title: Chaperone-assisted overexpression of an active D-carbamoylase from Agrobacterium tumefaciens AM 10.
creator: Sareen, D
creator: Sharma, R
creator: Vohra, R M
subject: QR Microbiology
description: The N-carbamoyl-D-amino acid amidohydrolase (D-carbamoylase) gene (dcb) from Agrobacterium tumefaciens AM 10 was cloned by polymerase chain reaction in plasmid pET28a and was overexpressed in Escherichia coli JM109 (DE3). However, almost 80% of the enzyme remained trapped in inclusion bodies. To facilitate the expression of the properly folded active enzyme, the chaperones GroEL/ES were coexpressed in plasmid pKY206. This resulted in a 43-fold increase in active enzyme production compared to the wild-type strain. The histidyl-tagged D-carbamoylase was purified by a single step nickel-affinity chromatography to a specific activity of 9.5 U/mg protein.
publisher: Elsevier Science
date: 2001-12
type: Article
type: PeerReviewed
format: application/pdf
identifier: http://crdd.osdd.net/open/282/1/vohra2001.pdf
relation: http://www.sciencedirect.com/science/article/pii/S1046592801915323
identifier:   Sareen, D and Sharma, R and Vohra, R M  (2001) Chaperone-assisted overexpression of an active D-carbamoylase from Agrobacterium tumefaciens AM 10.  Protein expression and purification, 23 (3).  pp. 374-9.  ISSN 1046-5928     
relation: http://crdd.osdd.net/open/282/