• Title/Summary/Keyword: Enzyme polymerization

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Acceptor reaction of a novel transfructosylating enzyme from Bacillus sp.

  • Kim, Yeong-Mi;Sinha, Jayanta;Park, Jong-Pil;Yun, Jong-Won
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.583-586
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    • 2000
  • Many different oligosaccharides were produced by transferring the fructose residue of sucrose to maltose, cellobiose, lactose and sucrose (self-transfer), where their yields of fructosylated acceptor products accounted for $26{\sim}30%$ (w/w). The maximum conversion yield (30%) was obtained in fructosyl cellobioside formation with 500 g sucrose/l (substrate) and 200 g cellobiose/l (acceptor). These four acceptors gave various products having DP (degree of polymerization) $2{\sim}7$ by successive transfer reactions.

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The Preparation and Identification of Hydrolysis Oligosaccharide from White Copra Meal by Yeast Fermentation and Sunflower Seed Enzymes

  • Park, Gwi-Gun
    • Preventive Nutrition and Food Science
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    • v.5 no.4
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    • pp.179-183
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    • 2000
  • $\beta$-1,4-Mannotriose was prepared b he enzymatic hydrolysis of white copra meal (WCM) and the subsequent elimination of monosaccharides from the resultant hydrolysate with a yeast. The enzyme system from sunflower seed hydrolyzed WCM and produced monosaccharides and $\beta$-1,4-mannotriose without other oligomers at the final stage of the reaction. WCM(50g) was hydrolyzed at 5$0^{\circ}C$ and pH 4.5 for 24 hr with crude enzyme solution (500 mL) from sunflower seed. By the elimination of monosaccharides from the hydrolysis products with a yeast (Candida glaebosa), 8.1 g of crystalline mannotriose was obtained without the use of chromatographic techniques. After 48hr of yeast cultivation, the total sugar content decreased from 4.6% to 3.5%, whereas the average degree of polymerization increased from 2.3 to 3.1.

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Studies on Naringinase Produced from Aspergillus nidulance -Part III. Preparation and Properties of Immobilized Naringinase- (Aspergillus nidulance가 생산하는 Naringinase에 관한 연구 -제 3 보 고정화(固定化) Naringinase의 제조 및 그 성질-)

  • Bai, Dong-Hoon;Pyun, Yu-Ryang;Yu, Ju-Hyun
    • Korean Journal of Food Science and Technology
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    • v.10 no.2
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    • pp.209-214
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    • 1978
  • Naringinase produced from Aspergillus nidulans was immobilized in acrylamide gel by the entrapping method and its characteristics were studied. Optimum acrylamide concentration was 10%, but N.N'-methylene bisacrylamide concentration had no influence on the final enzyme gel activity. The suitable amount of enzyme dissolved in the polymerization reaction mixture was 126 units/ml. Optimum pH of immobilized enzyme was 5.0 which was the same as that of free enzyme. However, immobilized enzyme showed a higher optumum reaction temperature, markedly increased pH and temperature stability. In a packed-column reactor, the observed reaction rate was increased proportionally to flow rate up to 5ml/min., but independent above 6ml/min.. Activation energy of the immobilized enzyme was 13.01 Kcal/mole, and the energy required for the thermal inactivation was 39.4 Kcal/mole. The apparent Km for 100 mesh gel was $7.23{\times}10^{-3}$ mole.

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Synthesis and Characterization of Fructooligosaccharides Using Levansucrase with a High Concentration of Sucrose

