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

Fingerprint

Proteolysis
Granulocyte Colony-Stimulating Factor
Escherichia coli
Fusion reactions
Carboxyl and Carbamoyl Transferases
Peptide Hydrolases
Period Circadian Proteins
Degradation
Inclusion Bodies
Masks
Aspartic Acid
Solubility
Cytoplasm
Proteins
Agglomeration

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

Cite this

Proteolysis and synthetic strategy of human G-CSF in Escherichia coli BL21(DE3). / Song, Jong A.; Han, Kyung Yeon; Ahn, Keum Young; Park, Jin Seung; Seo, Hyuk Seong; Lee, Jeewon.

In: Enzyme and Microbial Technology, Vol. 45, No. 1, 08.07.2009, p. 7-14.

Research output: Contribution to journalArticle

Song, Jong A. ; Han, Kyung Yeon ; Ahn, Keum Young ; Park, Jin Seung ; Seo, Hyuk Seong ; Lee, Jeewon. / Proteolysis and synthetic strategy of human G-CSF in Escherichia coli BL21(DE3). In: Enzyme and Microbial Technology. 2009 ; Vol. 45, No. 1. pp. 7-14.
@article{b4ca61f00277471db6b59fca359efb59,
title = "Proteolysis and synthetic strategy of human G-CSF in Escherichia coli BL21(DE3)",
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.",
keywords = "C-terminal proteolysis, E. coli BL21(DE3), Human granulocyte colony-stimulating factor (hG-CSF), N-terminal fusion partner",
author = "Song, {Jong A.} and Han, {Kyung Yeon} and Ahn, {Keum Young} and Park, {Jin Seung} and Seo, {Hyuk Seong} and Jeewon Lee",
year = "2009",
month = "7",
day = "8",
doi = "10.1016/j.enzmictec.2009.02.010",
language = "English",
volume = "45",
pages = "7--14",
journal = "Enzyme and Microbial Technology",
issn = "0141-0229",
publisher = "Elsevier Inc.",
number = "1",

}

TY - JOUR

T1 - Proteolysis and synthetic strategy of human G-CSF in Escherichia coli BL21(DE3)

AU - Song, Jong A.

AU - Han, Kyung Yeon

AU - Ahn, Keum Young

AU - Park, Jin Seung

AU - Seo, Hyuk Seong

AU - Lee, Jeewon

PY - 2009/7/8

Y1 - 2009/7/8

N2 - 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.

AB - 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.

KW - C-terminal proteolysis

KW - E. coli BL21(DE3)

KW - Human granulocyte colony-stimulating factor (hG-CSF)

KW - N-terminal fusion partner

UR - http://www.scopus.com/inward/record.url?scp=67349157187&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67349157187&partnerID=8YFLogxK

U2 - 10.1016/j.enzmictec.2009.02.010

DO - 10.1016/j.enzmictec.2009.02.010

M3 - Article

AN - SCOPUS:67349157187

VL - 45

SP - 7

EP - 14

JO - Enzyme and Microbial Technology

JF - Enzyme and Microbial Technology

SN - 0141-0229

IS - 1

ER -