Tools and strategies for constructing cell-free enzyme pathways.

Petroll, K; Kopp, D; Care, A; Bergquist, PL; Sunna, A

Tools and strategies for constructing cell-free enzyme pathways.

Petroll, K Kopp, D Care, A

Bergquist, PL Sunna, A

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Publisher:: PERGAMON-ELSEVIER SCIENCE LTD
Publication Type:: Journal Article
Citation:: Biotechnol Adv, 2019, 37, (1), pp. 91-108
Issue Date:: 2019-01

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Field	Value	Language
dc.contributor.author	Petroll, K
dc.contributor.author	Kopp, D
dc.contributor.author	Care, A https://orcid.org/0000-0002-0035-7961
dc.contributor.author	Bergquist, PL
dc.contributor.author	Sunna, A
dc.date.accessioned	2022-03-25T04:27:58Z
dc.date.available	2018-11-20
dc.date.available	2022-03-25T04:27:58Z
dc.date.issued	2019-01
dc.identifier.citation	Biotechnol Adv, 2019, 37, (1), pp. 91-108
dc.identifier.issn	0734-9750
dc.identifier.issn	1873-1899
dc.identifier.uri	http://hdl.handle.net/10453/155557
dc.description.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.
dc.format	Print-Electronic
dc.language	eng
dc.publisher	PERGAMON-ELSEVIER SCIENCE LTD
dc.relation.ispartof	Biotechnol Adv
dc.relation.isbasedon	10.1016/j.biotechadv.2018.11.007
dc.rights	info:eu-repo/semantics/closedAccess
dc.subject	06 Biological Sciences, 09 Engineering, 10 Technology
dc.subject.classification	Biotechnology
dc.subject.mesh	Biocatalysis
dc.subject.mesh	Cell-Free System
dc.subject.mesh	Computer Simulation
dc.subject.mesh	Enzymes
dc.subject.mesh	Humans
dc.subject.mesh	Kinetics
dc.subject.mesh	Metabolic Engineering
dc.subject.mesh	Organic Chemicals
dc.subject.mesh	Synthetic Biology
dc.subject.mesh	Cell-Free System
dc.subject.mesh	Humans
dc.subject.mesh	Organic Chemicals
dc.subject.mesh	Enzymes
dc.subject.mesh	Kinetics
dc.subject.mesh	Computer Simulation
dc.subject.mesh	Biocatalysis
dc.subject.mesh	Synthetic Biology
dc.subject.mesh	Metabolic Engineering
dc.title	Tools and strategies for constructing cell-free enzyme pathways.
dc.type	Journal Article
utslib.citation.volume	37
utslib.location.activity	England
utslib.for	06 Biological Sciences
utslib.for	09 Engineering
utslib.for	10 Technology
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	closed_access	*
dc.date.updated	2022-03-25T04:27:56Z
pubs.issue	1
pubs.publication-status	Published
pubs.volume	37
utslib.citation.issue	1

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.

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http://hdl.handle.net/10453/155557