• Title/Summary/Keyword: Biomaterial

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Experimental Study on Temperature Dependence of Nitrate Sensing using an ISE-based On-site Water Monitoring System

  • Jung, Dae-Hyun;Kim, Dong-Wook;Cho, Woo Jae;Kim, Hak-Jin
    • Proceedings of the Korean Society for Agricultural Machinery Conference
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    • 2017.04a
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    • pp.122-122
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    • 2017
  • Recently, environmental problems have become an area of growing interests. In-situ monitoring of water quality is fundamental to most environmental applications. The accurate measurement of nitrate concentrations is fundamental to understanding biogeochemistry in aquatic ecosystems. Several studies have reported that one of the most feasible methods to measure nitrate concentration is the use of Ion Selective-electrodes (ISEs). The ISE application to water monitoring has several advantages, such as direct measurement methodology, high sensitivity, wide measurement range, low cost, and portability. However, the ISE methods may yield inconsistent results where there was a difference in temperature between the calibration and measurement solutions, which is associated with the temperature dependence of ionic activity coefficients in solution. In this study, to investigate the potential of using the combination of a temperature sensor and nitrate ISEs for minimizing the effect of temperature on real-time nitrate sensing in natural water, a prototype of on-site water monitoring system was built, mainly consisting of a sensor chamber, an array of 3 ISEs, an waterproof temperature sensor, an automatic sampling system, and an arduino MCU board. The analog signals of ISEs were obtained using the second-order Sallen-key filter for performing voltage following, differential amplification, and low pass filtering. The performance test of the developed water nitrate sensing system was conducted in a monitoring station of drinking water located in Jeongseon, Kangwon. A temperature compensation method based on two-point normalization was proposed, which incorporated the determination of temperature coefficient values using regression equations relating solution temperature and electrode signal determined in our previous studies.

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Efficient Target Site Selection for an RNA-cleaving DNAzyme through Combinatorial Library Screening

  • Kim, Ki-Sun;Choi, Woo-Hyung;Gong, Soo-Jeong;Oh, Sang-taek;Kim, Jae-Hyun;Kim, Dong-Eun
    • Bulletin of the Korean Chemical Society
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    • v.27 no.5
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    • pp.657-662
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    • 2006
  • Identification of accessible sites in targeted RNAs is a major limitation to the effectiveness of antisense oligonucleotides. A class of antisense oligodeoxynucleotides, known as the “10-23” DNA enzyme or DNAzyme, which is a small catalytic DNA, has been shown to efficiently cleave target RNA at purine-pyrimidine junctions in vitro. We have designed a strategy to identify accessible cleavage sites in the target RNA, which is hepatitis C virus nonstructural gene 3 (HCV NS3) RNA that encodes viral helicase and protease, from a pool of random DNAzyme library. A pool of DNAzymes of 58 nucleotides-length that possess randomized annealing arms, catalytic core sequence, and fixed 5'/3'-end flanking sequences was designed and screened for their ability to cleave the target RNA. The screening procedure, which includes binding of DNAzyme pool to the target RNA under inactive condition, selection and amplification of active DNAzymes, incubation of the selected DNAzymes with the target RNA, and target site identification on sequencing gels, identified 16 potential cleavage sites in the target RNA. Corresponding DNAzymes were constructed for the selected target sites and were tested for RNA-cleavage in terms of kinetics and accessibility. These selected DNAzymes were effective in cleaving the target RNA in the presence of $Mg^{2+}$. This strategy can be applicable to identify accessible sites in any target RNA for antisense oligonucleotides-based gene inactivation methods.

Production of Xylooligosaccharides by Yeast Cell Surface-Displayed Endoxylanase (효모 세포 표면 발현된 Endoxylanase를 이용한 Xylooligosaccharides의 생산)

  • Kim, Hyun-Jin;Lee, Jae-Hyung;Kim, Yeon-Hee;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.307-313
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    • 2008
  • The yeast surface expression system, pCTXYN (6.8 kb), of Bacillus endoxylanase gene (xynB, 642 bp) was constructed and introduced into Saccharomyces cerevisiae EBY100 cell. The transformed yeast cell showing the highest endoxylanase activity was selected through the active staining of colonies grown on YPDG medium containing xylan. With the yeast transformant, EBY100/pCTXYN, grown on galactose containing medium, it was found that the endoxylanase was successfully displayed on the yeast cell surface and the xylooligosaccharides were efficiently produced from xylan. The most of endoxylanase activity was detected in the cell fraction and reached about 1.9 unit/mL after 48 h cultivation. The optimized conditions for xylooligosaccharides production from xylan were determined as follows: substrate and its concentration, oat spelt xylan 6%; concentration of yeast whole-cell, 5 unit/mL; temperature, $50^{\circ}C$, and reaction time $2{\sim}4\;h$. When the oat spelts xylan and corncob xylan were hydrolyzed by treatment with cell surface-displayed endoxylanase, xylotriose was formed as a main product.

