Evaluation of immobilized metal affinity chromatography for purification of penicillin acylase |
| |
| Institution: | 1. Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike Road 501, Shanghai 201203, China;2. Thermo Fisher Scientific, Jinqiao Road 27, Shanghai 201206, China;1. Department of Biochemistry, Faculty of Science, Masaryk University, Brno, Czech Republic;2. Mendel Centre for Plant Genomics and Proteomics, Central European Technology Institute, Brno, Czech Republic;1. College of Polymer Science and Engineering, State Key Laboratory of Polymer Materials Engineering, Sichuan University, Chengdu 610065, People’s Republic of China;2. State Key Laboratory for Modification of Chemical Fibers and Polymer Materials, Donghua University, Shanghai 201620, People’s Republic of China;1. Contipro a.s., Dolní Dobrouč 401, 56102, Dolní Dobrouč, Czech Republic;2. Department of Analytical Chemistry, Faculty of Science, Palacký University, Olomouc, Czech Republic;3. International Clinical Research Center, St. Anne''s University Hospital Brno, Brno, Czech Republic;4. Institute of Biophysics of the Czech Academy of Sciences, Brno, Czech Republic;5. Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic |
| |
| Abstract: | The aim of this work was to test immobilized metal affinity chromatography (IMAC) for the purification of penicillin acylase. After evaluation of different metals, Cu2+ was selected. Different samples were tested: pure penicillin acylase, industrial clarified feedstock and crude extract. After comparing two eluents, NH4Cl and imidazole, it appeared that although both gave good results for recovery and activity, NH4Cl was a more selective eluent with a higher fold purification than imidazole (4.64 versus 2.04). Moreover, we shown that a multistep gradient of NH4Cl, greatly increased the degree of purification (12.36) compared with the one-step process as control (4.64). In addition, good recovery was obtained (97–100%). |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
