Isolation of an enzyme complex with carbon monoxide dehydrogenase activity containing corrinoid and nickel from acetate-growth Methanosarcina thermophila

Journal Article · Mon Dec 01 04:00:00 EST 1986 · J. Bacteriol.; (United States)

OSTI ID:7119768

Terlesky, K C; Nelson, M J.K.; Ferry, J G

Fast protein liquid chromatography of cell extract from methanol- or acetate-grown Methanoscarcina thermophila resolved two peaks of CO dehydrogenase activity. The activity of one of the CO dehydrogenases was sixfold greater in acetate-grown compared with methanol-grown cells. This CO dehydrogenase was purified to apparent homogeneity (70 ..mu..mol of methyl viologen reduced per min per mg of protein) and made up greater than 10% of the cellular protein of acetate-grown cells. The native enzyme (M/sub r/ 250,000) formed aggregates with an M/sub r/ of approximately 1,000,000. The enzyme contained five subunits (M/sub r/s 89,000, 71,000, 60,000, 58,000, and 19,000), suggesting a multifunctional enzyme complex. Nickel, iron, cobalt, zinc, inorganic sulfide, and a corrinoid were present in the complex. The UV-visible spectrum suggested the presence of iron-sulfur centers. The electron paramagnetic resonance spectrum contained g values of 2.073, 2.049, and 2.028; these features were broadened in enzyme that was purified from cells grown in the presence of medium enriched with /sup 61/Ni, indicating the involvement of this metal in the spectrum. The pattern of potassium cyanide inhibition indicated that cyanide binds at or near the CO binding site. The properties of the enzyme imply an involvement in the dissimilation of acetate to methane, possibly by cleavage of acetate or activated acetate.

• Research Organization: Virginia Polytechnic Institute and State Univ., Blacksburg
• OSTI ID: 7119768
• Journal Information: J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 168:3; ISSN JOBAA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACETATES
BACTERIA
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBON COMPOUNDS
CARBON MONOXIDE
CARBON OXIDES
CARBOXYLIC ACID SALTS
CHALCOGENIDES
CHEMICAL PROPERTIES
COBALT
DAYS LIVING RADIOISOTOPES
ELEMENTS
ENZYME ACTIVITY
ENZYMES
EVEN-ODD NUCLEI
INTERMEDIATE MASS NUCLEI
IRON
IRON 59
IRON ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
METABOLISM
METALS
METHANOGENIC BACTERIA
MICROORGANISMS
MOLECULAR STRUCTURE
NICKEL
NUCLEI
ORGANIC COMPOUNDS
OXIDES
OXIDOREDUCTASES
OXYGEN COMPOUNDS
PROTEINS
RADIOISOTOPES
SULFIDES
SULFUR COMPOUNDS
TRACER TECHNIQUES
TRANSITION ELEMENTS
ZINC