Open AccessPurification and Properties of Formaldehyde Dehydrogenase and Formate Dehydrogenase from Candida boidinii
Author(s) -
SCHÜTTE Horst,
FLOSSDORF Josef,
SAHM Hermann,
KULA MariaRegina
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.tb10108.x
Subject(s) - formate dehydrogenase , formaldehyde dehydrogenase , formate , chemistry , formaldehyde , nad+ kinase , alcohol dehydrogenase , methylglyoxal , dehydrogenase , biochemistry , glutathione , enzyme , methanol , organic chemistry , catalysis
Formaldehyde hydrogenase and formate dehydrogenase were purified 130‐fold and 19‐fold respectively from Candida boidinii grown an methanol. The final enzyme preparations were homogeneous as judged by acrylamide gel electrophoresis and by sedimentation in an ultracentrifuge. The molecular weights of the enzymes were determined by sedimentation equilibrium studies and calculated as 80000 and 74000 respectively. Dissociation into subunits was observed by treatment with sodium dodecylsulfate. The molecular weights of the polypeptide chains were estimated to be 40000 and 36000 respectively. The NAD‐linked formaldehyde dehydrogenase specifically requires reduced glutathione for activity. Besides formaldehyde only methylglyoxal served as a substrate but no other aldehyde tested. The K m values were found to be 0.25 mM for formaldehyde, 1.2 mM for methylglyoxal, 0.09 mM for NAD and 0.13 mM for glutathione. Evidence is presented which demonstrates that the reaction product of the formaldehyde‐dehydrogenase‐catalyzed oxidation of formaldehyde is Sformvlglutathione rather than formate. The NAD‐linked formate dehydrogenase catalyzes specifically the oxidation of formate to carbon dioxide. The K m values were found tobe 13 mM for formate and 0.09 mM for NAD.