Expression and activation of an esterase from Pseudomonas aeruginosa 1001 in Escherichia coli

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Title
Expression and activation of an esterase from Pseudomonas aeruginosa 1001 in Escherichia coli
Author(s)
Je Hyuk Lee; Sang Ki Rhee; Chul Ho Kim
Bibliographic Citation
Enzyme and Microbial Technology, vol. 35, no. 6, pp. 563-567
Publication Year
2004
Abstract
An expression vector was constructed to overproduce a maltose binding protein (MBP)-esterase fusion protein in Escherichia coli. Soluble fusion protein was separated by centrifugation after cell disruption. The fusion protein was partially purified with amylose resin. The higher concentration of fusion protein (above 2 mg/ml) did not show any activity but about 0.3 mg/ml of fusion protein had the highest activity (142 U/ml). It is due to the difficulty of contact between substrate and active site of enzyme in compact form at high concentration. The fusion protein over-expressed could not be separated into MBP and esterase by the action of protease 'Factor Xa'. The esterase could be cleaved from MBP fusion protein by the treatment of SDS with the Factor Xa, and the resulting esterase activity was increased to 34% after cleavage.
Keyword
EsteraseMaltose binding proteinPseudomonas aeruginosa
ISSN
0141-0229
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.enzmictec.2004.08.029
Type
Article
Appears in Collections:
Jeonbuk Branch Institute > Microbial Biotechnology Research Center > 1. Journal Articles
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