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A new extant respirometric assay to estimate intrinsic growth parameters applied to study plasmid metabolic burden
Author(s) -
Seoane Jose,
Sin Gürkan,
Lardon Laurent,
Gernaey Krist V.,
Smets Barth F.
Publication year - 2009
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.22518
Subject(s) - pseudomonas putida , plasmid , extant taxon , biology , biological system , robustness (evolution) , biochemical engineering , organism , identifiability , recombinant dna , computational biology , biochemistry , enzyme , dna , mathematics , statistics , genetics , gene , evolutionary biology , engineering
Start‐up phenomena in microbial biokinetic assays are not captured by the most commonly used growth‐related equations. In this study we propose a new respirometric experimental design to estimate intrinsic growth parameters that allow us to avoid these limitations without data omission, separate mathematical treatment, or wake‐up pulses prior to the analysis. Identifiability and sensitivity analysis were performed to confirm the robustness of the new approach for obtaining unique and accurate estimates of growth kinetic parameters. The new experimental design was applied to establish the metabolic burden caused by the carriage of a pWW0 TOL plasmid in the model organism Pseudomonas putida KT2440. The metabolic burden associated was manifested as a reduction in the yield and the specific growth rate of the host, with both plasmid maintenance and the over‐expression of recombinant proteins from the plasmid contributing equally to the overall effect. Biotechnol. Bioeng. 2010;105: 141–149. © 2009 Wiley Periodicals, Inc.