A versatile and economical method for the release of recombinant proteins from Escherichia coli by 1-propanol cell disruption
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 an...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Royal Society of Chemistry
2016
|
| Online Access: | http://psasir.upm.edu.my/id/eprint/54955/1/A%20versatile%20and%20economical%20method%20for%20the%20release.pdf |
| _version_ | 1796976177432231936 |
|---|---|
| author | Sewn, Cen Lo Ramakrishnan, Nagasundara Ramanan Beng, Ti Tey Wen, Siang Tan Pau, Loke Show Tau, Chuan Ling Chien, Wei Ooi |
| author_facet | Sewn, Cen Lo Ramakrishnan, Nagasundara Ramanan Beng, Ti Tey Wen, Siang Tan Pau, Loke Show Tau, Chuan Ling Chien, Wei Ooi |
| author_sort | Sewn, Cen Lo |
| collection | UPM |
| description | 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. |
| first_indexed | 2024-03-06T09:22:03Z |
| format | Article |
| id | upm.eprints-54955 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T09:22:03Z |
| publishDate | 2016 |
| publisher | Royal Society of Chemistry |
| record_format | dspace |
| spelling | upm.eprints-549552018-07-11T03:27:27Z http://psasir.upm.edu.my/id/eprint/54955/ A versatile and economical method for the release of recombinant proteins from Escherichia coli by 1-propanol cell disruption Sewn, Cen Lo Ramakrishnan, Nagasundara Ramanan Beng, Ti Tey Wen, Siang Tan Pau, Loke Show Tau, Chuan Ling Chien, Wei Ooi 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. Royal Society of Chemistry 2016-06-23 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/54955/1/A%20versatile%20and%20economical%20method%20for%20the%20release.pdf 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 10.1039/c6ra10550e |
| spellingShingle | Sewn, Cen Lo Ramakrishnan, Nagasundara Ramanan Beng, Ti Tey Wen, Siang Tan Pau, Loke Show Tau, Chuan Ling Chien, Wei Ooi A versatile and economical method for the release of recombinant proteins from Escherichia coli by 1-propanol cell disruption |
| title | A versatile and economical method for the release of recombinant proteins from Escherichia coli by 1-propanol cell disruption |
| title_full | A versatile and economical method for the release of recombinant proteins from Escherichia coli by 1-propanol cell disruption |
| title_fullStr | A versatile and economical method for the release of recombinant proteins from Escherichia coli by 1-propanol cell disruption |
| title_full_unstemmed | A versatile and economical method for the release of recombinant proteins from Escherichia coli by 1-propanol cell disruption |
| title_short | A versatile and economical method for the release of recombinant proteins from Escherichia coli by 1-propanol cell disruption |
| title_sort | versatile and economical method for the release of recombinant proteins from escherichia coli by 1 propanol cell disruption |
| url | http://psasir.upm.edu.my/id/eprint/54955/1/A%20versatile%20and%20economical%20method%20for%20the%20release.pdf |
| work_keys_str_mv | AT sewncenlo aversatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT ramakrishnannagasundararamanan aversatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT bengtitey aversatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT wensiangtan aversatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT paulokeshow aversatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT tauchuanling aversatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT chienweiooi aversatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT sewncenlo versatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT ramakrishnannagasundararamanan versatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT bengtitey versatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT wensiangtan versatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT paulokeshow versatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT tauchuanling versatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption AT chienweiooi versatileandeconomicalmethodforthereleaseofrecombinantproteinsfromescherichiacoliby1propanolcelldisruption |