Fan Nan, Li Yan, Zhou Quan, Chen Guo-Qiang
Department of Food Science and Engineering, Jinan University, Guangzhou 510632, China.
Sheng Wu Gong Cheng Xue Bao. 2004 May;20(3):441-4.
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.
麦角固醇是真菌的主要固醇。它是生产维生素D2、氢化可的松、孕酮和油菜素内酯的原料。麦角固醇的合成需要分子氧,据报道低氧张力会显著降低麦角固醇的浓度。透明颤菌血红蛋白基因(vgb)是一种来自革兰氏阴性专性需氧细菌透明颤菌的同二聚体血红蛋白基因,它能提高细胞的比氧摄取率,还能改善氧的运输。在本研究中,构建了含有透明颤菌血红蛋白基因(vgb)和遗传霉素(G418)的重组质粒pVgb-kanMX4,并将其转化到酿酒酵母1190中以提高麦角固醇的产量。在充足的氧气供应下,摇瓶中生长的重组菌株和野生型菌株的麦角固醇含量分别为1.07%和0.573%。在氧气限制条件下,重组菌株和野生型菌株中的麦角固醇含量分别降至0.39%和0.25%。在5升发酵罐中进行的30小时发酵研究中,重组菌株生长得到了含有1.38%麦角固醇的15.1克/升细胞干重(CDW);野生型菌株仅产生了含有0.9%麦角固醇的14.8克/升CDW。这些结果表明vgb在提高麦角固醇产量中发挥了作用。