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Over-expression of parathion hydrolase of <i>Flavobacterium balustinum </i>in <i>E. coli:</i> Purification and characterization of His-tagged parathion hydrolase

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Title Over-expression of parathion hydrolase of Flavobacterium balustinum in E. coli: Purification and characterization of His-tagged parathion hydrolase
 
Creator Somara, Sita
Manavathi, Bramanandam
Tebbe, Christoph C
Siddavatam, Dayananda
 
Description 82-86
The organophosphorus pesticide degrading (opd)
gene was cloned downstream to the transcriptional and translational signals
of expression vectors pTrc99A and pET32A. The resulting recombinant expression
plasmids pNH2 and pHH2 were introduced into E. coli JM105 and E. coli
BL21 respectively. On induction the E. coli cells containing pNH2
produced high levels of parathion hydrolase. A 60 kD fusion protein was produced
in E. coli cultures containing recombinant plasmid pHH2. The molecular
mass of the fusion protein coincided with the molecular mass of 40 kD parathion
hydrolase and 20 kD N-terminal His tag encoded by the vector. Further, the
fusion protein was purified using Ni-column and the N-terminal Histag was
removed by digesting it with thrombin. The resulting protein folded properly in
presence of Zn2+ ions, and showed parathion hydrolase activity.
 
Date 2013-07-15T06:26:46Z
2013-07-15T06:26:46Z
2002-04
 
Type Article
 
Identifier 0975-0959 (Online); 0301-1208 (Print)
http://hdl.handle.net/123456789/19752
 
Language en_US
 
Rights CC Attribution-Noncommercial-No Derivative Works 2.5 India
 
Publisher NISCAIR-CSIR, India
 
Source IJBB Vol.39(2) [April 2002]