Abstract
Metabolic engineering has been an important approach for microbial bio-production. To produce bio-chemicals with engineered microorganisms, metabolic pathways have been edited using several common strategies, including gene disruption, gene overexpression, and gene attenuation. Here, we demonstrated metabolic channeling based on enzymatic metabolic enzyme ligation as a noteworthy approach for enhancing a desired metabolic flux. To achieve metabolic channeling , the metabolic enzymes should be in close proximity in cells. In the literature, several methodologies have been recently applied to achieve metabolic channeling . Meanwhile, we have proposed a strategy for possessing metabolic enzymes in close proximity, by utilizing sortase A as a stapler to tether such enzymes in Escherichia coli. By tethering metabolic enzymes that catalyze the reactions before and after a target metabolite, the metabolic flux may be enhanced. This chapter describes the approach for enhancing acetate-producing flux by sortase-A-assisted metabolic ligation in E. coli.
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Acknowledgment
This work was supported by Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Interdisciplinary Research Areas (Innovative Bioproduction Kobe), MEXT, Japan.
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Matsumoto, T., Tanaka, T., Kondo, A. (2018). Sortase A-Assisted Metabolic Enzyme Ligation in Escherichia coli for Enhancing Metabolic Flux. In: Braman, J. (eds) Synthetic Biology. Methods in Molecular Biology, vol 1772. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7795-6_6
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DOI: https://doi.org/10.1007/978-1-4939-7795-6_6
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