• Title/Summary/Keyword: organic two-phase reaction

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Optimization of Catechol Production Using Immobilized Resting Cells of Pseudomonas putida in Aqueous/organic Two-phase System

  • Chae, Hee-Jeong;Yoo, Young-Je
    • Journal of Microbiology and Biotechnology
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    • v.7 no.5
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    • pp.345-351
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    • 1997
  • An aqueous/organic two-phase reaction system was applied to the production of catechol using immobilized resting cells of Pseudomonas putida CY 400. Water/ethyl ether system was used because of high partition coefficient of catechol and thus to reduce the product inhibition and degradation. Among the tested immobilization carriers, polyacrylamide gel gave the highest catechol productivity. The immobilization seemed to protect the cells against solvent toxicity. From the simulation of reaction conditions based on two-phase models, it was found that there was an optimum acetate concentration at fixed benzoate and cell concentrations for the catechol productivity. A lower phase volume ratio (lower fraction of organic phase) gave a higher productivity. However, the substrate conversion was low at low phase volume ratio.

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Synthesis of an Aspartame Precursor Using Thermolysin in Organic Two-Phase System (유기용매 이상계에서 Thermolysin에 의한 아스파탐 전구체 생산)

  • 이인영;안경섭;이선복
    • Microbiology and Biotechnology Letters
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    • v.20 no.1
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    • pp.61-67
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    • 1992
  • The synthesis of N-benzyloxycarbonyl-L-aspartyl-L-phenylalanine methyl ester(ZAPM), a precursor of aspartame, from N-benzyloxycarbonyl-L-aspartic acid(Z-Asp) and L-phenylalanine methyl ester hydrochloride(L-PM-HCl) was investigated in ethylacetate-MES buffer two-phase system using thermolysin. In organic two-phase system, the degree of spontaneous hydrolysis of L-PM. HCl was significantly reduced with increasing the volume ratio of organic to aqueous phase. Stability of thermolysin in organic two-phase system was found to be higher than that in MES buffer solution. More than 90% of initial enzyme activity was maintained after 10 days of incubation in case that the volume of organic phase was equal to that of buffer phase, while the half life of thermolysin was about 2 days in aqueous buffer solution. The results of partitioning of substrates and product in organic two-phase system showed that the difference in partition coefficients between substrates and product was maximum at pH 5.5. The optimal pH for 2-APM synthesis in organic two-phase system was found to be 5.5-5.8, which is consistent with the value expected from the partition experiments. As the concentration of substrates was increased the conversion yield of Z-APM was increased with concomitant reduction of L-PMqHC1 hydrolysis. In case that the concentration of L-PM-HCl and Z-Asp were 160 mM and 80 mM respectively, the conversion yield of Z-APM reached 90% after 28 hrs of reaction. The yield obtained at different volume ratio of organic phase compares well with the predicted equilibrium constant in biphasic system.

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Development of centrifugal extractor for organic phase extraction using a height controlled separation weir and a divert plate (분리 웨어의 상하 조절과 전형판을 이용한 유기상 원심추출기 개발)

  • 김영환;윤지섭;정재후;홍동희;박기용
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 1997.10a
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    • pp.515-518
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    • 1997
  • Resident time of the centrifugal extractor for organic phase extraction using a height controlled separator weir and a divert plate is the important factor that affects significantly the chemical material extraction and the productivity in the chemical and mechanical processes. In this paper, it describes the design of the device for extraction of an organic phase from radioactive wastes, and considers phase separating weir and divert disk, both being designed to be adjustable in their positions, for effectively separating an organic phase. A height-adjustable separating weir unit used for separating the organic phase from the aqueous phase using a phase separating weir and designed to control the height of the separating weir as desired so as to allow the weir to be positioned at a boundary layer between two separated phases. The centrifugal extractor controls satisfactorily the mixed reaction time of two phases within the separator regardless of the variations of the mixing ratio of the two phases and the rotating speed of the extractor, is designed to be adjustable in its position in the vertical direction, thus allowing the user to appropriately select the mixed reaction time of the two phases within the extractor as desired. From development of a centrifugal extractor, it can effectively recover such usable elements, and preferably reducing the output quantity of radioactive wastes.

