New platform for cytochrome p450 reaction combining in situ immobilization on biopolymer
Jae Hyung Lee1, Dong Heon Nam, Sahng Ha Lee
1Department of Chemical and Biomolecular Engineering, and ‡Department of Materials Science and Engineering, Korea Advanced Institute of Science and Technology , 335 Science Road, Daejeon 305-701, Republic of Korea.
Abstract:
We describe an efficienct chemical conversion platform with in situ immobilization of P450-BM3 on poly(3-hydroxybutyrate) granules. Through fusion with phasin, P450-BM3 is easily immobilized on poly(3-hydroxybutyrate) granules in Escherichia coli. In our work, the immobilized P450 exhibited higher stability and catalytic activity compared to free P450 against changes of pH, temperature, and concentrations of urea and ions. Through quick recovery of immobilized enzyme, the P450-P(3HB) complex successfully catalyzed an O-dealkylation reaction several times with maintained activity. Using the robust P450-P(3HB) complex, we performed a P450-catalyzed reaction on a preparative reactor scale (100 mL) and high-level production (12.3 μM) of 7-hydroxycoumarine from 7-ethoxycoumarin could be achieved.
