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Cloning and Expression of Thermostable $\beta$-Glycosidase Gene from Thermus filiformis Wai33 A1 in Escherichia coli and Enzyme Characterization

  • Kang, Sang-Kee;Cho, Kwang-Keun;Ahn, Jong-Kun;Kang, Seung-Ha;Han, Kyung-Ho;Lee, Hong-Gu;Choi, Yun-Jaie
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.584-592
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    • 2004
  • A thermostable $\beta$-glycosidase gene, tfi $\beta$-gly, was cloned from the genomic library of Thermus filiformis Wai33 A1. ifi $\beta$-gly consists of 1,296 bp nucleotide sequence and encodes a polypeptide of 431 amino acids. It shares a strong amino acid sequence similarity with the $\beta$-glycosidases from other Thermus spp. belonging to the glycosyl hydrolase family 1. In the present study, the enzyme was overexpressed in Escherichia coli BL21 (DE3) using the pET21b(+) vector system. The recombinant enzyme was purified to homogeneity by heat treatment and a $Ni^{2+}$-affinity chromatography. Polyacrylamide gel electrophoresis (PAGE) showed that the recombinant Tfi $\beta$-glycosidase was a monomeric form with molecular mass of 49 kDa. The temperature and pH range for optimal activity of the purified enzyme were 80- $90^{\circ}C$ and 5.0-6.0, respectively. Ninety-three percent of the enzyme activity was remained at $70^{\circ}C$ after 12 h, and its half-life at $80^{\circ}C$ was 6 h, indicating that Tfi $\beta$-glycosidase is highly thermostable. Based on its K_m$, or $K_{cat}K_m$, ratio, Tfi $\beta$-glycosidase appeared to have higher affinity for $\beta$-D-glucoside than for $\beta$-D-galactoside, however, $K_{cat} for \beta$-D-galactoside was much higher than that for $\beta$-D-glucoside. The activity for lactose hydrolysis was proportionally increased at $70^{\circ}C$ and pH 7.0 without substrate inhibition until reaching 250 mM lactose concentration. The specific activity of Tfi TEX>$\beta$-glycosidase on 138 mM lactose at $70{^\circ}C$ and pH 7.0 was 134.9 U/mg. Consequently, this newly cloned enzyme appears to have a valuable advantage of conducting biotechnological processes at elevated temperature during milk pasteurization in the production of low-lactose milk.

Analyses of Studies on Swallowing Rehabilitation Program for Stroke patients in Korea (뇌졸중 환자 대상의 연하재활프로그램에 대한 국내 연구 논문 분석)

  • Kwak, Hye-Weon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.11
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    • pp.7394-7402
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    • 2015
  • The purpose of this study was to investigate the status of studies on swallowing rehabilitation interventions for stroke patients that had been conducted in the past 5 years (2010-2014) in Korea and to examine their findings. A literature search was conducted through RISS, National Assembly Library, and KISS, and 12 papers were analyzed. Four articles were from nursing science, and 8 from other disciplines. Qualitative evaluation of the articles was conducted by using an evaluation framework that was designed by the researcher, which took into account the checklist presented by the Scottish Intercollegiate Guideline Network (2013). In the results of the qualitative evaluation on the papers, 4 studies scored 7 out of 10 points, 6 studies scored between 5 and 6 points, and 2 studies scored 4 points. Intervention sessions were conducted for 25 minutes, on average, each at a frequency of 4.1 sessions per week, for a total of 33.7 sessions. This study found that direct swallowing training and comprehensive intervention of respiratory and oral, including pharyngeal were effective in relieving difficulties in swallowing. In the future, such findings are expected to be used in nursing interventions for establishment of the basis for evidence-based nursing practice.

An Evaluation of Plant Growth Promoting Activities and Salt Tolerance of Rhizobacteria Isolated from Plants Native to Coastal Sand Dunes (해안사구의 토착식물로부터 분리된 근권세균의 내염능과 식물성장촉진능 평가)

