Dissertation > Excellent graduate degree dissertation topics show

Penicillium Expansum TS414 Lipase in Pichia Pastori: Expression, Purification and Enantioselective Esterification of (R, S)-naproxen

Author: CaiZhiXiong
Tutor: TangLiangHua
School: Fujian Normal University
Course: Biochemistry and Molecular Biology
Keywords: Lipase Fermentation Purification Naproxen Immobilization Esterification Resolution
CLC: Q814
Type: Master's thesis
Year: 2011
Downloads: 14
Quote: 0
Read: Download Dissertation

Abstract


This thesis has taken the expression, purification of Penicillium expansum TS414 lipase in Pichia pastori and the recombinant Penicillium expansum TS414 lipase-catalyzed enantioselective esterification of (R,S)-naproxen in the orgnic media as the main lines and carried out the following researches:Firstly,the shake flask fermention conditons for X33-TS414-lip engineering bacterial were optimized.Conditions are as follows:pH:7.5;inoculated quantity:2OD,methanol concentration:0.75%,flask volume:10%,rotation speed:250rpm,temperature:30℃.Based on the conditons, three different methanol feeding strategies were carried out on the fermentation tank. The lipase’s activities of DO based strategy was 104U/ml,enzyme yield was 962U/l·h; chromotropic acid method to maintain a low concentration of methanol was 443U/ml,enzyme production rate was 4614.58U/l·h;Mixed feeding methanol-glycerol fed-batch under chromotropic acid method was 573 U/ml,enzyme production rate was 6821.43U/l·h.This optimized strategy resulted in the highest productivity (6821.43U/l·h),which is 5.51-fold higher than the DO-based strategy.Secondly, through three purification steps which are saturated ammonium sulfate precipitation, Sephacryl S-100 gel filtration chromatography and Phenyl Sepharose hydrophobic interaction chromatography, the recombinant Penicillium expansum TS414 lipase achieved electrophoretically pure,the ratio of enzyme activity from the initial 2988.03U/mg increased to 7909.74U/mg, which was increased 2.65 times, and its activity recovery was 39.69%.The results of the recombinant Penicillium expansum TS414 lipase were similar to the native lipase in kinetic resolution of (R S)-naproxen.Lastly,the esterification reaction for resolution of (R,S)-naproxen by the immobilized recombinant Penicillium expansum TS414 lipase was researched.The optimized conditions were as follows:1.Immobilization conditions:Tianjin Guangfu AB-8 macroporous resin,the resin and the enzyme mass ratio:1:8,pH:9.0,immobilized adsorption time:1.5h, lyophilizing:lh.2.Resolution of esterification reaction:the amount of immobilized enzyme was 26.7g/l,the amount of racemic naproxen was1.48mg, n-propanol concentration was 7.73μl,system’s water content was 0.05%.Under the optimum conditions reacted 120h, the percent conversion and enantiomeric selectivity respectively reached 48.27% and 98.01%. The effect of recombinant Penicillium expansum TS414 lipase-catalyzed enantioselective enantification of (R,S)-naproxen was better.

Related Dissertations

  1. Study on Protective Effects of Apple Polyphenol on Immune System Injury Caused by γ-Ray,S661.1
  2. Research on the Super-resolution Technologies of Polarimetric SAR Images,TN957.52
  3. Superresolution of Hyperspectral Images Based on Spatial-Spectral Information Coordination,TN911.73
  4. Research on Remote Sensing Image Processing Combining High Fidelity Compression and Resolution Enhancement,TP751
  5. Study on Wastewater Purification Effect of the Free Surface Wetland with Different Substrates and Plant Combination,X703
  6. Studies on Fermentation Optimization, Purification and Enzyme Characteristics of Lipase from Aspergillus Oryzae FS-1,TQ925.6
  7. Studies on Isolation, Purification and Chemical Structure for Polysaccharides from Camellia Chrysantha(Hu) Tuyama,S567.19
  8. Research of Glycosaminoglycan from Perna Virids Effect on Blood Lipid,R285.5
  9. Purification of Endogenous Proteases from (Litopenaeus Vannamei) Shrimp Head and Their Enzymology Properties,S985.21
  10. Directed Evolution of Lipase from Penicillium Expansum FS1884 by Error Prone PCR,Q78
  11. Study on the Preparation of the Low Molecular Weight Fuciodan from Sargassum Henslowianum (C.Agardh) and Anti-tumor Activity,TS254.9
  12. Under microaerobic conditions closed calcium carbide furnace exhaust COS gas adsorption purification,X781
  13. Studies on Extraction and Analysis of Fructooligosaccharides in Smallanthus Sonchifolius,TS255.1
  14. Research and Implementation of the Conflict Resolution in Process Modeling of Magic Platform,TP311.5
  15. Study on the Synthesis and Properties of Poly(1,4-Cyclohexyledimethylene Terephthalate) (PCT),TQ323.4
  16. Experiment Study of the Biogas Purification Process by Zeolite Filling Washing,TQ221.11
  17. Optimization of Fermentation Conditions, Purification, Cloning and Expression of a Cold-active Lipase from Pseudomonas Sp.RT-1,TQ925
  18. Breeding of Fungal α-amylase High-producing Strain by Genome Shuffling,TQ925
  19. Extraction, Purification and Antifungal Activity of Anthocyanins in Drop Fruits of Chinese Baberry,TQ914.1
  20. Research on Anaerobic Fermentation Processing Technology of Dead Tilapia,TS254.4

CLC: > Biological Sciences > Bioengineering ( Biotechnology ) > Enzyme Engineering
© 2012 www.DissertationTopic.Net  Mobile