Purification and some properties of malate synthase from the methylotrophic yeast Hansenula polymorpha

Abstract Malate synthase, one of the key enzymes in the glyoxylate cycle, was purified 122-fold to homogeneity from ethanol-grown Hansenula polymorpha . SDS-polyacrylamide gel electrophoresis showed that the enzyme has a subunit size of 62 000 daltons. The molecular mass of native malate synthase was determined to be 250 000 daltons by gel filtration, indicating that the enzyme is a tetramer. Cell fractionation studies and immunogold staining, carried out on ultrathin sections of ethanol-grown H. polymorpha , using malate synthase-specific antibodies, showed that malate synthase was localized in the matrix of peroxisomes.