Artificial self-sufficient P450 in reversed micelles.
Cytochrome P450s are heme-containing monooxygenases that require electron transfer proteins for their catalytic activities. They prefer hydrophobic compounds as substrates and it is, therefore, desirable to perform their reactions in non-aqueous media. Reversed micelles can stably encapsulate proteins in nano-scaled water pools in organic solvents. However, in the reversed micellar system, when multiple proteins are involved in a reaction they can be separated into different micelles and it is then difficult to transfer electrons between proteins. We show here that an artificial self-sufficient cytochrome P450, which is an enzymatically crosslinked fusion protein composed of P450 and electron transfer proteins, showed micelle-size dependent catalytic activity in a reversed micellar system. Furthermore, the presence of thermostable alcohol dehydrogenase promoted the P450-catalyzed reaction due to cofactor regeneration.
Hirakawa
Hidehiko
H
Department of Bioengineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Kamiya
Noriho
N
Kawarabayasi
Yutaka
Y
Nagamune
Teruyuki
T
D000426:Alcohol Dehydrogenase; D003432:Cross-Linking Reagents; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry*; D004579:Electron Transport; D008823:Micelles; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins
D003432:Cross-Linking Reagents; D008823:Micelles; D011993:Recombinant Fusion Proteins; D003577:Cytochrome P-450 Enzyme System; D000426:Alcohol Dehydrogenase