지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제21권 제6호
- 발행연도
- 2011.1
- 수록면
- 637 - 645 (9page)
이용수
초록· 키워드
오류제보하기
Production of recombinant proteins by excretory expression has many advantages over intracellular expression in Escherichia coli. Hyperexpression of a secretory exoglucanase,Exg, of Cellulomonas fimi was previously shown to saturate the SecYEG pathway and result in dramatic cell death of E. coli. In this study, we demonstrated that overexpression of the PspA in the JM101(pM1VegGcexL-pspA) strain enhanced excretion of Exg to 1.65 U/ml using shake-flask cultivation, which was 80% higher than the highest yield previously obtained from the optimized JM101(pM1VegGcexL)strain. A much higher excreted Exg activity of 4.5 U/ml was further achieved with high cell density cultivation using rich media. Furthermore, we showed that the PspA overexpression strain enjoyed an elevated critical value (CV), which was defined as the largest quotient between the intracellular unprocessed precursor and its secreted mature counterpart that was still tolerable by the host cells prior to the onset of cell death, improving from the previously determined CV of 20/80 to the currently achieved CV of 45/55 for Exg. The results suggested that the PspA overexpression strain might tolerate a higher level of precursor Exg making use of the SecYEG pathway for secretion. The reduced lethal effect might be attributable to the overexpressed PspA, which was postulated to be able to reduce membrane depolarization and damage. Our findings introduce a novel strategy of the combined application of metabolic engineering and construct optimization to the attainment of the best possible E. coli producers for secretory/excretory production of recombinant proteins, using Exg as the model protein.
목차
등록된 정보가 없습니다.
참고문헌 (22)
참고문헌 신청
Abramoff, M. D / 2004 / Image processing with ImageJ / Biophotonics Ind 11 : 36 ~ 42
DeLisa, M. P. / 2004 / Phage shock protein PspA of Escherichia coli relieves saturation of protein export via the Tat pathway / J. Bacteriol 186 : 366 ~ 373
Ferrer-Miralles, N. / 2009 / Microbial factories for recombinant pharmaceuticals / Microb. Cell Fact 8 : 17 ~
Fu, Z. B / 2005 / Cell death caused by hyper-expression of a secretory exoglucanase in Escherichia coli / Protein Expr. Purif 42 : 67 ~ 77
Fu, Z. B / 2006 / A two-stage refinement approach for the enhancement of excretory production of an exoglucanase from Escherichia coli / Protein Expr. Purif 48 : 205 ~ 214
DeLisa, M. P. / 2004 / Phage shock protein PspA of Escherichia coli relieves saturation of protein export via the Tat pathway / J. Bacteriol 186 : 366 ~ 373
Ferrer-Miralles, N. / 2009 / Microbial factories for recombinant pharmaceuticals / Microb. Cell Fact 8 : 17 ~
Fu, Z. B / 2005 / Cell death caused by hyper-expression of a secretory exoglucanase in Escherichia coli / Protein Expr. Purif 42 : 67 ~ 77
Fu, Z. B / 2006 / A two-stage refinement approach for the enhancement of excretory production of an exoglucanase from Escherichia coli / Protein Expr. Purif 48 : 205 ~ 214
이 논문의 저자 정보
최근 본 자료
댓글(0)
0