Combinatorial approach to study enzyme/surface interactions.
A fast combinatorial approach to access information about the immobilization behavior and kinetics of enzymes on a variation of surfaces is presented. As a test system, Candida Antarctica Lipase B was immobilized on a self-assembled monolayer bearing a gradient of surface energy. The respective immobilization behavior was monitored by Fourier transform infrared micro-spectroscopy. In addition, the activity of the immobilized enzyme was monitored over the entire film in real time with a specially developed fluorescence activity assay embedded into a siloxane gel. It was found that the highest amount of active protein was immobilized on the hydrophilic end of the gradient surface. This effect is associated with a higher surface roughness of this area resulting in hydrophobic micro-environments in which the enzyme gets immobilized.
Loos
Katja
K
Kennedy
Scott B
SB
Eidelman
Naomi
N
Tai
Yian
Y
Zharnikov
Michael
M
Amis
Eric J
EJ
Ulman
Abraham
A
Gross
Richard A
RA
D020650:Combinatorial Chemistry Techniques / Q000379:methods; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005656:Fungal Proteins; D005782:Gels; D007700:Kinetics; D008049:Lipase / Q000737:chemistry* / Q000502:physiology; D012833:Siloxanes / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared; D013499:Surface Properties
D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D005782:Gels; D012833:Siloxanes; D008049:Lipase; C431349:lipase B, Candida antarctica