Chaperone-Based Genetic Engineering and Fermentation Scale-up for High-Efficient ChCODH Production in Escherichia coli
- 주제(키워드) Escherichia coli , CODH , Carbon monoxide dehydrogenase , enzyme production , chaperone , fermenter
- 발행기관 서강대학교 일반대학원
- 지도교수 이진원
- 발행년도 2026
- 학위수여년월 2026. 2
- 학위명 석사
- 학과 및 전공 일반대학원 화공생명공학과
- 실제URI http://www.dcollection.net/handler/sogang/000000082369
- UCI I804:11029-000000082369
- 본문언어 영어
- 저작권 논문은 저작권에 의해 보호받습니다.
목차
1. Introduction 6
2. Materials and Methods 8
2.1. Bacterial strains and plasmids 8
2.2. Media composition 11
2.3. Culture conditions in flask cultivation for production of ChCODH 11
2.4. Culture conditions in lab-scale 5 L fermenter cultivation for production of ChCODH 12
2.5. ChCODH purification 12
2.6. ChCODH activity assay 14
2.7. Analytic method 14
2.8. RNA extraction and semi-quantitative RT-PCR for
transcriptional analysis 15
3. Results and Discussion 17
3.1. Effect of co-expressing chaperone protein for soluble ChCODH
production 17
3.1.1. Screening of an optimal chaperone system. 17
3.1.2. Validation of KJE system co-expression efficacy upon
transfer to IPTG-regulated plasmid 22
3.2. Scale-up to 5 L fermenter and identifying the optimal induction
cell density for efficient ChCODH production 25
3.3. Transcriptional analysis of chaperone genes to elucidate the
mechanism of enhanced ChCODH production at high cell density 35
4. Conclusion 38
5. Acknowledgement 39
6. References 40
