4.7 Article

Enhancement of polyhydroxybutyrate production by introduction of heterologous phasin combination in Escherichia coli

Journal

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Volume 225, Issue -, Pages 757-766

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ELSEVIER
DOI: 10.1016/j.ijbiomac.2022.11.138

Keywords

Phasin; phaP; Halomonas sp; YLGW01

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This study aims to investigate the effect of heterologous phasins on the production of polyhydroxybutyrate (PHB) in E. coli and determine the best combination of phaP genes. By introducing a combination of phaP genes from a high-producing strain, Halomonas sp. YLGW01, into E. coli, the resulting strain showed a 2.9-fold increase in PHB production, with increased cell mass and PHB content reaching 65%. This strain also exhibited increased tolerance to inhibitors and could utilize lignocellulosic biomass as a carbon source.
Phasin is a surface-binding protein of polyhydroxyalkanoate (PHA) granules that is encoded by the phaP gene. As its expression increases, PHA granules become smaller, to increase their surface area, and are densely packed inside the cell, thereby increasing the PHA content. A wide range of PHA-producing bacteria have phaP genes; however, their PHA productivity differs, although they are derived from the cognate bacterial host cell. Modulating phasin expression could be a new strategy to enhance PHA production. This study aimed to char-acterize the effect of heterologous phasins on the reconstitution of E. coli BL21(DE3) and determine the best synergistic phaP gene combination to produce polyhydroxybutyrate (PHB). We identified novel phasins from a PHB high-producer strain, Halomonas sp. YLGW01, and introduced a combination of phaP genes into Escherichia coli. The resulting E. coli phaP1,3 strain had enhanced PHB production by 2.9-fold, leading to increased cell mass and increased PHB content from 48 % to 65 %. This strain also showed increased tolerance to inhibitors, such as furfural and vanillin, enabling the utilization of lignocellulose biosugar as a carbon source. These results sug-gested that the combination of phaP1 and phaP3 genes from H. sp. YLGW01 could increase PHB production and robustness.

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