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Applied and Environmental Microbiology, April 2004, p. 1931-1934, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.1931-1934.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Fum3p, a 2-Ketoglutarate-Dependent Dioxygenase Required for C-5 Hydroxylation of Fumonisins in Fusarium verticillioides

Yousong Ding, Ravi S. Bojja, and Liangcheng Du*

Chemistry Department, University of Nebraska—Lincoln, Lincoln, Nebraska 68588-0304

Received 13 August 2003/ Accepted 7 January 2004

Fumonisins are polyketide-derived mycotoxins produced by several agriculturally important Fusarium species. The B series fumonisins, FB1, FB2, FB3, and FB4, are fumonisins produced by wild-type Fusarium verticillioides strains, differing in the number and location of hydroxyl groups attached to the carbon backbone. We characterized the protein encoded by FUM3, a gene in the fumonisin biosynthetic gene cluster. The 33-kDa FUM3 protein (Fum3p) was heterologously expressed and purified from Saccharomyces cerevisiae. Yeast cells expressing the Fum3p converted FB3 to FB1, indicating that Fum3p catalyzes the C-5 hydroxylation of fumonisins. This result was verified by assaying the activity of Fum3p purified from yeast cells. The C-5 hydroxylase activity of purified Fum3p required 2-ketoglutarate, Fe2+, ascorbic acid, and catalase, all of which are required for 2-ketoglutarate-dependent dioxygenases. The protein also contains two His motifs that are highly conserved in this family of dioxygenases. Thus, Fum3p is a 2-ketoglutarate-dependent dioxygenase required for the addition of the C-5 hydroxyl group of fumonisins.


* Corresponding author. Mailing address: Chemistry Department, University of Nebraska—Lincoln, Lincoln, NE 68588-0304. Phone: (402) 472-2998. Fax: (402) 472-9402. E-mail: ldu{at}unlserve.unl.edu.


Applied and Environmental Microbiology, April 2004, p. 1931-1934, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.1931-1934.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.







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