%0 Journal Article
%@ 1046-5928
%A Sareen, D
%A Sharma, R
%A Nandanwar, H S
%A Vohra, R M
%D 2001
%F open:840
%I Elsevier Science
%J Protein expression and purification
%K -Carbamoylase; Agrobacterium; dye affinity; purification
%N 1
%P 170-5
%T Two-step purification of d(-)-specific carbamoylase from Agrobacterium tumefaciens AM 10.
%U http://crdd.osdd.net/open/840/
%V 21
%X A simple, economical and rapid affinity chromatography procedure with red dye as a ligand has been described for the two-step purification of a relatively thermostable d(-)-carbamoylase from the cell-free extract of Agrobacterium tumefaciens AM 10. The enzyme was purified 232-fold to homogeneity with a recovery of 30% in the presence of 2 mM dithiothreitol. The specific activity of the enzyme was 7.88 U/mg protein. The enzyme is a dimer with a native molecular mass of 67 kDa and a subunit relative molecular mass of 38 kDa. The isoelectric point of the enzyme was found to be 5.83. The K(m) values for N-carbamoyl-dl-methionine and N-carbamoyl-d-phenylglycine were 3.84 and 5.0 mM, respectively.
%Z Copyright of this article belongs to Elsevier Science.