Purification and properties of alcohol oxidase from Candida methanosorbosa M-2003

Alcohol oxidase from Candida methanosorbosa was purified about sixfold with a yield of 37.6% from the culture broth of Candida methanosorbosa M-2003. The enzyme preparation was homogeneous on slab gel electrophoresis. The purified enzyme had an optimal pH from 6.0 to 9.0 and was stable in the range 6.0-8.5. Its optimal temperature of reaction was 50 degrees C, and it was stable below 50 degrees C. In the presence of NaN3, the enzyme retained its initial activity at 30 degrees C for 35 days, indicating stability for a long term, so far. The isoelectric point was pH 4.3. Its molecular weight was 620, 000 by gel filtration chromatography and 80,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results indicate that the enzyme consists of 8 subunits.