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

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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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Acknowledgements

The authors would like to thank the Australian Government and the University of Technology Sydney for the financial support provided.

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Authors and Affiliations

  1. Climate Change Cluster, School of Life Sciences, The University of Technology, Sydney, Australia

    Shawn Price, Unnikrishnan Kuzhiumparambil, Mathieu Pernice, Andrei Herdean & Peter Ralph

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  1. Shawn Price
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  2. Unnikrishnan Kuzhiumparambil
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  3. Mathieu Pernice
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  4. Andrei Herdean
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Correspondence to Shawn Price.

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Price, S., Kuzhiumparambil, U., Pernice, M. et al. Enhancement of cyanobacterial PHB production using random chemical mutagenesis with detection through FACS. Bioprocess Biosyst Eng 46, 297–306 (2023). https://doi.org/10.1007/s00449-022-02834-5

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  • DOI: https://doi.org/10.1007/s00449-022-02834-5

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