[Localization and nucleotide sequence of propionyl acylase gene of Streptomyces mycarofaciens].
Our research showed that propionyl acylase gene is on 4.16kb insert DNA fragment originated from S. mycarofaciens in recombinant plasmid pIJM9. For localization of this gene on 4.16kb insert DNA fragment, sub-cloning has been carried out with religation of BamHI digested plasmid pIJM9. Among the transformants, molecular weight of recombinant plasmid pIJM95 harbouring in No. 5 transformant is 5.0kb. Molecular weight of insert DNA fragment is 0.53kb in this plasmid. In bioconversion experiment for spiramycin of No. 5 transformant, the bioconversion product was analysed with TLC, bioautography, HPLC and mass spectrum (FAB). Results showed that the bioconvert product is propionylspiramycin. No. 5 transformant is able to transform spiramycin into propionylspiramycin. Propionyl acylase gene was locolized on 0.53kb insert DNA fragment of recombinant plasmid pIJM95. Analysis result of DNA sequence showed that content of G + C is 68.2% for 0.53kb insert DNA fragment. More than 70 kinds of restriction endonuclease have cut sit on this fragment. From No.54-No.393 nucleotide, there is an open reading frame, which codes a polypeptide consisted of 122 amino acids. Start codon is ATG, stop codon is TGA.
Cui
L
L
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing.
Li
Y
Y
Liu
B
B
D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D008969:Molecular Sequence Data; D010957:Plasmids; D015183:Restriction Mapping; D015572:Spiramycin / Q000378:metabolism*; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*
D015572:Spiramycin; D000217:Acyltransferases; C023906:spiramycin I 3-hydroxyl acylase