• Title, Summary, Keyword: immobilized enzyme

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Activity and Stability of Immobilized Enzyme on Silk Sericin Bead

  • Oh, Hanjin;Lee, Ki Hoon
    • International Journal of Industrial Entomology
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    • v.27 no.2
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    • pp.329-332
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    • 2013
  • In present preliminary report, we showed the possibility of silk sericin (SS) in enzyme immobilization. SS beads were prepared and enzymes were immobilized on it. The specific activity of immobilized a-chymotrypsin retained more than 87% compared to the free enzyme. The immobilized a-chymotrypsin has better stability against ethanol especially those immobilized on SS beads coagulated in methanol. Immobilized trypsin and lipase had also comparable apparent activity compared to free enzyme. Our result indicates that SS could be a good candidate for enzyme immobilization support due to its hydrophilicity.

Continuous Production of Fructooligosaccharides Using Fructosyltransferase Immobilized on Ion Exchange Resin

  • Yun, Jong-Won;Song, Seung-Koo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.1 no.1
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    • pp.18-21
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    • 1996
  • A continuous production of fructooligosaccharides from sucrose was investigated by fructosyltransferase immobilized on a high porous resin, Diaion HPA25. The optimum pH(5.5) and temperature(55$^{\circ}C$) of the enzyme for activity was unaltered by immobilization, and the immobilized enzyme became less sensitive to the pH change. The optimal operation conditions of the immobilized enzyme column for maximizing the productivity were as follows: 600g/L of sucrose feed concentration, flow rate of superficial space velocity 2.7h-1. When the enzyme column was run at 50$^{\circ}C$, about 8% loss of the initial activity of immobilized enzyme was observed after 30 days of continuous operation, during which high productivity of 1174g/L$.$h was achieved. The kinds of products obtained using the immobilized enzyme were almost the same as those using soluble enzymes or free cells.

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Manufacturing of Enzyme Immobilized Sheet Using Carboxymethylated RMP Substrate (카르복시메틸화 RMP를 이용한 효소 고정화시트의 제조)

  • 조남석
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.35 no.2
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    • pp.39-45
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    • 2003
  • This study was performed to develop the new type enzyme immobilization sheet from carboxymethylated refiner mechanical pulp (CRMP) substrate. Enzyme immobilization was attempted to couple carboxyl groups of CRMP with amino groups of the enzyme, trypsin, through the reaction of carbodiimide reagent, 1-ethyl-3-(3-dimethyl aminopropyl)-carbodimide (EDC ). Immobilization carrier, water insoluble CRMP fraction (CRMP-IS), was successfully reacted with the enzyme, formed peptide linkage like -CONH- at 1680$cm^{-1}$ / and new ester linkage like -COO$CH_3$, methylester at 1735$cm^{-1}$ /, and produced enzyme immobilized substrate (CRMP-IST). The enzyme immobilized handsheet was prepared by mixing the above chelated enzyme immobilized substrate(CRMP-IST) with kraft pulp by paper sheet machine like papermaking process. The sheet weight and strength were increased with increasing dosage of CRMP-IST, and decreased at more than 10% mixing of CRMP-IST, but higher than the controls. Concerning activities of immobilized trypsin(CRMP-IST) sheet by caseinolysis, the teared-off sheet with shaking was shown higher enzyme activities than sheet shape without shaking. In conclusion, this enzyme immobilized sheet would be expected easy handling for practical application and reutilization.

