  • Ma, Kyung Na;Cho, Hyun-Woo;Myung, Seung-Woon
    • Analytical Science and Technology
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    • v.26 no.1
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    • pp.19-26
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    • 2013
  • Gibberllic acid ($GA_3$) is one of gibberellins (GAs) that are a class of plant growth hormones that exert profound and diverse effects on plant growth and development. $GA_3$ is essentially non-UV absorbing and is difficult to assay by UV-detector. For effective extraction of gibberellic acid from fruits by using liquid-liquid extraction, optimized pH and extraction solvent were established. The selective and sensitive derivative of $GA_3$ for HPLC/UV-vis was derivatized using phenacyl bromide, and the experimental factors, including reaction time, reaction temperature and amount of derivatizing reagent and base were investigated for the effective synthesis. The derivatized $GA_3$ with phenacyl bromide was effectively analyzed by HPLC/UV-vis. The structure of derivatized $GA_3$ was confirmed by HPLC/ESI-MS. For apple, LOD and LOQ were 0.008 mg/kg and 0.027 mg/kg, respectively. For pear, LOD and LOQ were 0.003 mg/kg, 0.012 mg/kg, respectively. The established method can be applied to more effective analysis of $GA_3$ from plant and food.

Cloning and Characterization of Xylanase 11B Gene from Paenibacillus woosongensis (Paenibacillus woosongensis의 Xylanase 11B 유전자 클로닝과 특성분석)

  • Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.155-161
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    • 2017
  • A gene coding for the xylanase predicted from the partial genomic sequence of Paenibacillus woosongensis was cloned by PCR amplification and sequenced completely. This xylanase gene, designated xyn11B, consisted of 1,071 nucleotides encoding a polypeptide of 356 amino acid residues. Based on the deduced amino acid sequence, Xyn11B was identified to be a modular enzyme, including a single carbohydrate-binding module besides the catalytic domain, and was highly homologous to xylanases belonging to glycosyl hydrolase family 11. The SignalP4.1 server predicted a stretch of 26 residues in the N-terminus to be the signal peptide. Using DEAE-Sepharose and Phenyl-Sepharose column chromatography, Xyn11B was partially purified from the cell-free extract of recombinant Escherichia coli carrying a copy of the P. woosongensis xyn11B gene. The partially purified Xyn11B protein showed maximal activity at $50^{\circ}C$ and pH 6.5. The enzyme was more active on arabinoxylan than on oat spelt xylan and birchwood xylan, whereas it did not exhibit activity towards carboxymethylcellulose, mannan, and para-nitrophenyl-${\beta}$-xylopyranoside. The activity of Xyn11B was slightly increased by $Ca^{2+}$ and $Mg^{2+}$, but was significantly inhibited by $Cu^{2+}$, $Ni^{2+}$, $Fe^{3+}$, and $Mn^{2+}$, and completely inhibited by SDS.

In-situ Bioremediation of Total Petroleum Hydrocarbons-Contaminated Soil by Pseudomonas Species (토양 내 TPH(Total Petroleum Hydrocarbons)의 생물학적 분해 연구)

  • Kim, Jee-Young;Lee, Sang-Seob
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.161-167
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    • 2011
  • We previously showed that five strains belonging to Pseudomonas could remove TPH (Total Petroleum Hydrocarbons) efficiently when they are applied to TPH-contaminated soil. We optimized the bioremediation condition using different hydrocarbons and nutrients conditions to improve the efficiency. We setup lab-scale column bioreactor to monitor TPH and diesel removal efficiency. When we applied five Pseudomonas sp. mixtures to 25,000 $mg{\cdot}kg^{-1}$ TPH-contaminated soil (diesel 10,000 $mg{\cdot}kg^{-1}$, kerosene 10,000 $mg{\cdot}kg^{-1}$, gasoline 5,000 $mg{\cdot}kg^{-1}$) with the optimum condition, 76.3% of TPH removal efficiency was shown for 25 days. Meanwhile, in the application of five Pseudomonas sp. mixtures to 20,000 $mg{\cdot}kg^{-1}$ diesel-contaminated soil with the optimum condition, 99.2% of diesel removal efficiency was shown for 40 days. In the application to lab-scale bioreactor with five high efficiency bacteria, 88.5% of TPH removal efficiency was shown for 45 days. Based on the results from this study, we confirmed that this mixed Pseudomonas sp. consortium might improve the bioremediation of TPH in contaminated soil, the efficacy can be controlled by improving the nutrients. We also confirmed that the nutrients and oxygen for biodegradation of TPH could contribute on the management and control of applications of these strains for the study of bioremediation of TPH-contaminated soil.

