• Title/Summary/Keyword: Microbial reaction

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Production of 4-Ethyl Malate through Position-Specific Hydrolysis of Photobacterium lipolyticum M37 Lipase

  • Lim, Chae Ryeong;Lee, Ha young;Uhm, Ki-Nam;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.32 no.5
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    • pp.672-679
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    • 2022
  • Microbial lipases are used widely in the synthesis of various compounds due to their substrate specificity and position specificity. 4-Ethyl malate (4-EM) made from diethyl malate (DEM) is an important starting material used to make argon fluoride (ArF) photoresist. We tested several microbial lipases and found that Photobacterium lipolyticum M37 lipase position-specifically hydrolyzed DEM to produce 4-EM. We purified the reaction product through silica gel chromatography and confirmed that it was 4-EM through nuclear magnetic resonance analysis. To mass-produce 4-EM, DEM hydrolysis reaction was performed using an enzyme reactor system that could automatically control the temperature and pH. Effects of temperature and pH on the reaction process were investigated. As a result, 50℃ and pH 4.0 were confirmed as optimal reaction conditions, meaning that M37 was specifically an acid lipase. When the substrate concentration was increased to 6% corresponding to 0.32 M, the reaction yield reached almost 100%. When the substrate concentration was further increased to 12%, the reaction yield was 81%. This enzyme reactor system and position-specific M37 lipase can be used to mass-produce 4-EM, which is required to synthesize ArF photoresist.

Microbial Activity of Ammonia Oxidizing Bacteria and Ammonia Oxidizing Archaea in the Rice Paddy Soil in Wang-gung Area of Iksan, Korea (익산 왕궁지역 논 토양에서의 질산화 세균과 질산화 고세균의 미생물학적 작용)

  • Kim, Hyun-su
    • Journal of Soil and Groundwater Environment
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    • v.21 no.4
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    • pp.50-59
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    • 2016
  • Spatial and temporal changes in nitrification activities and distribution of microbial population of ammonia oxidizing bacteria (AOB) and ammonia oxidizing archaea (AOA) in paddy soils were investigated. Soil samples were collected in March and October 2015 from rice paddy with and without the presence of confined animal feeding operations. Incubation experiments and quantitative polymerase chain reaction showed that AOA's contribution to nitrification kinetics was much higher in locations where organic nitrogen in animal waste is expected to significantly contribute to overall nitrogen budget, and temporal variations in nitrification kinetics were much smaller for AOA than AOB. These differences were interpreted to indicate that different microbial responses of two microbial populations to the types and concentrations of nitrogen substrates were the main determining factors of nitrification processes in the paddy soils. The copy numbers of ammonium monooxygenase gene showed that AOA colonized the paddy soils in higher numbers than AOB with stable distribution while AOB showed variation especially in March. Although small in numbers, AOB population turned out to exert more influence on nitrification potential than AOA, which was attributed to higher fluctuation in AOB cell numbers and nitrification reaction rate per cells.

Continuous removal of heavy metals by coupling a microbial fuel cell and a microbial electrolytic cell

  • Xie, Guo R.;Choi, Chan S.;Lim, Bong S.;Chu, Shao X.
    • Membrane and Water Treatment
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    • v.11 no.4
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    • pp.283-294
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    • 2020
  • This work aims at studying the feasibility of continuous removal of mixed heavy metal ions from simulated zinc plating wastewaters by coupling a microbial fuel cell and a microbial electrolysis cell in batch and continuous modes. The discharging voltage of MFC increased initially from 0.4621 ± 0.0005 V to 0.4864 ± 0.0006 V as the initial concentration of Cr6+ increased from 10 ppm to 60 ppm. Almost complete removal of Cr6+ and low removal of Cu2+ occurred in MFC of the MFC-MEC-coupled system after 8 hours under the batch mode; removal efficiencies (REs) of Cr6+ and Cu2+ were 99.76% and 30.49%. After the same reaction time, REs of nickel and zinc ions were 55.15% and 76.21% in its MEC. Cu2+, Ni2+, and Zn2+ removal efficiencies of 54.98%, 30.63%, 55.04%, and 75.35% were achieved in the effluent within optimum HRT of 2 hours under the continuous mode. The incomplete removal of Cu2+, Ni2+ and Zn2+ ions in the effluent was due to the fact that the Cr6+ was almost completely consumed at the end of MFC reaction. After HRT of 12 hours, at the different sampling locations, Cr6+ and Cu2+ removal efficiencies in the cathodic chamber of MFC were 89.95% and 34.69%, respectively. 94.58%, 33.95%, 56.57%, and 75.76% were achieved for Cr6+, Cu2+, Ni2+ and Zn2+ in the cathodic chamber of MEC. It can be concluded that those metal ions can be removed completely by repeatedly passing high concentration of Cr6+ through the cathode chamber of MFC of the MFC-MEC-coupled system.

