Biosynthesis of D-ribose by transketolase-deficient Bacillus subtilis PLK1B = Transketolase 결핍 균주인 Bacillus subtilis PLK1B에 의한 D-ribose 생합성

목차 (Table of Contents)

• Ⅰ. INTRODUCTION 1
• Ⅱ. MATERIALS AND METHODS 5
• 1. Bacterial strains 5
• 2. NTG chemical mutagenesis and selection of mutants 6
• 2.1 Method of NTG chemical mutagenesis 6
• 2.2 Selection of mutants 7
• 2.3 Stabilization of the final mutant 8
• 3. Culture medium and fermentations 11
• 3.1 Seed culture 11
• 3.2 Fermentation comparison of the final mutant with the parental strain 11
• 3.3 Effects of carbon source on D-ribose production 12
• 3.4 Effects of nitrogen source on D-ribose production 13
• 3.5 Effects of metal ions on D-ribose production 13
• 3.6 Determining the optimal concentration of CuCl2 in culture medium 14
• 3.7 Effects of ribitol on D-ribose production 15
• 3.8 Determining the optimal concentration of ribitol in culture medium 15
• 3.9 Conditions of jar-scale fermentation 16
• Ⅲ. RESULTS 18
• 1. Construction of mutant library through random mutagenesis 18
• 2. Stabilization of the final mutant B. subtilis PLK1B 24
• 3. Fermentation comparison of the final mutant and parental strain 26
• 4. Determining the optimal carbon source and nitrogen source 31
• 5. Effects of metal ion on D-ribose production 38
• 6. Determining the optimal concentration of CuCl2 for D-ribose production 42
• 7. Effects of ribitol on D-ribose production and determining the optimal concentration of ribitol in culture medium 46
• 8. Batch fermentation of B. subtilis PLK1B in jar fermentor 55
• 9. Fed-batch fermentation of B. subtilis PLK1B in jar fermentor 59
• IV. CONCLUSION 62
• V. REFERENCES 64
• 국문 요약 68
• APPENDIX 70
• Abstract 70
• 1. Introduction 72
• 2. Materials and Methods 75
• 2.1 Strains and plasmids 75
• 2.2 Primers 78
• 2.3 Genetic manipulation 81
• 3. Results 86
• 3.1 Comparison of glucose-PTS gene sequencing of B. subtilis PLK1B and B. subtilis 168 86
• 3.2 Construction of recombinant vector 92
• 4. Conclusion 95
• 5. References 96
• 국문요약 98