TY  - THES
T1  - Development of efficient cytochrome P450-dependent whole-cell biotransformation reactions for steroid hydroxylation and drug discovery
A1  - Hakki,Tarek
Y1  - 2009/01/16
N2  - Cytochromes P450 play a vital role in the steroid biosynthesis in the human adrenal gland, e.g. the production of hydrocortisone and aldosterone by CYP11B1 and CYP11B2, respectively. The steroid hydroxylases of the CYP11B family are important targets for drug development. Since they are very closely related, the discovery of selective inhibitors has been a focus of interest. Furthermore, hydrocortisone is a precursor for drugs with high therapeutic potential. Therefore, the purpose of this work was the development of an efficient system for CYP11B-dependent whole-cell biotransformation to facilitate the bioproduction of hydrocortisone and the discovery of selective inhibitors. The present work shows clearly that hydrocortisone production can be dramatically enhanced (3.4-fold) by coexpression of the natural redox partners of CYP11B1. Moreover, a high production efficiency has been achieved by optimisation of the reaction conditions. Additionally, in the course of this work an automated screening technology plate-format has been developed using a CYP11B2-expressing fission yeast strain. Additionally, the conditions for HPLC analysis of steroids were optimised and a high throughput screening system for the discovery of CYP11B2 inhibitors has been established. The new screening system was successfully used for the investigation of a library of pharmacologically active compounds resulting in the identification of several novel potential inhibitors of CYP11B2. 
KW  - Cytochrom P-450
KW  - Arzneimittelforschung
KW  - Steroidstoffwechsel
KW  - Screening
KW  - Hypertonie
CY  - Saarbrücken
PB  - Universitäts- und Landesbibliothek
AD  - Postfach 151141, 66041 Saarbrücken
UR  - http://scidok.sulb.uni-saarland.de/volltexte/2009/2054
ER  -