Cloning of glyceraldehyde-3-phosphate dehydrogenase-encoding genes in Mucor circinelloides (Syn. racemosus) and use of the gpd1 promoter for recombinant protein production.

Three genes (gpd1, gpd2, and gpd3) encoding glyceraldehyde-3-phosphate dehydrogenase were isolated from the dimorphic zygomycete Mucor circinelloides by PCR using degenerated primers. Transcription of gpd1 could be detected during vegetative growth under both aerobic and anaerobic conditions, whereas neither gpd2 nor gpd3 transcription was detected, indicating that gpd1 is the major transcribed gpd gene. The transcription of gpd1 was regulated by carbon source. The gpd1 promoter was successfully used for recombinant expression of genes of both homologous (crgA encoding a regulator of carotene biosynthesis) and heterologous (gox1 from Aspergillus niger encoding glucose oxidase; GOX) nature. Growth of a gox1 transformant strain resulted in the secretion of enzymatically active GOX. The potential advantages of using a dimorphic fungus for heterologous protein production are discussed.