Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/68163
標題: Simultaneous purification and immobilization of D-hydantoinase on the immobilized metal affinity membrane via coordination bonds
作者: Ko, Y.M.
Chen, C.I.
Shieh, C.J.
Liu, Y.C.
關鍵字: D-Hydantoinase;Immobilized metal affinity membrane;Enzyme;immobilization;Enzyme purification;Enzyme stability;d-amino acids;d-p-hydroxyphenylglycine;microbial transformation;agrobacterium-tumefaciens;hydrolyzing activity;escherichia-coli;ion;proteins;enzymes;binding
Project: Biochemical Engineering Journal
期刊/報告no:: Biochemical Engineering Journal, Volume 61, Page(s) 20-27.
摘要: 
This study constructs the immobilized metal affinity membrane (IMAM) via coupling of epichlorohydrin, iminodiacetic acid, and nickel ion on the regenerated cellulose membrane. The D-hydantoin-hydrolyzing enzyme (DHTase) harboring a poly-His tagged residue was used as a model protein immobilized on the prepared IMAM. Various immobilization conditions were examined based on the yield of N-carbamoyl-D-p-hydroxyphenylglycine in batch reactions. The immobilization conditions were studied and the optimal conditions are as follows. By employing an IMAM with nickel ion of 155.5 +/- 5 mu mol/disc immersed in 0.1 M Tris-HCl buffer pH 8 (with 0.8M sodium chloride) and immobilized time of 14h. a DHTase activity of 4.2 +/- 0.3 U/disc was obtained. The immobilized DHTase membrane can achieve a larger pH and thermal tolerant range than that of free enzyme. Meanwhile, the stability test showed that 99% of enzyme activity could be retained after being repeated 15-times. The storage test also displayed 99% enzyme preservation after 7 weeks of storage. (C) 2011 Elsevier B.V. All rights reserved.
URI: http://hdl.handle.net/11455/68163
ISSN: 1369-703X
DOI: 10.1016/j.bej.2011.11.013
Appears in Collections:期刊論文

Show full item record
 

Google ScholarTM

Check

Altmetric

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.