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A versatile and economical method for the release of recombinant proteins from Escherichia coli by 1-propanol cell disruption


Sewn, Cen Lo and Ramakrishnan, Nagasundara Ramanan and Beng, Ti Tey and Wen, Siang Tan and Pau, Loke Show and Tau, Chuan Ling and Chien, Wei Ooi (2016) A versatile and economical method for the release of recombinant proteins from Escherichia coli by 1-propanol cell disruption. RSC Advances, 6. pp. 62291-62297. ISSN 2046-2069


Cell disruption is a crucial step in the downstream processing of intracellular proteins from microbial cells. In general, mechanical methods of cell disruption are energy intensive and non-selective in protein release. Thus, chemical cell disruption offering a high selectivity in protein release and a low operating cost is highly sought after. In this study, a versatile cell disruption method based on alcohol permeabilization was developed and applied to a model system, namely Escherichia coli expressing an enhanced green fluorescent protein (EGFP). The disruption process was optimized using a response surface methodology. The optimum condition of cell disruption was attained at 32.2% (v/v) 1-propanol (1-PrOH), pH 8.8 and 25 °C. The active EGFP release obtained from the optimized 1-PrOH cell disruption (1.27 mg mL−1) was comparable to that from ultrasonication treatment (1.35 mg mL−1). In addition, the selective EGFP release achieved by 1-PrOH cell disruption (0.38 mg mg−1) was 2-fold higher than that by ultrasonication treatment (0.19 mg mg−1). The power consumption of 1-PrOH cell disruption (0.018 kW) was the lowest in comparison to that of ultrasonication treatment (0.08 kW) and glass bead vortexing (0.03 kW). Moreover, a high relative EGFP release can be achieved by 1-PrOH cell disruption conducted over a wide range of working parameters [3–62% (v/v) of 1-PrOH, pH 8–11 and 15–30 °C]. The versatility of this 1-PrOH cell disruption method is useful for the release of labile proteins through a fine adjustment of the working parameters based on the target protein's alcohol tolerance limit. This versatile and economically viable method holds great promise for lab- or large-scale microbial cell disruption due to the wide availability, inexpensiveness and recyclability of alcohol.

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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
DOI Number: https://doi.org/10.1039/c6ra10550e
Publisher: Royal Society of Chemistry
Keywords: Escherichia coli; Microbial cells; Recombinant proteins
Depositing User: Mohd Hafiz Che Mahasan
Date Deposited: 11 Jul 2018 03:27
Last Modified: 11 Jul 2018 03:27
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1039/c6ra10550e
URI: http://psasir.upm.edu.my/id/eprint/54955
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