Enhancing photosynthetic biomass productivity of industrial oleaginous microalgae by overexpression of RuBisCO activase

Eukaryotic microalgae are promising cell factories for photosynthesis-driven CO2 sequestration and oil production, however one key challenge has been to genetically enhance their biomass productivity. Here we showed that the biomass productivity of a model industrial oleaginous microalga Nannochloropsis oceanica can be elevated by overexpression of a nuclear-encoded, cbbX-homologous, candidate RuBisCO activase (g1915 or 'nNoRca-like') that is transcriptionally induced under air-level CO2 (i.e., 0.04% v/v) as compared to 5% CO2 in wild-type cells. In the mutants, under air-level CO2, growth rate was enhanced by similar to 32%, biomass accumulation by similar to 46% and lipid productivity by similar to 41%, which were also accompanied by similar to 28% elevation in photosynthesis; furthermore, abundance of RuBisCO large subunit protein increased by similar to 45%, suggesting nNoRca-like overexpression enhanced microalgal photosynthesis by upregulating the level and activity of RuBisCO. Interestingly, under 5% CO2, no significant enhancement of RuBisCO protein expression was observed. Thus a genetic strategy that activates RuBisCO by augmenting its functional partners was proposed to increase biomass production of industrial microalgae. Our work expands the reservoir of potential photosynthetic gene targets for plant biotechnology and provides a basis for constructing advanced microalgal cell factories for carbon sequestration and biofuel production.