We have isolated an enolase gene (enoA) from Aspergillus oryzae by heterologous hybridization using the corresponding Saccharomyces cerevisiae ENO2 gene as a probe. A 2.9-kb BglII-fragment contained the entire structural gene enoA including 5'- and 3'- flanking regions. The homology between A. oryzae enoA and S. cerevisiae ENO2 genes is 66.9% when introns are removed. Genomic Southern analysis indicated that there is only one enolase gene in A. oryzae.