Proteolysis and synthetic strategy of human G-CSF in Escherichia coli BL21(DE3)

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

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We report for the first time that the C-terminal region of hG-CSF suffers from proteolytic degradation when human granulocyte colony-stimulating factor (hG-CSF) is directly expressed in Escherichia coli BL21(DE3). It is believed that the rapid proteolysis occurs at the C-terminus of hG-CSF that is very easily exposed to E. coli protease(s) during a short period following protein synthesis and prior to completion of the formation of the inclusion body. The recombinant hG-CSF that is expressed with an N-terminal fusion partner is effectively protected from the proteolysis. It seems that since the N-terminus of hG-CSF is located very close to the C-terminus, the presence of the N-terminal fusion partner masks the C-terminal region of hG-CSF and protects it from proteolytic degradation by E. coli protease(s). Furthermore, the solubility of hG-CSF markedly increased in E. coli cytoplasm when a stress-responsive and aggregation-resistant protein, i.e. aspartate carbamoyl-transferase catalytic chain (PyrB) was used as a novel N-terminal fusion partner proteins.

Original languageEnglish
Pages (from-to)7-14
Number of pages8
JournalEnzyme and Microbial Technology
Volume45
Issue number1
DOIs
Publication statusPublished - 2009 Jul 8

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Keywords

  • C-terminal proteolysis
  • E. coli BL21(DE3)
  • Human granulocyte colony-stimulating factor (hG-CSF)
  • N-terminal fusion partner

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Applied Microbiology and Biotechnology

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