Development of anaerobically inducible nar promoter expression vectors for the expression of recombinant proteins in Escherichia coli |
| |
Authors: | Kim Nag-Jong Choi Jong Hyun Kim Yeon Chul Lee Jongwon Lee Sang Yup Chang Ho Nam Lee Pyung Cheon |
| |
Institution: | a Department of Chemical and Biomolecular Engineering (BK21 Program), Bioinformatics Research Center, BioProcess Engineering Research Center and Center for Ultramicrochemical Process Systems, KAIST, 335 Gwahangno, Yuseong-gu, Daejeon 305-701, Republic of Korea b Metabolic and Biomolecular Engineering National Research Laboratory, Bioinformatics Research Center, BioProcess Engineering Research Center and Center for Ultramicrochemical Process Systems, KAIST, 335 Gwahangno, Yuseong-gu, Daejeon 305-701, Republic of Korea c The Institute of Biopharmaceuticals, LG Life Sciences, Ltd./R&D, 104-1, Munji-dong, Yuseong-gu, Daejeon 305-380, Republic of Korea d Department of Biochemistry, School of Medicine, Catholic University of Daegu, Daegu 705-034, Republic of Korea e Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology, 335 Gwahangno, Yuseong-gu, Daejeon 305-701, Republic of Korea f Institute for the BioCentury, Korea Advanced Institute of Science and Technology, 335 Gwahangno, Yuseong-gu, Daejeon 305-701, Republic of Korea g Department of Molecular Science and Technology, Ajou University, Woncheon-dong, Yeongtong-gu, Suwon 443-749, Republic of Korea |
| |
Abstract: | Dissolved oxygen (DO)-controlled nar promoter expression vectors were constructed, and their expression efficiency was compared with that of the T7 promoter pET22 expression vector by expressing human growth hormone (hGH), enhanced green fluorescence protein (EGFP), and β-tyrosinase in Escherichia coli cells. The nar promoter expression vector pRBS, which was engineered with a 5′-untranslated region and ribosomal binding site for the T7 promoter, expressed hGH at a rate of up to 32% of the total cellular proteins (TCP) in E. coli W3110narL−. The expression level of hGH was further enhanced, up to ∼42% of the TCP, by adding the N-terminal peptide tag of β-galactosidase to hGH, which was comparable to the expression of ∼43% of the TCP in pET-lac:hGH/BL21(DE3). A further engineered expression vector, pRBS(fnr), which coexpressed fumarate/nitrate reductase (fnr), expressed more EGFP than pET22 in BL21(DE3). In addition, recombinant β-tyrosinase was successfully expressed at a rate of up to ∼45% of the TCP in pRBS(fnr) in W3110narL−. From these results, the DO-controlled nar promoter system developed in this study can be considered a reliable and cost-effective expression system for protein production, especially in large-scale fermentation, as an alternative to the pET/BL(DE3) system. |
| |
Keywords: | Anaerobic expression nar promoter Expression vector |
本文献已被 ScienceDirect PubMed 等数据库收录! |
|