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Expression of enzymatically active rat liver and human placental catechol-O-methyltransferase in Escherichia coli; purification and partial characterization of the enzyme

Lundström, Kenneth; Tilgmann, Carola LU ; Peränen, Johan; Kalkkinen, Nisse and Ulmanen, Ismo (1992) In Biochimica et Biophysica Acta, Gene Structure and Expression 1129(2). p.149-154
Abstract

To produce sufficient amounts of recombinant catechol-O-methyltransferase (COMT) for structural and functional studies the coding regions of the rat liver and human placental COMT genes have been introduced into a bacterial expression vector pKEX14. Recombinant COMT was produced in Escherichia coli up to 10% of total bacterial protein after the induction of the T7 RNA polymerase gene with isopropyl-β-d-thiogalactopyranoside. Both the rat and human enzymes were enzymatically active, soluble and reacted with anti-COMT antiserum in Western blotting. Both enzymes were purified from E. coli cells and partially characterized by determining their specific activity, apparent molecular weight and pI. © 1992.

Please use this url to cite or link to this publication:
author
publishing date
type
Contribution to journal
publication status
published
keywords
(E. coli), Active recombinant protein, Bacterial expression, Catechol-O-methyltransferase, Protein purification
in
Biochimica et Biophysica Acta, Gene Structure and Expression
volume
1129
issue
2
pages
6 pages
publisher
Elsevier
external identifiers
  • Scopus:0026595563
ISSN
0167-4781
DOI
10.1016/0167-4781(92)90479-J
language
English
LU publication?
no
id
bf6de83c-2abb-4d94-9593-52c839e6ee28
date added to LUP
2016-04-11 13:24:05
date last changed
2016-12-04 04:50:19
@misc{bf6de83c-2abb-4d94-9593-52c839e6ee28,
  abstract     = {<p>To produce sufficient amounts of recombinant catechol-O-methyltransferase (COMT) for structural and functional studies the coding regions of the rat liver and human placental COMT genes have been introduced into a bacterial expression vector pKEX14. Recombinant COMT was produced in Escherichia coli up to 10% of total bacterial protein after the induction of the T7 RNA polymerase gene with isopropyl-β-d-thiogalactopyranoside. Both the rat and human enzymes were enzymatically active, soluble and reacted with anti-COMT antiserum in Western blotting. Both enzymes were purified from E. coli cells and partially characterized by determining their specific activity, apparent molecular weight and pI. © 1992.</p>},
  author       = {Lundström, Kenneth and Tilgmann, Carola and Peränen, Johan and Kalkkinen, Nisse and Ulmanen, Ismo},
  issn         = {0167-4781},
  keyword      = {(E. coli),Active recombinant protein,Bacterial expression,Catechol-O-methyltransferase,Protein purification},
  language     = {eng},
  month        = {01},
  number       = {2},
  pages        = {149--154},
  publisher    = {ARRAY(0xb40ce38)},
  series       = {Biochimica et Biophysica Acta, Gene Structure and Expression},
  title        = {Expression of enzymatically active rat liver and human placental catechol-O-methyltransferase in Escherichia coli; purification and partial characterization of the enzyme},
  url          = {http://dx.doi.org/10.1016/0167-4781(92)90479-J},
  volume       = {1129},
  year         = {1992},
}