Open AccessAcetyl‐Coenzyme‐A Carboxylase of Candida lipolytica
Author(s) -
MISHINA Masayoshi,
KAMIRYO Tatsuyuki,
NUMA Shosaku,
TANAKA Atsuo,
FUKUI Saburo
Publication year - 1976
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1976.tb11115.x
Subject(s) - biochemistry , enzyme , polyacrylamide gel electrophoresis , chemistry , gel electrophoresis , enzyme assay , cofactor , pyruvate carboxylase , ouchterlony double immunodiffusion , biology , antiserum , antibody , immunology
Acetyl‐coenzyme‐A carboxylase has been isolated in homogeneous form from Candida lipolytica . The homogeneity of the enzyme preparation is evidenced by analytical ultracentrifugation, dodecylsulfate‐polyacrylamide gel electrophoresis and Ouchterlony double‐diffusion analysis. The purified enzyme exhibits a specific activity of 8.0 U/mg protein at 25°C and contains 1 mol biotin/263000 g protein. The sedimentation coefficient ( S 20, w ) of the enzyme is 18 S. It has been shown by dodecyl‐sulfate‐polyacrylamide gel electrophoresis that the enzyme possesses only one kind of subunit with a molecular weight of 230000. This finding, together with the biotin content, indicates that the C. lipolytica enzyme has a highly integrated subunit structure. The C. lipolytica enzyme is very labile, but is stabilized by glycerol. The enzyme is markedly activated by poly(ethyleneglycol), the activation being due principally to a decrease in the K m values for substrates. Even in the presence of this activator, the K m value for acetyl‐CoA of the C. lipolytica enzyme is much higher than that of the enzyme from Saccharomyces cerevisiae and animal tissues. The C. lipolytica enzyme, unlike the enzyme from animal tissues, is not activated by citrate.