Impact of surfactants on solubilization and activity of the carotenoid cleavage dioxygenase, AtCCD1, in an aqueous micellar reaction system.
The effect of various surfactants on both the solubilization of the carotenoid cleavage dioxygenase, AtCCD1, from cell lysates and the enzymatic activity in an aqueous micellar system was investigated. Solubilization with sodium cholate more than doubled the specific activity. Lag phases were observed when Tween surfactants were used for substrate delivery and were dependent on the surfactant and enzyme modification. In contrast to His(6)- and GST-tagged AtCCD1, unmodified AtCCD1 showed a 45% increased maximum rate in the Tween 20 system compared to Triton X-100 based reference system. The results emphasize the importance of engineering the interface for the in vitro application of this enzyme family.
Schilling
M
M
Biochemical Engineering, DECHEMA e.V., Karl-Winnacker-Institut, Theodor-Heuss-Allee 25, 60486 Frankfurt/Main, Germany.
Haetzelt
F
F
Schwab
W
W
Schrader
J
J
D002338:Carotenoids / Q000378:metabolism; D049308:Dioxygenases / Q000737:chemistry* / Q000378:metabolism*; D008823:Micelles; D012995:Solubility; D013501:Surface-Active Agents / Q000737:chemistry*; D014867:Water / Q000737:chemistry
D008823:Micelles; D013501:Surface-Active Agents; D014867:Water; D002338:Carotenoids; D049308:Dioxygenases; C431939:carotenoid cleavage dioxygenase 1