Cloning, nucleotide sequence determination and expression of the genes encoding cytochrome P-450soy and ferredoxinsoy (soyB) from Streptomyces griseus |
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Authors: | M K Trower † R Lenstra C Omer ‡ S E Buchholz§ F S Sariaslani |
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Institution: | Central Research and Development Department, E. I. du Pont de Nemours and Company, Inc., Experimental Station, Wilmington, Delaware 19880-0228. |
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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. |
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