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Malla, Sailesh,Niraula, Narayan Prasad,Liou, Kwangkyoung,Sohng, Jae Kyung Elsevier 2009 Journal of bioscience and bioengineering Vol.108 No.2
<P><B>Abstract</B></P><P>To enhance doxorubicin (DXR) production, the structural sugar biosynthesis genes <I>desIII</I> and <I>desIV</I> from <I>Streptomyces venezuelae</I> ATCC 15439 and the glycosyltransferase pair <I>dnrS/dnrQ</I> from <I>Streptomyces peucetius</I> ATCC 27952 were cloned into the expression vector pIBR25, which contains a strong <I>ermE⁎</I> promoter. The recombinant plasmids pDnrS25 and pDnrQS25 were constructed for overexpression of <I>dnrS</I> and the <I>dnrS/dnrQ</I> pair, whereas pDesSD25 and pDesQS25 were constructed to express <I>desIII/desIV</I> and <I>dnrS/dnrQ–desIII/desIV</I>, respectively. All of these recombinant plasmids were introduced into <I>S. peucetius</I> ATCC 27952. The recombinant strains produced more DXR than the <I>S. peucetius</I> parental strain: a 1.2-fold increase with pDnrS25, a 2.8-fold increase with pDnrQS25, a 2.6-fold increase with pDesSD25, and a 5.6-fold increase with pDesQS25. This study showed that DXR production was significantly enhanced by overexpression of potential biosynthetic sugar genes and glycosyltransferase.</P>
Generation of sugar modified-doxorubicin from engineered strain of Streptomyces peucetius
Sailesh Malla,Kwangkyoung Liou,Kyung Sohng 한국당과학회 2008 한국당과학회 학술대회 Vol.2008 No.1
Combinatorial biosynthesis is an alternative way for accessing naturally unavailable natural products or improving activity of already existing biomolecules by their modification. Biosynthesis of different deoxy-aminosugar and its attachment to the same or different anthracyclinone aglycones in vivo would lead to the formation of novel anthracycline compounds. Unlike doxorubicin, anthracyclines with N-alkylated sugar moieties were weakly mutagenic or not mutagenic at all in both bacterial and mammalian cells. Disruption of glycosyltransferase gene, dnrS, involved in the biosynthesis of the doxorubicin from Streptomyces peucetius ATCC 27952 led to accumulation of a non-glycosylated intermediate ε-rhodomycinone. Complementation experiment was carried out to introduce L-rhodosamine sugar for the production of rhodosaminyl-doxorubicin. Chromosomal integration of desVI encoding N,N-dimethyltransferase from Streptomyces venezuelae and aknS encoding glycosyltransferase along with its auxiliary gene aknT from Streptomyces galilaeus in the dnrS disruptant of S. peucetius led to formation of rhodosaminyl-doxorubicin.