| Chapter 1 General Introduction | 第1-12页 |
| Chapter 2 Research Background | 第12-27页 |
| ·chiral Drug | 第12-15页 |
| ·Definition of Chirality and Chiral drug | 第12页 |
| ·Stereoisomer Discrimination of chiral drug | 第12-14页 |
| ·Prospect of Chiral Drug | 第14-15页 |
| ·Synthesis of Chiral Drug by Non-Biological Method | 第15-17页 |
| ·Asymmetric Chemo-Synthesis | 第15页 |
| ·Chiral Poll Synthesis | 第15-16页 |
| ·Crystallization for Chiral Separation | 第16页 |
| ·Membrane-Based Separations | 第16页 |
| ·Chromatographic Techniques | 第16-17页 |
| ·Synthesis of Chiral Drug by Biological Method | 第17-20页 |
| ·Enzyme Resolution | 第17-18页 |
| ·Biotransformation | 第18-20页 |
| ·Importance of HPA | 第20-21页 |
| ·Synthesis of HPA | 第21-25页 |
| ·Synthesis of HPA in Industrial | 第21页 |
| ·Chromatographic Techniques | 第21-22页 |
| ·Crystallization for Separation of Racemic Mandelic | 第22页 |
| ·Enzyme Resolution | 第22-25页 |
| ·Biosynthesis of HPA through biotransformation | 第25页 |
| ·Innovation of the Subject | 第25-27页 |
| Chapter 3 Materials and Methods | 第27-37页 |
| ·Materials | 第27-28页 |
| ·Chemicals | 第27页 |
| ·Strains | 第27页 |
| ·Culture medium | 第27-28页 |
| ·Apparatus or Instruments | 第28页 |
| ·Biotransformation Methods | 第28-31页 |
| ·Screening of Microorganisms | 第28-29页 |
| ·UV Mutagenesis and Microvave Mutagenesis | 第29-30页 |
| ·Optimization of Culture Medium | 第30页 |
| ·Biosynthesis of HPA by Using Growing Cells | 第30-31页 |
| ·Biosynthesis of HPA by Using Permeabilized Cells | 第31页 |
| ·Optimization of Biotransformation Conditions | 第31页 |
| ·Dehydrogenase Purification Methods | 第31-33页 |
| ·Preparation of crude enzyme solution | 第31-32页 |
| ·Enzyme Purification | 第32-33页 |
| ·Analytic Methods | 第33-37页 |
| ·Determination of Cell Concentration by Spectrophotometer | 第33页 |
| ·Determination of Cell Density by Plate Counting | 第33页 |
| ·Determination of HPA Concentration by HPLC | 第33-34页 |
| ·Determination of Optical Rotation of HPA by Polarimeter | 第34页 |
| ·Determination of Optical Purity of HPA by HPCE | 第34-35页 |
| ·Determination of molecular weight of product by HPLC-MS | 第35页 |
| ·Determination of (R)/(S)-HPA Dehydrogenase Activity | 第35页 |
| ·Determination of Protein | 第35-37页 |
| Chapter 4 Asymmetric Biosynthesis of R-HPA by S.cerevisia | 第37-53页 |
| ·Introduction | 第37页 |
| ·Screening of Strains | 第37-38页 |
| ·Mutagenesis and Mutant Select | 第38-43页 |
| ·UV Mutagenesis | 第38-41页 |
| ·Microwave Mutagenesis | 第41-42页 |
| ·Genetic Stability | 第42-43页 |
| ·Optimization of Culture Medium | 第43-45页 |
| ·Influence of Different Carbon Sources | 第43页 |
| ·Influence of Glucose Concentration | 第43-44页 |
| ·Influence of Different Nitrogen Sources | 第44-45页 |
| ·Optimization of Bioreduction Conditions | 第45-49页 |
| ·Growth Curve | 第45页 |
| ·Influence of Substrate Concentration | 第45-46页 |
| ·Influence of Product Concentration | 第46页 |
| ·Influence of Shake Speed | 第46-48页 |
| ·Influence of Temperature | 第48页 |
| ·Influence of pH | 第48-49页 |
| ·Biotransformation Process in Optimal Conditions | 第49-52页 |
| ·Conclusions | 第52-53页 |
| Chapter 5 Asymmetric Synthesis of S-HPA by L.kefir | 第53-66页 |
| ·Introduction | 第53-54页 |
| ·Domestication of Strains with High Transforming Ability | 第54页 |
| ·Optimization of Culture Medium | 第54-58页 |
| ·Effect of Carbon Sources | 第54-56页 |
| ·Effect of Nitrogen Sources | 第56页 |
| ·Optimization Ratio of Carbon and Nitrogen Source | 第56-58页 |
| ·Optimization of Bioreduction Conditions | 第58-62页 |
| ·Growth Curve for Lk5.1 | 第58页 |
| ·Effect of Substrate Concentration | 第58-59页 |
| ·Effect of Product Concentration | 第59-60页 |
| ·Effect of Shake Speed | 第60-61页 |
| ·Effect of Temperature | 第61页 |
| ·Effect of pH | 第61-62页 |
| ·Biotransformation Process in Optimal Conditions | 第62-65页 |
| ·Conclusions | 第65-66页 |
| Chapter 6 Asymmetric Synthesis of R-HPA by Co-culture Sc10.9.48 with Lk5.1 | 第66-76页 |
| ·Introduction | 第66页 |
| ·Comparation of Co-culture and Pure Culture | 第66-68页 |
| ·Growth Characteristics | 第66-67页 |
| ·Inference of Substrate Concentrations | 第67-68页 |
| ·Mechanism Exploration | 第68-71页 |
| ·S-HPA isomerisation | 第69页 |
| ·Bioreduction Catalyzed by Permeabilized Cells | 第69-70页 |
| ·Addition of lactic acid | 第70-71页 |
| ·Optimisation of Bioreduction Conditions | 第71-74页 |
| ·Different Proportion of Inoculum s | 第71-72页 |
| ·Comparison of Aerobic/Anaerobic Condition | 第72页 |
| ·Influence of Temperature | 第72-73页 |
| ·Influence of pH Value | 第73-74页 |
| ·Biotransformation Process in Optimal Conditions | 第74-75页 |
| ·Conclusions | 第75-76页 |
| Chapter 7 Purification of HPA Dehydrogenase | 第76-96页 |
| ·Introduction | 第76页 |
| ·Standard Curve of Bradford | 第76页 |
| ·Enzyme Assay | 第76-79页 |
| ·R- HPA Dehydrogenase Purification | 第79-81页 |
| ·Preparation of Crude Enzyme Solution | 第79页 |
| ·Ammonium Sulphate Precipitation | 第79-80页 |
| ·DEAE-Sepharose | 第80-81页 |
| ·S- HPA Dehydrogenase Purification | 第81-84页 |
| ·Preparation of Crude Enzyme Solution | 第81-82页 |
| ·Ammonium Sulphate Precipitation | 第82-83页 |
| ·DEAE-Sepharose | 第83-84页 |
| ·Conclusions | 第84-96页 |
| Appendix | 第96-101页 |
| Appendix I HPLE for the separation of HPA and PGA in fermentation liquid | 第96-97页 |
| Appendix II HPLE for the separation of HPA and PGA in crude enzyme solution | 第97-98页 |
| Appendix III HPCE for the separation of R-HPA and S-HPA | 第98-99页 |
| Appendix IV Mass spectrum of HPA and PGA | 第99-101页 |
| Acknowledgements | 第101-102页 |
| Curriculum Vitae | 第102页 |
