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Selective and efficient extraction of recombinant proteins from the periplasm of Escherichia coli using low concentrations of chemicals

  • Fermentation, Cell Culture and Bioengineering
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Experiments were conducted to determine chemicals at low concentrations, which can be utilized for selective release of periplasmic proteins. It was revealed that 80–100 % of the activity of alpha-amylase, beta-lactamase, and Fab D1.3 was retained in the presence of 0.05 and 0.1 % Triton X-100, 0.1 % Tween 20, 0.1 % DOC, 0.01 % BAC, 0.01 % CTAB, 10 mM EDTA, 1 mM and 10 mM DEA, 10 mM NTA, 0.1 and 1 % SHMP, 200 mM urea, 100–500 mM GndCl, and 1 % solvents (hexane, xylene, toluene, benzene, pyridine and isoamyl alcohol). Performance of these chemicals, recognized as generally safe, for selective release of proteins from the periplasm of Escherichia coli was investigated. DOC was a general and very efficient agent, and at concentrations as low as 0.05, 0.1, and 0.025 %, released beta-lactamase, alpha-amylase, and Fab D1.3 selectively with yield factors of 2.7, 2.3, and 3.6 times greater than osmotic shock procedure, respectively. EDTA (1 and 10 mM) discharged Fab D1.3 with efficiency more than osmotic shock (target protein yield of 110 and 138 %, correspondingly). Isoamyl alcohol (10 % v/v) was effective for periplasmic release of alpha-amylase and particularly Fab D1.3, with target protein yields of 75 and 168 %, respectively.

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Acknowledgments

The author would like to thank Owen Thomas and Eirini Theodosiou (The University of Birmingham, UK) for their valuable assistance. The contribution of Timothy Dafforn (The University of Birmingham, UK) in CD spectropolarimetry is gratefully acknowledged. Financial support received from the Ministry of Health and Medical Education of Iran for this study is highly appreciated.

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  1. Reza Jalalirad

    Present address: Production and Research Complex, Pasteur Institute of Iran, Tehran-Karaj Highway, 31599, Karaj, Iran

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  1. School of Chemical Engineering, College of Engineering and Physical Sciences, The University of Birmingham, Edgbaston, B15 2TT, UK

    Reza Jalalirad

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Correspondence to Reza Jalalirad.

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Jalalirad, R. Selective and efficient extraction of recombinant proteins from the periplasm of Escherichia coli using low concentrations of chemicals. J Ind Microbiol Biotechnol 40, 1117–1129 (2013). https://doi.org/10.1007/s10295-013-1307-1

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