Precursor for biosynthesis of sugar moiety of doxorubicin depends on rhamnose biosynthetic pathway in Streptomyces peucetius ATCC 27952.
The doxorubicin biosynthetic gene cluster in Streptomyces peucetius ATCC 27952 contains a TDP-D-glucose 4,6-dehydratase gene, dnmM, that is putatively involved in the biosynthesis of daunosamine, but the gene contains a frameshift in the DNA sequence that would cause premature termination of translation. In pursuit of another TDP-D-glucose 4,6-dehydratase in S. peucetius, a homologue gene, rmbB, was found, whose deduced product exhibits high sequence similarity to a number of TDP-D-glucose 4,6-dehydratases. The gene was located within a putative rhamnose biosynthetic gene cluster at another locus in the genome. RmbB was verified to be a functional TDP-D-glucose 4,6-dehydratase by enzyme assay as it catalyzed the conversion of TDP-D-glucose into TDP-4-keto-6-deoxy-D-glucose. Inactivation of rmbB in the S. peucetius genome abolished the production of doxorubicin while complementation of the same gene in an rmbB knockout mutant restored the doxorubicin production. Hence, rmbB provides TDP-4-keto-6-deoxy-D-glucose as a nucleotide sugar precursor for the biosynthesis of doxorubicin.
Department of Pharmaceutical Engineering, Institute of Biomolecule Reconstruction (iBR), Sun Moon University, Tangjeong-myeon, Asansi, Chungnam 336-708, South Korea.
Bijay
B
Singh
Chang-Beom
CB
Lee
Jae Kyung
JK
Sohng
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D053898:Biosynthetic Pathways / Q000235:genetics; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D004317:Doxorubicin / Q000096:biosynthesis* / Q000378:metabolism; D057075:Enzyme Assays; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016368:Frameshift Mutation; D020868:Gene Silencing; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D005947:Glucose / Q000031:analogs & derivatives* / Q000378:metabolism; D006595:Hexosamines / Q000096:biosynthesis; D006836:Hydro-Lyases / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D005810:Multigene Family; D012210:Rhamnose / Q000096:biosynthesis* / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D013942:Thymine Nucleotides / Q000378:metabolism*; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D006595:Hexosamines; D013942:Thymine Nucleotides; C112447:dTDP-6-deoxy-4-ketoglucose; C007217:daunosamine; D004317:Doxorubicin; D006836:Hydro-Lyases; C022771:dTDPglucose 4,6-dehydratase; D005947:Glucose; D012210:Rhamnose