Phototrophic growth of a Rubisco-deficient mesophilic purple nonsulfur bacterium harboring a Type III Rubisco from a hyperthermophilic archaeon

Journal of Biotechnology
Shosuke YoshidaTadayuki Imanaka

Abstract

The hyperthermophilic archaeon, Thermococcus kodakaraensis KOD1 harbors a structurally novel, Type III Rubisco (Rbc(Tk)). In terms of protein engineering of Rubiscos, the enzyme may provide an alternative target to the conventional Type I and Type II enzymes. With a future aim to improve the catalytic properties of Rbc(Tk), here we examined whether or not the enzyme could support growth of a mesophilic organism dependent on CO2 fixation. Via double-crossover homologous recombination, we first deleted three Rubisco genes present on the chromosome of the photosynthetic mesophile Rhodopseudomonas palustris No. 7. The mutant strain (delta3) could neither grow under photoautotrophic nor photoheterotrophic conditions. We introduced the rbc(Tk) gene into strain delta3 either on a plasmid, or by integrating the gene onto the chromosome. The two transformant strains harboring rbc(Tk) displayed growth under photoautotrophic and photoheterotrophic conditions, both dependent on CO2 fixation. Specific growth rates and Rubisco activity levels were compared under photoheterotrophic conditions among the two transformants and the wild-type strain. We observed that the levels of Rubisco activity in the respective cell-free extracts correlated we...Continue Reading

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Citations

Oct 12, 2010·The Journal of Biological Chemistry·Yuichi NishitaniKunio Miki
Dec 24, 2010·Genes & Genetic Systems·Ryosuke FujikanePatrick Forterre
Apr 29, 2015·International Journal of Molecular Sciences·Yoon-Jung MoonYoung-Ho Chung
Aug 28, 2019·Proceedings of the National Academy of Sciences of the United States of America·Evgenii N FrolovNikolay A Chernyh

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