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Synthetic Biology underpins advances in the bioeconomy
Biological systems - including the simplest cells - exhibit a broad range of functions to thrive in their environment. Research in the Imperial College Centre for Synthetic Biology is focused on the possibility of engineering the underlying biochemical processes to solve many of the challenges facing society, from healthcare to sustainable energy. In particular, we model, analyse, design and build biological and biochemical systems in living cells and/or in cell extracts, both exploring and enhancing the engineering potential of biology.
As part of our research we develop novel methods to accelerate the celebrated Design-Build-Test-Learn synthetic biology cycle. As such research in the Centre for Synthetic Biology highly multi- and interdisciplinary covering computational modelling and machine learning approaches; automated platform development and genetic circuit engineering ; multi-cellular and multi-organismal interactions, including gene drive and genome engineering; metabolic engineering; in vitro/cell-free synthetic biology; engineered phages and directed evolution; and biomimetics, biomaterials and biological engineering.
@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}
}
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 -