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Improved partitioning in aqueous two-phase system of tyrosine-tagged recombinant lactate dehydrogenase

Fexby, Sara LU and Bülow, Leif LU (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.

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organization
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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
  • 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
2017-01-01 08:23:11
@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},
  keyword      = {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},
  volume       = {25},
  year         = {2002},
}