Direct evidence for the function of FUM13 in 3-ketoreduction of mycotoxin fumonisins in Fusarium verticillioides.
Fumonisins are mycotoxins produced by Fusarium verticillioides, a widespread pathogen of corn. Although the gene cluster for the biosynthesis of fumonisins has been cloned, the majority of the genes have not been biochemically characterized. Here, we report the biochemical characterization of FUM13, a gene that encodes a short-chain dehydrogenase/reductase required for fumonisin biosynthesis. FUM13 has been expressed in E. coli, and the produced protein, Fum13p, has been purified. When the protein was incubated with 3-keto fumonisin B(3) (FB(3)) in the presence of NADPH, FB(3) was produced. The data provide direct evidence for the role of FUM13 in the 3-ketoreduction of fumonisins. In a functional complementation experiment, FUM13 gene was introduced into tsc10 mutants of the yeast Saccharomyces cerevisiae, which carry a mutation in the 3-ketosphinganine reductase gene in the sphingolipid pathway. The tsc10 mutants were not able to grow on the selection medium, but the same mutants transformed with FUM13 were able to grow. The results further confirm the function of FUM13 in 3-ketoreduction in vivo.
Yi
Han
H
Chemistry Department, University of Nebraska-Lincoln, Lincoln, 68588-0304, USA.
Bojja
Ravi S
RS
Fu
Jayzonn
J
Du
Liangcheng
L
D004926:Escherichia coli / Q000235:genetics; D037341:Fumonisins / Q000378:metabolism*; D005670:Fusarium / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D009154:Mutation; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014162:Transfection
D037341:Fumonisins; D011994:Recombinant Proteins; D010088:Oxidoreductases