Enhanced ergosterol production by recombinant Saccharomyces cerevisiae 1190 harboring Vitreoscilla hemoglobin gene (vgb) / 生物工程学报

ABSTRACT

Ergosterol is a principal sterol of fungi. It is a raw material for production of vitamin D2, hydrocortisone, progesterone and brassinolide. Synthesis of ergosterol requires molecular oxygen, and low oxygen tensions was reported to dramatically reduce ergosterol concentration. Vitreoscilla Hemoglobin Gene (vgb), a homodimeric hemoglobin gene from Gram-negative obligate aerobic bacterium Vitreoscilla, enables a higher specific cellular oxygen uptake rate, it also improves the oxygen transportation. In this study, recombinant plasmid pVgb-kanMX4 containing Vitreoscilla Hemoglobin Gene (vgb) and geneticin (G418) was constructed and transformed into Saccharomyces cerevisiae 1190 for enhanced ergosterol production. With sufficient oxygen supply, the ergosterol contents of recombinant and wild type strains grown in shake flasks were 1.07% and 0.573%, respectively. Under oxygen limitation condition, ergosterol contents in recombinant and wild type strains were reduced to 0.39% and 0.25%, respectively. In a 30 hours fermentation study conducted in a 5 liter fermentor, 15.1 g/L Cell Dry Weight (CDW) containing 1.38% ergosterol was obtained from growth of the recombinant strains; Only 14.8 g/L CDW containing 0.9% ergosterol was produced by the wild type strain. These results demonstrated that vgb played a role in enhancing ergosterol production.