Engineered bidirectional promoters enable rapid multi-gene co-expression optimization

Thomas Vogl, Thomas Kickenweiz, Julia Pitzer, Lukas Sturmberger, Astrid Weninger, Bradley W. Biggs, Eva-Maria Köhler, Armin Baumschlager, Jasmin Elgin Fischer, Patrick Hyden, Marlies Wagner, Martina Baumann, Nicole Borth, Martina Geier, Parayil Kumaran Ajikumar and Anton Glieder ()
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Thomas Vogl: Graz University of Technology
Thomas Kickenweiz: Graz University of Technology
Julia Pitzer: Austrian Centre of Industrial Biotechnology (ACIB GmbH)
Lukas Sturmberger: Austrian Centre of Industrial Biotechnology (ACIB GmbH)
Astrid Weninger: Graz University of Technology
Bradley W. Biggs: Manus Biosynthesis
Eva-Maria Köhler: Graz University of Technology
Armin Baumschlager: Graz University of Technology
Jasmin Elgin Fischer: Graz University of Technology
Patrick Hyden: Graz University of Technology
Marlies Wagner: Graz University of Technology
Martina Baumann: Austrian Centre of Industrial Biotechnology (ACIB GmbH)
Nicole Borth: Austrian Centre of Industrial Biotechnology (ACIB GmbH)
Martina Geier: Austrian Centre of Industrial Biotechnology (ACIB GmbH)
Parayil Kumaran Ajikumar: Manus Biosynthesis
Anton Glieder: Graz University of Technology

Nature Communications, 2018, vol. 9, issue 1, 1-13

Abstract: Abstract Numerous synthetic biology endeavors require well-tuned co-expression of functional components for success. Classically, monodirectional promoters (MDPs) have been used for such applications, but MDPs are limited in terms of multi-gene co-expression capabilities. Consequently, there is a pressing need for new tools with improved flexibility in terms of genetic circuit design, metabolic pathway assembly, and optimization. Here, motivated by nature’s use of bidirectional promoters (BDPs) as a solution for efficient gene co-expression, we generate a library of 168 synthetic BDPs in the yeast Komagataella phaffii (syn. Pichia pastoris), leveraging naturally occurring BDPs as a parts repository. This library of synthetic BDPs allows for rapid screening of diverse expression profiles and ratios to optimize gene co-expression, including for metabolic pathways (taxadiene, β-carotene). The modular design strategies applied for creating the BDP library could be relevant in other eukaryotic hosts, enabling a myriad of metabolic engineering and synthetic biology applications.

Date: 2018
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DOI: 10.1038/s41467-018-05915-w

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