TY  - JOUR
N1  - Copyright of this article belongs to Elsevier Science.
ID  - open282
UR  - http://www.sciencedirect.com/science/article/pii/S1046592801915323
IS  - 3
A1  - Sareen, D
A1  - Sharma, R
A1  - Vohra, R M
Y1  - 2001/12//
N2  - 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.
PB  - Elsevier Science
JF  - Protein expression and purification
VL  - 23
SN  - 1046-5928
TI  - Chaperone-assisted overexpression of an active D-carbamoylase from Agrobacterium tumefaciens AM 10.
SP  - 374
AV  - restricted
EP  - 9
ER  -