Sayed, Asad Ali; (1995) Monitoring of intracellular components during fermentation and some applications for control. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

At present most of fermentation monitoring is concerned with collecting information obtained from the environment in which the cell is growing; for example pH, dissolved oxygen tension, carbon dioxide evolution rate, oxygen uptake rate, extracellular enzymes, sugars and metabolites. When the product of interest is located intracellularly, cell breakage forms the major difficulty being laborious and time consuming (Balankenstein and Kula, 1991). This research project focused on rapid and reproducible cell breakage, and the demonstration of some uses for the data generated. The analysis of cell disruption kinetics within the Micron Lab 40 were found to be as pilot scale homogenisers described by Hetherington et. al, 1971. Cell disruption within the Micron Lab 40 has been shown to be reproducible. Statistical analysis of the stages from cell breakage, through to intracellular component measurement (assay) yielded an SD of <7%. The intracellular total soluble protein and G6PDH profiles for P. putida ML2, E. coli, and S. cerevisiae have been obtained with the emphasis on rapid & frequent monitoring. ADH profiles have also been obtained for the yeast fermentations. A prototype high pressure homogeniser (<10mL working volume) developed within the department was found to produce high levels of cell breakage (>50%) at moderate pressures (300-500 bar) in a single pass.

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