SGR Control

Introduction
In microbial cultivation processes, specifically with recombinant Escherichia coli, one

essential procedure is to monitor and control the growth characteristics of the culture. The

specific growth rate (SGR) is an essential cultivation process variable because it represents a

characteristic of the physiological state of the cell culture. The SGR is also related to the

biosynthesis of the target product [4,5]. In addition, the quality of the desired product and the

entire cultivation process can be defined by the specific growth rate of the biomass [6–8]. The

physiological state of the culture is often determined by a concrete specific growth rate. In

some cases the maximal specific product formation rates π appear at specific growth

rates μ that are lower than the maximal growth rate μmax. In other cases, the specific product

formation rate π increases monotonically with μ.

In the case of Crabtree-positive microorganisms when the specific growth rates exceed the

critical values the metabolism shifts to diauxic growth due to exceeding the respiratory

capacity. This in turn leads to production of overflow metabolites which hamper the growth,

decrease the productivity and deters the product quality.

Indirect control of specific growth rate

The control of specific growth especially in the case of industrial fermentations is quite

challenging due to the problem of batch-to-batch variability. The root cause for this

phenomenon is the fluctuation in the biomass concentration (with respect to the changes in

inoculum size). Controlling the biomass concentration itself will elude the problem of batch-

to-batch variability as the specific growth rate is a derived quantity of biomass concentration

(Lubbert et al. 2008). Regulation of substrate concentration especially on industrial

fermenters will not completely eliminate the problem of batch-to-batch reproducibility. This

is primarily due to the fed-batch fermentations being operated at lower μ than μmax which

results in substrate concentration being less than 0.1 g/kg. Such concentration ranges are very

much below the detection levels of commercially available online sensors. On other hand

these costly sensors are installed at a single site in large fermenters and the measurements are

represented as local measurements. Due to gradients in mixing the true value of substrate

concentration is often not reflected which leads to improper control of substrate

concentration.

Control of specific growth rate using dielectric spectroscopy

During the fed-batch fermentation, when the growth rates exceed the critical values the

crabtree-positive microorganisms enter diauxic growth and the sugars are converted into

secondary metabolites due to the overflow metabolism. To forestall such scenarios M. Dabros

et al., 2018 designed an estimator for specific growth rate based on biomass concentration

values obtained from dielectric spectroscopy in real-time. The specific growth rates for both

S. cerevisiae and E. coli cultivations were maintained by a feedforward-feedback controller.

The controller was coupled with an online FTIR probe for the early detection of overflow

metabolites. In another study, Zitzman et al used the combination of dielectric spectroscopy

and optical density probe to cultivate Drosophila melanogaster S2 cells in both fed-batch and

perfusion mode.

Specific growth rate estimator based on OUR/CER

As conversion rates form the basis for the computation of yield coefficients, specific rates are

computed using the relationship as shown in Eqn. 1

ri
q i=
X
Real-time estimation of specific growth using off-gas analysis was performed on two

microbial systems (P. pastoris and E. coli) producing recombinant proteins. The systems

were assumed to follow Luedking Piret model and the same was implemented to estimate

specific growth rate. (Weschelberger 2013 still to write). A