PubMed · 11712728
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M Turner, A Affonso, J Fudin, W Rivera. Clinical case study.. https://pubmed.ncbi.nlm.nih.gov/11712728/
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Calorimetry has shown real potential at bench-scale for chemical and biochemical processes. The aim of this work was therefore to scale-up the system by adaptation of a standard commercially available 300-L pilot-scale bioreactor. To achieve this, all heat flows entering or leaving the bioreactor were identified and the necessary instrumentation implemented to enable on-line monitoring and dynamic heat balance estimation. Providing that the signals are sufficiently precise, such a heat balance would enable calculation of the heat released or taken up during an operational (bio)process. Two electrical Wattmeters were developed, the first for determination of the power consumption by the stirrer motor and the second for determination of the power released by an internal calibration heater. Experiments were designed to optimize the temperature controller of the bioreactor such that it was sufficiently rapid so as to enable the heat accumulation terms to be neglected. Further calibration experiments were designed to correlate the measured stirring power to frictional heat losses of the stirrer into the reaction mass. This allows the quantitative measurement of all background heat flows and the on-line quantitative calculation of the (bio)process power. Three test fermentations were then performed with B. sphaericus 1593M, a spore-forming bacterium pathogenic to mosquitoes. A first batch culture was performed on a complex medium, to enable optimization of the calorimeter system. A second batch culture, on defined medium containing three carbon sources, was used to show the fast, accurate response of the heat signal and the ability to perfectly monitor the different growth phases associated with growth on mixed substrates, in particular when carbon sources became depleted. A maximum heat output of 1100 W was measured at the end of the log-phase. A fed-batch culture on the same defined medium was then carried out with the feed rate controlled as a function of the calorimeter signal. A maximum heat output of 2250 W was measured at the end of the first log-phase. This work demonstrates that real-time quantitative calorimetry is not only possible at pilot-scale, but could be readily applied at even larger scales. The technique requires simple, readily available devices for determination of the few necessary heat flows, making it a robust, cost-effective technique for process development and routine monitoring and control of production processes.
Experimental data are presented that resulted from aerobic batch degradation experiments in activated sludge with simple carbon sources (acetate and dextrose) as substrates. Data collection was done using combined respirometric-titrimetric measurements. The respirometer consists of an open aerated vessel and a closed non-aerated respiration chamber for monitoring the oxygen uptake rate related to substrate degradation. The respirometer is combined with a titrimetric unit that keeps the pH of the activated sludge sample at a constant value by addition of acid and/or base. The experimental data clearly showed that the activated sludge bacteria react with consumption or production of protons during aerobic degradation of the two carbon sources under study. Thus, the cumulative amount of added acid and/or base could serve as a complementary information source on the degradation processes. For acetate, protons were consumed during aerobic degradation, whereas for dextrose protons were produced. For both carbon sources, a linear relationship was found between the amount of carbon source added and the amount of protons consumed (in case of acetate: 0.38 meq/mmol) or produced (in case of dextrose: 1.33 meq/mmol) during substrate degradation. A model taking into account substrate uptake, CO(2) production, and NH(3) uptake for biomass growth is proposed to describe the aerobic degradation of a C(x)H(y)O(z)-type carbon source. Theoretical evaluation of this model for reference parameters showed that the proton effect due to aerobic substrate degradation is a function of the pH of the liquid phase. The proposed model could describe the experimental observations with both carbon sources.
The structural and practical identifiability of a model for description of respirometric-titrimetric data derived from aerobic batch substrate degradation experiments of a C(x)H(y)O(z) carbon source with activated sludge was evaluated. The model processes needed to describe titrimetric data included substrate uptake, CO(2) production, and NH(3) uptake for biomass growth. The structural identifiability was studied using the Taylor series method and a recently proposed generalization method. It showed that combining respirometric and titrimetric data allows structural identifiability of one extra parameter combination, the biomass yield, Y(H), compared to estimation on separate data sets, on condition that the nitrogen fraction in biomass (i(XB)) is known. However, data from short-term batch substrate degradation experiments were not sufficiently informative to allow practical identification of all structurally identifiable parameters. Combining respirometry and titrimetry resulted in improvements of parameter confidence intervals compared to estimation on separate respirometric or titrimetric data sets. However, the level of the improvement seems to be substrate dependent: parameter confidence intervals improved considerably more for dextrose than for acetate degradation models. Noteworthy is the finding that the half-saturation substrate concentrations can be different depending on whether they are estimated from respirometric or titrimetric data. Moreover, this difference appears to be dependent on the carbon source considered: for dextrose, titrimetry-based K(S) values are higher than respirometry-based values while for acetate the opposite was found. It was hypothesized that this can be explained by the different point in cell metabolism where the proton production or consumption takes place, leading to a corresponding difference in timing between pH effect and oxygen consumption. Finally, the biomass yield Y(H) and the nitrogen content of the biomass i(XB) could be estimated from combined respirometric-titrimetric data obtained with addition of a known amount of carbon source. Y(H) can also be estimated from r(O) data when the initial substrate concentration S(S)(0) is known. The values found correspond to values reported in literature, but, interestingly, also seem able to reflect the occurrence of storage processes when pulses of acetate and dextrose are added.