重庆理工大学学报（自然科学版）2016，Vol.30Issue(4)：73-78,6.DOI:10.3969/j.issn.1674-8425(z).2016.04.013
重组精氨酸脱亚胺酶的制备工艺
Study of Preparation Technology of Recombinant Arginine Deiminase
何云凤 1郭晋霞 1范开2
作者信息
- 1. 重庆理工大学 药学与生物工程学院，重庆 400054
- 2. 重庆富进生物医药有限公司，重庆 400050
- 折叠
摘要
Abstract
The 30 L scale fermentation, preparation technology and biological activities of recombinant arginine deiminase( rADI-ES)were investigated. The seed solution was obtained by shaxing flasx activation of the constructed engineering bacteria,and then transferred it to 30 L fermenter for enlarging cultivation,and IPTG was used for its induction expression. The expression products broxe bacterium by high-pressure homogenizer,and inclusion bodies were washed by buffer, and after dissolved refolding,the product was purified with DEAE anion exchange chromatography and Butyl hydrophobic interaction chromatography column,and we detected its expression and purity by SDS-PAGE and RP-HPLC analysis. The activity of rADI-ES was determined in vitro. We successfully amplified and induced expression of the engineering bacteria,and obtained the rADIES with the molecular weight of 46 xDa. The expression quantity of recombinant protein counts about more than 25% total protein,and the purity of the purified rADI-ES is more than 95% and its enzymatic activity is 42 IU. The expeiment show that the process is feasible to obtain a higher purity and activity of recombinant arginine deiminase.关键词
精氨酸脱亚胺酶/发酵/复性/分离纯化Key words
arginine deiminase/fermentation/refold/purification分类
生物科学引用本文复制引用
何云凤,郭晋霞,范开..重组精氨酸脱亚胺酶的制备工艺[J].重庆理工大学学报（自然科学版）,2016,30(4):73-78,6.
