Setting up and modelling of overflowing fed-batch cultures of Bacillus subtilis for the production and continuous removal of lipopeptides

TL;DR

This study develops a dynamic Monod-based model for overflowing fed-batch cultures of Bacillus subtilis, enabling continuous mycosubtilin removal and correlating growth parameters with product productivity.

Contribution

It introduces a novel overflow fed-batch setup and a detailed growth model that accurately predicts biomass and product dynamics, including foam overflow effects.

Findings

Model accurately simulates growth rate evolution.

Optimal feed rate of 0.086 h--1 yields 1.27 mg/g/h productivity.

Foam biomass concentration is lower than in the bioreactor.

Abstract

This work is related to the setup of overflowing exponential fed-batch cultures (O-EFBC) derived from carbon limited EFBC dedicated to the production of mycosubtilin, an antifungal lipopeptide belonging to the iturin family. O-EFBC permits the continuous removal of the product from the bioreactor achieving a complete extraction of mycosubtilin. This paper also provides a dynamical Monod-based growth model of this process that is accurate enough to simulate the evolution of the specific growth rate and to correlate it to the mycosubtilin specific productivity. Two particular and dependant phenomena related to the foam overflow are taken into account by the model: the outgoing flow rate of a broth volume and the loss of biomass. Interestingly, the biomass concentration in the foam was found to be lower than the biomass concentration in the bioreactor relating this process to a recycling one. Parameters of this model are the growth yield on substrate and the maximal specific growth rate estimated from experiments led at feed rates of 0.062, 0.071 and 0.086 h --1. The model was extrapolated to five additional experiments carried out at feed rates of 0.008, 0.022, 0.040, 0.042 and 0.062 h --1 enabling the correlation of the mean specific growth rates with productivity results. Finally, a feed rate of 0.086 h --1 corresponding to a mean specific growth rate of 0.070 h --1 allowed a specific productivity of 1.27 mg of mycosubtilin g --1 of dried biomass h --1 .