Myo-Inositol dehydrogenase(s) from Acetomonas oxydans. Optimization of conditions for solubilization of membrane-bound enzyme.
Methods are described for the first reported successful isolation of the soluble form of the membrane-bound myo-inositol dehydrogenase(s) from Acetomonas oxydans. Conditions for optimum yields of active enzyme in a crude membrane-free protein extract were established. The relevant conditions are (a) cell rupture by solid-shearing, (b) solubilization of the complete 60000g pellet with sodium deoxycholate at pH7.2, (c) rapid separation of the released protein from sodium deoxycholate by gel chromatography.
D000091:Acetobacter / Q000201:enzymology*; D000429:Alcohol Oxidoreductases / Q000302:isolation & purification*; D002458:Cell Fractionation; D002462:Cell Membrane / Q000201:enzymology*; D002498:Centrifugation; D002850:Chromatography, Gel; D003840:Deoxycholic Acid; D004492:Edetic Acid; D006863:Hydrogen-Ion Concentration; D007294:Inositol; D009113:Muramidase; D011506:Proteins / Q000302:isolation & purification; D012964:Sodium; D012995:Solubility; D014461:Ultracentrifugation; D014465:Ultrasonics
D011506:Proteins; D003840:Deoxycholic Acid; D007294:Inositol; D004492:Edetic Acid; D012964:Sodium; D000429:Alcohol Oxidoreductases; D009113:Muramidase