生物技术通报 ›› 2013, Vol. 0 ›› Issue (11): 181-185.

• 研究报告 • 上一篇    下一篇

还原型谷胱甘肽发酵及分离纯化条件的优化

王硕1, 谢跃武1, 张惠文2, 阎浩林1, 徐明恺2,   

  1. 1.沈阳药科大学,沈阳 110016;2.中国科学院沈阳应用生态研究所 森林与土壤生态国家重点实验室,沈阳 110064
  • 收稿日期:2013-04-27 出版日期:2013-11-14 发布日期:2013-11-14
  • 作者简介:王硕,男,硕士,研究方向:发酵法生产还原型谷胱甘肽;E-mail:shuowang87@126.com

Optimization Conditions of Fermentation and Separation and Purification of Reduced-Glutathione

Wang Shuo1, Xie Yuewu1, Zhang Huiwen2, Yan Haolin1, Xu Mingkai2   

  1. (1. Shenyang Pharmaceutical University,Shenyang 110016;2. State Key Laboratory of Forest and Soil Ecology,Institute of Applied Ecology,Chinese Academy of Sciences, Shenyang 110064)
  • Received:2013-04-27 Published:2013-11-14 Online:2013-11-14

摘要: 旨在通过优化还原型谷胱甘肽的发酵及分离纯化条件,为GSH大规模的工厂化生产提供理论基础。首先通过筛选获得一株GSH高产菌酿酒酵母Y18,其胞内GSH含量为9.05 mg/g干菌体。进一步研究表明,在培养基中添加L-半胱氨酸和缬氨酸,以及发酵过程中补加葡萄糖可使GSH的总产量分别提高62.0%和146.1%。此外,对GSH在水溶液中稳定性的研究发现偏酸性环境(pH4.5)和氮气保护可显著降低GSH的损失率。最终通过对大孔树脂D001×7和Sephadex G-10的分离纯化条件的优化,获得纯度为98.1%的GSH,收率为73.2%。

关键词: 酿酒酵母, 还原型谷胱甘肽, 发酵, 分离纯化

Abstract: The aim of the study is to provide theoretical basis in GSH large-scale production, by optimizing the conditions of its fermentation and purification. Firstly, a GSH high-yield Saccharomyces cerevisiae Y18 was obtained, with the production of 9.05 mg GSH per gram dry cells. Secondly, our results indicated that adding L-cysteine and Valine to the medium, or additional glucose during fermentation process could increase GSH production by 62.0% and 146.1%, respectively. Moreover, the weak acidity condition(pH4.5)and nitrogen input protected GSH from oxidation, and improved its stability. Finally, we obtained high-purity(98.1%)GSH and a promising recovery(73.2%)after optimizing the conditions for separation and purification GSH in Macroporous resin D001×7 and Sephadex G-10 gel chromatography.

Key words: Saccharomyces cerevisiae, Reduced glutathione, Fermentation, Separation and purification