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{Aw:2019:10.1002/bit.26901,
author = {Aw, R and Polizzi, KM},
doi = {10.1002/bit.26901},
journal = {Biotechnology and Bioengineering},
pages = {656--666},
title = {Biosensorassisted engineering of a highyield Pichia pastoris cellfree protein synthesis platform},
url = {http://dx.doi.org/10.1002/bit.26901},
volume = {116},
year = {2019}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Cellfree protein synthesis (CFPS) has recently undergone a resurgence partly due to the proliferation of synthetic biology. The variety of hosts used for cellfree extract production has increased, which harnesses the diversity of cellular biosynthetic, protein folding, and posttranslational modification capabilities available. Here we describe a CFPS platform derived from Pichia pastoris, a popular recombinant protein expression host both in academia and the biopharmaceutical industry. A novel ribosome biosensor was developed to optimize the cell extract harvest time. Using this biosensor we identified a potential bottleneck in ribosome content. Therefore, we undertook strain engineering to overexpress global regulators of ribosome biogenesis to increase in vitro protein production. CFPS extracts from the strain overexpressing FHL1 had a 3fold increase in recombinant protein yield compared to those from the wildtype X33 strain. Furthermore, our novel CFPS platform can produce complex therapeutic proteins, as exemplified by the production of human serum albumin to a final yield of 48.1 μg mL1. Therefore, this work not only adds to the growing number of CFPS systems from diverse organisms, but also provides a blueprint for rapidly engineering new strains with increased productivity in vitro that could be applied to other organisms.
AU  - Aw,R
AU  - Polizzi,KM
DO  - 10.1002/bit.26901
EP  - 666
PY  - 2019///
SN  - 0006-3592
SP  - 656
TI  - Biosensorassisted engineering of a highyield Pichia pastoris cellfree protein synthesis platform
T2  - Biotechnology and Bioengineering
UR  - http://dx.doi.org/10.1002/bit.26901
UR  - https://onlinelibrary.wiley.com/doi/full/10.1002/bit.26901
UR  - http://hdl.handle.net/10044/1/65179
VL  - 116
ER  -
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