  • Seo Eun-Seong;Lee Jin-Ha;Choi Jae-Young;Seo Mi-Young;Lee Hee-Sun;Chang Seuk-Sang;Lee Hyung-Jong;Choi Jeong-Sik;Kim Doman
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.5
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    • pp.339-344
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    • 2004
  • A method for synthesizing branched fructo-oligosaccharides (BFOS) with a high concentration of sucrose ($1{\~}3$ M) was developed using levansucrase prepared from Leuconortoc mesenteroides B-1355C. The degree of polymerization of oligosaccharides synthesized according to the present method ranged from 2 to over 15. The synthesized BFOS were stable at a pH ranges of 2 to 4 under $120^{\circ}C$. The percentage of BFOS in the reaction digest was $95.7\%$ (excluding monosaccharides; $4.3\%$ was levan). BFOS reduced the insoluble glucan formation by Streptococcus sobrinus on the surfaces of glass vials or stainless steel wires in the presence of sucrose. They also reduced the growth and acid productions of S, sobrinus. Oligosaccharides can be used as sweeteners for foods such as beverages requiring thermo- and acid-stable properties and 3s potential inhibitors of dental caries.

Application of Polyaniline to an Enzyme-Amplified Electrochemical Immunosensor as an Electroactive Report Molecule

  • Kwon, Seong-Jung;Seo, Myung-Eun;Yang, Hae-Sik;Kim, Sang-Youl;Kwak, Ju-Hyoun
    • Bulletin of the Korean Chemical Society
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    • v.31 no.11
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    • pp.3103-3108
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    • 2010
  • Conducting polymers (CPs) are widely used as matrixes for the entrapment of enzymes in analytical chemistry and biosensing devices. However, enzyme-catalyzed polymerization of CPs is rarely used for immunosensing due to the difficulties involved in the quantitative analysis of colloidal CPs in solution phase. In this study, an enzyme-amplified electrocatalytic immunosensor employing a CP as a redox marker has been developed. A polyanionic polymer matrix, $\alpha$-amino-$\omega$-thiol terminated poly(acrylic acid), was employed for precipitation of CP. The acrylic acid group acts as a polyanionic template. The thiol terminus of the polymer was used to produce self-assembled monolayers (SAMs) on Au electrodes and the amine terminus was employed for immobilization of biomolecules. In an enzymeamplified sandwich type immunosensor, the polyaniline (PANI) produced enzymatically is attracted by the electrostatic force of the matrix polymer. The precipitated PANI was characterized by electrochemical methods.

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.

Molecular weight Control of Polyhydroxybutyrate (PHB) in Recombinant Escherichia coli (재조합 대장균에서의 Polyhydroxybutyrate (PHB)의 분자량 조절)

  • 심상준;안토니신스키
    • KSBB Journal
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    • v.13 no.1
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    • pp.96-100
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    • 1998
  • Two promoters (trc and P$\rho$) were inserted in PHA operon derived from Alcaligenes eutrophus to obtain high chain molecules of polyhydroxybutyrate (PHB) in recombinant Escherichia coli. Newly designed PHA operon was used to control the gene expressions of hydroxybutyric CoA and PHA polymerization, separately. Plasmids containing new synthetic operon was transformed into E. coli DH5$\alpha$ and analyzed for PHB production. Without induction of the PHA biosynthetic operon, PHA synthase which has low activity might supply low concentration of initiator during the polymerization reaction, resulting very high molecular weight of polymer. An increase of PHB average molecular weight was observed with decreased IPTG (isopropyl $\beta$ -Dithiogalactosidase) concentration. When no IPTG was added to the culture of E. coli DH5$\alpha$ /$\rho$ SJS1 which contained two promoters in PHA operon, high chain polymer having an average molecular weight of $2.5{\times}10^7$ was achieved. Analysis of the enzyme activities of PHA biosynthetic enzymes suggests that PHA synthase, the enzyme responsible for polymerizing 3-hydroxybutyric CoA, controls the molecular weight of PHB produced in vivo.

