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

Paper ID Volume ID Publish Year Pages File Format Full-Text
17167 42648 2014 4 PDF Available
Title
Synthesis of Mycoplasma arginine deiminase in E. coli using stress-responsive proteins
Abstract

•E. coli proteins Mdh and Tsf were used as fusion partners for synthesis of mADI.•The solubility of Mdh or Tsf fused mADI in E. coli dramatically increased.•The fusion partner-free mADI retain their solubility and biological activity.

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; Stress-responsive protein; Solubility enhancer
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Synthesis of Mycoplasma arginine deiminase in E. coli using stress-responsive proteins
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 63, September 2014, Pages 46–49
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us