Abstract
Single enzyme systems or engineered microbial hosts have been used for decades but the notion of assembling multiple enzymes into cell-free synthetic pathways is a relatively new development. The extensive possibilities that stem from this synthetic concept makes it a fast growing and potentially high impact field for biomanufacturing fine and platform chemicals, pharmaceuticals and biofuels. However, the translation of individual single enzymatic reactions into cell-free multi-enzyme pathways is not trivial. In reality, the kinetics of an enzyme pathway can be very inadequate and the production of multiple enzymes can impose a great burden on the economics of the process. We examine here strategies for designing synthetic pathways and draw attention to the requirements of substrates, enzymes and cofactor regeneration systems for improving the effectiveness and sustainability of cell-free biocatalysis. In addition, we comment on methods for the immobilisation of members of a multi-enzyme pathway to enhance the viability of the system. Finally, we focus on the recent development of integrative tools such as in silico pathway modelling and high throughput flux analysis with the aim of reinforcing their indispensable role in the future of cell-free biocatalytic pathways for biomanufacturing.
| Language | English |
|---|---|
| Pages | 91-108 |
| Number of pages | 18 |
| Journal | Biotechnology Advances |
| Volume | 37 |
| Issue number | 1 |
| Early online date | 3 Dec 2018 |
| DOIs | |
| Publication status | Published - Jan 2019 |
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Keywords
- Synthetic biology
- In vitro biocatalysis
- Cell-free enzyme pathways
- Multi-enzyme cascades
- Pathway design
- Enzyme immobilisation
- Kinetic modelling
- Metabolite analysis
Cite this
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Tools and strategies for constructing cell-free enzyme pathways. / Petroll, Kerstin; Kopp, Dominik; Care, Andrew; Bergquist, Peter L.; Sunna, Anwar.
In: Biotechnology Advances, Vol. 37, No. 1, 01.2019, p. 91-108.Research output: Contribution to journal › Review article › Research › peer-review
TY - JOUR
T1 - Tools and strategies for constructing cell-free enzyme pathways
AU - Petroll,Kerstin
AU - Kopp,Dominik
AU - Care,Andrew
AU - Bergquist,Peter L.
AU - Sunna,Anwar
PY - 2019/1
Y1 - 2019/1
N2 - Single enzyme systems or engineered microbial hosts have been used for decades but the notion of assembling multiple enzymes into cell-free synthetic pathways is a relatively new development. The extensive possibilities that stem from this synthetic concept makes it a fast growing and potentially high impact field for biomanufacturing fine and platform chemicals, pharmaceuticals and biofuels. However, the translation of individual single enzymatic reactions into cell-free multi-enzyme pathways is not trivial. In reality, the kinetics of an enzyme pathway can be very inadequate and the production of multiple enzymes can impose a great burden on the economics of the process. We examine here strategies for designing synthetic pathways and draw attention to the requirements of substrates, enzymes and cofactor regeneration systems for improving the effectiveness and sustainability of cell-free biocatalysis. In addition, we comment on methods for the immobilisation of members of a multi-enzyme pathway to enhance the viability of the system. Finally, we focus on the recent development of integrative tools such as in silico pathway modelling and high throughput flux analysis with the aim of reinforcing their indispensable role in the future of cell-free biocatalytic pathways for biomanufacturing.
AB - Single enzyme systems or engineered microbial hosts have been used for decades but the notion of assembling multiple enzymes into cell-free synthetic pathways is a relatively new development. The extensive possibilities that stem from this synthetic concept makes it a fast growing and potentially high impact field for biomanufacturing fine and platform chemicals, pharmaceuticals and biofuels. However, the translation of individual single enzymatic reactions into cell-free multi-enzyme pathways is not trivial. In reality, the kinetics of an enzyme pathway can be very inadequate and the production of multiple enzymes can impose a great burden on the economics of the process. We examine here strategies for designing synthetic pathways and draw attention to the requirements of substrates, enzymes and cofactor regeneration systems for improving the effectiveness and sustainability of cell-free biocatalysis. In addition, we comment on methods for the immobilisation of members of a multi-enzyme pathway to enhance the viability of the system. Finally, we focus on the recent development of integrative tools such as in silico pathway modelling and high throughput flux analysis with the aim of reinforcing their indispensable role in the future of cell-free biocatalytic pathways for biomanufacturing.
KW - Synthetic biology
KW - In vitro biocatalysis
KW - Cell-free enzyme pathways
KW - Multi-enzyme cascades
KW - Pathway design
KW - Enzyme immobilisation
KW - Kinetic modelling
KW - Metabolite analysis
UR - http://www.scopus.com/inward/record.url?scp=85057740616&partnerID=8YFLogxK
U2 - 10.1016/j.biotechadv.2018.11.007
DO - 10.1016/j.biotechadv.2018.11.007
M3 - Review article
VL - 37
SP - 91
EP - 108
JO - Biotechnology Advances
T2 - Biotechnology Advances
JF - Biotechnology Advances
SN - 0734-9750
IS - 1
ER -