Enzyme and protein engineering: engineering a self-sufficient human cytochrome P450 monooxygenase system

2011 ◽  
Vol 22 ◽  
pp. S18
Author(s):  
Yusuf Deeni ◽  
Roland Wolf
Keyword(s):  
Cytochrome P450 ◽  
Protein Engineering ◽  
Cytochrome P450 Monooxygenase ◽  
Monooxygenase System ◽  
P450 Monooxygenase ◽  
Human Cytochrome

scholarly journals Purification of Cytochrome P450 and Ferredoxin, Involved in Bisphenol A Degradation, from Sphingomonas sp. Strain AO1

2005 ◽  
Vol 71 (12) ◽  
pp. 8024-8030 ◽  
Author(s):  
Miho Sasaki ◽  
Ayako Akahira ◽  
Ko-ichi Oshiman ◽  
Tetsuaki Tsuchido ◽  
Yoshinobu Matsumura
Keyword(s):  
Mass Spectrometry ◽  
Cytochrome P450 ◽  
Bisphenol A ◽  
Cytochrome P450 Monooxygenase ◽  
Mass Spectrometry Analysis ◽  
Monooxygenase System ◽  
P450 Monooxygenase ◽  
Monooxygenase Activity ◽  
Chromatography Analysis ◽  
Ferredoxin Reductase

ABSTRACT In a previous study (M. Sasaki, J. Maki, K. Oshiman, Y. Matsumura, and T. Tsuchido, Biodegradation 16:449-459, 2005), the cytochrome P450 monooxygenase system was shown to be involved in bisphenol A (BPA) degradation by Sphingomonas sp. strain AO1. In the present investigation, we purified the components of this monooxygenase, cytochrome P450 (P450bisd), ferredoxin (Fdbisd), and ferredoxin reductase (Redbisd). We demonstrated that P450bisd and Fdbisd are homodimeric proteins with molecular masses of 102.3 and 19.1 kDa, respectively, by gel filtration chromatography analysis. Spectroscopic analysis of Fdbisd revealed the presence of a putidaredoxin-type [2Fe-2S] cluster. P450bisd, in the presence of Fdbisd, Redbisd, and NADH, was able to convert BPA. The Km and k cat values for BPA degradation were 85 ± 4.7 μM and 3.9 ± 0.04 min−1, respectively. NADPH, spinach ferredoxin, and spinach ferredoxin reductase resulted in weak monooxygenase activity. These results indicated that the electron transport system of P450bisd might exhibit strict specificity. Two BPA degradation products of the P450bisd system were detected by high-performance liquid chromatography analysis and were thought to be 1,2-bis(4-hydroxyphenyl)-2-propanol and 2,2-bis(4-hydroxyphenyl)-1-propanol based on mass spectrometry-mass spectrometry analysis. This is the first report demonstrating that the cytochrome P450 monooxygenase system in bacteria is involved in BPA degradation.

Molecular genetics of the human cytochrome P450 monooxygenase superfamily

Xenobiotica ◽  
1998 ◽  
Vol 28 (12) ◽  
pp. 1129-1165 ◽  
Author(s):  
G. SMITH ◽  
M. J. STUBBINS ◽  
L. W. HARRIES ◽  
C. R. WOLF
Keyword(s):  
Cytochrome P450 ◽  
Molecular Genetics ◽  
Cytochrome P450 Monooxygenase ◽  
P450 Monooxygenase ◽  
Human Cytochrome

Biodegradation of bisphenol A and 4-alkylphenols by Novosphingobium sp. strain TYA-1 and its potential for treatment of polluted water

2012 ◽  
Vol 66 (10) ◽  
pp. 2202-2208 ◽  
Author(s):  
T. Toyama ◽  
Y. Kainuma ◽  
S. Kikuchi ◽  
K. Mori
Keyword(s):  
Cytochrome P450 ◽  
Bisphenol A ◽  
Cytochrome P450 Monooxygenase ◽  
Polluted Water ◽  
Monooxygenase System ◽  
Simultaneous Removal ◽  
P450 Monooxygenase ◽  
Alkyl Chains ◽  
Batch Bioreactor ◽  
Polluted Waters

We investigated the use of Novosphingobium sp. strain TYA-1 for the simultaneous removal of bisphenol A (BPA) and 4-alkylphenols (4-APs) from complex polluted waters. Strain TYA-1 degraded BPA and utilized it as a sole carbon and energy source via oxidative skeletal rearrangement involving the cytochrome p450 monooxygenase system. Strain TYA-1 also degraded 4-APs with branched side alkyl chains (4-tert-butylphenol [4-tert-BP], 4-sec-butylphenol, 4-tert-pentylphenol, 4-tert-octylphenol [4-tert-OP], and branched nonylphenol mixture) via 4-alkylcatechols but could not degrade 4-APs with linear side alkyl chains. Degradation of 4-APs, like that of BPA, involved the cytochrome p450 monooxygenase system in strain TYA-1. A sequencing batch bioreactor (100 mL of polluted water [50 mg/L BPA, 50 mg/L 4-tert-BP, and 5 mg/L 4-tert-OP]; 6 h of reaction time/cycle; 12 cycles in total) containing alginate-immobilized TYA-1 cells (15 mg dry cells) simultaneously removed BPA, 4-tert-BP, and 4-tert-OP from complex polluted waters. These immobilized TYA-1 cells could be reused for a total of 9 cycles without any loss of degradation activity. Our results support the potential of using immobilized TYA-1 cells for the simultaneous removal of BPA and 4-APs from complex polluted waters.

Immobilization of a Bacterial Cytochrome P450 Monooxygenase System on a Solid Support

2016 ◽  
Vol 128 (48) ◽  
pp. 15226-15230 ◽  
Author(s):  
Cheau Yuaan Tan ◽  
Hidehiko Hirakawa ◽  
Risa Suzuki ◽  
Tomoaki Haga ◽  
Fumiya Iwata ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Solid Support ◽  
Cytochrome P450 Monooxygenase ◽  
Monooxygenase System ◽  
P450 Monooxygenase
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