In PI3K inhibitor addition, the sterols produced by the cyp61 – mutant strains corresponding to peaks 2 and 3 were at ratios of 55% and 44%, 32% and 68%, 48% and 52% in the 385-cyp61 hph /cyp61 zeo , CBS-cyp61 hph , Av2-cyp61 zeo strains, respectively. In the heterozygous strain 385-CYP61/cyp61 hph , the main sterol produced was ergosterol. A visible

change in the color of the cyp61 – mutants was evident when compared to their corresponding parental strain (Figure  7). The first ones had a more intense red color, suggesting that the mutant strains produced more carotenoids. In all cases, the maximum carotenoid content was reached after 120 h of Small molecule library chemical structure cultivation, which coincides with the late stationary phase of growth (Figure  8). The total carotenoid contents relative to the parental strains after 24, 72 and 120 h of cultivation, respectively, were as follows: 126%, 132% and 101% in

strain 385-CYP61/cyp61 hph ; 179%, 217% and 191% in strain 385-cyp61 click here hph /cyp61 zeo ; 116%, 153% and 138% in strain CBS-cyp61 hph and 100%, 141% and 134 % in strain Av2-cyp61 zeo (Table  4). dendrorhous mutant strain (in ppm)   Strains   UCD 67-385 385-cyp61 (+/−) 385-cyp61 (−/−) Cultivation time (h) 24 72 120 24 72 120 24 72 120 Astaxanthin 52.6±22.3 26.3±2.7 224.0±42.1 89.1±13.4 34.9±5.1 223.7±8.6

126.5±31.0 Protirelin 49.8±18.2 434.7±56.2 Phoenicoxanthin ND ND ND ND ND ND ND ND ND Cantaxanthin ND ND 13.4±3.3 ND ND ND ND ND ND HO-keto-γ-carotene ND 1.0±0.5 ND ND 1.9±0.3 ND ND 2.2±1.3 ND HO-keto-torulene 2.6±1.1 1.1±0.2 30.1±6.7 ND ND 35.5±1.0 ND ND 62.1±7.3 Keto-γ-carotene 8.0±4.9 2.7±1.4 7.8±1.9 ND 1.2±0.6 9.7±1.0 ND 5.7±2.9 21.4±7.9 HO-echinenone 1.8±0.6 1.2±0.9 2.6±0.5 ND 2.6±0.5 9.2±0.4 ND 3.6±1.6 15.6±4.4 Echinenone ND ND 2.0±0.4 ND ND ND ND ND ND Lycopene 4.0±2.0 ND ND ND 1.4±0.7 1.1±1.0 ND 4.3±1.9 ND γ-carotene ND 0.2±0.03 2.7±0.5 ND ND ND ND 0.8±0.4 ND β-carotene 1.1±0.5 0.8±0.3 2.7±1.1 ND 1.7±1.0 6.3±0.8 ND 4.8±3.5 15.8±9.1 Total carotenoids 70.7±26.9 36.1±8.6 290.1±53.4 89.1±13.4 47.6±7.1 293.7±9.1 126.5±31.0 78.2±26.2 555.1±75.2   Strains         CBS 6938 CBS – cyp61 (−)       Cultivation time (h) 24 72 120 24 72 120       Astaxanthin 32.1±11.2 202.0±17.7 324.2±6.7 62.8±5.4 313.5±24.1 429.3±26.