An engineered Escherichia coli having a high intracellular level of ATP and enhanced recombinant protein production |
| |
Authors: | Kim Hye-Jung Kwon Yeong Deok Lee Sang Yup Kim Pil |
| |
Institution: | (1) Department of Biotechnology, Catholic University of Korea, Bucheon, Gyeonggi, 420-743, Korea;(2) Department of Chemical and Biomolecular Engineering (BK21 Program), Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, KAIST, Daejon, 305-701, Korea;(3) Present address: Biotech R&D Center, Paik-Kwang Industrial Co., Kunsan, Cheonbuk, 573-879, Republic of Korea; |
| |
Abstract: | Artificial amplification of gluconeogenic phosphoenolpyruvate carboxykinase (PCK) under glycolytic conditions enables Escherichia coli to maintain a greater intracellular ATP concentration during its growth phase. To demonstrate the biotechnological benefit
of E. coli harboring a high intracellular ATP concentration, we compared the recombinant protein synthesis of a soluble protein (enhanced
green fluorescence protein, GFP) with that of a secretory protein (alkaline protease, AP), under control of the T7 promoter
in E. coli BL21(DE3) overexpressing PCK. According to the batch fermentations, the strain overexpressing PCK produced more GFP and AP
with a lower increase in biomass than the control strain. In a chemostat culture (D = 0.7 h−1), the GFP production in the PCK overexpressing strain was 99.0 ± 4.31 mg/g cell, with a biomass of 0.22 g/L, while that of
the control strain was 53.5 ± 3.07 mg/g cell, with a biomass of 0.35 g/L. These results indicate that the PCK overexpressing
E. coli strain harboring high intracellular levels of ATP can be useful as a protein-synthesizing host. The potential uses of the
strain and associated rationale are discussed. |
| |
Keywords: | |
本文献已被 PubMed SpringerLink 等数据库收录! |
|