Purification and characterization of four catechol 1,2-dioxygenase isozymes from the benzamide-assimilating bacterium Arthrobacter species BA-5-17.

When Arthrobacter sp. BA-5-17 was grown on benzamide, the bacterium synthesized four different catechol 1,2-dioxygenase (CD, EC 1.13.11.1) isozymes (CD-I, II, III-1, and III-2). We purified each CD to homogeneity by a series of column chromatography. The molecular masses of the four CDs were between 68 and 72 kDa. The enzymes were made up of two identical subunits each with the molecular mass of 33 kDa. CD-I and II were indistinguishable in enzymatic properties tested. Most properties of CD-III-1 were similar to those of CD-III-2. However, CD-III-1 had a marked adsorption peak at 325 nm, which disappeared in CD-III-2 as well as in CD-I and II. CD-III-1 and III-2 were much more resistant to heating and inhibitors than CD-I and II.