The amino acid sequences in the C-terminal region of glucose-1-phosphate thymidylyltransferases determine their soluble expression in Escherichia coli

Bijay Singh, Chang Beom Lee, Je Won Park, Jae Kyung Sohng

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Two similar genes, dnmL and rmbA in Streptomyces peucetius, which encode for glucose-1-phosphate (G-1-P) thymidylyltransferases were expressed in Escherichia coli under similar conditions. While RmbA was expressed in soluble form, DnmL was found as insoluble aggregates in inclusion bodies. The difference in expression of these similar proteins led to investigate into the amino acid sequences of these proteins by sequence alignment, hydrophobicity scale and homology modeling. These analyses showed that the two proteins are different only in the C-terminal sequences. Deletion of C-terminal sequence of DnmL increased the expression level of truncated DnmL. Substitution of C-terminal sequence of DnmL with RmbA also expressed the recombinant protein in soluble form. Finally, mutation of six amino acids in DnmL rendered the protein expressed in soluble form. These results suggested that the soluble expression of the thymidylyltransferases lies in the C-terminal sequences. In conclusion, these methods of protein engineering will be a rational tool for enhancing solubility of proteins expressed in E.coli.

Original languageEnglish
Pages (from-to)179-187
Number of pages9
JournalProtein Engineering, Design and Selection
Volume25
Issue number4
DOIs
Publication statusPublished - 2012 Apr 1
Externally publishedYes

Fingerprint

glucose-1-phosphate thymidylyltransferase
Escherichia coli
Glucose
Amino acids
Amino Acid Sequence
Phosphates
Proteins
Amino Acids
Protein Engineering
Sequence Alignment
Inclusion Bodies
Streptomyces
Recombinant proteins
Hydrophobic and Hydrophilic Interactions
Recombinant Proteins
Solubility
Hydrophobicity
Substitution reactions
Genes
Mutation

Keywords

  • hydrophobicity
  • inclusion bodies
  • recombinant DNA technology
  • thymidylyltransferase

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Molecular Biology

Cite this

The amino acid sequences in the C-terminal region of glucose-1-phosphate thymidylyltransferases determine their soluble expression in Escherichia coli. / Singh, Bijay; Lee, Chang Beom; Park, Je Won; Sohng, Jae Kyung.

In: Protein Engineering, Design and Selection, Vol. 25, No. 4, 01.04.2012, p. 179-187.

Research output: Contribution to journalArticle

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