Construction, overexpression, and purification of Arthrobacter globiformisamine oxidase - Strep-Tag II fusion protein

The copper-containing amine oxidase from Arthrobacter globiformis has been expressed and purified as a fusion protein with a C-terminal Strep-tag II p eptide. This tag facilitates the rapid purification of the enzyme on a larg e scale using the StrepTactin POROS medium. For example, we have demonstrat ed that 50 mg of protein can be obtained in 2 days from 2 L of Escherichia coli. The purified fusion protein displays turnover and spectroscopic prope rties that are essentially identical to those of the wild-type enzyme. Give n the location of the C-terminus in four amine oxidase crystal structures, this strategy should be quite general for the rapid purification of amine o xidases from multiple sources. (C) 2001 Academic Press.