Heterologous production of paromamine in Streptomyces lividans TK24 using kanamycin biosynthetic genes from Streptomyces kanamyceticus ATCC12853.
The 2-deoxystreptamine and paromamine are two key intermediates in kanamycin biosynthesis. In the present study, pSK-2 and pSK-7 recombinant plasmids were constructed with two combinations of genes: kanABK and kanABKF and kacA respectively from kanamycin producer Streptomyces kanamyceticus ATCC12853. These plasmids were heterologously expressed into Streptomyces lividans TK24 independently and generated two recombinant strains named S. lividans Sk-2/SL and S. lividans SK-7/SL, respectively. ESI/ MS and ESI-LC/MS analysis of the metabolite from S. lividans SK-2/SL showed that the compound had a molecular mass of 163 [M + H]+, which corresponds to that of 2-deoxystreptamine. ESI/MS and MS/MS analysis of metabolites from S. lividans SK-7/SL demonstrated the production of paromamine with a molecular mass of 324 [M + H]+. In this study, we report the production of paromamine in a heterologous host for the first time. This study will evoke to explore complete biosynthetic pathways of kanamycin and related aminoglycoside antibiotics.
Nepal
Keshav Kumar
KK
Sun Moon University, Asan 336-708, Korea.
Oh
Tae-Jin
TJ
Sohng
Jae Kyung
JK
D000617:Aminoglycosides / Q000235:genetics / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry; D001709:Biotechnology; D005818:Genetic Engineering; D006595:Hexosamines / Q000235:genetics / Q000378:metabolism; D066298:In Vitro Techniques; D007612:Kanamycin / Q000096:biosynthesis* / Q000737:chemistry; D013058:Mass Spectrometry; D008970:Molecular Weight; D013045:Species Specificity; D048170:Streptomyces lividans / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D006595:Hexosamines; C005936:paromamine; D007612:Kanamycin; C007265:2-deoxystreptamine