Earticle

초록

영어

Among 27 cytochrome P450s (CYPs) of Nocardia farcinica IFM10152, three CYPs were identified to have O-dealkylation catalytic activity. Between two of them encoding CYP154 subfamilies, CYP154 encoded by nfa22930 gene showed distinct O-dealkylation and subsequent hydroxylation
activities for formononetin. Firstly, formononetin was O-dealkylated into daidzein, and daidzein was subsequently mono-hydroxylated at 3’-position of B-ring into ortho-dihydroxy-isoflavone. Apparent kcat/Km values of CYP154 for formononetin O-dealkylation and daidzein hydroxylation reaction were 3.57 and 1.84 μM-1min-1, respectively. The dissociation constants based on
spectral changes for binding with each substrate were 5.16 and 3.11 μM, respectively. Homology modeling and docking simulation predicted that Thr247 is a key residue responsible for the 3’-position hydroxylation reaction by forming hydrogen bond with 4’-hydroxyl group of daidzein and
making the proton atom at 3’-position face to heme center. And site-directed mutagenesis of Thr247 into alanine led to a drastic decrease in the binding affinity for daidzein (9.73 μM) and 3’-position hydroxylation catalytic activity down to 3 folds (0.48 μM-1min-1).

키워드

저자

• Kwon-young CHOI [ Dept. of Chemical and Biological Engineering, Seoul National University, Seoul. ]
• Hyung-yeon PARK [ Korea Bio-Hub Center, Bio-MAX Institue, Seoul National University, Seoul, South Korea. ]
• Nahum LEE [ Dept. of Chemical and Biological Engineering, Seoul National University, Seoul. ]
• Bishnu PANDEY [ Dept. of Chemical and Biological Engineering, Seoul National University, Seoul. ]
• Eunok JUNG [ Dept. of Chemical and Biological Engineering, Seoul National University, Seoul. ]
• Dahye JUNG [ Dept. of Chemical and Biological Engineering, Seoul National University, Seoul. ]
• Byunggee KIM [ Dept. of Chemical and Biological Engineering, Seoul National University, Seoul. ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576