Tunable Gene Expression System Independent of Downstream Coding Sequence

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Fine control of the expression levels of proteins constitutes a major challenge in synthetic biology and metabolic engineering. However, the dependence of translation initiation on the downstream coding sequence (CDS) obscures accurate prediction of the protein expression levels from mRNA sequences. Here, we present a tunable gene-expression system comprising 24 expression cassettes that produce predefined relative expression levels of proteins ranging from 0.001 to 1 without being influenced by the downstream CDS. To validate the practical utility of the tunable expression system, it was applied to a synthetic circuit displaying three states of fluorescence depending on the difference in protein expression levels. To demonstrate the suitability of application to metabolic engineering, this system was used to diversify the levels of key metabolic enzymes. As a result, expression-optimized strains were capable of producing 2.25 g/L of cadaverine, 2.59 g/L of L-proline, and 95.7 mg/L of 1-propanol. Collectively, the tunable expression system could be utilized to optimize genetic circuits for desired operation and to optimize metabolic fluxes through biosynthetic pathways for enhancing production yields of bioproducts. This tunable system will be useful for studying basic and applied biological sciences in addition to applications in synthetic biology and metabolic engineering.
Publisher
AMER CHEMICAL SOC
Issue Date
2020-11
Language
English
Article Type
Article
Citation

ACS SYNTHETIC BIOLOGY, v.9, no.11, pp.2998 - 3007

ISSN
2161-5063
DOI
10.1021/acssynbio.0c00029
URI
http://hdl.handle.net/10203/277877
Appears in Collection
CBE-Journal Papers(저널논문)
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