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Optimization of an Extracellular Dextranase Production from Lipomyces starkeyi KCTC 17343 and Analysis of Its Dextran Hydrolysates (Lipomyces starkeyi KCTC 17343에 의한 extracellular dextranase 최적생산과 덱스트란 hydrolysates 분석)

  • Chang, Yoon-Hyuck;Yeom, Joong-Hyun;Jung, Kyung-Hwan;Chang, Byung-Chul;Shin, Jung-Hee;Yoo, Sun-Kyun
    • Journal of Life Science
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    • v.19 no.4
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    • pp.457-461
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    • 2009
  • We optimized dextranase culture conditions by batch fermentation using Lipomyces starkeyi KCTC 17343. Furthermore, dextranase was purified by an ultra-membrane, and then dextran hydrolyzates were characterized. Cell growth and dextranase production varied depending on the initial culture pH and temperature. The conditions of optimal dextranase production were met in a pH range of 4-5 and temperature between $25-30^{\circ}C$. At optimal fermentation conditions, total enzyme activity and specific enzyme activity were about 4.85 IU/ml and 0.79 IU/g cells, respectively. The specific growth rate was examined to be $0.076\;hr^{-1}$. The production of dextranase in culture broth was very stably maintained after mid-log phase of growth. The enzyme hydrolyzed dextran into DP (degree of polymerization) 2 to 8 oligodextran series. Analysis of the composition of hydrolysates suggested that the enzyme produced is an endo-dextranase.

Degradation Characteristics of Ligninsulfonate by Laccase and Mn-peroxidase (Laccase와 Mn-peroxidase에 의한 Ligninsulfonate의 분해 특성)

  • Bae, Hyeun-Jong;Kim, Yoon-Soo;Mun, Sung-Phil
    • Journal of the Korean Wood Science and Technology
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    • v.23 no.4
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    • pp.27-32
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    • 1995
  • To understand whether ligninolytic enzyme catalyze polymerization or depolymerization of the high molecular weight (HMW) lignin, the action of laccase and Mn-peroxidase (MnP) towards commercial ligninsulfonates (LS) was examined in various conditions of pH and cosubstrates. Polymerization occurred when LS was incubated with laccase at pH 6.0. In contrast, the high molecular weight portions were significant1y reduced at pH 4.5, especially when glucose was added. When LS was treated with MnP at pH 4.5, compounds of low molecular weight were produced. In particular, when cellobiose was added to Mn-P reaction mixture, the breakdown of LS was observed. In conclusion, degradation of LS by laccase and MnP occurred primarily at pH 4.5 where-as polymerization of LS was dominant at pH 6.0. Color index, however, was not greatly changed in the degradation mixtures of LS.

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In vitro Polymerization and Copolymerization of Poly-3-hydroxypropionyl-CoA with the PHB Synthase from Ralstonia eutropha

  • Song, Jae-Jun;Goodwin, Steve;Lenz, Robert W.
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.48-51
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    • 2000
  • The poly(3-hydroxybutyrate) (PHB) synthase of Ralstonia. eutropha, which was produced by a recombinant strain E. coli and purified in one-step with a methyl-HIC column to a purity of more than 90%, was used to polymerize 3-hydroxypropionyl-CoA (3HPCoA) and to copolymerize 3HPCoA with 3-hydroxybutyryl-CoA (3HBCoA) in vitro. A $K_m$ of $189\;{\mu}M$ and a $k_{cat}$ of $10\;sec^{-1}$ were determined for the activity of the enzyme in the polymerization reaction of 3HPCoA based on the assumption that the dimer form of PHB synthase was the active form. Free coenzyme A was found to be a very effective competitive inhibitor for the polymerization of 3HPCoA with a $K_i$ of $85\;{\mu}M$. The maximum degree of conversion of 3HPCoA to polymer was less than 40 %. In the simultaneous copolymerization reactions of these two monomers, both the turnover number for the copolymerization reaction and the maximum degree of conversion of 3HPCoA and 3HBCoA to copolymers increased with an increase in the amount of 3HBCoA in the monomer mixture. However, the maximum conversion of 3HPCoA to a copolymer was less than 35 % regardless of the ratio of 3HPCoA to 3HBCoA. Block copolymers were obtained by the sequential copolymerization of the two monomers and these copolymers had a much narrower molecular weight distribution than those obtained by the simultaneous copolymerization of the same molar ratio of 3HPCoA and 3HBCoA.

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