• Title/Summary/Keyword: Lipase reaction

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Enzyme-Catalyzed Transesterification Processes in Organic Solvents (미생물 계면활성제에 관한 연구(제3보);유기용매에서 효소를 촉매로 한 에스테르교환반응)

  • Kim, Sang-Chun;Nam, Ki-Dae
    • Journal of the Korean Applied Science and Technology
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    • v.9 no.1
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    • pp.7-13
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    • 1992
  • Lipases catalyzed the transesterification reaction between esters and various primary and secondary alcohols in a 99% organic medium, porcine pancreatic, yeast, mold lipases can vigorously act as catalysts in a number of nearly anhydrous organic solvents. Various transesterification reactions catalyzed by porcine pancreatic lipase in hexane obey Michaelis-Menten kinetics. The dependence of the catalytic activity of the enzyme in organic media on the pH of the aqueous solution from which it was recovered is bell-shaped, with the maximum coinciding with the pH optimum of the enzymatic activity in water. The catalytic power exhibited by the lipases in organic solvents is comparable to that displayed in water. In addition to transesterification, lipases Can catalyze several other processes in organic media.

Rapid Detection of Virulence Factors of Aeromonas Isolated from a Trout Farm by Hexaplex-PCR

  • Nam, In-Young;Joh, Ki-Seong
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.297-304
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    • 2007
  • The detection of virulence factors of Aeromonas is a key component in determining potential pathogenicity because these factors act multifunctionally and multifactorially. In this study water samples were collected from a trout farm on a seasonal basis, and diseased fish and Aeromonas species were isolated and identified. For rapid detection of six virulence factors of isolated Aeromonas, a hexaplex-polymerase chain reaction (hexaplex-PCR) assay was used. The detected virulence factors include aerolysin (aer), GCAT (gcat), serine protease (ser), nuclease (nuc) lipase (lip) and lateral flagella (laf). The dominant strain found in our isolates was Aeromonas sobria, and the dominant virulence factors were aer and nuc for all seasons. We confirmed that A. sobria and two of the virulence genes (aer and nuc) are related. We proposed a method by which one can identify the major strains of Aeromonas: A. hydrophila, A. sobria, A. caviae, and A. veronii, using hexaplex-PCR.

Solid Bases as Racemization Catalyst for Lipase-catalyzed Dynamic Kinetic Resolution of Naproxen 2,2,2-Trifluoroethyl Thioester (리파아제에 의한 나프록센 2,2,2-트리플로로에틸 씨오에스터의 Dynamic Kinetic Resolution을 위한 라세미화 촉매로서의 고체 염기)

  • 김상범;원기훈;문상진;김광제;박홍우
    • KSBB Journal
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    • v.19 no.3
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    • pp.215-220
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    • 2004
  • A variety of solid bases such as inorganic bases, basic anion exchange resins, and resin-bound bases were tested as a catalyst for racemization of (S)-naproxen 2,2,2-trifluoroethyl thioester in isooctane at 45$^{\circ}C$. Among the various bases, DIAIOM WA30, which is a weakly basic anion exchange resin with a tertiary amine based on a highly porous type styrene-divinylbenzene copolymer, showed the highest catalytic activity. The second-order interconversion constant of DIAION WA30 was 8.6${\times}$10$\^$-4/ mM$\^$-1/h$\^$-1/ and about 3 times higher than that of trioctylamine under the same conditions. The rate of DIAION WA30-catalyzed racemization decreased with increasing an amount of water added to the reaction medium. Lipase-catalyzed kinetic resolution of racemic naproxen 2,2,2-trifluoroethyl thioester was successfully carried out under in situ racemization of substrate with DIAION WA30 in isooctane at 45$^{\circ}C$. More than 60% conversion and 99% enantiomeric excess for the desired (S)-naproxen product were obtained. Furthermore, such a solid base catalyst could be easily separated and reused in contrast to trioctylamine.

Scale- Up of Water-Oil Hydrolysis System

  • Hur, Byung-Ki;Kim, Eun-Ki
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.773-777
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    • 1999
  • Scale-up experiments for hydrolysis of beef tallow, fat, and palm kernel with lipase derived from Candida cylindracea were carried out in 1-1, 100-1, and 10,000-1 reactors. The optimum agitation speed for the hydrolysis of the 1-1 reactor was investigated and found to be 350rpm, and this was a basis for the scale-up of agitation speed. The hydrolysis system in this work was the oil-water system in which the hydrolysis seems to process a heterogeneous reaction. An emulsion condition was the most important factor for determining the reaction rate of hydrolysis. Therefore, the scale-up of agitation speed was performed by using the power n = 1/3 in an equation of the rules of thumb method. The geometrical similarity for scaling-up turned out to be unsatisfactory in this study. Thus, the working volume per one agitator was used for the scale-up. In the case of scale-up from a 1-1 reactor to a 100-1 reactor, the hydrolysis of palm kernel was very much scaled-up by initiating the rules of thumb method. However, the hydrolysis of fat and beef tallow in a 100-1 reactor was a little higher than that of the 1-1 reactor because of the difference of geometrical similarity. The scale-up of hydrolysis from the 100-1 reactor to the 10,000-1 reactor was improved compared to that of the 1-1 to 100-1 reactor. The present results indicated that the scale-up of hydrolysis in the oil-water system by the rules of thumb method was more satisfactory under the condition of geometrical similarity. Even in the case where geometrical similarity was not satisfactory, the working volume per one agitator could be used for the scale-up of a heterogeneous enzyme reaction.

