Human cytochrome P450 expression in bacteria: Whole-cell high-throughput activity assay for CYP1A2, 2A6 and 3A4

2018 ◽  
Vol 158 ◽  
pp. 134-140 ◽  
Author(s):  
Francisco Esteves ◽  
Diana Campelo ◽  
Philippe Urban ◽  
Sophie Bozonnet ◽  
Thomas Lautier ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
High Throughput ◽  
Activity Assay ◽  
Whole Cell ◽  
Human Cytochrome ◽  
P450 Expression

scholarly journals A High‐Throughput Mass Spectrometric Enzyme Activity Assay Enabling the Discovery of Cytochrome P450 Biocatalysts

2019 ◽  
Vol 58 (30) ◽  
pp. 10114-10119 ◽  
Author(s):  
Tristan de Rond ◽  
Jian Gao ◽  
Amin Zargar ◽  
Markus de Raad ◽  
Jack Cunha ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Cytochrome P450 ◽  
High Throughput ◽  
Mass Spectrometric ◽  
Activity Assay ◽  
Enzyme Activity Assay

Development of the “Inje Cocktail” for High-throughput Evaluation of Five Human Cytochrome P450 Isoforms in vivo

2007 ◽  
Vol 82 (5) ◽  
pp. 531-540 ◽  
Author(s):  
J Y Ryu ◽  
I S Song ◽  
Y E Sunwoo ◽  
J H Shon ◽  
K H Liu ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
High Throughput ◽  
Human Cytochrome ◽  
Cytochrome P450 Isoforms

scholarly journals A High‐Throughput Mass Spectrometric Enzyme Activity Assay Enabling the Discovery of Cytochrome P450 Biocatalysts

2019 ◽  
Vol 131 (30) ◽  
pp. 10220-10225 ◽  
Author(s):  
Tristan de Rond ◽  
Jian Gao ◽  
Amin Zargar ◽  
Markus de Raad ◽  
Jack Cunha ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Cytochrome P450 ◽  
High Throughput ◽  
Mass Spectrometric ◽  
Activity Assay ◽  
Enzyme Activity Assay

scholarly journals Quantitative Whole-Cell Cytochrome P450 Measurement Suitable for High-Throughput Application

2008 ◽  
Vol 13 (2) ◽  
pp. 135-141 ◽  
Author(s):  
Wayne A. Johnston ◽  
Weiliang Huang ◽  
James J. De Voss ◽  
Martin A. Hayes ◽  
Elizabeth M.J. Gillam
Keyword(s):  
Cytochrome P450 ◽  
High Throughput ◽  
Signal To Noise Ratio ◽  
Recombinant Expression ◽  
Spectral Determination ◽  
Cytochrome P450 Enzymes ◽  
Whole Cell ◽  
Expression Levels ◽  
Whole Cells ◽  
Modified Method

The recombinant expression of cytochrome P450 enzymes involved in drug metabolism is of interest to the pharmaceutical and biotechnological industries due to the versatile catalytic properties of these enzymes. Accurate quantification of cytochrome P450 enzymes expressed in bacterial culture generally depends on disruption and fractionation of cells to prepare membranes for spectral analysis. Although whole-cell methods for spectral determination have been reported, problems with poor reproducibility and low signal-to-noise ratio confound the use of such techniques where P450 hemoprotein expression levels are relatively low, such as in cultures of certain mammalian forms. In particular, interference from bacterial hemoproteins often obscures the P450 peak. In the current study, the combination of culture concentration, incubation under microaerobic conditions, and a modified method of baseline correction enabled reproducible quantification of cytochrome P450s in whole cells. This whole-cell method is well suited to high-throughput application, as large sets or libraries of enzymes can be expressed in parallel and relative expression levels measured without downstream cell processing. ( Journal of Biomolecular Screening 2008:135-141)

scholarly journals High-throughput Screening of Inhibitory Effects of Bo-yang-hwan-o-tang on Human Cytochrome P450 Isoforms in vitro Using UPLC/MS/MS

2012 ◽  
Vol 28 (12) ◽  
pp. 1197-1201 ◽  
Author(s):  
Miran LEE ◽  
Jeonghyeon PARK ◽  
Mi-sun LIM ◽  
Sook Jin SEONG ◽  
Joomi LEE ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
High Throughput ◽  
High Throughput Screening ◽  
Inhibitory Effects ◽  
Human Cytochrome ◽  
Cytochrome P450 Isoforms
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