An improved method for purification of recombinant truncated heme oxygenase-1 by expanded bed adsorption and gel filtration

作者：Hu, Hong Bo^*; Wang, Wei; Han, Ling; Zhou, Wen Pu; Zhang, Xue Hong

来源：Bioprocess and Biosystems Engineering, 2007, 30(2): 87-90.

DOI：10.1007/s00449-006-0103-y

• expanded bed adsorption
• gel filtration
• heme oxygenase-1
• purification

Recombinant truncated human heme oxygenase-1 (hHO-1) expressed in Escherichia coli was efficiently separated and purified from feedstock by DEAE-ion exchange expanded bed adsorption. Protocol optimization of hHO-1 on DEAE adsorbent resulted in adsorption in 0 M NaCl and elution in 150 mM NaCl at a pH of 8.5. The active enzyme fractions separated from the expanded bed column were further purified by a Superdex 75 gel filtration step. The specific hHO-1 activity increased from 0.82 +/- 0.05 to 24.8 +/- 1.8 U/mg during the whole purification steps. The recovery and purification factor of truncated hHO-1 of the whole purification were 72.7 +/- 4.7 and 30.2 +/- 2.3%, respectively. This purification process can decrease the demand on the preparation of feedstock and simplify the purification process.

• 出版日期2007-3
• 单位上海交通大学