Presentation Information
[P01-072]Polyhydroxybutyrate production from sucrose by the introduction of invertase to Cupriavidus necator
○Hana Nadhifah1, Prihardi Kahar1, Yutaro Mori1, Chiaki Ogino1 (1. Graduate School of Engineering, Kobe University (Japan))
Keywords:
biodegradable polymer,PHB accumulation,Cupriavidus necator,enzyme
[Purpose]
Cupriavidus necator is a well-studied model organism for producing polyhydroxybutyrate (PHB), a biodegradable polymer with high potential as an alternative to petrochemical-based plastic. However, large-scale production remains costly due to the high expense of carbon sources. This study focuses on a genetic engineering strategy to enable PHB production from sucrose as a low-cost, high-supply carbon source.
[Method]
The introduction of the neutral-alkaline invertase gene was explored to enable sucrose utilization, a strategy not previously reported. This creates opportunities for direct sucrose-based biomass fermentation by engineered C. necator, reducing the overall biomass pretreatment costs.
[Results]
The invertase B isolated from Anabaena sp. PCC 7120 and an arabinose-inducible promoter were introduced into C. necator PHB-4 (DSM 541). The invertase gene was chosen for its compatibility with C. necator’s optimal pH (6-8) and high catalytic efficiency with sucrose. Cultivation in 50 g/L sucrose minimal mineral medium resulted in a decrease in sucrose content during the cultivation, confirming the successful introduction of the invertase gene.
[Consideration]
After 48 hours, the sucrose was hydrolyzed into glucose and fructose, with the fructose being immediately used to produce PHB, reaching up to 22.89 ± 0.18% after 163 hours of cultivation. Meanwhile, glucose remained in the medium. Further improvements are needed to enable the glucose assimilation pathway and enhance the efficiency of sugar conversion into PHB.
[Conclusion]
PHB from C. necator is highly demanded and potential candidate for biodegdarable and sustainable plastic materials. The succession of sucrose hydrolysis ability in C. necator recombinant strain will increase the potential to utilize all of the sugar composition in sorghum as new inexpensive biomass source for future PHB production.
Cupriavidus necator is a well-studied model organism for producing polyhydroxybutyrate (PHB), a biodegradable polymer with high potential as an alternative to petrochemical-based plastic. However, large-scale production remains costly due to the high expense of carbon sources. This study focuses on a genetic engineering strategy to enable PHB production from sucrose as a low-cost, high-supply carbon source.
[Method]
The introduction of the neutral-alkaline invertase gene was explored to enable sucrose utilization, a strategy not previously reported. This creates opportunities for direct sucrose-based biomass fermentation by engineered C. necator, reducing the overall biomass pretreatment costs.
[Results]
The invertase B isolated from Anabaena sp. PCC 7120 and an arabinose-inducible promoter were introduced into C. necator PHB-4 (DSM 541). The invertase gene was chosen for its compatibility with C. necator’s optimal pH (6-8) and high catalytic efficiency with sucrose. Cultivation in 50 g/L sucrose minimal mineral medium resulted in a decrease in sucrose content during the cultivation, confirming the successful introduction of the invertase gene.
[Consideration]
After 48 hours, the sucrose was hydrolyzed into glucose and fructose, with the fructose being immediately used to produce PHB, reaching up to 22.89 ± 0.18% after 163 hours of cultivation. Meanwhile, glucose remained in the medium. Further improvements are needed to enable the glucose assimilation pathway and enhance the efficiency of sugar conversion into PHB.
[Conclusion]
PHB from C. necator is highly demanded and potential candidate for biodegdarable and sustainable plastic materials. The succession of sucrose hydrolysis ability in C. necator recombinant strain will increase the potential to utilize all of the sugar composition in sorghum as new inexpensive biomass source for future PHB production.
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