Cloning and expression of propionyl acylase gene of Streptomyces mycarofaciens mutant.
The midecamycin producer, S. mycarofaciens mutant, which has propionyl acylase activity, can convert spiramycin into propionyl spiramycin. Plasmid pIJ702 was used as a vector for the cloning of propionyl acylase gene. After shot-gun cloning, the DNA fragments of the mutant were cloned in S. lividans TK54. The results of TLC and HPLC showed that No.9 transformant can convert spiramycin into propionyl spiramycin. It demonstrated that the propionyl acylase gene was cloned and expressed in S. lividans TK54. The insert fragment of No.9 recombinant plasmid is about 4.16 kb. Southern hybridization showed that the fragment originated from S. mycarofaciens mutant. The restriction endonuclease map of No.9 recombinant plasmid was constructed.
Li
Y A
YA
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing.
Sun
Y P
YP
Shi
L Y
LY
Li
X P
XP
Li
H L
HL
D000217:Acyltransferases / Q000235:genetics*; D015139:Blotting, Southern; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D010957:Plasmids; D015183:Restriction Mapping; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*; D014169:Transformation, Bacterial
D000217:Acyltransferases; C023906:spiramycin I 3-hydroxyl acylase