Improved partitioning in aqueous two-phase system of tyrosine-tagged recombinant lactate dehydrogenase
(2002) In Protein Expression and Purification 25(2). p.9-263- Abstract
The partitioning of Bacillus stearothermophilus lactate dehydrogenase (LDH) in an aqueous two-phase system was studied. Particularly, the influence of tyrosine tags on the partitioning was evaluated. The hydrophobic effect, caused by the addition of tyrosine residues, was determined in a system based on dextran and the thermoseparating ethylene oxide-propylene oxide random copolymer (EO30PO70). Five different LDH variants were constructed with N-terminal tags containing tyrosines (Y3 and Y6), tyrosines and prolines (Y3P2 and Y6P2), and only prolines (P2). LDH fused with tags containing tyrosines increased the partitioning coefficient, and the more tyrosines added to the protein, the larger improvement in partitioning. When prolines were... (More)
The partitioning of Bacillus stearothermophilus lactate dehydrogenase (LDH) in an aqueous two-phase system was studied. Particularly, the influence of tyrosine tags on the partitioning was evaluated. The hydrophobic effect, caused by the addition of tyrosine residues, was determined in a system based on dextran and the thermoseparating ethylene oxide-propylene oxide random copolymer (EO30PO70). Five different LDH variants were constructed with N-terminal tags containing tyrosines (Y3 and Y6), tyrosines and prolines (Y3P2 and Y6P2), and only prolines (P2). LDH fused with tags containing tyrosines increased the partitioning coefficient, and the more tyrosines added to the protein, the larger improvement in partitioning. When prolines were added between the tyrosine-rich tag and the protein, a further increased partitioning was obtained. The enhanced partitioning was attributed to the rigid structure of the proline, which in turn led to an increase in the exposure of the tag to the surroundings. The best tyrosine tag, Y6P2, increased the partition coefficient four times and additionally, a higher thermostability was observed.
(Less)
- author
- Fexby, Sara LU and Bülow, Leif LU
- organization
- publishing date
- 2002-07
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Amino Acid Sequence, Enzyme Stability, Geobacillus stearothermophilus, L-Lactate Dehydrogenase, Models, Molecular, Molecular Sequence Data, Mutation, Polyethylenes, Polypropylenes, Protein Conformation, Recombinant Fusion Proteins, Solutions, Tyrosine, Water
- in
- Protein Expression and Purification
- volume
- 25
- issue
- 2
- pages
- 7 pages
- publisher
- Academic Press
- external identifiers
-
- pmid:12135559
- scopus:0036421610
- ISSN
- 1046-5928
- DOI
- 10.1016/S1046-5928(02)00008-6
- language
- English
- LU publication?
- yes
- id
- 1a26efdd-3938-436d-83a7-7fffb1b21f19
- date added to LUP
- 2016-04-18 15:55:22
- date last changed
- 2024-01-04 02:08:51
@article{1a26efdd-3938-436d-83a7-7fffb1b21f19,
abstract = {{<p>The partitioning of Bacillus stearothermophilus lactate dehydrogenase (LDH) in an aqueous two-phase system was studied. Particularly, the influence of tyrosine tags on the partitioning was evaluated. The hydrophobic effect, caused by the addition of tyrosine residues, was determined in a system based on dextran and the thermoseparating ethylene oxide-propylene oxide random copolymer (EO30PO70). Five different LDH variants were constructed with N-terminal tags containing tyrosines (Y3 and Y6), tyrosines and prolines (Y3P2 and Y6P2), and only prolines (P2). LDH fused with tags containing tyrosines increased the partitioning coefficient, and the more tyrosines added to the protein, the larger improvement in partitioning. When prolines were added between the tyrosine-rich tag and the protein, a further increased partitioning was obtained. The enhanced partitioning was attributed to the rigid structure of the proline, which in turn led to an increase in the exposure of the tag to the surroundings. The best tyrosine tag, Y6P2, increased the partition coefficient four times and additionally, a higher thermostability was observed.</p>}},
author = {{Fexby, Sara and Bülow, Leif}},
issn = {{1046-5928}},
keywords = {{Amino Acid Sequence; Enzyme Stability; Geobacillus stearothermophilus; L-Lactate Dehydrogenase; Models, Molecular; Molecular Sequence Data; Mutation; Polyethylenes; Polypropylenes; Protein Conformation; Recombinant Fusion Proteins; Solutions; Tyrosine; Water}},
language = {{eng}},
number = {{2}},
pages = {{9--263}},
publisher = {{Academic Press}},
series = {{Protein Expression and Purification}},
title = {{Improved partitioning in aqueous two-phase system of tyrosine-tagged recombinant lactate dehydrogenase}},
url = {{http://dx.doi.org/10.1016/S1046-5928(02)00008-6}},
doi = {{10.1016/S1046-5928(02)00008-6}},
volume = {{25}},
year = {{2002}},
}