Two-step purification of d(-)-specific carbamoylase from Agrobacterium tumefaciens AM 10.
DIR@IMTECH: CSIR-Institute of Microbial Technology
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Title |
Two-step purification of d(-)-specific carbamoylase from Agrobacterium tumefaciens AM 10.
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Creator |
Sareen, D
Sharma, R Nandanwar, H S Vohra, R M |
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Subject |
QR Microbiology
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Description |
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.
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Publisher |
Elsevier Science
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Date |
2001-02
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Type |
Article
PeerReviewed |
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Format |
application/pdf
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Identifier |
http://crdd.osdd.net/open/304/1/vohra2000.pdf
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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Relation |
http://www.sciencedirect.com/science/article/pii/S1046592800913366
http://crdd.osdd.net/open/304/ |
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