Imperial College London
Alternate

Professor Karen Polizzi

Faculty of Engineering, Department of Chemical Engineering

Professor of Biotechnology
 
 
 
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Contact

 

+44 (0)20 7594 2851k.polizzi

 
 
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Location

 

411ACE ExtensionSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Freemont:2021:10.1186/s12934-021-01604-4,
author = {Freemont, P},
doi = {10.1186/s12934-021-01604-4},
journal = {Microbial Cell Factories},
pages = {1--11},
title = {Refactoring of a synthetic raspberry ketone pathway with EcoFlex},
url = {http://dx.doi.org/10.1186/s12934-021-01604-4},
volume = {20},
year = {2021}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Background A key focus of synthetic biology is to develop microbial or cell-free based biobased routes to value-added chemicals such as fragrances. Originally, we developed the EcoFlex system, a Golden Gate toolkit, to study genes/pathways flexibly using Escherichia coli heterologous expression. In this current work, we sought to use EcoFlex to optimise a synthetic raspberry ketone biosynthetic pathway. Raspberry ketone is a high-value (~ £20,000 kg−1) fine chemical farmed from raspberry (Rubeus rubrum) fruit.Results By applying a synthetic biology led design-build-test-learn cycle approach, we refactor the raspberry ketone pathway from a low level of productivity (0.2 mg/L), to achieve a 65-fold (12.9 mg/L) improvement in production. We perform this optimisation at the prototype level (using microtiter plate cultures) with E. coli DH10β, as a routine cloning host. The use of E. coli DH10β facilitates the Golden Gate cloning process for the screening of combinatorial libraries. In addition, we also newly establish a novel colour-based phenotypic screen to identify productive clones quickly from solid/liquid culture.Conclusions Our findings provide a stable raspberry ketone pathway that relies upon a natural feedstock (L-tyrosine) and uses only constitutive promoters to control gene expression. In conclusion we demonstrate the capability of EcoFlex for fine-tuning a model fine chemical pathway and provide a range of newly characterised promoter tools gene expression in E. coli.
AU  - Freemont,P
DO  - 10.1186/s12934-021-01604-4
EP  - 11
PY  - 2021///
SN  - 1475-2859
SP  - 1
TI  - Refactoring of a synthetic raspberry ketone pathway with EcoFlex
T2  - Microbial Cell Factories
UR  - http://dx.doi.org/10.1186/s12934-021-01604-4
UR  - https://microbialcellfactories.biomedcentral.com/articles/10.1186/s12934-021-01604-4
UR  - http://hdl.handle.net/10044/1/90542
VL  - 20
ER  -
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