Projects per year
Results: This is the first investigation using Near Infrared Spectroscopy (NIRS) in situ combined with metabolic flux modelling which is both a significant challenge and considerable extension of these techniques. We investigated the feasibility of our approach using the industrial workhorse Pichia pastoris in a simplified model system. A parental P. pastoris strain (i.e. which does not synthesize recombinant protein) was used to allow definition of distinct metabolic states focusing solely upon the prediction of intracellular fluxes in central carbon metabolism. Extracellular fluxes were determined using off-line conventional reference methods and on-line NIR predictions (calculated by multivariate analysis using the partial least squares algorithm, PLS). The results showed that the PLS-NIRS models for biomass and glycerol were accurate: correlation coefficients, R2, above 0.90 and the root mean square error of prediction, RMSEP, of 1.17 and 2.90 g/L, respectively. The analytical quality of the NIR models was demonstrated by direct comparison with the standard error of the laboratory (SEL), which showed that performance of the NIR models was suitable for quantifying biomass and glycerol for calculating extracellular metabolite rates and used as independent inputs for the MFA (RMSEP lower than 1.5 × SEL). Furthermore, the results for the MFA from both datasets passed consistency tests performed for each steady state, showing that the precision of on-line NIRS is equivalent to that obtained by the off-line measurements.
Conclusions: The findings of this study show for the first time the potential of NIRS as an input generating for MFA models, contributing to the optimization of cell factory metabolism in real-time.
- realtime flux modelling
- industrial cell factory
- real-time metabolic flux analysis
- pichia pastoris
1/10/10 → 30/09/11
McNeil, B., 25 Oct 2012.
Research output: Contribution to conference › Speech
Applications of metabolomics to microbial "Cell Factories" for biomanufacturing: current trends and future prospectsMousdale, D. & Mcneil, B., 12 Dec 2011, Fermentation Microbiology and Biotechnology. El-Mansi, E. M. T., Bryce, C. F. A. & Demain, A. L. (eds.). 3 ed. London, 20 p.
Research output: Chapter in Book/Report/Conference proceeding › Chapter
The roles of the alternative NADH dehydrogenases during oxidative stress in cultures of the filamentous fungus Aspergillus nigerO'Donnell, A., Harvey, L. M. & Mcneil, B., May 2011, In: Fungal biology. 115, 4-5, p. 359-369 11 p.
Research output: Contribution to journal › Article › peer-review