Determination of Ametryn Residue in Agricultural Commodities Using HPLC-UVD/MS (HPLC-UVD/MS를 이용한 농산물 중 ametryn의 분석)

  • Lee, Su-Jin;Kim, Young-Hak;Song, Lee-Seul;Choung, Myoung-Gun
    • The Korean Journal of Pesticide Science
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    • v.15 no.2
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    • pp.125-133
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    • 2011
  • Ametryn is used in USA, China, and Japan, but not introduced in Korea yet. So, MRL (Maximum Residue Level), and analytical method of ametryn were not establishment in Korea. Therefore, this experiment was conducted to establish a determination method for ametryn residue in crops using HPLC-UVD/MS. Ametryn residue was extracted with acetone from representative samples of five raw products which comprised hulled rice, soybean, apple, green pepper, and Chinese cabbage. The extract was diluted with saline water, and dichloromethane partition was followed to recover ametryn from the aqueous phase. Florisil column chromatography was additionally employed for final clean up of the extract. The ametryn was quantitated by HPLC with UVD, using a Tosoh ODS 120T ($4.6{\times}250$ mm) column. The crops were fortified with ametryn at 2 levels per crop. Mean recovery ratio were ranged from 83.7% for a 0.2 mg/kg in soybean to 91.1% for a 1.0 mg/kg in hulled rice. The coefficients of variation were ranged from 1.2% for a 1.0 mg/kg in hulled rice to 3.6% for a 1.0 mg/kg in soybean. Quantitative limit of amatryn was 0.02 mg/kg in representative 5 crop samples. A LC/MS with selected-ion monitoring was also provided to confirm the suspected residue. Therefore, this analytical method was reproducible and sensitive enough to determine the residue of ametryne in agricultural commodities.

Technique for Concurrent Processing Graph Structure and Transaction Using Topic Maps and Cassandra (토픽맵과 카산드라를 이용한 그래프 구조와 트랜잭션 동시 처리 기법)

  • Shin, Jae-Hyun
    • KIPS Transactions on Software and Data Engineering
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    • v.1 no.3
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    • pp.159-168
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    • 2012
  • Relation in the new IT environment, such as the SNS, Cloud, Web3.0, has become an important factor. And these relations generate a transaction. However, existing relational database and graph database does not processe graph structure representing the relationships and transactions. This paper, we propose the technique that can be processed concurrently graph structures and transactions in a scalable complex network system. The proposed technique simultaneously save and navigate graph structures and transactions using the Topic Maps data model. Topic Maps is one of ontology language to implement the semantic web(Web 3.0). It has been used as the navigator of the information through the association of the information resources. In this paper, the architecture of the proposed technique was implemented and design using Cassandra - one of column type NoSQL. It is to ensure that can handle up to Big Data-level data using distributed processing. Finally, the experiments showed about the process of storage and query about typical RDBMS Oracle and the proposed technique to the same data source and the same questions. It can show that is expressed by the relationship without the 'join' enough alternative to the role of the RDBMS.

Digital Forensic Investigation of HBase (HBase에 대한 디지털 포렌식 조사 기법 연구)

  • Park, Aran;Jeong, Doowon;Lee, Sang Jin
    • KIPS Transactions on Computer and Communication Systems
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    • v.6 no.2
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    • pp.95-104
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    • 2017
  • As the technology in smart device is growing and Social Network Services(SNS) are becoming more common, the data which is difficult to be processed by existing RDBMS are increasing. As a result of this, NoSQL databases are getting popular as an alternative for processing massive and unstructured data generated in real time. The demand for the technique of digital investigation of NoSQL databases is increasing as the businesses introducing NoSQL database in their system are increasing, although the technique of digital investigation of databases has been researched centered on RDMBS. New techniques of digital forensic investigation are needed as NoSQL Database has no schema to normalize and the storage method differs depending on the type of database and operation environment. Research on document-based database of NoSQL has been done but it is not applicable as itself to other types of NoSQL Database. Therefore, the way of operation and data model, grasp of operation environment, collection and analysis of artifacts and recovery technique of deleted data in HBase which is a NoSQL column-based database are presented in this paper. Also the proposed technique of digital forensic investigation to HBase is verified by an experimental scenario.