  • Hong, Sun Hwa;Lee, Mi Hyang;Kim, Ji Seul;Lee, Eun Young
    • Microbiology and Biotechnology Letters
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    • v.40 no.3
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    • pp.261-267
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    • 2012
  • Coastal sand dunes are important for ecosystems due to the variety of rare species that can be found in this kind of habitat, and the beautiful landscapes they create. For environmental remediation, a potential strategy is phytoremediation using the symbiotic relationship of plants and microbes in the rhizosphere, which has proven ecologically sound, safe, and cost effective. Ninety-five colonies were isolated from the rhizosphere soil (RS) or rhizoplane (RP) of Rorippa islandica, Rumex crispus, Artemisia princeps var. orientalis, Lilium sp Stellaria media, and Gramineae. These colonies were then tested for plant growth promoting activities (PGPAs) such as 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity, and siderphores synthesis ability. In addition, salt tolerance was evaluated at 4% and 8% salt concentrations. It was observed that amongst the test subjects about 50% of the strains had a high resistance to salinity. Many of them could produce indole-3-acetic acid (IAA) IAA (in RS 13.9% and in RP 7.6%), exhibited ACC deaminase activity (55.8% in RS and 36.6% in RP), and could synthesize siderphores (62.7% in RS and 50% in RP). Correlation coefficient analyses were carried out for the three kinds of plant growth promoting abilities (PGPA) and salt tolerance. A positive correlation was found between an ability to synthesize siderphores and ACC deaminase activity (r=0.605, p<0.037). Similarly, positive correlations were noted between salt tolerance and ACC deaminase activity (r=0.762, p<0.004, r=0.771), and salt tolerance and an ability to synthesize siderphores (r=0.771, p<0.003).

Fine-Scale Population Structure of Accumulibacter phosphatis in Enhanced Biological Phosphorus Removal Sludge

  • Wang, Qian;Shao, Yongqi;Huong, Vu Thi Thu;Park, Woo-Jun;Park, Jong-Moon;Jeon, Che-Ok
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1290-1297
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    • 2008
  • To investigate the diversities of Accumulibacter phosphatis and its polyhydroxyalkanoate (PHA) synthase gene (phaC) in enhanced biological phosphorus removal (EBPR) sludge, an acetate-fed sequencing batch reactor was operated. Analysis of microbial communities using fluorescence in situ hybridization and 16S rRNA gene clone libraries showed that the population of Accumulibacter phosphatis in the EBPR sludge comprised more than 50% of total bacteria, and was clearly divided into two subgroups with about 97.5% sequence identity of the 16S rRNA genes. PAO phaC primers targeting the phaC genes of Accumulibacter phosphatis were designed and applied to retrieve fragments of putative phaC homologs of Accumulibacter phosphatis from EBPR sludge. PAO phaC primers targeting $G_{1PAO},\;G_{2PAO},\;and\;G_{3PAO}$ groups produced PCR amplicons successfully; the resulting sequences of the phaC gene homologs were diverse, and were distantly related to metagenomic phaC sequences of Accumulibacter phosphatis with 75-98% DNA sequence identities. Degenerate NPAO (non-PAO) phaC primers targeting phaC genes of non-Accumulibacter phosphatis bacteria were also designed and applied to the EBPR sludge. Twenty-four phaC homologs retrieved from NPAO phaC primers were different from the phaC gene homologs derived from Accumulibacter phosphatis, which suggests that the PAO phaC primers were specific for the amplification of phaC gene homologs of Accumulibacter phosphatis, and the putative phaC gene homologs by PAO phaC primers were derived from Accumulibacter phosphatis in the EBPR sludge. Among 24 phaC homologs, a phaC homolog (GINPAO-2), which was dominant in the NPAO phaC clone library, showed the strongest signal in slot hybridization and shared approximately 60% nucleotide identity with the $G_{4PAO}$ group of Accumulibacter phosphatis, which suggests that GINPAO-2 might be derived from Accumulibacter phosphatis. In conclusion, analyses of the 16S rRNA and phaC genes showed that Accumulibacter phosphatis might be phylogenetically and metabolically diverse.

King's Status Reflected in The Joseon Dynasty's Document transmission System (조선 문서행이체제에 반영된 국왕의 위상)

  • Lee, Hyeongjung
    • The Korean Journal of Archival Studies
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    • no.66
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    • pp.203-227
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    • 2020
  • This article explores the influence of the king in the Joseon dynasty's document transmission system, focusing on some exceptional cases. According to the Joseon's law, the form of official documents depended on rank differences between receiver and sender. However, there were cases of not following the general principles such as Byungjo(兵曹), Seungjeongwon(承政院) and Kyujanggak(奎章閣). Byungjo was a ministry in charge of military administration. Seungjeongwon was a royal secretary institution which assisted the king and delivered king's orders that existed from the early Joseon. Kyujanggak was a royal library and an assistant institution of the king that was established in the JeongJo(正祖) era. Byungjo was regarded as a relatively high-ranking institution when it sent and received military-related documents. Seungjeongwon and Kyujanggak could use Kwanmoon(關文) to upper rank institution. Kwanmoon was the document form used for institutions of the same or lower rank than itself. Conversely, higher rank institutions used Cheobjeong(牒呈) which was stipulated as a document form to using upper rank institution in law to send them. The reason that they could have privileges in transmission document system was that Joseon had an administrative system centered on the king. Byungjo was an institution entrusted with military power from King. Seungjeonwon and Kyujanggak took charge of the assistance and the delivery of King's order. so they could have a different system of receiving and sending document than the others. In conclusion, the Joseon Dynasty operated exceptions in document administration based on the existence of the king, it means Joseon's transmission document system was basically operated under the Confucian bureaucracy with the king as its peak.

