Cloning and in vitro characterization of dTDP-6-deoxy-L-talose biosynthetic genes from Kitasatospora kifunensis featuring the dTDP-6-deoxy-L-lyxo-4-hexulose reductase that synthesizes dTDP-6-deoxy-L-talose.
Kitasatospora kifunensis, the talosin producer, was used as a source for the dTDP-6-deoxy-l-talose (dTDP-6dTal) biosynthetic gene cluster, serving as a template for four recombinant proteins of RmlA(Kkf), RmlB(Kkf), RmlC(Kkf), and Tal, which complete the biosynthesis of dTDP-6dTal from dTTP, alpha-d-glucose-1-phosphate, and NAD(P)H. The identity of dTDP-6dTal was validated using (1)H and (13)C NMR spectroscopy. K. kifunensistal and tll, the known dTDP-6dTal synthase gene of Actinobacillus actinomycetemcomitans origin, have low sequence similarity and are distantly related within the NDP-6-deoxy-4-ketohexose reductase family, providing an example of the genetic diversity within the dTDP-6dTal biosynthetic pathway.
Karki
Suman
S
Department of Biological Science, Myongji University, Yongin, Republic of Korea.
Yoo
Hye-Gyeong
HG
Kwon
So-Yeon
SY
Suh
Joo-Won
JW
Kwon
Hyung-Jin
HJ
D003001:Cloning, Molecular; D014644:Genetic Variation; D007783:Lactones / Q000378:metabolism*; D008969:Molecular Sequence Data; D009702:Nucleoside Diphosphate Sugars / Q000378:metabolism*; D013306:Streptomycetaceae / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D013401:Sugar Alcohol Dehydrogenases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
C510913:L-talo-gamma-lactone; D007783:Lactones; D009702:Nucleoside Diphosphate Sugars; C553917:dTDP-6-deoxytalose; D013401:Sugar Alcohol Dehydrogenases; C405941:thymidine diphosphate-6-deoxy-L-lyxo-4-hexulose reductase