Cooperativity of ${\alpha}$- and ${\beta}$-Subunits of Group II Chaperonin from the Hyperthermophilic Archaeum Aeropyrum pernix K1

  • Kim, Jeong-Hwan;Lee, Jin-Woo;Shin, Eun-Jung;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.212-217
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    • 2011
  • ${\alpha}$ and ${\beta}$-subunits (ApCpnA and ApCpnB) are group II chaperonins from the hyperthermophilic archaeum Aeropyrum pernix K1, specialized in preventing the aggregation and inactivation of substrate proteins under conditions of transient heat stress. In the present study, the cooperativity of ${\alpha}$- and ${\beta}$-subunits from the A. pernix K1 was investigated. The ApCpnA and ApCpnB chaperonin genes were overexpressed in E. coli Rosetta and Codonplus (DE3), respectively. Each of the recombinant ${\alpha}$- and ${\beta}$-subunits was purified to 92% and 94% by using anionexchange chromatography. The cooperative activity between purified ${\alpha}$- and ${\beta}$-subunits was examined using citrate synthase (CS), alcohol dehydrogenase (ADH), and malate dehydrogenase (MDH) as substrate proteins. The addition of both ${\alpha}$- and ${\beta}$-subunits could effectively protect CS and ADH from thermal aggregation and inactivation at $43^{\circ}C$ and $50^{\circ}C$, respectively, and MDH from thermal inactivation at $80^{\circ}C$C and $85^{\circ}C$. Moreover, in the presence of ATP, the protective effects of ${\alpha}$- and ${\beta}$-subunits on CS from thermal aggregation and inactivation, and ADH from thermal aggregation, were more enhanced, whereas cooperation between chaperonins and ATP in protection activity on ADH and MDH (at $85^{\circ}C$) from thermal inactivation was not observed. Specifically, the presence of both ${\alpha}$- and ${\beta}$- subunits could effectively protect MDH from thermal inactivation at $80^{\circ}C$ in an ATP-dependent manner.

Purification and Anticoagulant Activity of a Fucoidan from Korean Undaria pinnatifida Sporophyll

  • Kim , Woo-Jung;Kim, Sung-Min;Kim, Hyun-Guell;Oh, Hye-Rim;Lee, Kyung-Bok;Lee, Yoo-Kyung;Park, Yong-Il
    • ALGAE
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    • v.22 no.3
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    • pp.247-252
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    • 2007
  • Crude fucoidan was extracted from the sporophyll of Korean Undaria pinnatifida collected at a coastal area ofWando, Korea, mainly by dilute acid extraction, ethanol precipitation, CaCU Precipitation, with an yield of approxi-mately 3.9% in mass. It was further purified by DEAE-cellulose column chromatography and its chemical composi-don and in vitro anticoagulant activity was determined. The average molecular mass of the purified fucoidan wasestimated about 2.1 x 103 kDa by size-fractionation HPLC and it consisted of neutral sugar (52.34% in mass), uronicacid (26.2%), and sulfate esters (7.4%). From the HPAEC-PAD analysis, the monosaccharide composition of thepurified fucoidan was shown to be fucose, galactose, xylose, and mannose, with a molar ratio of 1, 0.2, 0.02, 0.15,respectively, demonstrating that major monosacd-iande was fucose (72.3% in mol percentage) and other sugars,xylose (1.5%), galactose (14.6%), and mannose (10.9%) were present as minor component. The results suggested thatthis fucoidan is a sulfated, U-type fucoidan. The activated partial thrombloplastin time (APTT) assay of the purifiedfucoidan showed that the purified fucoidan elicited anticoagulant activity in a dose-dependent manner. Five jUg ofsporophyll fucoidan delayed the blood clotting time up to 5 times than untreated control and also up to 1.5 timesthan the same amount of the commercial fucoidan, respectively. Although it is preliminary, these results suggestthat the fucoidan of Korean Undaria vinnatifida sporophyll would be promising candidates for the development ofan anticoaeulant.

