Biofuel derived from microalgae have several advantages over other oleaginous crops, however, still needs to be improved with its cost aspect and can be achieved by developing of a strain with improved lipid productivity. In this study, the CRISPR-Cas9 system was incorporated to carry out a target-specific knockout of the phospholipase A2 gene in Chlamydomonas reinhardtii. The targeted gene encodes a key enzyme in the Lands cycle. As a result, the mutants showed a characteristic of increased diacylglycerol pool, followed by a higher accumulation of triacylglycerol without being significantly compensated with the cell growth. As a result, the overall lipid productivities of phospholipase A2 knockout mutants have increased by up to 64.25% (to 80.92 g L-1 d-1). This study can provide crucial information for the biodiesel industry.
Keywords: Biofuel; CRISPR-CAS9; Chlamydomonas reinhardtii; Phospholipase; Triacylglycerol.
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