• Title/Summary/Keyword: natural substrates

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Minor Coat Protein pIII Domain (N1N2) of Bacteriophage CTXф Confers a Novel Surface Plasmon Resonance Biosensor for Rapid Detection of Vibrio cholerae

  • Shin, Hae Ja;Hyeon, Seok Hywan;Cho, Jae Ho;Lim, Woon Ki
    • Microbiology and Biotechnology Letters
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    • v.49 no.4
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    • pp.510-518
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    • 2021
  • Bacteriophages are considered excellent sensing elements for platforms detecting bacteria. However, their lytic cycle has restricted their efficacy. Here, we used the minor coat protein pIII domain (N1N2) of phage CTXφ to construct a novel surface plasmon resonance (SPR) biosensor that could detect Vibrio cholerae. N1N2 harboring the domains required for phage adsorption and entry was obtained from Escherichia coli using recombinant protein expression and purification. SDS-PAGE revealed an approximate size of 30 kDa for N1N2. Dot blot and transmission electron microscopy analyses revealed that the protein bound to the host V. cholerae but not to non-host E. coli K-12 cells. Next, we used amine-coupling to develop a novel recombinant N1N2 (rN1N2)-functionalized SPR biosensor by immobilizing rN1N2 proteins on gold substrates and using SPR to monitor the binding kinetics of the proteins with target bacteria. We observed rapid detection of V. cholerae in the range of approximately 103 to 109 CFU/ml but not of E. coli at any tested concentration, thereby confirming that the biosensor exhibited differential recognition and binding. The results indicate that the novel biosensor can rapidly monitor a target pathogenic microorganism in the environment and is very useful for monitoring food safety and facilitating early disease prevention.

Effects of the Mixing Ratio of the Different Substrates and the Concentration of Fertigation in Nutrient Solution on the Growth of Tomato Plug Seedlings (배지의 혼합비율과 관비 양액 농도가 토마토 플러그묘의 생장에 미치는 영향)

  • Kim, Hong-Gi;Cho, Ja-Yong;Yu, Sung-Oh;Yang, Seung-Yul;Kang, Jong-Gu;Heo, Buk-Gu
    • Journal of Bio-Environment Control
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    • v.16 no.2
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    • pp.108-114
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    • 2007
  • This study was conducted to clarify the effects of the different mixing ratios of substrate mixtures based on peat moss and the concentration of nutrient solution on the growth of tomato (Lycopersicon esculentum Mill.) seedlings. Substrates such as peat moss, rice hull, carbonized rice hull, decomposed sawdust, perlite and granular rock wool were mixed and used. The concentration of nutrient solution were adjusted to EC $0.5{\sim}1.5mS/cm$. The volumetric moisture contents became higher as peat moss mixed were much more. Total porosities in all substrate mixtures were over 80%, and pH in substrate mixtures became lower as the volume of peat moss mixed higher. Mixing ratios of substrates suitable for the production of tomato seedlings with the higher quality were peat moss:rice hull:carbonized rice hull:decomposed sawdust:perlite=25:10:25:20:20(v/v). The plant growth was not significant among the different substrate mixtures. However, plant growth such as plant height, leaf area, and total dry weight became significantly increased as EC increasing.

Channeling of Intermediates Derived from Medium-Chain Fatty Acids and De novo-SYnthesized Fatty Acids to Polyhydroxyalkanoic Acid by 2-Bromooctanoic Acid in Pseudomonas fluorescens BM07

