Energetics of bacterial growth: balance of anabolic and catabolic reactions.

Physiological responses to mixing in large scale bioreactors

Gradients of glucose in time and space are shown in a 30 m3 cultivation of Saccharomyces cerevisiae grown in minimal medium to a cell density of 20 gl−1. The fed-batch concept was used with glucose as the limiting component which was fed continuously to the process. As the mean glucose concentration declined throughout the process, the level of glucose was at all times different in three sampling ports (bottom/middle/top) of the reactor. These gradients were furthermore shown to depend on the feed position. This means that if the feed was supplied in the relatively stagnant mixing zone above the top impeller, the gradients were more pronounced than by feed in the well mixed bottom impeller zone. A rapid sampling system was constructed, and continuous glucose samples of every 0.15 s were analysed from a point of the reactor. Fifty samples were collected with this system, but the amount and frequency is possible to change. The results of these series show a variance of the glucose concentration where at one stage, a peak appeared of a relative difference in concentration of 40 mgl−1. The pattern of these rapid glucose fluctuations was shown to depend on the turbulence level at the location of the feed. It was shown, that the fluctuations were more pronounced when the feed was localised in a relatively stagnant area than in the well-mixed impeller area, where the deviation from the mean was negligible. The fluid flow, in the impeller (gassed and ungassed) and bulk area (ungassed) of the reactor, was characterised by turbulence measurements using thermal anemometry. These types of areas resembles well the different areas of sampling as mentioned above. The turbulent frequencies in these areas were in the range of 10−1 to 104 Hz with the highest amplitudes at low frequencies. The spectra depicts a uniform time scale for all zones, especially at the low frequencies. The dominance of low frequency, high amplitude flow variations and the observed short-time oscillations in substrate concentration support the hypothesis of substrate transport over fairly long distances without substantial mixing both in the impeller, but especially, in the bulk zone of the reactor.

Substrate gradients in bioreactors: origin and consequences

(1974). Anaerobic digestion of organic matter. C R C Critical Reviews in Environmental Control: Vol. 4, No. 1-4, pp. 131-191.

Anaerobic digestion of organic matter

A Large bioreactor is an inhomogenous system with concentration gradients which depend on the fluid dynamics and the mass transfer of the reactor, the feeding strategy, the saturation constant, and the cell density. The responses of Escherichia coli cells to short-term oscillations of the carbon/energy substrate in glucose limited fed-batch cultivations were studied in a two-compartment reactor system consisting of a stirred tank reactor (STR) and an aerated plug flow reactor (PFR) as a recycle loop. Short-term glucose excess or starvation in the PFR was simulated by feeding of glucose to the PFR or to the STR alternatively. The cellular response to repeated short-term glucose excess was a transient increase of glucose consumption and acetate formation. But, there was no accumulation of acetate in the culture, because it was consumed in the STR part where the glucose concentration was growth limiting. However, acetate accumulated during the cultivation if the oxygen supply in the PFR was insufficient, causing higher acetate formation. The biomass yield was then negatively influenced, which was also the case if the PFR was used to simulate a glucose starvation zone. The results suggest that short-term heterogeneities influence the cellular physiology and growth, and can be of major importance for the process performance. (c) 1995 John Wiley & Sons, Inc.

Influence of substrate oscillations on acetate formation and growth yield in Escherichia coli glucose limited fed-batch cultivations.

Summary: As the interval between methanol additions to methanol-limited cultures of Pseudomonas methylotropha was increased beyond 20 s the yield of bacterial dry wt/g of methanol fell significantly. Discontinuous methanol additions also caused cycling in the values of a number of parameters, including pH, dissolved oxygen tension, CO2 production and amino acid pool concentration. It is suggested that after each addition of methanol a burst of growth occurred, followed by a period of starvation. These observations are discussed in the light of continuous culture practice and theory.

J. D. Brooks

Papers

The Effect of Discontinuous Methanol Addition on the Growth of a Carbon-limited Culture of Pseudomonas