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The gene dosage effect of carbonic anhydrase on the biosynthesis of poly(3-hydroxybutyrate) under autotrophic and mixotrophic culture conditions

Abstract

Ralstonia eutropha is a hydrogen-oxidizing chemolithotrophic bacterium that can synthesize poly[(R)-3-hydroxybutyrate] [P(3HB)], which is used as a biodegradable plastic. Various genes coding for carbonic anhydrase (CA), which are involved in CO2 fixation and metabolism, are present in the genome of R. eutropha. Although the significance of CA for CO2 metabolism in R. eutropha is well known, CAs have never been deliberately employed to enhance P(3HB) production from CO2. Thus, we investigated the gene dosage effect of the β-carbonic anhydrase (can) gene on P(3HB) accumulation in R. eutropha H16 under autotrophic and mixotrophic conditions. The multicopy plasmid-based dosage of the can gene showed a 1.5-fold increase in P(3HB) content in cells grown under nitrogen-source-limited autotrophic conditions. A similar result was observed under nitrogen-source-limited mixotrophic conditions in the presence of glycerol. Thus, these findings suggest a viable strategy to increase P(3HB) yield under autotrophic and mixotrophic culture conditions.

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Acknowledgements

The authors gratefully acknowledge the helpful discussions and comments of Dr. Akiko Komiyama (Kyushu Electric Power Co., Inc., Fukuoka, Japan). This work was supported by a Grant-in-Aid for Scientific Research (KAKENHI 19K22923), Japan.

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Correspondence to Takeharu Tsuge.

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Thorbecke, R., Yamamoto, M., Miyahara, Y. et al. The gene dosage effect of carbonic anhydrase on the biosynthesis of poly(3-hydroxybutyrate) under autotrophic and mixotrophic culture conditions. Polym J 53, 209–213 (2021). https://doi.org/10.1038/s41428-020-00409-3

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