Purification and characterization of an amidase from an XXXX -degrading Rhodococcus sp.

@entity10694 @entity1522 @entity8798 @entity1025 @entity3208 @entity8863 @entity14941 @entity71229 @entity3412 @entity515 @entity2733 @entity3323 @entity8594 @entity1432 @entity2975 @entity3411 @entity71230 @entity71231 @entity3287 @entity68
A constitutively expressed aliphatic amidase from a Rhodococcus sp. catalyzing @entity3323 deamination was purified to electrophoretic homogeneity. The molecular weight of the native enzyme was estimated to be 360,000. Upon @entity3208 - @entity2975 gel electrophoresis, the purified preparation yielded a homogeneous protein band having an apparent molecular weight of about 44,500. The amidase had pH and temperature optima of 8.5 and 40 degrees C, respectively, and its isoelectric point was pH 4.0. The amidase had apparent K(m) values of 1.2, 2.6, 3.0, 2.7, and 5.0 mM for @entity3323 , @entity14941 , @entity71229 , @entity71230 , and @entity71231 , respectively. Inductively coupled plasma-atomic emission spectometry analysis indicated that the enzyme contains 8 mol of @entity68 per mol of the native enzyme. No labile @entity3287 was detected. The amidase activity was enhanced by, but not dependent on @entity8798 , @entity8594 , and @entity3412 . However, the enzyme activity was partially inhibited by @entity1025 and totally inhibited in the presence of @entity3411 , @entity10694 , @entity8863 , @entity515 , specific @entity68 chelators, and @entity1432 blocking reagents. The @entity2733 -terminal sequence of the first 18 @entity1522 displayed 88% homology to the aliphatic amidase of Brevibacterium sp. strain R312.
