Breeding a novel chlorophyll-deficient mutant of Auxenochlorella pyrenoidosa for high-quality protein production by atmospheric room temperature plasma mutagenesis

In the present work, a novel chlorophyll-deficient mutant of Auxenochlorella pyrenoidosa named A4-1 was generated by atmospheric room temperature plasma (ARTP) mutagenesis. Compared to the green wild type (WT) strain, the A4-1 mutant cultured in the dark displayed yellow colour with a 118-fold decrease of chlorophyll a and no detected chlorophyll b. Higher contents of protein (44.22 % DW), total amino acids (AAs, 34.84 % DW) and essential AAs (17.50 % DW) were also achieved, showing 31 %, 22 % and 30 % increases compared to the WT, respectively (p < 0.05). Metabolite profile analysis revealed that the chlorophyll biosynthesis pathway in the A4-1 mutant was probably inhibited in the dark, while more carbon skeletons might be utilized for de novo AAs synthesis. These results demonstrated that the A4-1 mutant not only has extremely low chlorophyll content, but also has higher protein content, making it a very promising candidate to produce microalgal protein for future foods.