Open AccessAcyl‐Coenzyme‐A Synthetase I from Candida lipolytica
Author(s) -
HOSAKA Kohei,
MISHINA Masayoshi,
TANAKA Takao,
KAMIRYO Tatsuyuki,
NUMA Shosaku
Publication year - 1979
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1979.tb12811.x
Subject(s) - ouchterlony double immunodiffusion , enzyme , sephadex , biochemistry , polyacrylamide gel electrophoresis , degree of unsaturation , chemistry , gel electrophoresis , tributyrin , coenzyme a , chromatography , biology , antibody , lipase , antiserum , reductase , immunology
Acyl‐coenzyme‐A synthetase I from Candida lipolytica has been purified to homogeneity as evidenced by polyacrylamide gel electrophoresis in the presence and absence of dodecylsulfate as well as by Ouchterlony double‐diffusion analysis. The purification procedure involves resolution of cellular particles with Triton X‐100 and chromatography on phosphocellulose, Blue‐Sepharose and Sephadex G‐100. The purified enzyme exhibits a specific activity of 20–24 U/mg protein at 25°C, which is about 100‐fold higher than those of long‐chain acyl‐CoA synthetases hitherto reported. The molecular weight of the enzyme has been estimated by polyacrylamide gel electrophoresis in the presence of dodecylsulfate to be approximately 84000. The enzyme is specific for fatty acids with 14–18 carbon atoms regardless of the degree of unsaturation. Studies with the use of specific antibody to acyl‐CoA synthetase I have indicated that this enzyme is immunochemically distinguishable from acyl‐CoA synthetase II.