  • LEE, HO-JOO;RHO, JONG-KOOK;KAMBIZ AKBARI NOGHABI,;LEE, SEUNG-EUN;CHOI, MUN-HWAN;YOON, SUNG-CHUL
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1256-1266
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    • 2004
  • 2-Bromooctanoic acid (2-BrOA) is known to block the formation of polyhydroxyalkanoic acid (PHA) in Pseudomonasfluorescens BM07 without any influence on the cell growth when grown on fructose, but it inhibits the cell growth when grown on octanoate (OA) (Lee et al., Appl. Environ. Microbiol. 67: 4963- 4974, 2001). We investigated the role of 2-BrOA in the PHA synthesis of the bacterium grown with mixtures of fructose and fatty acids. OA, 11­phenoxyundecanoic acid (1 1-POU), and 5-phenylvaleric acid (5-PV) were selected as model substrates. When supplemented with 50 mM fructose, all these carboxylic acids suppressed the formation of PHA from fructose, however, the ~-oxidation coenzyme A monomers derived from the carboxylic acids were efficiently polymerized, but the conversion yield [(mol of carboxylate substrate converted into PHA)/(mol of carboxylate substrate in the feed)] was low (e.g., maximally $\~53\%$ for 5 mM 11-POU). Addition of 2-BrOA (up to 5 mM) to the mixed carbon sources raised the conversion yield sensitively and effectively only at low levels of the acid substrates (e.g., 2 mM 1 1-POU or 5 mM OA): For instance, $100\%$ of 2 mM ll-POU were converted into PHA in the presence of 5 mM 2-BrOA, whereas only $\~10\%$ of the 1 1-POU were converted in the absence of 2-BrOA. However, at highly saturated suppressing levels (e.g., 5 mM ll-POU), 2-BrOA inhibitor showed no significant additional effect on the conversion ($60- 70\%$ conversion irrespective of 2-BrOA level). The existence of competitive and compensative relationship between 2­BrOA and all the carboxylic acid substrates used may indicate 'Present address: Section on Brain Physiology and Metabolism, Bldg. 10, Rm. 6N202, National Institute on Agmg, National Institute of Health, Bethesda, MD 20892, U.S.A. that all the acid substrate-derived inhibiting species bind to the same site as the 2-BrOA inhibiting species does. We, therefore, suggest that 2-BrOA can be used for efficiently increasing the yield of conversion of expensive substituted fatty acids into PHA and then substituted 3-hydroxyacids by hydrolyzing it.

Effects of the Different Substrates on the Plant Growth and Mineral Contents of Hydroponically Grown Plantago asiatica in Aggregate Culture (배지종류가 추식 수경재배 질경이의 생장과 무기성분 함량에 미치는 영향)

  • Cho, Ja-Yong;Kim, Hong-Gi;Yu, Sung-Oh;Yang, Seung-Yul;Yang, Won-Mo;Kim, Byoung-Woo;Heo, Buk-Gu
    • Journal of Bio-Environment Control
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    • v.16 no.1
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    • pp.40-46
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    • 2007
  • This study was conducted to clarify the effects of the different substrates in terms of perlite (100%), peat moss (100%), granular rock wool (100%) and mixing perlite (50%) with peat moss (50%) on the growth responses and mineral contents of hydroponically grown Plantago asiatic in aggregate culture. Overall early plant growth such as plant height, stem diameter, number of leaves, root length, fresh and dry weight of shoot and root were increased in the order of that miked perlite and peat moss (50%:50%, v/v)>peat moss (100%)>granular rock wool (100%)>perlite (100%). Ca and Mg contents in plants became highest in the plants grown in the peat moss, however, Mg and Na in the granular rock wool. $P_2O_5$ content in plants were most increased in proportion to the plant growth increment in the peat moss (100%) and the mixing substrates of perlite and peat moss (50:50, v/v).

Preparation of Pure Cellulose Substrate from Low-Grade Woods by Autohydrolysis (저급목재의 자기가수분해 전처리에 의한 고순도 셀룰로오스 기질의 제조)

