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Abstract

(R,S)-(±)-linalool was the source of carbon and energy during the isolation of Thauera linaloolentis 47LolT, a denitrifying Betaproteobacterium. Preliminary studies have shown the formation of linalool from geraniol, which is the first evidence of the activity of a new enzyme, the linalool-isomerase (9). Until now only the linalool-dehydrataseisomerase of Castellaniela defragrans 65Phen was known. This enzyme only transforms (S)-(+)-linalool. This Bachelor thesis documents the first in vitro measurement of the the activity of the linalool-isomerase of T. linaloolentis 47LolT. A tenfold enrichment of the enzyme succeeded with protein anion exchange chromatography, but a further purification failed. The enzyme does not bind to anion exchange material and is hydrophobic. Two major protein bands and several less intense bands were shown at the reached purification level. The extract as well as the purified fractions needed the reduction agent dithionite to reach the maximum enzyme activity. The activity has an optimum pH-value in the alkaline region and the reaction from geraniol to linalool has an activation energy of 79 kJ mol-1. Substrate conversion and the first enrichment of the linalool-isomerase proved that there does exist an enzyme for the isomerisation of tertiary alcohols. This thesis is a contribution to the knowledge of diversity of enzymatic reactions, which is desirable for biotechnological applications.