A rapid method for the purification of methanol dehydrogenase from Methylobacterium extorquens.
Methanol dehydrogenase (MDH) is a water soluble quinoprotein that catalyzes the oxidation of methanol as an important carbon source in methylotrophic bacteria. A rapid method for the purification of MDH from Methylobacterium extorquens AM1 was developed using a single cation exchange chromatographic step, followed by ultrafiltration for final purification, enzyme concentration, and buffer exchange. MDH was obtained in an excellent overall yield with a final enzyme purity of greater than 97%. Storage at -80 degrees C in 20mM phosphate buffer, pH 7.0, showed only a negligible loss of enzyme activity after six months.
Liu
Qinfeng
Q
Department of Chemistry, The University of Toledo, Toledo, OH 43606, USA.
Kirchhoff
Jon R
JR
Faehnle
Christopher R
CR
Viola
Ronald E
RE
Hudson
Richard A
RA
D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002852:Chromatography, Ion Exchange / Q000379:methods; D020601:Methylobacterium extorquens / Q000201:enzymology*
D001426:Bacterial Proteins; D000429:Alcohol Oxidoreductases; C019641:alcohol dehydrogenase (acceptor)