Amplicilin biosynthesis by immobilized enzyme

  • Kim, Young-Sik;Ryu, Dewy-D.Y.
    • Archives of Pharmacal Research
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    • v.3 no.1
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    • pp.7-12
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    • 1980
  • Ampliciline was synthesized from 6-amino-pencillanic acid (6-APA) and D-.alpha. phenylglycine methyl ester by using amplicilin synthesizing enzyme from Peudomonas melanogenum (IAM 1655). The whole cell enzyme was immobilized by entrapping it in the polyacrylamide gel lattices. The polymer used in the enzyme entrapment was made from 150 mg per ml of acrylamide monomer and 8 mg per ml of N, N'-methylenebisacrylamide. About 200 mg/whole cell enzyme was mixed in the polymer for entrapment. The maximal activity retention after immobilization was 56%. The optimal pH values for the whole cell enzyme and the immobilized whole cell enzyme were 6.0 and 5.9, respectively. The optimal temperature for the enzyme activity were the same for both type of preparations. The enzyme stabilities against pH and heat increased for immobilized whole cell enzyme. Immobilized cell was more stable especially in the acidic condition while both type were found to be very suceptible to thermal inactivation at a temperature above 4.deg.C. The kinetic constants obtained from Lineweaver-Burk plot based on two substate reaction mechanism showed somewhat higher value for immobilized whole cell enzyme as compared to the whole cell enzyme : the Km value for 6-APA were 7.0 mM and 12.5 mM while Km values for phenylglycine methyl ester were 4.5 mM and 8.2 mM, respectively. Using the immobilized whole cell enzyme packed in a column reactor, the productivity of ampiciline was studied by varying the flow rate of substrate solution. At the space velocity, SV, 0.14 hr$^{-1}$ the conversion was 45%. Operational stability found in terms of half life was 30 hr at SV = 0.2 hr.

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Preparation and characterization of polymer-coated mesoporous silica nanoparticles and their application in Subtilisin immobilization

  • Ozbek, Belma;Unal, Sule
    • The Korean Journal of Chemical Engineering
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    • v.34 no.7
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    • pp.1992-2001
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    • 2017
  • The preparation and characterization of polymer-coated mesoporous silica nanoparticles (MSNs) and their application in Subtilisin (Alcalase$^{(R)}$) immobilization were investigated. For the synthesis of polymer-coated MSNs, acrylic acid (AA) and chitosan (CS) mixture were blended as poly(acrylic acid) (PAA) and CS polymer layer onto MSNs via in-situ polymerization technique. Then, both uncoated MSNs and polymer-coated mesoporous silica nanoparticles (CS-PAA/MSNs) were characterized by taking into account properties such as morphologic pattern, size distribution, surface charge of the particles as well as thermogravimetric stability with SEM, TEM, Zetasizer and TGA analyses. Subtilisin was immobilized onto polymer-coated mesoporous silica nanoparticles via adsorption technique. For optimizing the enzyme immobilization process, the percent enzyme loading depending on the matrix amount, immobilization time and pH were investigated. Then, the activity values of immobilized enzyme and free enzyme were compared at various pH and temperature values. The maximum enzyme activity was achieved at pH 9.0 for both immobilized and free enzyme. Immobilized enzyme showed more stability at higher temperatures compared with free enzyme. Furthermore, the operational and storage stability of immobilized enzyme were determined. The activity of immobilized enzyme was reduced from 100% to 45.83% after five repeated uses. The storage stability of immobilized enzyme was found to be higher than that of free enzyme. The activity of immobilized enzyme was reduced from 100% to 60% after 28 days of storage time. We concluded that the polymer-coated MSNs were a suitable matrix for Subtilisin immobilization compared to uncoated MSNs.

Properties of Penicillin Amidohydrolase Immobilized on Nylon Fiber

  • B. L. Seng;Iw-Han Cho;J. S. Rhee;Dewey D. Y. Ryu
    • Bulletin of the Korean Chemical Society
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    • v.1 no.1
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    • pp.10-17
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    • 1980
  • Penicillin amidohydrolase was partially purified from the fermented broth of Bacillus megaterium, and was immobilized on nylon fiber. The surface area of nylon fiber was increased by roughening it with fine sand and activated by acid treatment. The free amino groups on the nylon fiber exposed by such treatment were then utilized to immobilize the penicillin amidase. Enzymatic properties of penicillin amidohydrolase immobilized on the nylon fiber by covalent bonding and cross linking with glutaraldehyde were studied and compared with those of soluble enzyme. The optimal pH and temperature profile of immobilized enzyme showed only slightly broader peaks, and the values of kinetic constants, $K_m$, $K_{ia}$, and $K_{ip}$, of the immobilized enzyme are only slightly greater than those of the soluble enzyme. These results suggest that the mass transfer effect on the reaction rate for the penicillin amidase immobilized on nylon fiber is not so significant as the enzyme immobilized on some other support material like bentonite. The experimental results of batch reaction agreed well with the results of computer simulation for both the immobilized and soluble enzyme systems, confirming the validity of the rate equation derived which was based on the combined double inhibition by two reaction products.

