Cloning, nucleotide sequence determination and expression of the genes encoding cytochrome P-450soyand ferredoxinsoy(soyB) from Streptomyces griseus
- 1 August 1992
- journal article
- research article
- Published by Wiley in Molecular Microbiology
- Vol. 6 (15) , 2125-2134
- https://doi.org/10.1111/j.1365-2958.1992.tb01386.x
Abstract
Xenobiotic transformation by Streptomyces griseus (ATCC13273) is catalysed by a cytochrome P-450, designated cytochrome P-450soy. A DNA segment carrying the structural gene encoding P-450soy (soyC) was cloned using an oligonucleotide probe constructed from the protein sequence of a tryptic peptide. Following DNA sequencing the deduced amino acid sequence of P-450soy was compared with that for P-450cam, revealing conservation of important structural components including the haem pocket. Expression of the cloned soyC gene product was demonstrated in Streptomyces lividans by reduced CO:difference spectral analysis and Western blotting. Downstream of soyC, a gene encoding a putative [3Fe-4S] ferredoxin (soyB), named ferredoxinsoy, was identified.Keywords
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