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Applied and Environmental Microbiology, November 2004, p. 6518-6524, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6518-6524.2004

Aflatoxin Biosynthesis Cluster Gene cypA Is Required for G Aflatoxin Formation

Kenneth C. Ehrlich,1,{dagger}* Perng-Kuang Chang,1,{dagger} Jiujiang Yu,1 and Peter J. Cotty1,2

Southern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, New Orleans, Louisiana,1 Division of Plant Pathology and Microbiology, Department of Plant Sciences, University of Arizona, Tucson, Arizona2

Received 15 March 2004/ Accepted 24 June 2004

Aspergillus flavus isolates produce only aflatoxins B1 and B2, while Aspergillus parasiticus and Aspergillus nomius produce aflatoxins B1, B2, G1, and G2. Sequence comparison of the aflatoxin biosynthesis pathway gene cluster upstream from the polyketide synthase gene, pksA, revealed that A. flavus isolates are missing portions of genes (cypA and norB) predicted to encode, respectively, a cytochrome P450 monooxygenase and an aryl alcohol dehydrogenase. Insertional disruption of cypA in A. parasiticus yielded transformants that lack the ability to produce G aflatoxins but not B aflatoxins. The enzyme encoded by cypA has highest amino acid identity to Gibberella zeae Tri4 (38%), a P450 monooxygenase previously shown to be involved in trichodiene epoxidation. The substrate for CypA may be an intermediate formed by oxidative cleavage of the A ring of O-methylsterigmatocystin by OrdA, the P450 monooxygenase required for formation of aflatoxins B1 and B2.

* Corresponding author. Mailing address: SRRC/ARS/USDA, 1100 RE Lee Blvd., P.O. Box 19687, New Orleans, LA 70179. Phone: (504) 286 4369. Fax: (504) 286 4419. E-mail: ehrlich{at}srrc.ars.usda.gov.

{dagger} K.C.E. and P.-K.C. contributed equally to this work.

Applied and Environmental Microbiology, November 2004, p. 6518-6524, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6518-6524.2004




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