Computational identification of gene over-expression targets for metabolic engineering of taxadiene production
- 983 Downloads
Taxadiene is the first dedicated intermediate in the biosynthetic pathway of the anticancer compound Taxol. Recent studies have taken advantage of heterologous hosts to produce taxadiene and other isoprenoid compounds, and such ventures now offer research opportunities that take advantage of the engineering tools associated with the surrogate host. In this study, metabolic engineering was applied in the context of over-expression targets predicted to improve taxadiene production. Identified targets included genes both within and outside of the isoprenoid precursor pathway. These targets were then tested for experimental over-expression in a heterologous Escherichia coli host designed to support isoprenoid biosynthesis. Results confirmed the computationally predicted improvements and indicated a synergy between targets within the expected isoprenoid precursor pathway and those outside this pathway. The presented algorithm is broadly applicable to other host systems and/or product choices.
KeywordsTaxol Taxadiene Taxadiene synthase Over-expression E. coli Heterologous biosynthesis Metabolic engineering
- Boghigian BA, Myint M, Wu J, Pfeifer BA (2011a) Simultaneous production and partitioning of heterologous polyketide and isoprenoid natural products in an Escherichia coli two-phase bioprocess. J Ind Microbiol Biotechnol. doi:10.1007/s10295-011-0969-9
- Boghigian BA, Salas DF, Ajikumar PK, Stephanopoulos G, Pfeifer BA (2011b) Analysis of heterologous taxadiene production in K- and B-derived Escherichia coli. Appl Microbiol Biotechnol. doi:10.1007/s00253-011-3528-4
- Hahn FM, Hurlburt AP, Poulter CD (1999) Escherichia coli open reading frame 696 is idi, a nonessential gene encoding isopentenyl diphosphate isomerase. J Bacteriol 181:4499–4504Google Scholar
- Kacser H, Burns JA (1973) The control of flux. Symp Soc Exp Biol 27:65–104Google Scholar