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A study of the lipoprotein lipase inhibitory mechanism of Poncirus trifoliata water extracts (탱자 (Poncirus trifoliata)의 lipoprotein lipase 억제메커니즘)

  • Lee, Sung Mee;Kang, Yun Hwan;Kim, Kyoung Kon;Kim, Tae Woo;Choe, Myeon
    • Journal of Nutrition and Health
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    • v.48 no.1
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    • pp.9-18
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    • 2015
  • Purpose: Poncirus trifoliata has been reported to have anti-inflammatory, antioxidant, and immune activities. However, its anti-obesity activity and the mechanism by which the water extract of dried, immature fruit of Poncirus trifoliata (PF-W) acts are not clear. This study suggests a potential mechanism associated with the anti-obesity activity of PF-W. Methods: We measured the effect of PF-W on lipoprotein lipase (LPL) regulation using enzyme-linked immunosorbent assay (ELISA) and an activity assay. The LPL regulation mechanism was examined by reverse transcription polymerase chain reaction (RT-PCR) to measure the mRNA expression of biomarkers related to protein transport and by western blot for analysis of the protein expression of the transcription factor CCAAT-enhancer-binding protein ($C/EBP{\beta}$). Results: The total polyphenol and flavonoid content of PF-W was $52.15{\pm}4.02$ and $6.56{\pm}0.47mg/g$, respectively. PF-W treatment decreased LPL content in media to $58{\pm}5%$ of that in control adipocyte media, and increased LPL content to $117{\pm}3.5%$ of that in control adipocytes, but did not affect the mRNA expression of LPL. PF-W also increased the mRNA expression of sortilin-related receptor (SorLA), a receptor that induces endocytosis and intracellular trafficking of LPL, in a concentration- and time-dependent manner. Finally, cell fractionation revealed that PF-W treatment induced the expression of $C/EBP{\beta}$, a SorLA transcription factor, in the nuclei of 3T3-L1 adipocytes. Conclusion: The LPL secretion and activity assay showed PF-W to be an LPL secretion inhibitor, and these results suggest the potential mechanism of PF-W involving inhibition of LPL secretion through $C/EBP{\beta}$-mediated induction of SorLA expression.

Lipase-Catalyzed Reactions for Fats and Oils in Non-Polar Solvent (유기용매 내에서의 유지의 리파제 촉매반응)

  • Daeseok Han;Kwon, Dae-Young;Rhee, Joon-Shick
    • Microbiology and Biotechnology Letters
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    • v.16 no.3
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    • pp.250-258
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    • 1988
  • Lipases are well known as the enzymes which catalyze the hydrolysis of ester bonds combining aliphatic chains and glycerol on mono-, di- and triglycerides. Their reactions are characterized by be-ing heterogeneous and catalyzing the water-insoluble substrates. This property has been one of the Hurdles which delayed the application of lipases in fats and oils industry, However, with the development of biological reaction system of which organic solvent is introduced in part or whole as the reaction media, enzymatic manipulation of fats and oils is attracting increasing attention from the academic and industrial sectors. Trials in two-phase system and reversed micellar system to produce fatty acids through enzymatic hydrolysis of triglycerides preyed to be efficient in respect to volumetric productivity, fat hydrolysis rate, product separation, etc. In organic solvent system lipases have been found to have the ability to catalyze aminolysis, transesterification, esterification, thiotransesterification and oximolysis that are virtually impossible to catalyze in water. The organic solvent system is being extensively used in interesterifying glycerides to produce a fat with the modified physical and chemical nature.

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Synthesis of Cocoa Butter Alternative from Coconut Oil Fraction and Palm Oil Fractions by Lipase-Catalyzed Interesterification (Coconut Oil 분별유와 Palm 분별유로부터 효소적 Interesterification에 의한 코코아 버터 대체유지 합성)

  • Bae, Sang-Kyun;Lee, Kyung-Su;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.10
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    • pp.1487-1494
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    • 2010
  • Structured lipid (SL) for cocoa butter alternative was synthesized by interesterification of coconut oil fraction and palm stearin (6:4 and 8:2, by weight) in a shaking water bath at $60^{\circ}C$ and 180 rpm. It was performed for various reaction times (1, 2, 3, and 6 hr). The reaction was catalyzed by sn-1,3 specific Lipozyme TLIM (immobilized lipase from Thermomyces lanuginosus). SL-solid part was obtained from acetone fractionation at $0^{\circ}C$. SL-solid part was blended with other palm oils and fractions for desirable property of cocoa butter alternative (SL-solid part : palm middle fraction : palm stearin solid : palm oil, 70.4:18.4:2.9:8.3, by weight). In reversed-phase HPLC analysis, triacylglycerol species of cocoa butter alternative had partition number of 40 (10.77%), 42 (13.06%), 44~46 (17.38%) and 48 (51.88%). Major fatty acids of cocoa butter alternative were lauric acid (16.5%), myristic acid (12.28%), palmitic acid (46.03%), and linoleic acid (14.75%). Solid fat content (SFC) and polymorphic form (${\beta}'$ form) of cocoa butter alternative prepared were similar to those of commercial cocoa butter replacer (CBR).

