Genomic clustering of functionally related genes is rare in yeasts and other eukaryotes with only few examples available. Here, we summarize our data on a nontelomeric  MAL  cluster of a non-conventional methylotrophic yeast  Ogataea  ( Hansenula )  polymorpha  containing genes for α-glucosidase MAL1, α-glucoside permease MAL2 and two hypothetical transcriptional activators. Using genome mining, we detected  MAL  clusters of varied number, position and composition in many other maltose-assimilating non-conventional yeasts from different phylogenetic groups. The highest number of  MAL  clusters was detected in  Lipomyces starkeyi  while no  MAL  clusters were found in  Schizosaccharomyces pombe  and  Blastobotrys adeninivorans . Phylograms of α-glucosidases and α-glucoside transporters of yeasts agreed with phylogenesis of the respective yeast species. Substrate specificity of unstudied α-glucosidases was predicted from protein sequence analysis. Specific activities of  Scheffersomyces  stipitis  α-glucosidases MAL7, MAL8, and MAL9 heterologously expressed in  Escherichia coli  confirmed the correctness of the prediction-these proteins were verified promiscuous maltase-isomaltases. α-Glucosidases of earlier diverged yeasts  L. starkeyi ,  B. adeninivorans  and  S. pombe  showed sequence relatedness with α-glucosidases of filamentous fungi and bacilli.