  • Cho, Nam-Seok;Kim, Byoung-Ro;Paik, Ki-Hyon
    • Journal of the Korean Wood Science and Technology
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    • v.30 no.4
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    • pp.8-16
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    • 2002
  • This study was performed to produce the high reactive lignin zero substrates from autohydrolyzed wood resources. In chemical compositions of used raw-materials, there were significant differences between two species, Japanese larch (Larix leptolepis) and oak (Quercus mongolica) woods. Japanese larch contained 25 to 3.5 times higher amounts of extractives than oak wood, which is mainly derived from high content of arabinogalactan in Japanese larch wood. Oak wood has 5% lower lignin content and 3% higher holocellulose and pentosans than larch wood. Concerned to changes in wood components during autohydrolysis pretreatment at 22 kg/cm2 steaming pressure for 5~60 min, glucose content was constant during pretreatment, while hemicellulose and lignin were abruptly changed. Hemicellulose fraction was decreased significantly and lignin contents increased because of its condensation reaction with hemicellulose degradation products. The pH of hydrolyzates during pretreatment was decreased, reached upto pH 3 and since then leveled off. In the case of oak wood, same tendency was observed as in Japanese larch. Autohydrolysis followed by sodium chlorite and sulfite or bisulfite pretreatment was very effective in delignification of the substrates. In particular, two-stage delignification of autohydrolyzed woods with alkali and O2-alkali resulted in very low lignin content substrates, such as 0~0.2% lignin substrate.

Isolation and Identification of Hydrolytic Enzyme-producing Bacteria from Spent Mushroom Substrate (버섯부산물유래 가수분해효소분비 박테리아의 분리 및 동정)

  • Kim, Young-Il;Jeong, Se-Hyung;Seok, Joon-Sang;Yang, Si-Yong;Huh, Jung-Won;Kwak, Wan-Sup
    • Journal of Animal Science and Technology
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    • v.50 no.5
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    • pp.713-720
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    • 2008
  • This study was conducted to isolate and identify xylanase- and cellulase-producing thermophilic bacteria from stacked spent mushroom substrates and to determine the optimal medium conditions for their growth. Bacteria with the highest xylanase and CMCase activities were strain 3 and 201-7. Both of them were identified as Bacillus spp. and named Bacillus subtilis KU3 and Bacillus subtilis KU201-7. The optimal medium condition of Bacillus subtilis KU3 was obtained when 3%(w/v) of yeast extract and 1%(w/v) of maltose were used as nitrogen and carbon sources, respectively. That of Bacillus subtilis KU201-7 was obtained when 0.5%(w/v) of yeast extract and 0.5%(w/v) of CMC were used as nitrogen and carbon sources, respectively.

Isolation and Identification of High Cellulolytic Bacteria from Spent Mushroom Substrate and Determination of Optimal Medium Conditions for the Growth (버섯폐배지로부터 섬유소분해력이 높은 중온성 균의 분리 및 균주생산을 위한 배지조건의 최적화)

  • Kim, Young-Il;Jung, Se-Hyung;Seok, Joon-San;Yang, Si-Yong;Huh, Jeong-Weon;Kwak, Wan-Sup
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.255-260
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    • 2007
  • This study was conducted to isolate and identify bacteria producing xylanase and cellulase from spent mushroom substrates and to determine the optimal medium conditions for their growth. Bacteria showing high xylanase and carboxymethyl cellulase activities and low protease and amylase activities were strain 201-3 and strain 206-3. Strain 201-3 was identified as Enterobacter ludwigii and named Ent. ludwigii KU201-3. 206-3 was identified as Bacillus cereus and named B. cereus KU206-3. The optimal medium condition of Ent. ludwigii KU201-3 was obtained when 1%(w/v) of soybean meal and 3%(w/v) of sucrose were used as nitrogen and carbon source, respectively. That of B. cereus KU206-3 was obtained when 3%(w/v) of soybean meal and 1%(w/v) of molasses were used as nitrogen and carbon sources, respectively.

Enzymatic and Energetic Properties of an Aerobic Respiratory Chain­Linked NADH Oxidase System in Marine Bacterium Vibrio natriegens