A Quality Comparison of Traditional Korean Papers: Mixtures of Bast-Fiber with Straw pulp(Rice straw paper) in Different Composition Ratio (고정(藁精) 혼합비율에 따른 한지의 물성 비교)

  • Jung, Sun-Young
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.39 no.1 s.119
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    • pp.48-55
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    • 2007
  • The aim of this article is to review the general property of the famous traditional Korean paper, "rice straw paper"(Bast-Fiber mixed with straw pulp), and to compare the paper qualities varying with the composition ratio of straw pulp, in order to revive the traditional paper in modem Korea. The experiment was carried out by making first the two different mixture papers, i. e., one is the paper of 50% bast fiber mixed with 50% straw pulp, and the other is the paper of 75% bast fiber with 25% oats straw pulp, and finally the 100% bast fiber paper was made for the purpose of comparison. The qualitative properties of these three kinds of paper with the different mixing ratio of the straw pulp were evaluated, and the findings of the experiment can be summarized as follow: 1. As to the quality aspects of the paper strength like tensile strength, breaking length, elongation, and tear strength, the test proved the 100% bast fiber paper as best, and the 50% mixed paper as the next good one. 2. In aspect of the printing adaptability such as density, opacity, brightness, whiteness, lab colors, air permeability, and roughness, the 50% mixed paper proved to be the best, due to the short cells in the straw pulp. 3. As to the air permeability, the larger ratio of straw pulp was found to be the lesser, and 75% bast fiber with 25% straw pulp mixture paper and the 100% bast fiber one were found 1/5 degree efficient. 4. In terms of the water absorption degree, 100% bast fiber paper was the fastest, but in case of mixture paper, 50% mixed one was a little faster up to the point of 1cm, while the two kinds of mixed ones appear to be almost similar to each other beyond the point. 5. The straw pulp mixed paper, especially the 50% mixed one was evaluated as the highest by the calligraphers who had experienced using the papers in terms of movement and feeling of the caligraphy and painting. In addition, although the 25% mixed paper is judged to be good for book printing because of the strength, the 50% mixed paper can be thought to be more desirable for painting and calligraphy. In conclusion, we will be able to make the quality paper with durability, by mixing the straw pulp with the bast fiber in proper ratio, following the tradition of Korean paper making.

Molecular Analysis of Colonized Bacteria in a Human Newborn Infant Gut

  • Park Hee-Kyung;Shim Sung-Sub;Kim Su-Yung;Park Jae-Hong;Park Su-Eun;Kim Hak-Jung;Kang Byeong-Chul;Kim Cheol-Min
    • Journal of Microbiology
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    • v.43 no.4
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    • pp.345-353
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    • 2005
  • The complex ecosystem of intestinal micro flora is estimated to harbor approximately 400 different microbial species, mostly bacteria. However, studies on bacterial colonization have mostly been based on culturing methods, which only detect a small fraction of the whole microbiotic ecosystem of the gut. To clarify the initial acquisition and subsequent colonization of bacteria in an infant within the few days after birth, phylogenetic analysis was performed using 16S rDNA sequences from the DNA iso-lated from feces on the 1st, 3rd, and 6th day. 16S rDNA libraries were constructed with the amplicons of PCR conditions at 30 cycles and $50^{\circ}C$ annealing temperature. Nine independent libraries were produced by the application of three sets of primers (set A, set B, and set C) combined with three fecal samples for day 1, day 3, and day 6 of life. Approximately 220 clones ($76.7\%$) of all 325 isolated clones were characterized as known species, while other 105 clones ($32.3\%$) were characterized as unknown species. The library clone with set A universal primers amplifying 350 bp displayed increased diversity by days. Thus, set A primers were better suited for this type of molecular ecological analysis. On the first day of the life of the infant, Enterobacter, Lactococcus lactis, Leuconostoc citreum, and Streptococcus mitis were present. The largest taxonomic group was L. lactis. On the third day of the life of the infant, Enterobacter, Enterococcus faecalis, Escherichia coli, S. mitis, and Streptococcus salivarius were present. On the sixth day of the life of the infant, Citrobacter, Clostridium difficile, Enterobacter sp., Enterobacter cloacae, and E. coli were present. The largest taxonomic group was E. coli. These results showed that microbiotic diversity changes very rapidly in the few days after birth, and the acquisition of unculturable bacteria expanded rapidly after the third day.

