Figure 2.

Biochemical analyses.A, conversion of phthalate by PDO_KF1 and PDR_KF1. Purified enzymes (10 μM each) were incubated with 100 μM phthalate in the presence of 100 μM NADH. The reaction product was analyzed by HPLC. B, conversion of terephthalate by purified PDO_KF1 and PDR_KF1. Purified enzymes (10 μM each) were incubated with 500 μM terephthalate in the presence of 200 μM NADH. C, dependence of initial velocity of oxygen consumption on the phthalate concentration in air-saturated buffer. Red lines represent fits of the Michaelis–Menten equation to the data. D, dependence of initial velocity of oxygen consumption on the terephthalate concentration in air-saturated buffer. Red lines represent fits of the Michaelis–Menten equation to the data. E, conversion of phthalate and terephthalate to protocatechuate and cis-dihydrodiol terephthalate, respectively with recombinant E. coli BL21, named R1 cells containing phthalate catabolic genes (phtABCD encoding PDO, PDR, phthalate cis-4,5-dihydrodiol dehydrogenase, and cis-4,5-diol phthalate decarboxylase, respectively).