Technologies for Biosystems Engineering

Billerbeck, Sonja; Dietz, Sven; Morgado, Gaspar; Panke, Sven

doi:10.1007/978-94-007-4534-6_4

Cited by 2 publications

(1 citation statement)

References 107 publications

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“…In contrast, Escherichia coli is easy to engineer and has proven a flexible host for the production of commodity chemicals and fuels (Lee & Kim, ). The use of E. coli for heterologous production of propionic acid presents several advantages, including low nutrient requirements, a plethora of bibliomic data, and established engineering tools (Billerbeck, Dietz, Morgado, & Panke, ).…”

Section: Introductionmentioning

confidence: 99%

Engineering Escherichia coli for propionic acid production through the Wood–Werkman cycle

Gonzalez-Garcia

McCubbin

Turner

et al. 2019

Biotech & Bioengineering

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Native to propionibacteria, the Wood–Werkman cycle enables propionate production via succinate decarboxylation. Current limitations in engineering propionibacteria strains have redirected attention toward the heterologous production in model organisms. Here, we report the functional expression of the Wood–Werkman cycle in Escherichia coli to enable propionate and 1‐propanol production. The initial proof‐of‐concept attempt showed that the cycle can be used for production. However, production levels were low (0.17 mM). In silico optimization of the expression system by operon rearrangement and ribosomal‐binding site tuning improved performance by fivefold. Adaptive laboratory evolution further improved performance redirecting almost 30% of total carbon through the Wood–Werkman cycle, achieving propionate and propanol titers of 9 and 5 mM, respectively. Rational engineering to reduce the generation of byproducts showed that lactate (∆ldhA) and formate (∆pflB) knockout strains exhibit an improved propionate and 1‐propanol production, while the ethanol (∆adhE) knockout strain only showed improved propionate production.

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Section: Introductionmentioning

confidence: 99%

Engineering Escherichia coli for propionic acid production through the Wood–Werkman cycle

Gonzalez-Garcia

McCubbin

Turner

et al. 2019

Biotech & Bioengineering

View full text Add to dashboard Cite

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Synthetic Biology for Cell-Free Biosynthesis: Fundamentals of Designing Novel In Vitro Multi-Enzyme Reaction Networks

Morgado

Gerngross

Roberts

et al. 2016

Advances in Biochemical Engineering/Biotechnology

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Cell-free biosynthesis in the form of in vitro multi-enzyme reaction networks or enzyme cascade reactions emerges as a promising tool to carry out complex catalysis in one-step, one-vessel settings. It combines the advantages of well-established in vitro biocatalysis with the power of multi-step in vivo pathways. Such cascades have been successfully applied to the synthesis of fine and bulk chemicals, monomers and complex polymers of chemical importance, and energy molecules from renewable resources as well as electricity. The scale of these initial attempts remains small, suggesting that more robust control of such systems and more efficient optimization are currently major bottlenecks. To this end, the very nature of enzyme cascade reactions as multi-membered systems requires novel approaches for implementation and optimization, some of which can be obtained from in vivo disciplines (such as pathway refactoring and DNA assembly), and some of which can be built on the unique, cell-free properties of cascade reactions (such as easy analytical access to all system intermediates to facilitate modeling).

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Technologies for Biosystems Engineering

Cited by 2 publications

References 107 publications

Engineering Escherichia coli for propionic acid production through the Wood–Werkman cycle

Engineering Escherichia coli for propionic acid production through the Wood–Werkman cycle

Synthetic Biology for Cell-Free Biosynthesis: Fundamentals of Designing Novel In Vitro Multi-Enzyme Reaction Networks

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