A Systematic Review on the Reporting Quality of Acupuncture Treatment for Carpal Tunnel Syndrome (손목터널증후군에 사용된 침 치료 보고의 질 평가)

  • Hyun, Ji-Yoon;Shin, Joo-eun;Im, Chae-Jeong;Park, Ji-Yeun
    • Korean Journal of Acupuncture
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    • v.37 no.3
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    • pp.131-144
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    • 2020
  • Objectives : The aim of this study is to analyze the details of acupuncture treatment methods and the reporting quality of acupuncture on Carpal Tunnel Syndrome (CTS). Methods : Search was conducted in Pubmed, EMBASE, and Cochrane Library for acupuncture studies on CTS. The reporting quality of acupuncture treatment was assessed using the following guidelines: Standards for Reporting Interventions in Clinical Trials of Acupuncture (STRICTA) for analyzing the method of acupuncture treatment, Consolidated Standards of Reporting Trials (CONSORT) for analyzing study design and study process, and Risk of Bias (ROB) for analyzing bias. The number of reported items was calculated and evaluated as a proportion. The reported proportion of each study was classified into three grades: Grade A (% score ≥75), Grade B (50≤ % score <75), and Grade C (% score <50). Results : A total of 9 Randomized Controlled Trials (RCTs) were included in this study. All trials reported 12 items (66.67%) on average in STRICTA guidelines. Five studies were conducted with manual acupuncture and 3 studies were conducted with electroacupuncture. PC7 (Daereung) was most frequently used to treat CTS. In STRICTA guideline evaluation, 3 studies were classified as Grade A, 5 studies were classified as Grade B, and 1 study was classified as Grade C. In the CONSORT statement assessment, all trials reported an average of 20.56 items. Of the 9 RCTs, 6 studies were classified as Grade B and 3 studies were classified as Grade C. In ROB assessment, most studies showed a low (63.49%) or unclear (26.98%) risk of bias. The selective reporting bias and the incomplete outcome data bias were found to have the lowest risk of bias, and the allocation concealment of selection bias was found to have the most unclear risk of bias. Conclusions : Recent acupuncture studies on CTS showed moderate reporting quality. However, more detailed reports on acupuncture are still needed to establish more solid evidence of acupuncture treatment.

A Web-based Simulation Environment based on the Client/Server Architecture for Distance Education: SimDraw (원격교육을 위한 클라이언트/서버구조의 웹 기반 시뮬레이션 환경 : SimDraw)

  • 서현곤;사공봉;김기형
    • Journal of KIISE:Software and Applications
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    • v.30 no.11
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    • pp.1080-1091
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    • 2003
  • Recently, the distance education has been rapidly proliferated with the rapid growth of the Internet and high speed networks. There has been relatively much research with regard to online lecture (teaching and studying) tools for the distance education, compared to the virtual laboratory tools (for self-study and experiments). In this paper, we design and implement a web-based simulation tool, named as SimDraw, for the virtual laboratory in the distance education. To apply the web-based simulation technology into the distance education, some requirements should be met; firstly, the user interface of the simulation should be very easy for students. Secondly, the simulation should be very portable to be run on various computer systems of remote students. Finally, the simulation program on remote computers should be very thin so that students can easily install the program onto their computers. To meet these requirements, SimDraw adopts the client/server architecture; the client program contains only model development and animation functions so that no installation of a client program onto student's system is required, and it can be implemented by a Java applet in Web browsers. The server program supports client programs by offering the functions such as remote compiling, model storing, library management, and user management. For the evaluation of SimDraw, we show the simulation process using the example experimentation of the RIP(Routing Information Protocol) Internet routing protocol.

An Open API Proxy Server System for Widget Services (위젯 서비스를 위한 오픈 API 프록시 서버 시스템)

  • Ahn, Byung-Hyun;Lee, Hyuk-Joon;Choi, Yong-Hoon;Chung, Young-Uk
    • Journal of KIISE:Computing Practices and Letters
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    • v.16 no.9
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    • pp.918-926
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    • 2010
  • A widget is a small application running by the users' favorite services, so they are provided with web contents without explicitly visiting the web site. Although widgets can be easily implemented with Open APIs, only a few web sites provide them because of refactoring the structures of web resource to supply Open APIs to the widget developers. This paper presents an Open API Proxy Server System for widget services. The system consists of two components: an Open API Source Code Generator and an Open API Proxy Server. The Open API Source Code Generator provides a Graphical User Interface (GUI) for users to generate the Open APIs of user's choice and sends the Open API source code generation request to the Open API Proxy Server. The Open API Proxy Server using the HTML Table Processing Library receives the HTML web page from web site and extracts useful information from the target HTML table. The proxy server converts the extracted data into the corresponding XML document which becomes available through the Open API. We verify the operation of the proposed system through experiments with the HTML tables in the example web sites.