Expression of enzymatically active rat liver and human placental catechol-O-methyltransferase in Escherichia coli; purification and partial characterization of the enzyme
(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.
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/bf6de83c-2abb-4d94-9593-52c839e6ee28
- author
- Lundström, Kenneth ; Tilgmann, Carola LU ; Peränen, Johan ; Kalkkinen, Nisse and Ulmanen, Ismo
- publishing date
- 1992-01-06
- 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
-
- pmid:1730052
- 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
- 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}}, }