Enhancement of cyanobacterial PHB production using random chemical mutagenesis with detection through FACS.

Price, S; Kuzhiumparambil, U; Pernice, M; Herdean, A; Ralph, P

Enhancement of cyanobacterial PHB production using random chemical mutagenesis with detection through FACS.

Price, S Kuzhiumparambil, U Pernice, M Herdean, A

Ralph, P

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Publisher:: Springer Science and Business Media LLC
Publication Type:: Journal Article
Citation:: Bioprocess Biosyst Eng, 2023, 46, (2), pp. 297-306
Issue Date:: 2023-02

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Field	Value	Language
dc.contributor.author	Price, S
dc.contributor.author	Kuzhiumparambil, U
dc.contributor.author	Pernice, M
dc.contributor.author	Herdean, A https://orcid.org/0000-0003-2143-0213
dc.contributor.author	Ralph, P
dc.date.accessioned	2023-02-15T21:12:11Z
dc.date.available	2022-12-07
dc.date.available	2023-02-15T21:12:11Z
dc.date.issued	2023-02
dc.identifier.citation	Bioprocess Biosyst Eng, 2023, 46, (2), pp. 297-306
dc.identifier.issn	1615-7591
dc.identifier.issn	1615-7605
dc.identifier.uri	http://hdl.handle.net/10453/166170
dc.description.abstract	Poly-hydroxy-butyrate (PHB) bioplastic resin can be made directly from atmospheric CO2 using cyanobacteria. However, higher PHB productivities are required before large-scale production is economically viable. Random mutagenesis offers a way to create new production strains with increased PHB yields and increased biomass densities without complex technical manipulation associated with genetically modified organisms. This study used staining with lipid fluorescent dye (BODIPY 493/593) and fluorescence-activated cell sorting (FACS) to select high lipid content mutants and followed this with a well plate growth screen. Thirteen mutants were selected for flask cultivation and two strains produced significantly higher PHB yields (29% and 26% higher than wild type), biomass accumulation (36% and 33% higher than wild type) and volumetric PHB density (75% and 67% higher than wild type). The maximum PHB yielding strain (% dcw) was 12.0%, which was 43% higher than the wild type (8.3% in this study). The highest volumetric PHB density was 18.8 mg PHB/L compared to 10.7 mg PHB/L by the wild type. To develop cyanobacterial strain with higher PHB productivities, the combination of random chemical mutagenesis and FACS holds great potential to promote cyanobacteria bioplastic production becoming economically viable.
dc.format	Print-Electronic
dc.language	eng
dc.publisher	Springer Science and Business Media LLC
dc.relation.ispartof	Bioprocess Biosyst Eng
dc.relation.isbasedon	10.1007/s00449-022-02834-5
dc.rights	info:eu-repo/semantics/closedAccess
dc.subject	0903 Biomedical Engineering, 0904 Chemical Engineering, 1003 Industrial Biotechnology
dc.subject.classification	Biotechnology
dc.subject.mesh	Flow Cytometry
dc.subject.mesh	Polyesters
dc.subject.mesh	Hydroxybutyrates
dc.subject.mesh	Cyanobacteria
dc.subject.mesh	Mutagenesis
dc.subject.mesh	Cyanobacteria
dc.subject.mesh	Hydroxybutyrates
dc.subject.mesh	Polyesters
dc.subject.mesh	Flow Cytometry
dc.subject.mesh	Mutagenesis
dc.title	Enhancement of cyanobacterial PHB production using random chemical mutagenesis with detection through FACS.
dc.type	Journal Article
utslib.citation.volume	46
utslib.location.activity	Germany
utslib.for	0903 Biomedical Engineering
utslib.for	0904 Chemical Engineering
utslib.for	1003 Industrial 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	closed_access	*
dc.date.updated	2023-02-15T21:12:08Z
pubs.issue	2
pubs.publication-status	Published
pubs.volume	46
utslib.citation.issue	2

Abstract:

Poly-hydroxy-butyrate (PHB) bioplastic resin can be made directly from atmospheric CO2 using cyanobacteria. However, higher PHB productivities are required before large-scale production is economically viable. Random mutagenesis offers a way to create new production strains with increased PHB yields and increased biomass densities without complex technical manipulation associated with genetically modified organisms. This study used staining with lipid fluorescent dye (BODIPY 493/593) and fluorescence-activated cell sorting (FACS) to select high lipid content mutants and followed this with a well plate growth screen. Thirteen mutants were selected for flask cultivation and two strains produced significantly higher PHB yields (29% and 26% higher than wild type), biomass accumulation (36% and 33% higher than wild type) and volumetric PHB density (75% and 67% higher than wild type). The maximum PHB yielding strain (% dcw) was 12.0%, which was 43% higher than the wild type (8.3% in this study). The highest volumetric PHB density was 18.8 mg PHB/L compared to 10.7 mg PHB/L by the wild type. To develop cyanobacterial strain with higher PHB productivities, the combination of random chemical mutagenesis and FACS holds great potential to promote cyanobacteria bioplastic production becoming economically viable.

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http://hdl.handle.net/10453/166170