Cloning and nucleotide sequences of two genes involved in the 4''-O-acylation of macrolide antibiotics from Streptomyces thermotolerans.
A DNA fragment responsible for the 4''-O-acylation of macrolide antibiotics was cloned from a mutant strain of the carbomycin producer Streptomyces thermotolerans. The gene encoding the macrolide 4''-O-acyltransferase was within a 2.7-kb region of the cloned fragment (15-kb). Streptomyces lividans carrying the region converted exogenously added tylosin to 4''-O-acyltylosins. Nucleotide sequencing of the region showed two open reading frames (ORFs). Expression assay using deleted plasmids showed that both ORFs were essential for optimal expression of the acyltransferase activity. One of them (acyB1) was identical with carE reported previously as a gene encoding 4''-mycarosyl isovaleryl-CoA transferase. The other (acyB2) was assumed to encode a novel regulatory protein that could active acyB1 expression. acyB1 and acyB2 were highly conserved among streptomycetes with macrolide 4''-O-acyl transferase activity.
Arisawa
A
A
Mercian Corporation, Central Research Laboratories, Kanagawa, Japan.
Kawamura
N
N
Tsunekawa
H
H
Okamura
K
K
Tone
H
H
Okamoto
R
R
D000215:Acylation; D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003470:Culture Media; D015870:Gene Expression; D005798:Genes, Bacterial; D005809:Genes, Regulator; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism; D014166:Transferases / Q000737:chemistry / Q000235:genetics*; D015645:Tylosin / Q000378:metabolism
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D003470:Culture Media; C085543:acyB2 protein, Streptomyces thermotolerans; D014166:Transferases; D000217:Acyltransferases; C085542:4''-mycarosyl isovaleryl-coenzyme A transferase; D015645:Tylosin