Cloning, nucleotide sequence determination and expression of the genes encoding cytochrome P-450soy (soyC) and ferredoxinsoy (soyB) from Streptomyces griseus.
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.
Trower
M K
MK
Central Research and Development Department, E. I. du Pont de Nemours and Company, Inc., Experimental Station, Wilmington, Delaware 19880-0228.
Lenstra
R
R
Omer
C
C
Buchholz
S E
SE
Sariaslani
F S
FS
D000595:Amino Acid Sequence; D001483:Base Sequence; D001673:Biodegradation, Environmental; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D004269:DNA, Bacterial / Q000302:isolation & purification; D005288:Ferredoxins / Q000096:biosynthesis / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010446:Peptide Fragments / Q000302:isolation & purification; D014674:Plant Proteins, Dietary / Q000235:genetics; D017385:Sequence Homology; D013025:Glycine max / Q000201:enzymology / Q000235:genetics*; D013305:Streptomyces griseus / Q000201:enzymology / Q000235:genetics*
D004269:DNA, Bacterial; D005288:Ferredoxins; D015345:Oligonucleotide Probes; D010446:Peptide Fragments; D014674:Plant Proteins, Dietary; D003577:Cytochrome P-450 Enzyme System