Filamentous fungi have increasingly been used as hosts for heterologous protein production because of their high secretion capability and ability to add eukaryotic post-translational modifications. In this study, a novel uracil-deficient Aspergillus transformation system, which was based on an orotate phosphoribosyltransferase (pyrF) nutritional selection marker, was discovered. Additionally, a universal, purify-able vector that directed genes into the Aspergillus host strain was engineered. A genomic DNA segment encoding a novel a-amylase was isolated from the psychrotolerant fungus Geomyces pannorum and the open reading frame was determined, deduced 497 amino acids. G. pannorum alpha-amylase was then expressed in the newly constructed Aspergillus oryzae system, with an amylase activity reaching 958 U/ml. It was purified to electrophoretic homogeneity and has a molecular mass of approximately 54 kDa. The enzyme exhibited an optimal activity at pH 5.0 and 40 degrees C and retained over 20% of maximal activity over the temperature range 0-20 degrees C. To our knowledge, this report is the first of the heterologous expression of a cold-adapted enzyme in filamentous fungi. G. pannorum alpha-amylase is an economical amylase with many potential applications. (C) 2015 Elsevier Ltd. All rights reserved.