Combinational biosynthesis of isoprene by engineering the MEP pathway in Escherichia coli |
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Institution: | 1. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Avda. Rivadavia 1917, C1033AAJ Cuidad Autónoma de Buenos Aires, Argentina;2. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, Dr. Hector Torres, (INGEBI-CONICET), Vuelta de Obligado 2490, C1428ADN Buenos Aires, Argentina;3. School of Biosciences and Biotechnology, University of Camerino, 62032 Camerino (MC) Italy;4. Instituto de Genética Ewald A. Favret (CICVyA-INTA), De los reseros S/N, Castelar C25 (1712), Buenos Aires, Argentina |
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Abstract: | As an important feedstock in petrochemistry, isoprene is used in a wide range of industrial applications. It is produced almost entirely from petrochemical sources; however, these sources are being progressively depleted. A reliable biological process for isoprene production utilizing renewable feedstocks would be an industry-redefining development. There are two biosynthetic pathways producing isoprene: the mevalonate (MVA) pathway and the methyl erythritol 1-phosphate (MEP) pathway. In this study, the MEP pathway was modified in Escherichia coli BL21 (DE3) to produce isoprene. The isoprene synthase (IspS) gene chemically synthesized from Populus alba after codon optimization for expression in E. coli was heterologously expressed. The endogenous genes of 1-deoxy-d-xylulose-5-phosphate synthase (DXS) and 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR) were over-expressed. The isopentenyl pyrophosphate isomerase (Idi) gene from Streptococcus pneumoniae was exogenously over-expressed, and farnesyl diphosphate synthase (ispA) was weakened to enhance the yield. The control strain harboring empty plasmids did not emit any isoprene. The over-expression of the DXR gene only had little impact on the yield of isoprene. Idi from S. pneumoniae played a significant role in the improvement of isoprene production. The highest yield was achieved by an ispA-weakened DXS-IDI-IspS recombinant with 19.9 mg/l isoprene, which resulted in a 33-fold enhancement of the isoprene yield from the IspS recombinant. |
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Keywords: | Isoprene MEP pathway Isoprene synthase |
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