Next Article in Journal
Germinal Center Optimization Applied to Neural Inverse Optimal Control for an All-Terrain Tracked Robot
Previous Article in Journal
Robust Control Examples Applied to a Wind Turbine Simulated Model
Article Menu
Issue 1 (January) cover image

Export Article

Open AccessArticle
Appl. Sci. 2018, 8(1), 30; doi:10.3390/app8010030

Efficient Disruption of Escherichia coli for Plasmid DNA Recovery in a Bead Mill

1
Department of Chemical Engineering and Metallurgy, University of Sonora, Hermosillo 83000, Mexico
2
Department of Biotechnology and Bioengineering, Center for Research and Advanced Studies of the National Polytechnic Institute, Mexico City 07360, Mexico
3
Department of Scientific and Technological Research, University of Sonora, Hermosillo 83000, Mexico
*
Author to whom correspondence should be addressed.
Received: 5 November 2017 / Revised: 20 December 2017 / Accepted: 22 December 2017 / Published: 26 December 2017
(This article belongs to the Section Chemistry)
View Full-Text   |   Download PDF [1833 KB, uploaded 26 December 2017]   |  

Abstract

The release kinetics of pDNA in a bead mill was studied. Samples taken during the process were analyzed to determine total pDNA (pDNA(t)) and supercoiled pDNA (pDNA(sc)) concentration. In order to identify important variables of the process and to develop an empirical model for optimal pDNA(t) and pDNA(sc) release, a two level 23 factorial design was used with variables: mill frequency, cell concentration, and bead size. The results were analyzed by response surface methodology. The optimized conditions for pDNA(t) yield 13.26 mg/g dcw (93.41% recovery), with a mill frequency of 30 Hz, a bead size of 0.10–0.25 mm, and a cell concentration of 20 g wcw/L. However, the optimized conditions for pDNA(sc) yield 7.65 mg/g dcw (92.05% recovery), with a mill frequency of 15 Hz, a bead size of 0.10–0.25 mm, and a cell concentration of 10 g wcw/L. Cell disruption in a bead mill was proved efficient for the release of pDNA(t) and pDNA(sc) compared to the alkaline treatment. The results obtained suggest a compromise between pDNA(sc) purity and recuperation in the process development. View Full-Text
Keywords: mechanical cell lysis; bead mill; plasmid DNA recovery; release kinetics of pDNA mechanical cell lysis; bead mill; plasmid DNA recovery; release kinetics of pDNA
Figures

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Share & Cite This Article

MDPI and ACS Style

Padilla-Zamudio, A.; Lucero-Acuña, J.A.; Guerrero-Germán, P.; Ortega-López, J.; Tejeda-Mansir, A. Efficient Disruption of Escherichia coli for Plasmid DNA Recovery in a Bead Mill. Appl. Sci. 2018, 8, 30.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Appl. Sci. EISSN 2076-3417 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top