Study on the 2,4,6-Trinitrotoluene reduction rate by mediation of extracellular material of mixed culture (혼합배양 미생물의 세포외 물질이 2,4,6-Trinitrotoluene 변환에 미치는 영향에 관한 연구)

  • 한기봉
    • Journal of environmental and Sanitary engineering
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    • v.11 no.2
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    • pp.27-31
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    • 1996
  • 2,4,6-Trinitrotoluene(TNT) was reduced into intermediate products by mixed culture incubated in anaerobic condition. To test the effects of extracellular material to electron transfer between sulfide and TNT, filtered medium of mixed culture was loaded in the test tubes with TNT and sulfide. The transformation rate was measured under four different conditions. The rate under microbial activity was the fastest among under different conditions. With sulfide or filtrate alone and TNT, the reactions were measured as the slowest reactions or no reactions occured, respectively. The reaction rate coefficient were calculated by linear regression and the first order kinetic was fitted best. Also, the plot of rate coefficients (K$_{f}$) showed linear relationships when at time zero TNT and sulfide concentration were 20 mg/1 and 6.0 mM, respectively. By extrapolation, reaction rate coefficient of 100% filtrate could be calculated as 0.0054/minute. However, reaction rate was affected by different concentration of sulfide, so it is a dependent of sulfide concentration. The results of this test showed TNT reduction rate can be limited more by microbial reaction than by mediation of filtrate or sulfide and filtrate alone.

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Studies on Constituents of Higher Fungi of Korea(XL) - A Sterol of Phallus impudicus - (한국산(韓國産) 고등(高等) 균류(菌類)의 성분(成分) 연구(硏究)(제40보)(第40報) - 말뚝버섯의 스테롤 성분(成分) -)

  • Choi, Eung-Chil;Chung, Kyeong-Soo;Kim, Jong-Chan;Kim, Byong-kak
    • The Korean Journal of Mycology
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    • v.11 no.2
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    • pp.97-98
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    • 1983
  • The carpophores of Phallus impudicus collected at Gal-mae-ri in Gyeong-gi Province were subjected to chemical analysis for sterol constituents. The dried carpophores were homogenized and extracted with chloroform-methanol(2 : 1). After saponification of the dried extract, unsaponified components were obtained by ether extraction of the reaction mixture. A sterol fraction was separated by preparative TLC and from this fraction, ergosterol was identified by gas­chromatographic analysis with authentic sterols.

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Assessment of the Changes in the Microbial Community in Alkaline Soils using Biolog Ecoplate and DGGE (Biolog Ecoplate와 DGGE 방법을 이용한 알칼리화 토양의 미생물군집 변화 평가)

  • Lee, Eun Young;Hong, Sun Hwa
    • KSBB Journal
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    • v.28 no.5
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    • pp.275-281
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    • 2013
  • Soil microbial community analysis of farmland soil sprayed with lye in order to use fertilizer in Nigeria was performed. As a control, two kinds of soils not sprayed with lye, located in Eungo and Lagos with general practice in agriculture was selected. Soil sprayed with lye was pH 8.25 through alkalization reaction, while the other soil samples were pH 6.22 and 5.94. Substrate utilization and species diversity index of soil sprayed with lye were low than that of the other soils with the analysis of Biolog ecoplate. As a result of principal component analysis, the relationship between three samples was low. Microbial community analysis was performed by DGGE and most of them were soil uncultured bacterium. Especially, Uncultured Acidobacteria and Uncultured Methylocystis sp., which had been isolated from the rhizosphere of soybean grown in that site were discovered in the soil sprayed with lye.

