Combination of Entner-Doudoroff Pathway with MEP Increases Isoprene Production in Engineered Escherichia coli
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| Authors: | Huaiwei Liu Yuanzhang Sun Kristine Rose M Ramos Grace M Nisola Kris Ni?o G Valdehuesa Won–Keun Lee Si Jae Park Wook-Jin Chung |
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| Institution: | 1. Energy and Environment Fusion Technology Center, Department of Energy and Biotechnology, Myongji University, Cheoin-gu, Yongin-si, Gyeonggi-do, Republic of Korea.; 2. Division of Bioscience and Bioinformatics, Myongji University, Cheoin-gu, Yongin-si, Gyeonggi-do, Republic of Korea.; Infectious Disease Research Institute, United States of America, |
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| Abstract: | Embden-Meyerhof pathway (EMP) in tandem with 2-C-methyl-D-erythritol 4-phosphate pathway (MEP) is commonly used for isoprenoid biosynthesis in E. coli. However, this combination has limitations as EMP generates an imbalanced distribution of pyruvate and glyceraldehyde-3-phosphate (G3P). Herein, four glycolytic pathways—EMP, Entner-Doudoroff Pathway (EDP), Pentose Phosphate Pathway (PPP) and Dahms pathway were tested as MEP feeding modules for isoprene production. Results revealed the highest isoprene production from EDP containing modules, wherein pyruvate and G3P were generated simultaneously; isoprene titer and yield were more than three and six times higher than those of the EMP module, respectively. Additionally, the PPP module that generates G3P prior to pyruvate was significantly more effective than the Dahms pathway, in which pyruvate production precedes G3P. In terms of precursor generation and energy/reducing-equivalent supply, EDP+PPP was found to be the ideal feeding module for MEP. These findings may launch a new direction for the optimization of MEP-dependent isoprenoid biosynthesis pathways. |
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