• Title/Summary/Keyword: Sequential-Culture

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Purification and Characterization of a Thermostable Xylanase from Fomitopsis pinicola

  • Shin, Keum;Jeya, Marimuthu;Lee, Jung-Kul;Kim, Yeong-Suk
    • Journal of Microbiology and Biotechnology
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    • v.20 no.10
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    • pp.1415-1423
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    • 2010
  • An extracellular xylanase was purified to homogeneity by sequential chromatography of Fomitopsis pinicola culture supernatants on a DEAE-Sepharose column, a gel filtration column, and then on a MonoQ column with fast protein liquid chromatography. The relative molecular mass of the F. pinicola xylanase was determined to be 58 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis and by size-exclusion chromatography, indicating that the enzyme is a monomer. The hydrolytic activity of the xylanase had a pH optimum of 4.5 and a temperature optimum of $70^{\circ}C$. The enzyme showed a $t_{1/2}$ value of 33 h at $70^{\circ}C$ and catalytic efficiency ($k_{cat}=77.4\;s^{-1}$, $k_{cat}/K_m$=22.7 mg/ml/s) for oatspelt xylan. Its internal amino acid sequences showed a significant homology with hydrolases from glycoside hydrolase (GH) family 10, indicating that the F. pinicola xylanase is a member of GH family 10.

Purification and Characterization of a Thermostable Cellobiohydrolase from Fomitopsis pinicola

  • Shin, Keum;Kim, Yoon-Hee;Jeya, Marimuthu;Lee, Jung-Kul;Kim, Yeong-Suk
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1681-1688
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    • 2010
  • A screening for cellobiohydrolase (CBH) activity was performed and Fomitopsis pinicola KMJ812 was selected for further characterization as it produced a high level of CBH activity. An extracellular CBH was purified to homogeneity by sequential chromatography of F. pinicola culture supernatants. The molecular mass of the F. pinicola CBH was determined to be 64 kDa by SDS-PAGE and by size-exclusion chromatography, indicating that the enzyme is a monomer. The F. pinicola CBH showed a $t_{1/2}$ value of 42 h at $70^{\circ}C$ and catalytic efficiency of $15.8mM^{-1}s^{-1}(k_{cat}/K_m)$ for p-nitrophenyl-${\beta}$-D-cellobioside, one of the highest levels seen for CBH-producing microorganisms. Its internal amino acid sequences showed a significant homology with hydrolases from glycoside hydrolase family 7. Although CBHs have been purified and characterized from other sources, the F. pinicola CBH is distinguished from other CBHs by its high catalytic efficiency and thermostability.

A Study on Developing the Standardized Index of Evaluation for Upgrading the School Facilities (학교시설개선을 위한 평가지표개발에 관한 연구)

  • Song, Byung-Joon;Kim, Young-Suk;Chu, Beom
    • Korean Institute of Interior Design Journal
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    • v.21 no.6
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    • pp.260-270
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    • 2012
  • The demographic and environmental changes such as the decrease of the population growth rate and the increase of the population of the senior citizens, which become issues of the modern society, are bringing numerous changes of many aspects like national economy, community, and culture. These social phenomena give impacts on the school facilities as well. Thus owing to the decrease of the number of the students, the number of the newly constructed schools has been decreased drastically, and moreover the merger of the schools cause many schools to be default. Meanwhile, the school facilities have been expanded into the functions of the local community centers, and the various educational activities demand the diversity and flexibility of the spaces. These factors add more burdens to the current schools. Therefore the school facilities are needed to adopt a flexible and relevant change necessary for using the spaces in accordance with the changed needs of the times, and are needed to reinvented continuously not to limit the educational and social activities because of the degradation of the school facilities. Yet because there are no proper criteria for doing the task, problems related to the relevance of the task and the unnecessary investment are raised. So through this study I tried to demonstrate the standardized index in terms of the sequential priority of upgrading the facilities among the schools.

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Fabrication of Hydrophilic Poly(dimethylsiloxane)with Periodic Wrinkling Surface and Its Application (일정한 주름을 갖는 친수성 PDMS 제작 및 응용)

  • Lee, Dong-Guk;Oh, Chang-Kyu;Yang, Sung-Ho;Han, Seung-Jin;Jeong, Ok-Chan
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.63 no.5
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    • pp.671-675
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    • 2014
  • This paper presents a fabrication of hydrophilic Poly(dimethylsiloxane) (PDMS) with periodic wrinkling surface. The proposed periodic wrinkling surface was fabricated using the sequential processes of typical curing process of PDMS, cutting process, platinum deposition process, and wrinkling transfer process. The surface morphology of the fabricated wrinkling surface was observed by using optical and dynamic atomic force microscopy and discussed. The measured period and amplitude of wrinkling was about $2.2{\mu}m$ and $0.31{\mu}m$, respectively. And, the contact angles of water droplets on the wrinkled surface were measured in order to understand effect of the wrinkling surface on surface modification of hydrophobic PDMS. Our new finding was that the proposed wrinkling surface was hydrophilic and the measured contact angle was about $62^{\circ}$. Moreover, it was found out from the simple cell culture test that the fabricated wrinkling surface was more effective for cell spreading and adhesion than the case of native PDMS substrate.

