Purification and characterization of 40-kDa sterigmatocystin O-methyltransferase involved in aflatoxin biosynthesis.

Abstract

Sterigmatocystin-O-methyltransferase (ST-OMTase), an enzyme catalyzing O-methylation of sterigmatocystin with S-adenosylmethionine (SAM), was purified to electrophoretic homogeneity by immunoaffinity chromatography. A novel spectrofluorometric method was established to quantitatively determine the enzymatic activity of ST-OMTase. The purified protein, with a molecular weight of 40 kDa by SDS-PAGE, was sensitive to thiol reagents and low concentrations of heavy metal ions. Using a nutritional shift assay, the expression patterns for ST-OMTase and the transcripts of its corresponding gene, omtA, correlated well with that for aflatoxin B(1) formation. Neither methyltransferase activity nor omtA, mRNA was detected in the fungal cultures of nonaflatoxigenic isolates, including A. flavus, A. sojae, A. nidulans and A. versicolor under optimal growing conditions for aflatoxin B(1) production.