Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 372158
Title Toluene monooxygenase from the fungus Cladosporium sphaerospermum
Author(s) Luykx, D.M.A.M.; Prenafeta-Boldu, F.X.; Bont, J.A.M. de
Source Biochemical and Biophysical Research Communications 312 (2003)2. - ISSN 0006-291X - p. 373 - 379.
Department(s) RIKILT - Analyse & Ontwikkeling
AFSG Staff Departments (WUATV)
Publication type Refereed Article in a scientific journal
Publication year 2003
Keyword(s) cytochrome-c reductase - cytosolic cytochrome-p450 - aromatic-hydrocarbons - liver microsomes - energy-source - sole carbon - purification - metabolism - hydroxylation - pathway
Abstract Assimilation of toluene by Cladosporium sphaerospermum is initially catalyzed by toluene monooxygenase (TOMO). TOMO activity was induced by adding toluene to a glucose-pregrown culture of C. sphaerospermum. The corresponding microsomal enzyme needed NADPH and O2 to oxidize toluene and glycerol, EDTA, DTT, and PMSF for stabilization. TOMO activity was maximal at 35 °C and pH 7.5 and was inhibited by carbon monoxide, Metyrapone, and cytochrome c. TOMO preferred as substrates also other aromatic hydrocarbons with a short aliphatic side chain. Its reduced carbon monoxide difference spectrum showed a maximum at 451 nm. A substrate-induced Type I spectrum was observed on addition of toluene. These results indicated that TOMO is a cytochrome P450. TOMO and its corresponding reductase were eventually purified by a simultaneous purification revealing apparent molecular masses of 58 and 78 kDa, respectively.
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