Fine-tuned regulation of photosynthetic performance via γ-aminobutyric acid (GABA) supply coupled with high initial cell density culture for economic starch production in microalgae.

Microalgal starch is considered as renewable and sustainable feedstock for biofuels and biorefinery. High cell density culture is favourable for photoautotrophic starch production in microalgae in the aspects of productivity and economy, but it often encounters low starch content or extra stress exposure that limits the production. This study aimed to economically enhance photosynthetic starch production from CO 2 fixation in a green microalga Tetraselmis subcordiformis by regulating photosynthetic stress status with a signalling molecule γ-aminobutyric acid (GABA) combined with the application of high initial cell density culture. By increasing initial cell density (ICD) from the normal of 1.1 g L -1 (NICD) to as high as 2.8 g L -1 (HICD), the starch content, yield, and theoretical productivity were improved by 7%, 63%, and 42%, respectively. The addition of GABA under HICD resulted in 14%, 19%, and 26% of further enhancement in starch content, yield, and theoretical productivity, respectively. GABA exhibited distinct regulatory mechanisms on photosynthesis and stress status under HICD relative to NICD. GABA augmented excessive light energy absorption and electron transfer through photosystem II that reinforced the photoinhibition under NICD, while alleviated the stress reversely under HICD, both of which facilitated starch production by enabling a suitable stress status while simultaneously maintaining a sufficient photosynthetic activity. The increase of ICD and/or GABA supply particularly boosted amylopectin accumulation, leading to the changes in starch composition and was more favourable for fermentation-based biofuels production. Preliminary techno-economic analysis showed that the highest net extra benefit of 9.64 $ m -3 culture could be obtained under HICD with 2.5 mM GABA supply where high starch content (62%DW) and yield (2.5 g L -1 ) were achieved. The combined HICD-GABA regulation was a promising strategy for economic starch production from CO 2 by microalgae for sustainable biomanufacturing.