Construction of a stable bioengineered strain of biotechmycin.
A stable bioengineered strain of Biotechmycin (Streptomyces spiramyceticus WSJ-1) was constructed by integrating the 4"-isovaleryltransferase gene (ist) through homologous recombination into the chromosome of spiramycin-producing strain S.spiramyceticus F21. In this construction, a Streptomyces/E.coli shuttle plasmid pKC1139 (AmR) was used as the vector and tsr gene was inserted as the marker for selection of homologous recombination. This constructed strain, S.spiramyceticus WSJ-1, was genetically very stable in production titer and in the production of biotechmycin as well as in carrying tsr selective marker when grown without pressure. The fermentation of S.spiramyceticus WSJ-1 was also improved compared with the original strain harboring unintegrated plasmid. Southern hybridization confirmed the integrated status of the ist gene in the host genome.
Shang
G
G
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing, China.
Dai
J
J
Wang
Y
Y
D000217:Acyltransferases / Q000235:genetics*; D000900:Anti-Bacterial Agents / Q000378:metabolism; D015139:Blotting, Southern; D002855:Chromatography, Thin Layer; D005285:Fermentation; D011995:Recombination, Genetic; D015572:Spiramycin / Q000031:analogs & derivatives / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
D000900:Anti-Bacterial Agents; D015572:Spiramycin; D000217:Acyltransferases; C085542:4''-mycarosyl isovaleryl-coenzyme A transferase