Two-step purification of d(-)-specific carbamoylase from Agrobacterium tumefaciens AM 10.

Sareen, D and Sharma, R and Nandanwar, H S and Vohra, R M (2001) Two-step purification of d(-)-specific carbamoylase from Agrobacterium tumefaciens AM 10. Protein expression and purification, 21 (1). pp. 170-5. ISSN 1046-5928

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Abstract

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.

Item Type:	Article
Additional Information:	Copyright of this article belongs to Elsevier Science.
Uncontrolled Keywords:	-Carbamoylase; Agrobacterium; dye affinity; purification
Subjects:	Q Science > QR Microbiology
Depositing User:	Dr. K.P.S.Sengar
Date Deposited:	02 Feb 2012 05:02
Last Modified:	02 Feb 2012 05:02
URI:	http://crdd.osdd.net/open/id/eprint/840

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