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The Study of the Immobilization of Lipase Based on Biomimetic Mineralization

Author: MaCuiLi
Tutor: WangJingYun
School: Dalian University of Technology
Course: Biochemical Engineering
Keywords: Biomimetic Cytochrome C Protamine Dioxide Zirconia Entrapped lipase (RS) - phenylethanol
CLC: TQ925.6
Type: Master's thesis
Year: 2011
Downloads: 82
Quote: 0
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Abstract


Room temperature, atmospheric pressure, neutral pH conditions, the use of biological macromolecules template new way for enzyme immobilization induced the biomimetic synthetic inorganic oxide mineralization process. Lipases can not only in aqueous solution catalyzed hydrolysis of triglycerides, can be in the organic phase catalytic having a chiral selector of esterification and transesterification reactions, and having a wide range of applications in areas such as drug synthesis. In this paper, biomimetic process for lipase immobilized main content and the results are as follows: To investigate the role of different protein induce the synthesis of titanium oxide, zirconium the the Ti-BALDH and K2ZrF6 aqueous solution, which inducible protein cytochrome C carried out a detailed study, this is the first cytochrome C for biomimetic process synthesis of titania, zirconia. SEM, TEM, XRD showed that by the cytochrome C-induced synthesis of titania, zirconia, are amorphous spherical particles. By changing the morphology and size of the reaction system conditions such as pH, temperature regulation oxide particles on the cytochrome C induced a role in the synthesis of titanium oxide, zirconium oxide process. Bionic titanium process for lipase immobilized, immobilized the optimal conditions for: 5mg/mL fine protein inducing agent, pH 7.5, 0.05 mol / L phosphate buffer as a solvent, the amount of lipase 6mg/ml , 0.25mol/LTi-BALDH (titanium precursors), the immobilized enzyme encapsulation efficiency reached 70.1% and 20.3% of the activity recovery. The free enzyme and biomimetic titanium immobilized lipase optimum pH were pH 7.0 and pH 8.0, optimum temperature of 37 ℃ and 45 ° C, respectively, the immobilized enzyme at the same time shows good pH, temperature, storage and re-use stability. Biomimetic zirconium process for lipase immobilization, the optimum immobilization conditions for: 20mg/mL fine protein inducing agent, at pH6.5, 70mmol / L phosphate buffer, 5mg/mL lipase, 0.01 MK2ZrF6, enzyme embedded rate of 70%, immobilized enzyme activity 0.15U/mg. Zirconium bionic immobilized lipase temperature, pH and storage stability have been greatly improved, repeated 6 times, remains more than 60% of the initial enzyme activity of the immobilized lipase. Bionic zirconium immobilized lipase was used in the solvent of the organic phase catalytic (RS) - phenylethanol kinetic resolution. The optimal reaction conditions as follows: n-octane as a solvent, the substrate concentration of 0.1M (RS) - phenylethanol, 0.2M vinyl acetate, and the reaction volume 2mL, temperature 50 ℃, 70mg of the enzyme amount, speed 180rpm reaction 48h, this The conversion rate under the conditions and ees respectively to 49.9% and 99.9%, respectively.

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CLC: > Industrial Technology > Chemical Industry > Other chemical industries > Fermentation industry > Enzyme preparation ( enzyme ) > Lipase
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