Phosphate-inducible poly-hydroxy butyrate production dynamics in CO<inf>2</inf> supplemented upscaled cultivation of engineered Phaeodactylum tricornutum

Windhagauer, M; Abbriano, RM; Pittrich, DA; Doblin, MA

Phosphate-inducible poly-hydroxy butyrate production dynamics in CO<inf>2</inf> supplemented upscaled cultivation of engineered Phaeodactylum tricornutum

Windhagauer, M Abbriano, RM Pittrich, DA Doblin, MA

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Publisher:: SPRINGER
Publication Type:: Journal Article
Citation:: Journal of Applied Phycology, 2022, 34, (5), pp. 2259-2270
Issue Date:: 2022-10-01

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Field	Value	Language
dc.contributor.author	Windhagauer, M
dc.contributor.author	Abbriano, RM
dc.contributor.author	Pittrich, DA
dc.contributor.author	Doblin, MA
dc.date.accessioned	2023-06-05T03:35:11Z
dc.date.available	2023-06-05T03:35:11Z
dc.date.issued	2022-10-01
dc.identifier.citation	Journal of Applied Phycology, 2022, 34, (5), pp. 2259-2270
dc.identifier.issn	0921-8971
dc.identifier.issn	1573-5176
dc.identifier.uri	http://hdl.handle.net/10453/170627
dc.description.abstract	Diatoms such as Phaeodactylum tricornutum are emerging as sustainable alternatives to traditional eukaryotic microbial cell factories. In order to facilitate a viable process for production of heterologous metabolites, a rational genetic design specifically tailored to metabolic requirements as well as optimised culture conditions are required. In this study we investigated the effect of constitutive and inducible expression of the heterologous poly-3-hydroxybutyrate (PHB) pathway in P. tricornutum using non-integrative episomes in 3 different configurations. Constitutive expression led to downregulation of at least one individual gene out of three (phaA, phaB and phaC) and was outperformed by inducible expression. To further asses and optimise the dynamics of PHB accumulation driven by the inducible alkaline phosphatase 1 promoter, we upscaled the production to lab-scale bioreactors and tested the effect of supplemented CO2 on biomass and PHB accumulation. While ambient CO2 cultivation resulted in a maximum PHB yield of 2.3% cell dry weight (CDW) on day 11, under elevated CO2 concentrations PHB yield peaked at 1.7% CDW on day 8, coincident with PHB titres at 27.9 mg L−1 that were approximately threefold higher than ambient CO2. With other more valuable bio-products in mind, these results highlight the importance of the genetic design as well as substrate availability to supply additional reduction equivalents to boost biomass accumulation and relieve potential enzymatic bottlenecks for improved product accumulation.
dc.language	English
dc.publisher	SPRINGER
dc.relation.ispartof	Journal of Applied Phycology
dc.relation.isbasedon	10.1007/s10811-022-02795-y
dc.rights	info:eu-repo/semantics/openAccess
dc.subject	0607 Plant Biology, 0704 Fisheries Sciences, 1002 Environmental Biotechnology
dc.subject.classification	Marine Biology & Hydrobiology
dc.title	Phosphate-inducible poly-hydroxy butyrate production dynamics in CO<inf>2</inf> supplemented upscaled cultivation of engineered Phaeodactylum tricornutum
dc.type	Journal Article
utslib.citation.volume	34
utslib.for	0607 Plant Biology
utslib.for	0704 Fisheries Sciences
utslib.for	1002 Environmental Biotechnology
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - C3 - Climate Change Cluster
utslib.copyright.status	open_access	*
dc.date.updated	2023-06-05T03:35:10Z
pubs.issue	5
pubs.publication-status	Published
pubs.volume	34
utslib.citation.issue	5

Abstract:

Diatoms such as Phaeodactylum tricornutum are emerging as sustainable alternatives to traditional eukaryotic microbial cell factories. In order to facilitate a viable process for production of heterologous metabolites, a rational genetic design specifically tailored to metabolic requirements as well as optimised culture conditions are required. In this study we investigated the effect of constitutive and inducible expression of the heterologous poly-3-hydroxybutyrate (PHB) pathway in P. tricornutum using non-integrative episomes in 3 different configurations. Constitutive expression led to downregulation of at least one individual gene out of three (phaA, phaB and phaC) and was outperformed by inducible expression. To further asses and optimise the dynamics of PHB accumulation driven by the inducible alkaline phosphatase 1 promoter, we upscaled the production to lab-scale bioreactors and tested the effect of supplemented CO2 on biomass and PHB accumulation. While ambient CO2 cultivation resulted in a maximum PHB yield of 2.3% cell dry weight (CDW) on day 11, under elevated CO2 concentrations PHB yield peaked at 1.7% CDW on day 8, coincident with PHB titres at 27.9 mg L−1 that were approximately threefold higher than ambient CO2. With other more valuable bio-products in mind, these results highlight the importance of the genetic design as well as substrate availability to supply additional reduction equivalents to boost biomass accumulation and relieve potential enzymatic bottlenecks for improved product accumulation.

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http://hdl.handle.net/10453/170627