Characterization and expression of a P-450-like mycinamicin biosynthesis gene using a novel Micromonospora-Escherichia coli shuttle cosmid vector.
A 29 kb shuttle cosmid vector, pTYS507, was constructed from a cryptic Micromonospora griseorubida plasmid and the Escherichia coli cosmid pJB8. Subcloning of mycinamicin II biosynthesis genes in pTYS507 led to the identification of a DNA region that could complement a mutant of M. griseorubida that lacked both hydroxylase and epoxidase activities. Nucleotide sequence and mutational analysis suggested that a single P-450-like protein catalyzes both reactions.
Inouye
M
M
Institute for Life Science Research, Shizuoka, Japan.
Takada
Y
Y
Muto
N
N
Beppu
T
T
Horinouchi
S
S
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D001426:Bacterial Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D003360:Cosmids; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005822:Genetic Vectors; D006900:Hydroxylation; D018942:Macrolides; D008847:Micromonospora / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000378:metabolism; D015183:Restriction Mapping
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D018942:Macrolides; C026686:mycinamicins; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; C023111:epoxidase