  • Kang, Ji-Won;Kim, Young-Jae
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1080-1086
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    • 2005
  • Membranes prepared from Vibrio natriegens oxidized both NADH and deamino-NADH as substrates. The maximum activity of the membrane-bound NADH oxidase was obtained at about pH 8.5 in the presence of 0.2 M NaCl, whereas that of the NADH:ubiquinone oxidoreductase was obtained at about pH 7.5 in the presence of 0.2 M NaCl. Electron transfer from NADH or deamino-NADH to ubiquinone-l or oxygen generated a considerable membrane potential (${\Delta}{\psi}$), which occurred even in the presence of $20{\mu}M$ carbonylcyanide m-chlorophenylhydrazone (CCCP). However, the ${\Delta}{\psi}$ was completely collapsed by the combined addition of $10{\mu}M$ CCCP and $20{\mu}M$ monensin. On the other hand, the activity of the NADH oxidase and the ${\Delta}{\psi}$ generated by the NADH oxidase system were inhibited by about $90\%$ with $10{\mu}M$ HQNO, whereas the activity of the NADH:ubiquinone oxidoreductase and the ${\Delta}{\psi}$ generated at the NADH:ubiquinone oxidoreductase segment were inhibited by about $60\%$. Interestingly, the activity of the NADH:ubiquinone oxidoreductase and the ${\Delta}{\psi}$ generated at the NADH:ubiquinone oxidoreductase segment were resistant to the respiratory chain inhibitors such as rotenone, capsaicin, and $AgNO_3$, and the activity of the NADH oxidase and the ${\Delta}{\psi}$ generated by the NADH oxidase system were very sensitive only to $AgNO_3$. It was concluded, therefore, that V. natriegens cells possess a $AgNO_3$-resistant respiratory $Na^+$ pump that is different from the $AgNO_3$-sensitive respiratory $Na^+$ pump of a marine bacterium, Vibrio alginolyticus.

Polyhydroxyalkanoate (PHA) Production Using Waste Vegetable Oil by Pseudomonas sp. Strain DR2

  • Song, Jin-Hwan;Jeon, Che-Ok;Choi, Mun-Hwan;Yoon, Sung-Chul;Park, Woo-Jun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1408-1415
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    • 2008
  • To produce polyhydroxyalkanoate (PHA) from inexpensive substrates by bacteria, vegetable-oil-degrading bacteria were isolated from a rice field using enrichment cultivation. The isolated Pseudomonas sp. strain DR2 showed clear orange or red spots of accumulated PHA granules when grown on phosphate and nitrogen limited medium containing vegetable oil as the sole carbon source and stained with Nile blue A. Up to 37.34% (w/w) of intracellular PHA was produced from corn oil, which consisted of three major 3-hydroxyalkanoates; octanoic (C8:0, 37.75% of the total 3-hydroxyalkanoate content of PHA), decanoic (C10:0, 36.74%), and dodecanoic (C12:0, 11.36%). Pseudomonas sp. strain DR2 accumulated up to 23.52% (w/w) of $PHA_{MCL}$ from waste vegetable oil. The proportion of 3-hydroxyalkanoate of the waste vegetable-oil-derived PHA [hexanoic (5.86%), octanoic (45.67%), decanoic (34.88%), tetradecanoic (8.35%), and hexadecanoic (5.24%)] showed a composition ratio different from that of the corn-oil-derived PHA. Strain DR2 used three major fatty acids in the same ratio, and linoleic acid was the major source of PHA production. Interestingly, the production of PHA in Pseudomonas sp. strain DR2 could not occur in either acetate- or butyrate-amended media. Pseudomonas sp. strain DR2 accumulated a greater amount of PHA than other well-studied strains (Chromobacterium violaceum and Ralstonia eutropha H16) when grown on vegetable oil. The data showed that Pseudomonas sp. strain DR2 was capable of producing PHA from waste vegetable oil.

Biomineralization on Polydiacetylene Patterns Deposited by Using a Novel Mini Arrayer (신규 미니어레이어에 의한 폴리다이아세틸렌 패턴상의 생체유도결정화)

  • Lee, Won Doc;Lee, Gil Sun;Ahn, Dong June
    • Korean Chemical Engineering Research
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    • v.48 no.3
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    • pp.350-354
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    • 2010
  • In natural world various inorganic crystals are grown with controlled shape and size in hybrid forms with organics. Such natural processes have been attempted much to mimick artificially. One of the example is calcium carbonate which has been examined a lot in the field of biomineralization. In this study, we utilize well-organized surface of polydiacetylene(PDA) films as the crystal-growing template. We devised a novel mini-arrayer device that transfers PDA films at air/water interfaces of each array well and deposit them to hydrophobized glass substrates. This technical improvement will contribute to facilitate better understandings of biomineralization mechanism.