Optimizing initial plasmid copy number distribution for improved protein activity in a recombinant fermentation.

Patnaik, P R (2000) Optimizing initial plasmid copy number distribution for improved protein activity in a recombinant fermentation. Biochemical engineering journal, 5 (2). pp. 101-107. ISSN 1369-703X

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Abstract

Recombinant bacterial cells in a fermentation broth rarely contain the same number of plasmids, even though this simplification is often used. Recent work has however indicated limitations of the simplified approach. Based on these studies, the distribution of plasmid copy numbers per cell has been represented macroscopically here in a Gaussian form for the fraction of biomass as a function of the copy number. Applying this distribution and an experimentally validated kinetic model to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) synthesis by Escherichia coli containing the plasmid pBR Eco gap, it is seen that GAPDH production in a batch fermentation is maximized by a particular initial (non-zero) copy number variance and an optimal duration. To implement this distribution in a bioreactor, it is suggested that the profile may be discretized, inocula corresponding to the mean copy number of each fraction prepared, and then combined to obtain the seed culture.

Item Type:	Article
Additional Information:	Copyright of this article belongs to Elsevier Science
Uncontrolled Keywords:	Recombinant fermentation; Plasmid copy number; Optimal distribution; GAPDH productivity
Subjects:	Q Science > QR Microbiology
Depositing User:	Dr. K.P.S.Sengar
Date Deposited:	03 Jan 2012 16:24
Last Modified:	09 Jan 2015 09:51
URI:	http://crdd.osdd.net/open/id/eprint/318

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