| Acknowledgment | 第1-7页 |
| 摘要 | 第7-9页 |
| ABSTRACT | 第9-14页 |
| Chapter 1 General Introduction | 第14-28页 |
| ·Literature review | 第14-15页 |
| ·Carbon Utilization and Their Regulations in Methylotrophs | 第15-17页 |
| ·Three dimensional structures of pectinolytic enzymes | 第17页 |
| ·Protopectinases | 第17-21页 |
| ·Types of Protopectins | 第20页 |
| ·Characteristics of protopectins | 第20-21页 |
| ·Polygalacturonases | 第21-23页 |
| ·Types of Polygalacturonases | 第21-22页 |
| ·Characteristics of Polygalacturonases | 第22-23页 |
| ·Fermentation of Polygalacturonic acid | 第23-25页 |
| ·PL production | 第24-25页 |
| ·PGL production | 第25页 |
| ·Pichia pastoris | 第25-28页 |
| CHAPTER 2 Effect of two-stage pH control strategy on the polygalacturonic acid lyase productivity | 第28-40页 |
| ·Introduction | 第28-29页 |
| ·MATERIALS AND METHODS | 第29-32页 |
| ·Strain and plasmid | 第29页 |
| ·Media | 第29页 |
| ·Fermentation Conditions | 第29-30页 |
| ·Determination of dry cell weight | 第30页 |
| ·Determination of polygalacturonic acid lyase activity | 第30-31页 |
| ·Determination of residual methanol | 第31页 |
| ·Cell viability | 第31页 |
| ·Protease concentration | 第31-32页 |
| ·Determination of AOX activity | 第32页 |
| ·RESULTS | 第32-36页 |
| ·Impact of two stage pH control strategy on dry cell weight | 第32-33页 |
| ·Impact of two-stage pH control strategy on cell viability | 第33-34页 |
| ·Impact of two stage pH control on the PGL activity | 第34页 |
| ·Impact of two-stage pH control strategy on the residual methanol | 第34-35页 |
| ·Effect of pH on the protease activity in the fermentation culture | 第35-36页 |
| ·Effect of pH on the AOX activity of the yeast in fermentation culture | 第36页 |
| ·Discussions | 第36-39页 |
| ·Conclusion | 第39-40页 |
| CHAPTER 3 Effect of online methanol feeding strategy on the polygalacturonic acid lyase productivity during the induction phase | 第40-51页 |
| ·Introduction | 第40-41页 |
| ·MATERIALS AND METHOD | 第41-43页 |
| ·Strain and Plasmid | 第41页 |
| ·Media | 第41页 |
| ·Fermentation conditions | 第41-42页 |
| ·Analytical Methods | 第42页 |
| ·Determination of ADP, ATP and AMP | 第42-43页 |
| ·Results and discussions | 第43-50页 |
| ·Impact of online methanol induction phase on the DCW and cell viability | 第43-44页 |
| ·Effect on the protease activity of the medium | 第44-45页 |
| ·Effect of online methanol control strategy on polygalacturonic acid activity | 第45-47页 |
| ·Effect of online methanol control strategy on the phosphate related compounds of the yeast cell | 第47-49页 |
| ·Effect of two stage induction phase on the AOX gene | 第49-50页 |
| ·Conclusions | 第50-51页 |
| Chapter 4 Combined effect of both strategies on the polygalacturonic acid lyase productivity during the induction phase with a shift in the transition phase | 第51-64页 |
| ·Introduction | 第51页 |
| ·MATERIALS AND METHODS | 第51-53页 |
| ·Strain and Plasmid | 第51页 |
| ·Media | 第51-52页 |
| ·Fermentation conditions | 第52页 |
| ·Analytical Methods | 第52-53页 |
| ·Results and discussion | 第53-63页 |
| ·Effect of combined strategy on the cell biomass of the culture | 第53-55页 |
| ·Effect of combined strategy on the AOX activity of the yeast | 第55-56页 |
| ·Effect of combined strategy on the protease activity of the fermentation | 第56-57页 |
| ·Effect of combined strategy on the viability of the cells | 第57-58页 |
| ·Effect of combination strategy on the polygalacturonic acid lyase | 第58-60页 |
| ·Effect of combined strategy on the ATP levels of the fermentation broth | 第60-63页 |
| ·Conclusions | 第63-64页 |
| General Conclusions | 第64-66页 |
| Reference | 第66-74页 |
| Publications | 第74页 |
