Site directed immobilization of glucose-6-phosphate dehydrogenase via thiol-disulfide interchange : influence on catalytic activity of cysteines introduced at different positions

Publikationstyp
Journal Article
Date Issued
2013-06-13
Sprache
English
Author(s)
Simons, Jan R.  
Mosisch, Martin  
Torda, Andrew  
Hilterhaus, Lutz  
Institut
Technische Biokatalyse V-6  
TORE-URI
http://hdl.handle.net/11420/6586
Journal
Journal of biotechnology  
Volume
167
Issue
1
Start Page
1
End Page
7
Citation
Journal of Biotechnology 1 (167): 1-7 (2013)
Publisher DOI
10.1016/j.jbiotec.2013.06.002
Scopus ID
2-s2.0-84880120033
Publisher
Elsevier Science
This study shows the effect of site-directed enzyme immobilization upon the enzyme activity of covalently bound glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides. Immobilization points were introduced at sterically accessible sites in order to control the protein's orientation and twice as much activity was recovered in comparison to conventionally immobilized enzyme. Immobilization of G6PDH via genetically engineered cysteine provided a simple, but effective method to control the immobilization process. G6PDH variants with cysteine close to the active center (L218C), close to the dimer interface (D205C) as well as far from the active center (D453C) showed changes in activity and the efficacy of immobilization.
Subjects
Glucose-6-phosphate dehydrogenase
Leuconostoc mesenteroides
Site directed enzyme immobilization
Site directed mutagenesis
Thiol disulfide interchange
DDC Class
570: Biowissenschaften, Biologie