Modification of Palm Mid Fraction with Stearic Acid by Enzymatic Acidolysis Reaction (효소적 Acidolysis를 이용한 Stearic Acid 함유 팜중부유의 개질)

  • Jeon, Mi-Sun;Lee, Yun-Jeung;Kang, Ji-Hyun;Lee, Jeung-Hee;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.4
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    • pp.479-485
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    • 2009
  • The acidolysis was performed to produce structured lipid with palm mid fraction (PMF) and stearic acid for 7, 24, and 36 hr at $70^{\circ}C$. The reaction was catalyzed by lipozyme TLIM (immobilized lipase from Thermonyces lanuginosa, amount of 10% and 20% by weight of total substrates) in the shaking water bath. The reaction conditions for maximum incorporation of stearic acid on the structured lipid were obtained when molar ratio of PMF and stearic acid was 1:2; concentration of lipozyme TLIM was 20wt%; reaction temperature was $70^{\circ}C$; and reaction time was 36 hr. After reaction under this condition, incorporation of stearic acid in the structured lipid was obtained up to 36.3% while the major components of triacylglycerol were 1,2-dipalmitoyl-3-stearoylglycerol (PPS, 28.19 area%), 1-palmitoyl-2-oleoyl-3-stearoylglycerol (POS/PSO, 20.70 area%) and 1-palmitoyl-2,3-distearoylglycerol (PSS, 18.13 area%). However, the fatty acid composition at the sn-2 position suggested that the positional specificity of lipozyme TLIM was not observed due to the acyl migration.

Recent Trends in Research of Polyglycerin Fatty Acid Esters (폴리글리세린지방산에스터의 최근 연구동향(제1보))

  • Rang, Moon Jeong
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.4
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    • pp.1443-1459
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    • 2019
  • Polyglycerol fatty acid ester nonionic surfactants have been used for a long time in foods and have been suggested as an alternative to the safety problems of PEG based nonionic surfactants. The polyglycerol fatty acid ester surfactants are synthesized by combining a hydrophilic polyglycerin and a lipophilic fatty acid. The hydrophilic polyglycerin is polymerized using glycerin, glycidol, epichlorohydrin, etc. The main issues of the polyglycerol polymerization reaction are to increase the content of the polyglycerol in the form of linear rather than branched or cyclic forms and to narrow the distribution of the degree of polymerization. The method of binding a lipophilic fatty acid group to a hydrophilic polyglycerin includes chemical synthesis such as esterification reaction and enzyme synthesis using lipase enzyme. The main issues of polyglycerin fatty acid ester synthesis are to increase the yield and to control the degree of esterification while reducing side reactions.

Assessment of Adipocyte Differentiation and Maturation-related Gene Expression in the Epididymal Fat of Estrogen Receptor α Knockout (ERαKO) Mouse during Postnatal Development Period

  • Cheon, Yong-Pil;Ko, CheMyong;Lee, Ki-Ho
    • Development and Reproduction
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    • v.24 no.4
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    • pp.287-296
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    • 2020
  • The absence of functional estrogen receptor α (Esr1) results in an overgrowth of the epididymal fat, as observed in estrogen receptor α knockout (ERαKO) mouse. The present research was aimed to evaluate expression of various molecules associated with adipocyte differentiation and maturation in the epididymal fat of ERαKO mouse at several postnatal ages by using quantitative real-time polymerase chain reaction. The highest transcript levels of all molecules were detected at 12 months of postnatal age, except leptin which the mRNA level was increased at 5 months of age and was unchanged until 12 months of age. The expression levels of CCAAT enhancer binding protein (Cebp) alpha, androgen receptor, and lipoprotein lipase were decreased at 5 months of age but increased at about 8 months of age. The mRNA levels of Cebp gamma and sterol regulatory element binding transcription factor 1 remained steady until 8 months of age. Continuous increases of transcript levels during postnatal period were found in Cebp beta, estrogen receptor (ER) beta, fatty acid binding protein 4, and delta like non-canonical Notch ligand 1. The increases of peroxisome proliferator-activated receptor gamma and adiponectin mRNA levels were detected as early as 8 months of age. The levels of fatty acid synthase and resistin transcript at 5 and 8 months of age were lower than that at 2 months of age. These findings show the aberrant expression patterns of genes related to adipocyte differentiation and maturation in the postnatal epididymal fat pad by the disruption of ER alpha function.