Selective Allylic Hydroxylation of Acyclic Terpenoids by CYP154E1 from Thermobifida Fusca YX

Overview

Journal Beilstein J Org Chem

Specialty Chemistry

Date 2014 Jul 4

PMID 24991288

Citations 7

Authors

Anna M Bogazkaya

Clemens J von Buhler

Sebastian Kriening

Alexandrine Busch

Alexander Seifert

Jurgen Pleiss

Sabine Laschat

Vlada B Urlacher

Affiliations

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Abstract

Allylic alcohols are valuable precursors in the synthesis of pharmaceutical intermediates, agrochemicals and natural products. Regioselective oxidation of parental alkenes is a challenging task for chemical catalysts and requires several steps including protection and deprotection. Many cytochrome P450 enzymes are known to catalyse selective allylic hydroxylation under mild conditions. Here, we describe CYP154E1 from Thermobifida fusca YX that enables this type of oxidation. Several acyclic terpenoids were tested as possible substrates for CYP154E1, and the regio- and chemoselectivity of their oxidation was investigated. Using a previously established bioinformatics approach we identified position 286 in the active site of CYP154E1 which is putatively involved in substrate binding and thereby might have an effect on enzyme selectivity. To tune regio- and chemoselectivity of the enzyme three mutants at position 286 were constructed and used for substrate oxidation. All formed products were analysed with GC-MS and identified using chemically synthesised authentic samples and known compounds as references. Best regioselectivity towards geraniol and nerol was observed with the wild type enzyme mainly leading to 8-hydroxy derivatives (8-hydroxygeraniol or 8-hydroxynerol) with high selectivity (100% and 96% respectively). Highest selectivities during the oxidation of geranylacetone and nerylacetone were observed with the following variants: V286F led mainly to 7-hydroxygeranylacetone (60% of the total product) and V286A produced predominantly 12-hydroxynerylacetone (75% of total product). Thus, CYP154E1 and its mutants expand the tool-box for allylic hydroxylation in synthetic chemistry.

Citing Articles

Enzyme-catalyzed allylic oxidation reactions: A mini-review.

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Synthesis of 8-Hydroxygeraniol.

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