Artificial Self-Sufficient P450 in Reversed Micelles

• Main Authors: Teruyuki Nagamune, Yutaka Kawarabayasi, Hidehiko Hirakawa, Noriho Kamiya
• Format: Article
• Language: English
• Published: MDPI AG 2010-04-01
• Series: Molecules
• Subjects: artificial self-sufficient cytochrome P450; cofactor regeneration; transglutaminase; reversed micelles
• Online Access: http://www.mdpi.com/1420-3049/15/5/2935/

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.