Characterization of Cellobiohydrolase from a Newly Isolated Strain of Agaricus arvencis

  • Lee, Kyung-Min;Moon, Hee-Jung;Kalyani, Dayanand;Kim, Hoon;Kim, In-Won;Jeya, Marimuthu;Lee, Jung-Kul
    • Journal of Microbiology and Biotechnology
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    • v.21 no.7
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    • pp.711-718
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    • 2011
  • A highly efficient cellobiohydrolase (CBH)-secreting basidiomycetous fungus, Agaricus arvensis KMJ623, was isolated and identified based on its morphological features and sequence analysis of internal transcribed spacer rDNA. An extracellular CBH was purified to homogeneity from A. arvencis culture supernatant using sequential chromatography. The relative molecular mass of A. arvencis CBH was determined to be 65 kDa by SDSPAGE and 130 kDa by size-exclusion chromatography, indicating that the enzyme is a dimer. A. arvencis CBH showed a catalytic efficiency ($k_{cat}/K_m$) of 31.8 $mM^{-1}\;s^{-1}$ for p-nitrophenyl-${\beta}$-D-cellobioside, the highest level seen for CBH-producing microorganisms. Its internal amino acid sequences showed significant homology with CBHs from glycoside hydrolase family 7. Although CBHs have been purified and characterized from other sources, A. arvencis CBH is distinguished from other CBHs by its high catalytic efficiency.

Purification and characterization of a 33 kDa serine protease from Acanthamoeba lugdunensis KA/E2 isolated from a Korean keratitis patient

  • Kim, Hyo-Kyung;Ha, Young-Ran;Yu, Hak-Sun;Kong, Hyun-Hee;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.41 no.4
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    • pp.189-196
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    • 2003
  • In order to evaluate the possible roles of secretory proteases in the pathogenesis of amoebic keratitis, we purified and characterized a serine protease secreted by Acanthamoeba lugdunensis KA/E2, isolated from a Korean keratitis patient The ammonium sulfate-precipitated culture supernatant of the isolate was purified by sequential chromatography on CM-Sepharose, Sephacryl S-200, and mono Q-anion exchange column. The purified 33 kDa protease had a pH optimum of 8.5 and a temperature optimum of $55^{\circ}C$. Phenylmethylsulfonylfluoride and 4-(2-Aminoethyl)-benzenesulfonyl-fluoride, both serine protease specific inhibitors, inhibited almost completely the activity of the 33 kDa protease whereas other classes of inhibitors did not affect its activity. The 33 kDa enzyme degraded various extracellular matrix proteins and serum proteins. Our results strongly suggest that the 33 kDa serine protease secreted from this keratopathogenic Acanthamoeba play important roles in the pathogenesis of amoebic keratitis, such as in corneal tissue invasion, immune evasion and nutrient uptake.

Comparison of Bioethanol Production by Candida molischiana and Saccharomyces cerevisiae from Glucose, Cellobiose, and Cellulose

  • Zheng, Jianning;Negi, Abhishek;Khomlaem, Chanin;Kim, Beom Soo
    • Journal of Microbiology and Biotechnology
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    • v.29 no.6
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    • pp.905-912
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    • 2019
  • Bioethanol has attracted much attention in recent decades as a sustainable and environmentally friendly alternative energy source. In this study, we compared the production of bioethanol by Candida molischiana and Saccharomyces cerevisiae at different initial concentrations of cellobiose and glucose. The results showed that C. molischiana can utilize both glucose and cellobiose, whereas S. cerevisiae can only utilize glucose. The ethanol yields were 43-51% from different initial concentrations of carbon source. In addition, different concentrations of microcrystalline cellulose (Avicel) were directly converted to ethanol by a combination of Trichoderma reesei and two yeasts. Cellulose was first hydrolyzed by a fully enzymatic saccharification process using T. reesei cellulases, and the reducing sugars and glucose produced during the process were further used as carbon source for bioethanol production by C. molischiana or S. cerevisiae. Sequential culture of T. reesei and two yeasts revealed that C. molischiana was more efficient for bioconversion of sugars to ethanol than S. cerevisiae. When 20 g/l Avicel was used as a carbon source, the maximum reducing sugar, glucose, and ethanol yields were 42%, 26%, and 20%, respectively. The maximum concentrations of reducing sugar, glucose, and ethanol were 10.9, 8.57, and 5.95 g/l, respectively, at 120 h by the combination of T. reesei and C. molischiana from 50 g/l Avicel.