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Biodegradation of Hydrocarbon Contamination by Immobilized Bacterial Cells

  • Rahman Raja Noor Zaliha Abd.;Ghazali Farinazleen Mohamad;Salleh Abu Bakar;Basri Mahiran
    • Journal of Microbiology
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    • v.44 no.3
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    • pp.354-359
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    • 2006
  • This study examined the capacity of immobilized bacteria to degrade petroleum hydrocarbons. A mixture of hydrocarbon-degrading bacterial strains was immobilized in alginate and incubated in crude oil-contaminated artificial seawater (ASW). Analysis of hydrocarbon residues following a 30-day incubation period demonstrated that the biodegradation capacity of the microorganisms was not compromised by the immobilization. Removal of n-alkanes was similar in immobilized cells and control cells. To test reusability, the immobilized bacteria were incubated for sequential increments of 30 days. No decline in biodegradation capacity of the immobilized consortium of bacterial cells was noted over its repeated use. We conclude that immobilized hydrocarbon-degrading bacteria represent a promising application in the bioremediation of hydrocarbon-contaminated areas.

Immobilization of Hansenula polymorpha Alcohol Oxidase for Alcohol Biosensor Applications

  • Chung, Hyun-Jung;Cho, Hyun-Young;Kong, Kwang-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.30 no.1
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    • pp.57-60
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    • 2009
  • Alcohol oxidase catalyzes the oxidation of short lines alcohol to aldehyde. In this study, alcohol oxidase from Hansenula polymorpha (HpAOD) was induced by addition of 0.5% methanol as the carbon source and purified to electrophoretic homogeneity by column chromatographies. The purified HpAOD was immobilized with DEAE-cellulose particles and its biochemical properties were compared with those of free enzyme. The substrate specificity and the optimum pH of immobilized enzyme were similar to those of free enzyme. On the other hand, the Km values of free and immobilized enzymes for ethanol were 6.66 and 14.65 mM, respectively. The optimum temperature for free enzyme was ${50^{\circ}C}$, whereas that for immobilized enzyme was ${65^{\circ}C}$. Immobilized enzyme showed high stability against long storage. Immobilized enzyme was also tested for the enzymatic determination of ethanol by the colorimetric method. We detected 1 mg/liter ethanol ($1{\times}10^{-4}$% ethanol) by 2,6- dichloroindophenol system. Therefore, the present study demonstrated that immobilized HpAOD has high substrate specificity toward ethanol and storage stability, which may be of considerable interest for alcohol biosensor and industrial application.

Operation Modes Can Affect the Activity of Immobilized Enzyme onto Silk Fibroin Nanofibrous Membrane

  • Oh, Hanjin;Lee, Ki Hoon
    • International Journal of Industrial Entomology
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    • v.27 no.2
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    • pp.322-325
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    • 2013
  • In the present study, we report that the selection of operation mode is important to take the full advantage of nanofibrous membrane in enzyme immobilization. Silk fibroin nanofibrous membrane has been prepared by electrospinning, and a-chymotrypsin was immobilized as a model enzyme. When the immobilized enzyme was operated in the membrane reactor mode, the Michaelis constant, Km, was lower and the Vmax was higher compared to the batch reactor mode. No concentration gradient was observed in the membrane reactor mode and the immobilized enzyme was stable even after 7 times of re-use. Our results suggests that the enzyme immobilized nanofibrous membrane should be operated in the membrane reactor mode rather than in the bath reactor mode.

Hydrolysis of Oils by Using Immobilized Lipase Enzyme : A Review

  • Murty, V.Ramachanda;Bhat, Jayadev;Muniswaran, P.K.A.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.2
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    • pp.57-66
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    • 2002
  • This review focuses on the use of immobilized lipase technology for the hydrolysis of oils. The importance of lipase catalyzed fat splitting process, the various immobilization procedures, kinetics, deactivation kinetics, New immobilized lipases for chiral resolution, reactor configurations, and process considerations are all reviewed and discussed.