Sunday, July 24, 2011
Grand Ballroom, 5th fl (Sheraton New Orleans)
Nitrile hydratase activity in Rhodococcus rhodochrous DAP 96253 can be induced with selected multiple inducers including urea, Co, Fe and Ni. When multiply induced with Co and urea, cells of R. rhodochrous DAP 96253 expressed the highest level of nitrile hydratase activity. Co only induced cells gave the second highest nitrile hydratase activity, whereas in the uninduced cells, nitrile hydratase activity was lower than 1% of cells induced with Co and urea. When Co-urea induced cell lysate was examined on gradient SDS PAGE and analyzed by Image Quant TL, the nitrile hydratase bands (both α and β subunits) accounted for approximately 56% of the total cytosolic protein. In Co only induced samples, the nitrile hydratase bands (both α and β subunits) accounted for approximately 15% of the total protein. Preliminary investigation of PEG precipitation of Co-urea and Co cell lysate samples showed possible differences in subunit assembly of nitrile hydratase.