Microbial production of methylketones: properties of purified yeast secondary alcohol dehydrogenase

Journal Article · Mon Jun 01 04:00:00 EDT 1981 · J. Appl. Biochem.; (United States)

OSTI ID:5734223

Patel, R N; Hou, C T; Laskin, A I; Derelanko, P

Secondary alcohol dehydrogenase (SADH) was purified from extracts of a methanol-grown yeast, Pichia sp. The purified enzyme was homogeneous as judged by ultracentrifugation and by polyacrylamide gel electrophoresis. The purified SADH has a molecular weight of 98,000 as determined by gel filtration and 102,000 as determined by sedimentation equilibrium analysis. The sedimentation constant s/sub 20,w/ was 6.0. The subunit size of the SADH was 48,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, indicating that it consists of two subunits. The purified SADH contained two atoms of zinc per mole of enzyme protein. SADH catalyzed the oxidation of secondary alcohols. Primary alcohols (C/sub 1/ to C/sub 8/ tested) were not oxidized. The purified SADH and extracts of various yeasts and bacteria also catalyzed the reduction of methylketones to the corresponding secondary alcohols in the presence of reduced NAD/sup +/ as an electron donor. Both reactions (oxidation of secondary alcohols in the presence of NAD/sup +/ and reduction of methylketones in the presence of reduced NAD/sup +/) catalyzed by the purified SADH were inhibited by metal-chelating agents, thio reagent, and by antisera prepared against the purified enzyme. The apparent K/sub m/ values for NAD/sup +/, reduced NAD/sup +/, reduced NAD/sup +/, 2-butanol, and 2-butanone are 0.05, 0.1, 0.4, and 1 mM, respectively. The purified enzyme preferentially oxidized (-)-2-butanol and (-)-2-octanol, the rate of oxidation of (+)-2-butanol and (+)-2-octanol was 36% and 13% of that of 100% with (-)-2-butanol and (-)-2-octanol, respectively. The K/sub m/ values for (-)-2-butanol and (+)-2-butanol were 3.0 and 0.75 mM, respectively. Antisera prepared against purified Pichia SADH cross-reacted with the SADH derived from bacteria. This suggests difference in immunological properties between yeast and bacterial SADH.

• Research Organization: Exxon Research and Engineering Co., Linden, NJ
• OSTI ID: 5734223
• Journal Information: J. Appl. Biochem.; (United States), Journal Name: J. Appl. Biochem.; (United States) Vol. 3:3; ISSN JABID
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ALCOHOLS
ALKYL RADICALS
BUTANOLS
CATALYSIS
CHEMICAL PROPERTIES
CHEMICAL REACTIONS
DATA
ENZYMES
EXPERIMENTAL DATA
FUNGI
GROWTH
HYDROXY COMPOUNDS
INFORMATION
KETONES
METHANOL
METHYL RADICALS
MICROORGANISMS
NUMERICAL DATA
OCTANOLS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
PLANTS
PRODUCTION
RADICALS
REDOX REACTIONS
STIMULATION
YEASTS