Identification of Amino-Acids Residues for Key Role in Dextransucrase Activity of Leuconostoc mesenteroides B-742CB

  • Ryu, Hwa-Ja;Kim, Do-Man;Seo, Eun-Seong;Kang, Hee-Kyung;Lee, Jin-Ha;Yoon, Seung-Heon;Cho, Jae-Young;Robyt, John-F.;Kim, Do-Won;Chang, Suk-Sang;Kim, Seung-Heuk;Kimura, Atsuo
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.1075-1080
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    • 2004
  • Dextransucrase (DSRB742) from Leuconostoc mesenteroides NRRL B-742CB is a glucosyltransferase that catalyzes the synthesis of dextran using sucrose, or the synthesis of oligosaccharides when acceptor molecules, like maltose, are present. The DSRB742 gene (dsrB742) was cloned and the properties were characterized. In order to identify critical amino acid residues, the DSRB742 amino acid sequence was aligned with glucosyltransferase sequences, and three amino acid residues reported as sucrose binding amino acids in Streptococcus glucosyltransferases were selected for site-directed mutagenesis experiments. Asp-533, Asp-536, and His-643 were independently replaced with Ala or Asn. D533A and D536A dextransucrases showed reduced dextran synthesis activities, 2.3% and 40.8% of DSRB742 dextransucrase, respectively, and D533N, D536N, H643A, end H643N dextransucrases showed complete suppression of dextran synthesis activities altogether. Additionally, D536N dextransucrase showed complete suppression of oligosaccharide synthesis activities. However, modifications at Asp-533 or at His-643 retained acceptor reaction activities in the range of 8.4% to 21.3% of DSRB742 acceptor reaction activity. Thus at least two carboxyl groups of Asp-533 and Asp-536, and His-643 as a proton donor, are essential for the catalysis process.

Discovery of D-Stereospecific Dipeptidase from Thermophilic Bacillus sp. BCS-l and Its Application for Synthesis of D-Amino Acid-Containing Peptide

  • Baek, Dae-Heoun;Kwon, Seok-Joon;Park, Jin-Seo;Lee, Seung-Goo;Mheen, Tae-Ick;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.646-649
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    • 1999
  • A thermophilic bacterium producing D-stereospecific dipeptidase was isolated from Korean soil samples. The enzyme hydrolyzed the peptide bond between D-alanyl-D-alanine (D-Ala-D-Ala). The isolated bacterial strain was rod shaped, gram-positive, motile, and formed an endospore. Morphological and physiological characteristics suggested this microorganism a thermophilic Bacillus species, and was named as Bacillus sp. BCS-l. The production of D-stereospecific dipeptidase was growth-associated and optimal at $55^{\circ}C$. The enzyme was applied for the synthesis of D-amino acid-containing peptide, N-benzyloxycarbonyl-L-aspartyl-D-alanine benzyl ester (Z-L-Asp-D-AlaOBzl), as a model reaction. A thermodynamically controlled synthesis of Z-L-Asp-D-AlaOBzl was achieved in an organic solvent.

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Influence of Different Operational pH Conditions to Microbial Community in Biological Sequencing Batch Phosphorus Removal Process (생물학적 회분식 인 제거 공정에서 pH 영향과 미생물 군집의 변화)

  • Ahn, Johwan;Seviour, Robert
    • Journal of Korean Society on Water Environment
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    • v.29 no.4
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    • pp.459-465
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    • 2013
  • A sequencing batch reactor was operated under different pH conditions to see the influence of pH to microbial community in enhanced biological phosphorus removal (EBPR) systems. Long term influences of different steady-state pH conditions on the microbial community composition were evaluated by polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH). The shift in populations from polyphosphate-accumulating organisms (PAOs) to Alphaproteobacteria was observed when pH was changed from 7.5 to 7.0. Alphaproteobacteria with the typical morphological traits of tetrad-forming organisms (TFOs) eventually became dominant members. The alphaproteobacterial TFOs were the phenotype expected for glycogen-accumulating organisms (GAOs), which accumulate large amount of glycogen into the cell. The results strongly suggested that low operational pH condition encourages the appearance of the GAOs in EBPR process, significantly reducing the EBPR capacity.

Inhibition of Transglutaminase and Microbial Transglutaminase Activity by Garlic

  • Lee, Nam-Hyouck;Takeuchi, Atsuyoshi;Konno, Kunihiko
    • Food Science and Biotechnology
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    • v.16 no.2
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    • pp.223-227
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    • 2007
  • Ground garlic inhibited the cross-linking reaction of myosin and incorporation of monodansylcadaverine (MDC) in salted Alaska pollack surimi catalyzed by transglutaminase (TGase). The component responsible for the inhibition was a thermostable, low molecular weight compound. The component also inhibited microbial transglutaminase (MTGase). The inhibition by garlic was reversibly recovered upon addition of 2-mercaptoethanol. The inhibitory component was therefore hypothesized to contain sulfhydryl groups within its structure. Alliin itself did not inhibit the cross-linking reaction. However, the addition of alliin together with garlic increased the inhibition. This result suggested that compounds derived from alliin was responsible for the inhibition of TGase activity.