Solubilization of aggregation-prone heterologous proteins by covalent fusion of stress-responsive Escherichia coli protein, SlyD

Kyung Yeon Han, Jong A. Song, Keum Young Ahn, Jin Seung Park, Hyuk Seong Seo, Jeewon Lee

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

The proteome profile of Escherichia coli BL21(DE3) generated in response to heat shock stress was analyzed by two-dimensional electrophoresis (2-DE), wherein we identified a FKBP-type peptidyl-prolyl cis-trans isomerse (PPIases), SlyD, as a stress-responsive (i.e. aggregation-resistant) protein. Even under an imposed severe stress condition where 29 out of 858 soluble proteins were totally eliminated and the synthesis levels of 171 proteins decreased over 5-fold, a 3.37-fold increase induced by heat shock treatment was observed in the synthesis level of SlyD compared with a non-stress condition. As a fusion partner, as well as solubility enhancer, SlyD facilitated folding and significantly increased the solubility of many aggregation-prone heterologous proteins in E. coli cytoplasm. SlyD was very effective in sequestering interactive surfaces of heterologous proteins associated with non-specific protein-protein interactions and the formation of inclusion bodies, most likely as a result of intrinsic folding efficiencies and/or chaperone-like activities. SlyD was also shown to be suitable for the production of a biologically active fusion mutant of Pseudomonas putida cutinase that is of considerable biotechnological and commercial interest.

Original languageEnglish
Pages (from-to)543-549
Number of pages7
JournalProtein Engineering, Design and Selection
Volume20
Issue number11
DOIs
Publication statusPublished - 2007 Nov 1

Fingerprint

Escherichia coli Proteins
Escherichia coli
Fusion reactions
Agglomeration
Proteins
Solubility
Tacrolimus Binding Proteins
Peptidylprolyl Isomerase
Heat-Shock Response
Pseudomonas putida
Inclusion Bodies
Proteome
Electrophoresis
Shock
Membrane Proteins
Cytoplasm
Hot Temperature

Keywords

  • Escherichia coli BL21(DE3)
  • Proteome
  • SlyD
  • Solubility enhancer
  • Stress response

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology

Cite this

Solubilization of aggregation-prone heterologous proteins by covalent fusion of stress-responsive Escherichia coli protein, SlyD. / Han, Kyung Yeon; Song, Jong A.; Ahn, Keum Young; Park, Jin Seung; Seo, Hyuk Seong; Lee, Jeewon.

In: Protein Engineering, Design and Selection, Vol. 20, No. 11, 01.11.2007, p. 543-549.

Research output: Contribution to journalArticle

Han, Kyung Yeon ; Song, Jong A. ; Ahn, Keum Young ; Park, Jin Seung ; Seo, Hyuk Seong ; Lee, Jeewon. / Solubilization of aggregation-prone heterologous proteins by covalent fusion of stress-responsive Escherichia coli protein, SlyD. In: Protein Engineering, Design and Selection. 2007 ; Vol. 20, No. 11. pp. 543-549.
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