Characterization of an evolved carotenoids hyper-producer of Saccharomyces cerevisiae through bioreactor parameter optimization and Raman spectroscopy.

ABSTRACT

An evolutionary engineering approach for enhancing heterologous carotenoids production in an engineered Saccharomyces cerevisiae strain was used previously to isolate several carotenoids hyper-producers from the evolved populations. ß-Carotene production was characterized in the parental and one of the evolved carotenoids hyper-producers (SM14) using bench-top bioreactors to assess the impact of pH, aeration, and media composition on ß-carotene production levels. The results show that with maintaining a low pH and increasing the carbon-to-nitrogen ratio (CN) from 8.8 to 50 in standard YNB medium, a higher ß-carotene production level at 25.52 ± 2.15 mg ß-carotene g(-1) (dry cell weight) in the carotenoids hyper-producer was obtained. The increase in CN ratio also significantly increased carotenoids production in the parental strain by 298 % [from 5.68 ± 1.24 to 22.58 ± 0.11 mg ß-carotene g(-1) (dcw)]. In this study, it was shown that Raman spectroscopy is capable of monitoring ß-carotene production in these cultures. Raman spectroscopy is adaptable to large-scale fermentations and can give results in near real-time. Furthermore, we found that Raman spectroscopy was also able to measure the relative lipid compositions and protein content of the parental and SM14 strains at two different CN ratios in the bioreactor. The Raman analysis showed a higher total fatty acid content in the SM14 compared with the parental strain and that an increased CN ratio resulted in significant increase in total fatty acid content of both strains. The data suggest a positive correlation between the yield of ß-carotene per biomass and total fatty acid content of the cell.