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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 orcid ; 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.

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author
; ; ; and
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
  • pmid:1730052
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
2024-01-04 01:04:49
@article{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.</p>}},
  author       = {{Lundström, Kenneth and Tilgmann, Carola and Peränen, Johan and Kalkkinen, Nisse and Ulmanen, Ismo}},
  issn         = {{0167-4781}},
  keywords     = {{(E. coli); Active recombinant protein; Bacterial expression; Catechol-O-methyltransferase; Protein purification}},
  language     = {{eng}},
  month        = {{01}},
  number       = {{2}},
  pages        = {{149--154}},
  publisher    = {{Elsevier}},
  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}},
  doi          = {{10.1016/0167-4781(92)90479-J}},
  volume       = {{1129}},
  year         = {{1992}},
}