| Abstract | 第5-6页 |
| Chapter 1.General introduction | 第10-28页 |
| 1.1 Objectives | 第10页 |
| 1.2 Project background and significance | 第10-11页 |
| 1.3 Literature review | 第11-27页 |
| 1.3.1 Aspergillus niger | 第19-21页 |
| 1.3.2 Biochemistry of enzyme production by Aspergillus niger | 第21-22页 |
| 1.3.3 Mechanism of enzyme accumulation | 第22-23页 |
| 1.3.4 Mutagenesis of Aspergillus niger by Atmospheric and Room Temperature Plasma machine | 第23-24页 |
| 1.3.5 High-Throughput Screening as the best way to screen high yield strains of Aspergillus niger | 第24-27页 |
| 1.4 Purpose of this study | 第27-28页 |
| Chapter 2. General material and methods | 第28-36页 |
| 2.1 Strains and Media | 第28-34页 |
| 2.1.1 Main reagents used during experimental part | 第29-31页 |
| 2.1.2 The main instruments used in the experiment | 第31-32页 |
| 2.1.3 Main media formulation | 第32-33页 |
| 2.1.4 Main solutions | 第33-34页 |
| 2.2 Construction of Mutant Library | 第34页 |
| 2.3 48-MTPs Cultivation of Mutagenic Aspergillus niger Without Omissions | 第34页 |
| 2.4 Analysis of glucoamylase using p-NPG as a substrate | 第34-35页 |
| 2.5 Rapid analysis the pH of broth using methyl-orange | 第35-36页 |
| Chapter 3.Construction of mutant library | 第36-42页 |
| 3.1 Preparation of Aspergillus niger suspensions | 第38页 |
| 3.2 Mutants on the Petri dishes | 第38-39页 |
| 3.3 Mutants on the shake flask fermentation | 第39-40页 |
| 3.4 Determination of glucoamylase enzyme activity | 第40-42页 |
| Chapter 4. Development of a screening platform for Aspergillus niger mutants having increased production of glucoamylase | 第42-54页 |
| 4.1 Optimization of Aspergillus niger cultivation parameters needed to settle for the stable growing | 第42-47页 |
| 4.1.1 Optimizing the time of pre-culturing of Aspergillus niger | 第42-43页 |
| 4.1.2 Inoculum size | 第43-44页 |
| 4.1.3 Glass beads amount | 第44-45页 |
| 4.1.4 Shaking speed optimization | 第45页 |
| 4.1.5 Medium volume | 第45-47页 |
| 4.2 Culture of microorganism. 48-MTPs Cultivation of Mutagenic Aspergillus niger Without Omissions | 第47-50页 |
| 4.2.1 Growing single clone on solid PDA medium | 第47-48页 |
| 4.2.2 Growing single clone in liquid MDO3 medium | 第48-50页 |
| 4.2.2.1 Cultivation spores using two steps of the media: Start-MDO_3 | 第48页 |
| 4.2.2.2 Reducing one step using start medium for the pre-culturing | 第48-49页 |
| 4.2.2.3 Reducing the time of fermentation | 第49-50页 |
| 4.3 High-throughput system for screening | 第50-54页 |
| 4.3.1 Constructing High-Throughput model suitable for Aspergillus niger cultivation | 第52页 |
| 4.3.2 Optimization of Micro-Culture in 48-Deep Well MTPs | 第52-54页 |
| Chapter 5.Novel High-Throughput analyzing method | 第54-64页 |
| 5.1 Construction of High-Throughput Analyzing Method | 第54-61页 |
| 5.1.1 The reason why methyl-orange method is preferable | 第54-57页 |
| 5.1.2 Methyl -orange standard curve production | 第57-60页 |
| 5.1.3 Comparison the methyl-orange method to p-NPG method of detecting the activity of glucoamylase | 第60-61页 |
| 5.2 High-Throughput screening of A.niger with high yield of glucoamylase | 第61-64页 |
| Chapter 6.An additional High-Throughput screening platform for GOD production using A.niger as a producer | 第64-71页 |
| 6.1 High-Throughput cultivation | 第64-65页 |
| 6.1.1 Optimization of cultivation conditions on 48-deep well MTP | 第64页 |
| 6.1.2 Optimization of cultivation conditions on the flask | 第64-65页 |
| 6.2 High Throughput analyzing | 第65-68页 |
| 6.2.1 Comparison between centrifuge and filtration methods to obtain GOD production | 第65-66页 |
| 6.2.2 Spectrophotometry and High-Throughput analysis | 第66-68页 |
| 6.3 High-throughput screening of Aspergillus niger with high yield of GOD | 第68-71页 |
| Chapter 7. General conclusion and perspectives | 第71-74页 |
| References | 第74-82页 |
| Acknowledgements | 第82-83页 |
| List of publications | 第83页 |
