✦ LIBER ✦

Altering the Regioselectivity of Cytochrome P450 CYP102A3 of Bacillus subtilis by Using a New Versatile Assay System

✍ Scribed by Oliver Lentz; Anton Feenstra; Tilo Habicher; Bernhard Hauer; Rolf D. Schmid; Vlada B. Urlacher

Publisher: John Wiley and Sons
Year: 2005
Tongue: English
Weight: 118 KB
Volume: 7
Category: Article
ISSN: 1439-4227
DOI: 10.1002/cbic.200500266

No coin nor oath required. For personal study only.

✦ Synopsis

Abstract

A novel monooxygenase (CYP102A3) has been discovered within the Bacillus subtilis genome that reveals a similarity of 76 % to the well‐known cytochrome P450 BM‐3 of B. megaterium (CYP102A1). Both enzymes are natural fusion proteins consisting of a heme domain and a FAD/FMN‐reductase domain. Because of their high turnover rates, these biocatalysts are of special interest for industrial applications, but show only limited regioselectivity. In this work, the regioselectivity of CYP102A3 was changed by directed evolution and protein design to hydroxylate substrates not only in different subterminal, but also to a high extent, in terminal carbon chain positions. To enable a high‐throughput screening procedure, a very versatile assay was developed that is capable of discriminating between terminal and subterminal hydroxylation of carbon chains. A double mutant of CYP102A3 was obtained that produces 48 % octan‐1‐ol as the main product of the reaction.

📜 SIMILAR VOLUMES

Studies on the Interactions between Drug

Studies on the Interactions between Drugs and Estrogen: Analytical Method for Prediction System of Gynecomastia Induced by Drugs on the Inhibitory Metabolism of Estradiol Using Escherichia coli Coexpressing Human CYP3A4 with Human NADPH-Cytochrome P450 Reductase

✍ Takashi Satoh; Ken-ichi Fujita; Haruka Munakata; Shinji Itoh; Katsunori Nakamura 📂 Article 📅 2000 🏛 Elsevier Science 🌐 English ⚖ 94 KB

To establish a prediction system for drug-induced gynecomastia in clinical fields, a model reaction system was developed to explain numerically this side effect. The principle is based on the assumption that 50% inhibition concentration (IC 50 ) of drugs on the in vitro metabolism of estradiol (E2)