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The Synthesis of Kyotorphin Derivative by $\alpha$-Chymotrypsin ($\alpha$-Chymotrypsin 을 이용한 Kyotorphin 유도체의 합성)

  • Jeon, Yu Jin;Kim, Se Gwon
    • Journal of the Korean Chemical Society
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    • v.38 no.6
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    • pp.449-455
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    • 1994
  • In order to obtain the basic data for synthetic studies of bioactive peptide using enzyme, Kyotorphin(analgesic peptide) derivative was synthesized from Ac-Tyr-OH and $Arg-NH_2$ by $\alpha-chymotrysin$ in two phase system(organic phase and aqueous phase). In effect of organic solvent on Kyotorphin derivative synthesis from Ac-Tyr-OH(10 mM) and $Arg-NH_2$ (20 mM), the synthesis in ethyl acetate system of organic solvents was higher than those in other organic solvents (n-butanol, n-hexane, dichloromethane and chloroform). The optimal conditions for the synthesis are as follows: enzyme conc., 10 ${\mu}M;$ reaction pH, 7.0; reaction temp., $35^{\circ}C$ ; the ratio of organic phase volume/aqueous phase volume $(\alpha)$, 15. Under the optimal conditions, the yield was 70.2%, and the reaction achieved to equilibrium after 24 hrs.

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Production of L-Tryptophan by Enzymatic Processes (효소공정에 의한 트립토판 생산)

  • 이인영;안경섭;김의환;이선복
    • Microbiology and Biotechnology Letters
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    • v.20 no.1
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    • pp.73-78
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    • 1992
  • - Enzymatic synthesis of L-tryptophan(Trp) using E. coli tryptophanase has been investigated. In order to reduce the substrate inhibition by indole and to increase the product yield of L-tryptophan three different approaches have been made in this work. First, indole was intermittently fed to the reaction mixture in order to control the indole concentration at lower level. When 15 mM of indole was used as a total amount of substrate, conversion yield of 80% has been obtained with intermittent feeding while only 20% of indole was converted into L-tryptophan by conventional batch operation, The second method employed in this work was the use of cyclohexane-phosphate buffer organic two-phase system. In this system, indole was mainly partitioned into the organic-solvent phase and therefore substrate inhibition was expected to be reduced. L-Tryptophan production in organic two-phase system was, however, unexpectedly lower than that obtained in aqueous buffer solution. As a third method cyclodextrins have been added to the aqueous reaction mixture. It was found that the addition of $\beta$-cyclodextrin enhanced the tryptophan synthesis noticeably while $\alpha$-cycfodextrin showed little effect on tryptophan production.

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Enzymatic Hydrolysis of Hydrophobic Triolein by Lipase in a Mone-phase Reaction System Containing Cyclodextrin; Reaction Characteristics

  • Lee, Yong-Hyun;Kim, Tae-Kwon;Shin, Hyun-Dong;Park, Dong-Chan
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.103-108
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    • 1998
  • A hydrophobic substrate triolein was hydrolyzed by lipase in a mono-phase reaction system containing cyclodextrin(CD) as emulsifier. The triolein was transformation to an emulsion-like state in the CD containing reaction system in contrast to the oil-droplet like state without CD due to the formation of an inclusion complex between the lipids and CDs. The hydyrolysis reaction increased substantially in the CD containing reaction system, and the optimum reaction conditions including the amount of lipase, ${\beta}$-CD concentration, and mixing ratio of triolein and ${\beta}$-CD, were determined. The performance of the enzyme reaction in a mono-phase reaction system was compared with that of a two-phase reaction system which used water immiscible hexane as the organic solvent. The role of a CD in the mono-phase reaction system was elucidated by comparing the degree of the inclusion complex formation with triolein and oleic acid, Km and Vmax values, and product inhibition by oleic aicd in aqueous and CD containing reaction systems. The resulting enhanced reaction seems to be caused by two phenomena; the increased accessibility of lipase to triolein and reduced product inhibition by oleic acid through the formation of an inclusion complex.

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Production of enantiopure epoxides by yeast epoxide hydrolase using a two-phase membrane bioreactor (한국생물공학회 정기총회 및 연구논문발표회 발표논문)

  • Choe, Won-Jae;Choe, Cha-Yong
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.145-148
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    • 2000
  • Large-scale resolution of epoxides by the yeast Rhodotorula glutinis was demonstrated in an aqueous/organic two-phase cascade membrane bioreactor. Due to the chemical instability and low solubility of epoxides in aqueous phases, an organic solvent was introduced into the reaction mixture in order to enhance resolution of epoxide. A cascade hollow-fiber membrane bioreactor was used (i) to minimize the toxicity of organic solvents towards the epoxide hydrolase of Rhodotorula glutinis, and (ii) to remove inhibitory amounts of formed diol from the yeast cell containing aqueous phase. Dodecane was selected as a suitable solvent and 1,2-epoxyhexane as a model substrate. By use of this membrane bioreactor, highly concentrated (0.9 M in dodecane) enantiopure (>98% ee) (S)-1,2-epoxyhexane (6.5 g, 30% yield) was obtained from its racemic mixture.

