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 language | English |
|---|---|
| Pages (from-to) | 46-49 |
| Number of pages | 4 |
| Journal | Enzyme and Microbial Technology |
| Volume | 63 |
| DOIs | |
| Publication status | Published - 2014 Jan 1 |
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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 journal › Article
}
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
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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 -