The performance of a glass bead shaking technique for the disruption of Escherichia coli cells |
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Authors: | Ramakrishnan Nagasundara Ramanan Tau Chuan Ling Arbakariya B Ariff |
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Institution: | (1) Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia;(2) Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia;(3) Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia |
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Abstract: | The efficacy of a simple laboratory method for cell disruption based on the shaking of glass beads on a rotary shaker was
assessed in this study, via measurements of the release of total protein and interferon-α2b from E. coli. The optimum conditions for cell disruption were detected after 30 min of shaking in Tris-HCl buffer (pH 8) at 300 rpm with
1.5 g of glass beads (diameter: 0.5 mm) per mL of cell suspension volume. Three test runs were conducted under the above conditions
and the maximum average protein release values were determined as 3.048, 3.564, and 3.015 mg/mL, respectively. The amount
of protein release was comparable to the amount of protein release in ultrasonica-tion and glass bead vortexing procedures.
The amount of interferon-α2b release in the ultrasonication, glass bead vortexing, and glass bead shaking trials were 240,
172, and 201 ng/mL, respectively. This method was shown to process between 1 and 10 mL of sample volume in a 50 mL Falcon
tube without a great deal of deviation, and was able to handle in excess of 60 samples simultaneously. |
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Keywords: | cell disruption downstream processing E coli glass beads shaker interferon-α 2b |
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