| 학회 | 한국화학공학회 |
| 학술대회 | 2005년 가을 (10/21 ~ 10/22, 인하대학교) |
| 권호 | 11권 2호, p.2052 |
| 발표분야 | 생물화공 |
| 제목 | A new method for cell-free production of recombinant proteins containing multiple disulfide bonds |
| 초록 | Due to its highly reducing environment, in general, cytoplasm of Escherichia coli does not allow the formation of disulfide bonds. Therefore, expression of recombinant proteins containing disulfide bonds has been a challenging task for biochemical engineers. In this study, we could produce active recombinant plasminogen activator in a modified cell-free protein synthesis system. A simple treatment of the S30 extract with oxidized glutathione abolished reductase activity while maintaining the translational activity. Addition of DsbC (an E.coli protein disulfide bond isomerase), GroEL/ES (an E.coli chaperonin) and the optimal concentration of glutathione buffer increased solubility and activity of the product. Approximately 15 μg/mL of active rPA was produced after a 3 hour incubation of the modified cell-free synthesis reaction. |
| 저자 | 김태완1, 오인석1, 금정원1, 안진호1, 박창길2, 권용찬2, 최차용1, 김동명2 |
| 소속 | 1서울대, 2충남대 |
| 키워드 | cell-free protein synthesis; disulfide bonds; recombinant plasminogen activator |
| 원문파일 | 초록 보기 |