The Significant Role of Industrial Organizational Psychologists to Increase Employee Job Satisfaction

  • Bomi PARK;Seong-Gon KIM
    • The Journal of Industrial Distribution & Business
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    • v.14 no.12
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    • pp.33-41
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    • 2023
  • Purpose: This study's primary goal is to investigate and evaluate the critical contributions made by industrial-organizational psychologists to raising worker job satisfaction. The study aims to offer insights that can enhance workplace environments, improve organizational processes, and increase employees' general well-being by looking at the particular contributions made by this field. Research design, data and methodology: The study design, data collection strategies using the literature content analysis allow for a thorough analysis and synthesis of the body of knowledge about industrial-organizational psychologists' contributions to improving worker job satisfaction. This method entails a sequential procedure that includes formulating research questions and locating pertinent studies. Results: Numerous previous works have indicated that industrial-organizational psychologists are essential in determining how an organization is run and how satisfied its workforce is with their jobs. The four major roles (Job Design, Leadership, Workplace Culture, and Employee Development) that were found after a thorough analysis of the body of current literature are presented in this section. Conclusions: In sum, the roles found in this study provide organizations with a framework for creating work environments that prioritize the well-being and overall growth of the people who make up the organization while also helping them negotiate the complexity of the modern workplace.

Characterization and Culture Optimization of an Glucosidase Inhibitor-producing Bacteria, Gluconobactor oxydans CK-2165 (α-Glucosidase 저해제 생산 균주, Gluconobacter oxydans CK-2165의 특성 및 배양 최적화)

  • Kim, Byoung-Kook;Suh, Min-Jung;Park, Ji-Su;Park, Jang-Woo;Suh, Jung-Woo;Kim, Jin-Yong;Lee, Sun-Young;Choi, Jongkeun;Suh, Joo-Won;Lee, In-Ae
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.13 no.11
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    • pp.5179-5186
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    • 2012
  • Miglitol, a well-known therapeutic intervention agents for diabetes, exhibits competitive inhibitory activity against ${\alpha}$-glucosidase and it is usually produced through three sequential steps including chemical and bioconversion processes. Gluconobactor oxydans (G. oxydans) belonging to acetic acid bacteria biologically, converts 1-deoxy-1-(2-hydroxyethylamino)-D-glucitol (P1) into a key intermidiate, 6-(2-hydroxyetyl) amino-6-deoxy-${\alpha}$-L-sorbofuranose (P2) by incomplete oxidation. In this study, we identified and optimized fermentation conditions of CK-2165, that was selected in soil samples by comparing the bioconversion yield. CK-2165 strain was found to be closely related to G. oxydans based on the result of phylogenetic analysis using 16S rDNA sequence. Utilization of API 20 kits revealed that this strain could use glucose, mannose, inositol, sorbitol, rhamnose, sucrose, melibiose, amygdalin and arabinose as carbon sources. The culture conditions were optimized for industrial production and several important factors affecting bioconversion rate were also tested using mycelial cake. Cell harvested at the late-stationary phase showed the highest bioconversion yield and $MgSO_4$ was critically required for the catalytic activity.

Biobleaching of Softwood Kraft Pulp Using Recombinant Xylanase and Cellulase (재조합 Xylanase와 Cellulase의 연속처리에 의한 침엽수 Kraft 펄프의 Biobleaching)

  • Kim, Hyun Joo;Wi, Seung Gon;Bae, Hyeun-Jong
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.6
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    • pp.166-174
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    • 2007
  • To know the effect of enzymatic pre-treatment on softwood Kraft pulp, two xylanse-encoding genes, named xynl and xynll were isolated from Thrichoderma ressei. Structural genes of xylanase (XYNI, XYNII) and cellulase (EGIV-CBDII) were isolated from T. ressei and Rumicoccus albus respectively, and expressed in E. coli. bacterial culture. The specific activity of purified recombinant XYNI is higher than XYNII. The brightness of XYNI treated softwood Kraft pulp increased to 29.9%. On further sequential treatment with EGIV-CBDII and XYNI the brightness of softwood Kraft pulp were improved to 9.1 and 73% respectively. As expected the Kappa number of softwood Kraft pulp also decreased 8.1, 4.6 and 3.2% respectively. Results further indicate that this sequential combination of enzyme treatment has synergic effect for improving bleaching of softwood Kraft pulp.