Synthesis of Mycoplasma arginine deiminase in E. coli using stress-responsive proteins

Keum Young Ahn, Boram Lee, Kyung Yeon Han, Jong Am Song, Doo Sung Lee, Jeewon Lee

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

We found Escherichia coli proteins, elongation factor Ts (Tsf), and malate dehydrogenase (Mdh) that can exist in the form of native and soluble proteins even under stress situation such as heat shock and protein denaturing condition. To examine their property as solubility enhancers, aggregation-prone Mycoplasma arginine deiminase (mADI), which has been suggested as anti-cancer agent, was fused to the C-terminal of each of them and cloned into pET28a to be expressed in the E. coli cytoplasm. When mADI was fused to fusion partners (Mdh, Tsf), a significant portion of the recombinant mADI was expressed in a soluble fraction (>90%) whereas the directly expressed mADI was aggregated to the inclusion body. In addition, recombinant mADI released from the fusion tag retained its soluble form and presented its specific enzymatic activity of converting l-arginine into citrulline (>10. U/mg). These results show that Tsf and Mdh could serve as effective solubility enhancers for aggregation-prone proteins (e.g. mADI in this case) when used as fusion expression partners in bacterial expression systems.

Original languageEnglish
Pages (from-to)46-49
Number of pages4
JournalEnzyme and Microbial Technology
Volume63
DOIs
Publication statusPublished - 2014 Jan 1

Fingerprint

Mycoplasma
Heat-Shock Proteins
Escherichia coli
Malate Dehydrogenase
Proteins
Fusion reactions
Solubility
Agglomeration
Citrulline
Escherichia coli Proteins
Inclusion Bodies
Arginine
arginine deiminase
Cytoplasm
Neoplasms

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Synthesis of Mycoplasma arginine deiminase in E. coli using stress-responsive proteins. / Ahn, Keum Young; Lee, Boram; Han, Kyung Yeon; Song, Jong Am; Lee, Doo Sung; Lee, Jeewon.

In: Enzyme and Microbial Technology, Vol. 63, 01.01.2014, p. 46-49.

Research output: Contribution to journalArticle

Ahn, Keum Young ; Lee, Boram ; Han, Kyung Yeon ; Song, Jong Am ; Lee, Doo Sung ; Lee, Jeewon. / Synthesis of Mycoplasma arginine deiminase in E. coli using stress-responsive proteins. In: Enzyme and Microbial Technology. 2014 ; Vol. 63. pp. 46-49.
@article{6315fa60f5cf4ac19f5ef7bb08a65707,
title = "Synthesis of Mycoplasma arginine deiminase in E. coli using stress-responsive proteins",
abstract = "We found Escherichia coli proteins, elongation factor Ts (Tsf), and malate dehydrogenase (Mdh) that can exist in the form of native and soluble proteins even under stress situation such as heat shock and protein denaturing condition. To examine their property as solubility enhancers, aggregation-prone Mycoplasma arginine deiminase (mADI), which has been suggested as anti-cancer agent, was fused to the C-terminal of each of them and cloned into pET28a to be expressed in the E. coli cytoplasm. When mADI was fused to fusion partners (Mdh, Tsf), a significant portion of the recombinant mADI was expressed in a soluble fraction (>90{\%}) whereas the directly expressed mADI was aggregated to the inclusion body. In addition, recombinant mADI released from the fusion tag retained its soluble form and presented its specific enzymatic activity of converting l-arginine into citrulline (>10. U/mg). These results show that Tsf and Mdh could serve as effective solubility enhancers for aggregation-prone proteins (e.g. mADI in this case) when used as fusion expression partners in bacterial expression systems.",
keywords = "Mycoplasma arginine deiminase, Solubility enhancer, Stress-responsive protein",
author = "Ahn, {Keum Young} and Boram Lee and Han, {Kyung Yeon} and Song, {Jong Am} and Lee, {Doo Sung} and Jeewon Lee",
year = "2014",
month = "1",
day = "1",
doi = "10.1016/j.enzmictec.2014.05.004",
language = "English",
volume = "63",
pages = "46--49",
journal = "Enzyme and Microbial Technology",
issn = "0141-0229",
publisher = "Elsevier Inc.",

}

TY - JOUR

T1 - Synthesis of Mycoplasma arginine deiminase in E. coli using stress-responsive proteins

AU - Ahn, Keum Young

AU - Lee, Boram

AU - Han, Kyung Yeon

AU - Song, Jong Am

AU - Lee, Doo Sung

AU - Lee, Jeewon

PY - 2014/1/1

Y1 - 2014/1/1

N2 - We found Escherichia coli proteins, elongation factor Ts (Tsf), and malate dehydrogenase (Mdh) that can exist in the form of native and soluble proteins even under stress situation such as heat shock and protein denaturing condition. To examine their property as solubility enhancers, aggregation-prone Mycoplasma arginine deiminase (mADI), which has been suggested as anti-cancer agent, was fused to the C-terminal of each of them and cloned into pET28a to be expressed in the E. coli cytoplasm. When mADI was fused to fusion partners (Mdh, Tsf), a significant portion of the recombinant mADI was expressed in a soluble fraction (>90%) whereas the directly expressed mADI was aggregated to the inclusion body. In addition, recombinant mADI released from the fusion tag retained its soluble form and presented its specific enzymatic activity of converting l-arginine into citrulline (>10. U/mg). These results show that Tsf and Mdh could serve as effective solubility enhancers for aggregation-prone proteins (e.g. mADI in this case) when used as fusion expression partners in bacterial expression systems.

AB - We found Escherichia coli proteins, elongation factor Ts (Tsf), and malate dehydrogenase (Mdh) that can exist in the form of native and soluble proteins even under stress situation such as heat shock and protein denaturing condition. To examine their property as solubility enhancers, aggregation-prone Mycoplasma arginine deiminase (mADI), which has been suggested as anti-cancer agent, was fused to the C-terminal of each of them and cloned into pET28a to be expressed in the E. coli cytoplasm. When mADI was fused to fusion partners (Mdh, Tsf), a significant portion of the recombinant mADI was expressed in a soluble fraction (>90%) whereas the directly expressed mADI was aggregated to the inclusion body. In addition, recombinant mADI released from the fusion tag retained its soluble form and presented its specific enzymatic activity of converting l-arginine into citrulline (>10. U/mg). These results show that Tsf and Mdh could serve as effective solubility enhancers for aggregation-prone proteins (e.g. mADI in this case) when used as fusion expression partners in bacterial expression systems.

KW - Mycoplasma arginine deiminase

KW - Solubility enhancer

KW - Stress-responsive protein

UR - http://www.scopus.com/inward/record.url?scp=84904672122&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84904672122&partnerID=8YFLogxK

U2 - 10.1016/j.enzmictec.2014.05.004

DO - 10.1016/j.enzmictec.2014.05.004

M3 - Article

C2 - 25039059

AN - SCOPUS:84904672122

VL - 63

SP - 46

EP - 49

JO - Enzyme and Microbial Technology

JF - Enzyme and Microbial Technology

SN - 0141-0229

ER -