Provided are a polypeptide having glycerol-3-phosphate dehydrogenase activity and a 80% or more homology to an amino acid sequence as set forth in SEQ ID NO: 1 and a polypeptide having glycerol-3-phosphate phosphatase activity and a 80% or more homology to an amino acid sequence as set forth in SEQ ID NO: 2. Provided is also a method for producing glycerol, which includes: culturing a host cell transformed with a vector containing a polynucleotide including a first polynucleotide encoding an amino acid sequence as set forth in SEQ ID NO: 1 and a second polynucleotide encoding an amino acid sequence as set forth in SEQ ID NO: 2 which are operably linked to a suitable regulatory sequence; and recovering glycerol from the culture.