Cytochrome P450 immobilisation as a route to bioremediation/biocatalysis

FEBS Letters

Kelly, Steven L.¹  Lamb, Stephen B.²  Lamb, David C.¹  Stuckey, David C.² 

[1]Institute of Biological Sciences, University of Wales Aberystwyth, Aberystwyth SY23 3DA, UK

[2]Department of Chemical Engineering, Imperial College, Prince Consort Road, London, SW7 2BY, UK

关键词: Cytochrome P450;    Colloidal liquid aphron (CLA);    Immobilization;    Fusion protein;   

DOI  :  10.1016/S0014-5793(98)00771-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The diverse substrate specificity of the cytochrome P450 (P450; CYP) enzyme superfamily offers the opportunity to develop enzymatic systems for environmental detoxification and biotransformations of drugs, pesticides and fine chemicals. Here we report on the immobilisation of a fusion protein between plant cytochrome P450-71B1 (CYP71B1) and its electron donor, plant NADPH cytochrome P450 reductase using an oil-in-water macro-emulsion, termed polyaphron, which contains a proportion of internal organic phase (φ) greater than 0.74. Efficiency of P450 immobilisation was greater than 85%, and in this state enzymatic activity could be measured for more than 24 h at 15°C. Chlortoluron, a recalcitrant herbicide pollutant in the environment, was shown to be metabolised, with the major metabolite (N-monodemethylated chlortoluron) being separated from the substrate due to partitioning into the aqueous phase. The turnovers exhibited superactivity compared with those obtained using free enzyme located in membranes prepared following heterologous expression in Saccharomyces cerevisiae and Escherichia coli. The potential to exploit the unprecedented catalytic diversity of the P450 superfamily in biocatalysis is discussed.

【 授权许可】

Unknown