Properties and Industrial Applications of Seaweed Polysaccharides-degrading Enzymes from the Marine Microorganisms (해양 미생물 유래 해조 다당류 분해 효소의 특성 및 산업적 응용)

  • Kim, Jeong-Hwan;Kim, Yeon-Hee;Kim, Sung-Koo;Kim, Byung-Woo;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.189-199
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    • 2011
  • Recently seaweed polysaccharides have been extensively studied due to their various biological functions including antitumor, antiviral, anticoagulant, and anti-inflammatory activities. Although seaweed polysaccharides are known to possess numerous beneficial properties, their industrial applications have been limited due to the low inclusion efficiency and high cost of manufacturing involved in chemical hydrolysis. In addition, the smell of seaweed has been a limiting factor in its application in the food and cosmetic industries. Therefore, novel hydrolysis methods and the deodorization of seaweed are required if the extensive application of seaweed polysaccharides is to be seen. A number of studies have examined various seaweed polysaccharide-degrading enzymes, which have been isolated from marine microorganisms, and enzymatic hydrolysis processes have been investigated for the improvement of production yields and the bioefficacy of seaweed polysaccharides. This review is a synopsis on the properties of seaweed polysaccharides-degrading enzymes from marine microorganisms and their industrial applications. The review reveals the pressing need for more concentrated research on the development of new biological materials from seaweed.

Imporoved Method for the Preparation of Silk Fibroin Hyoysates

  • Shukhrat Madyarov;Lee, kawng-Gill;Yeo, Joo-Hong;Jin Nam;Lee, Yong-Woo
    • Journal of Sericultural and Entomological Science
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    • v.41 no.2
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    • pp.108-115
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    • 1999
  • An improvement of methods in fibroin hydrolysates preparat significantly enlarges their applications to practical use. Acidic hydrolysis by hydrohloric acid is one of the methods for silk fibroin depolymeriza6tion. A low yield of final product and long time of the process are the demerits of this method. Possibility of preparation of water-soluble silk hydrolysates with more yield and less expenses is investigated in this study. Such possibility is occurred with the increasing tratment temperature and simultaneously decreasing treatment time, concentration of hydrochoric acid respectively, the concentration of sodium hydroxide used for neutraliza6tion of hydolysates after hydrolysis. Colour is decreased in this case and a small amount of activated, too. Protection of hydrolysates against precipitation after neutraliza6tion, and separation and during concentrating process is the other merit of this method. Creamy-coloured insoluble silk powder is the remainder of hydrolyzed fibroin. This is the only the immobiliza6tion of enzymes and other physiological active substances. Fine particles of this powder can be used as additives for artificial diets and cultural media, as well as raw materials for polymer membranes, etc.

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Autoimmune Regulator Gene (Aire) is Expressed in Lymph Node Fibroblastic Reticular Cell, BLS4 (Autoimmune regulator gene (Aire)의 마우스 림프절 FRC세포, BLS4에서 발현)

  • Moon, Kyoung-Mi;Seo, Hee-Ju;Oh, Ji-Youn;Lee, Jae-Seol;Kim, Hae-Young;Kim, Jeong-Hwan;Choi, Woo-Bong;Kim, Byung-Woo;Kim, Kwang-Hyeon;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.20 no.5
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    • pp.670-675
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    • 2010
  • Autoimmune regulator gene (Aire) is expressed in the thymus and controls the expression of peripheral self-antigens, known as promiscuous genes. Aire and promiscuous genes are involved in T cell tolerance and autoimmunity in the thymus. Here, we identified Aire-expressing fibroblastic reticular cell (FRC), which was derived from mouse lymph node and also expressed in insulin promiscuous antigen. The expression of insulin was increased in cultured FRC over-expressed with Aire. These data suggest that Aire regulates promiscuous gene expression in FRC, and that this function might be under peripheral selection control.

Enhancement of Saccharification Yield of Ulva pertusa Kjellman by High Pressure Homogenization Process for Bioethanol Production (구멍갈파래의 고압 균질 전처리 공정을 통한 바이오에탄올 생산용 당화수율 증진)

  • Choi, Woon-Yong;Lee, Choon-Geun;Ahn, Ju-Hee;Seo, Yong-Chang;Lee, Sang-Eun;Jung, Kyung-Hwan;Kang, Do-Hyung;Cho, Jeong-Sub;Choi, Geun-Pyo;Lee, Hyeon-Yong
    • KSBB Journal
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    • v.26 no.5
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    • pp.400-406
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    • 2011
  • This study was investigated to improve the saccharification yield of Ulva pertusa Kjellman by the high pressure homogenization process. It was found that the high pressure homogenization pretreatment effectively destructed the cell wall structures only by using water. The high pressure homogenization process was operated under various conditions such as 10000, 20000 or 30000 psi with different recycling numbers. The optimal condition was determined as 30000 psi and 2 pass of recycling numbers and the sugar conversion yields were 16.02 (%, w/w) of glucose and 14.70 (%,w/w) of xylose, respectively. In the case of enzymatic treating the hydrolyzates with 5 FPU/glucan of celullase and 100 units/mL of amyloglucosidase, 65.8% of carbohydrates was converted into glucose. Using the hydrolysates of Ulva pertusa Kjellman, 48.7% of ethanol was obtained in the culture S.cerevisiae. These results showed that the high pressure homogenization process could efficiently hydrolyze the marine resource by using only water for bioethanol production.