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Synthesis of Glycosides by Transglycosylation of $\alpha$-Amylase from Soluble Starch in Water-Organic Two Phase System (전분을 기질로 한 이상계에서 Amylase의 당전이반응에 의한 배당체의 합성)

  • 박종이;이재동;이태호;장경립
    • Korean Journal of Microbiology
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    • v.35 no.1
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    • pp.1-6
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    • 1999
  • Benzylalcohol-$\alpha$-glucoside (BG) was synthesized from soluble starch by transglycosylation of $\alpha$-amylase. Transglycosylation in water-organic two phase system containing 1% soluble starch as a glycosyl donor, 90% benzylalcohol as a glycosyl acceplor, 10% citrate buffer solulion (0.1 M, pH 5.0), and 10 unit of $\alpha$-amylase (Aspergilllw oryzae) was showed highcst efficiency. About 4 mg BG was obtained from 10 mg starch in reaction for 80 hrs at $40^{\circ}C$. Initially benzylalcohol-$\alpha$-maltoside Q3M) was major product, but as the reaction proceeded, it was hydrolyzed to glucose and BG. Finally the product of transglycosylation by $\alpha$-amylase was only BG. The both products did not show reducing powcr and hydrolyzed by $\alpha$-glucosidase and $\alpha$-amylase, respectively. The molecular wcights of both were estimated to be 270 and 432 by ES1-Mass, respectively.

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Biosynthesis of Unnatural Phospholipids by Phospholipase D: II. Effect of Organic Solvents on Transphosphatidylation (PhosBholipase D에 의한 비천연 인지방질의 합성: IIl 포스타티딜기 전이반응에 미치는 유기용매의 효과)

  • 정의호;이해익이상영
    • KSBB Journal
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    • v.6 no.3
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    • pp.281-288
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    • 1991
  • This research was carried to investigate the effects of several organic solvents on the enzymatic transphosphatidylation in emulsion and two-phase solvent systems. The solvents having a similar dielectric constant with diethylether were effective for the enzyme activity. Diethylether and butylacetate were the most effective solvents, when added 12-15%(v/v) and 10-40%(v/v), respectively, for the synthesis of phosphatidylglycerol, phosphatidylethyleneglycol and phosphatidylpropyleneglycol. In the emulsion system, the size of ovolecithin liposome was increased and the clearness of the phospholipid bilayer was reduced as increasing the diethylether concentration. In the twophase solvent system, the rapidest reaction was obtained when water-organic solvent ratio was close to 1. The ratio of aqueous phase. however, should be lowered to 37% to gain the sole product of transphosphatidy1ation, without phosphatidohydrolysis.

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Synthesis and Characterization of Photopolymerizable Liquid Crystalline Compounds Having Two Reactive Sites

  • Jang, Ki-Suk;Kang, Suk-Hoon;Chang, Ji-Young
    • Bulletin of the Korean Chemical Society
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    • v.28 no.10
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    • pp.1651-1655
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    • 2007
  • Rod-like polymerizable LC molecules having two different reactive groups, i.e. acryl and diacetylene groups were prepared. 4-Hydroxyphenyldiacetylenes were synthesized by the coupling reaction of 1-bromoalkynes with 4-ethynylphenol and then reacted with 4-(6-acryloyloxyalkyloxy)benzoic acid to give polymerizable LC molecules 4a-d. The mesomorphic properties of compounds 4a-d were investigated by differential scanning calorimetry, polarized optical microscopy and X-ray diffractometry. Compounds 4a-c exhibited smectic and nematic phases, but compound 4d having a longest alkyl tail among the series formed only a smectic phase. Photopolymerizability of acryl and diacetylene groups was investigated by IR spectroscopy. An anisotropic polymer film could be prepared by selective polymerization of acryl groups with 365 nm UV light in the presence of a photoinitiator (2,2-dimethoxy-2-phenylacetophenone). The subsequent reaction of diacetylene groups with 254 nm UV light disrupted the anisotropic structure, suggesting that these LC molecules could be used for imaging on the film.