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›› 2009, Vol. 9 ›› Issue (4): 781-785.

• 生化工程专栏 • 上一篇    下一篇

离子交换法分离S-苄基-g-L-谷氨酰-L-半胱氨酸

刘步云 周治 姚忠 吴明刚 宋希文 佘维娜 肖彦羚 韦萍   

  1. 南京工业大学制药与生命科学学院 南京工业大学制药与生命科学学院 南京工业大学制药与生命科学学院 南京工业大学制药与生命科学学院 南京工业大学制药与生命科学学院 南京工业大学制药与生命科学学院 南京工业大学制药与生命科学学院 南京工业大学制药与生命科学学院
  • 收稿日期:2009-02-09 修回日期:2009-05-13 出版日期:2009-08-20 发布日期:2009-08-20
  • 通讯作者: 姚忠

Separation of S-Benzyl-g-L-glutamyl-L-cysteine Using Ion-exchange Method

LIU Bu-yun, ZHOU Zhi, YAO Zhong, WU Ming-gang, SONG Xi-wen, SHE Wei-na, XIAO Yan-ling, WEI Ping,   

  1. State Key Laboratory of Material-Oriented Chemical Engineering, College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology College of Life Science and Pharmacy, Nanjing University of Technology College of Life Science and Pharmacy, Nanjing University of Technology College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology College of Pharmacy and Life Science,Nanjing University of Technology
  • Received:2009-02-09 Revised:2009-05-13 Online:2009-08-20 Published:2009-08-20
  • Contact: YAO Zhong,

摘要: 采用离子交换法对酶法合成的S-苄基-g-L-谷氨酰-L-半胱氨酸(S-Bzl-GGC)进行了分离纯化. 研究了Q Sephrose FF阴离子交换树脂对S-Bzl-GGC的吸附等温线,很好地符合Sips吸附等温线模型. 考察了pH值、洗脱梯度、流速和进样量等条件对纯化效果的影响,建立了一套基于Q Sephrose FF阴离子交换树脂的纯化方案. 结果表明,在pH 8.2、洗脱梯度0~30% B(Tris-HCl缓冲液+1 mol/L NaCl) 12 mL、流速2 mL/min、进样量500 mL的优化条件下,经一步分离,产物纯度可达98.5%,经1H-NMR鉴定产物结构正确.

关键词: S-苄基-g-L-谷氨酰-L-半胱氨酸, 吸附等温线, 离子交换, 纯化

Abstract: The synthesized S-benzyl-g-L-glutamyl-L-cysteine (S-Bzl-GGC) with enzymatic methods was purified with ion-exchange method by using Q sepharose FF as adsorbent. The adsorption isotherm was investigated, and Sips equation provided the best fitting of experimental data. The influences of pH value, elution gradient, flow rate and sample quantity on the separation process were also discussed. The results showed that S-Bzl-GGC could be well separated at the conditions of pH 8.2, salt elution gradient of 0~30% B, 12 mL, flow rate of 2 mL/min and sample size of 500 mL. Under the optimal conditions, the purity of product was 98.5% after one step purification and the structure of product was further identified by 1H-NMR.

Key words: S-benzyl-g-L-glutamyl-L-cysteine, adsorption isotherm, ion exchange, purification

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