Premium
Isolierung und Rekonstitution des Alkan‐Monooxygenase‐Systems der Hefe Lodderomyces elongisporus
Author(s) -
Schunck W.H.,
Riege P.,
Honeck H.,
Müller H.G.
Publication year - 1983
Publication title -
zeitschrift für allgemeine mikrobiologie
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0044-2208
DOI - 10.1002/jobm.19830231007
Subject(s) - chemistry , monooxygenase , hemeprotein , biochemistry , cytochrome , reductase , hydroxylation , flavoprotein , yeast , enzyme , stereochemistry , cytochrome p450 , heme
It is generally assumed that the first step of alkane degradation in several yeast strains and bacteria is catalyzed by a monooxygenase system containing cytochrome P‐450 as the terminal oxidase and a NADPH‐cytochrome P‐450 reductase as the electron transfer component. To prove this hypothesis both proteins were purified to homogeneity from the microsomal fraction of the alkane‐assimilating yeast L. elongisporus and reconstituted in vitro. The cytochrome P‐450, having a specific content of 12–17 nmoles/mg protein turned out to be a typical member of this class of hemoproteins with respect to the molecular weight (53 500) and spectral properties. The reductase component with a molecular weight of 79000 contains FAD and FMN as prosthetic groups. The recombination of the hemoprotein and the flavoprotein results in an enzyme system which catalyzes the monoterminal hydroxylation of hexadecane. The activity of the reconstituted enzyme system is remarkably stimulated by nonionic detergents, suggesting the requirement of an additional lipid factor. The possible importance of other electron transfer systems and of substrate transport is discussed with respect ot the function of the alkane monooxygenase system under in vivo conditions.