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

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

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)


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
Issue number1
Publication statusPublished - 2009 Jul 8

Bibliographical note

Funding Information:
We thank Professor Hang Chul Shin at Soongsil University for kindly providing the gene clones of hG-CSF. This study was supported by the National Research Laboratory Project of the Ministry of Education, Science and Technology (grant no. ROA-2007-000-20084-0) and by the Korea Health 21 R&D Project of the Ministry of Health & Welfare of the Republic of Korea (grant no. A050750). This work was also supported by the Second Brain Korea 21 Project. Additional support from Korea Research Foundation (grant no. KRF-2004-005-D00057) are also appreciated.


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

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology


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