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Lactate production in anaerobic carbohydrate fermentations with mixed cultures of microorganisms is generally observed only in very specific conditions: the reactor should be run discontinuously and peptides and B vitamins must be present in the culture medium as lactic acid bacteria (LAB) are typically auxotrophic for amino acids. State-of-the-art anaerobic fermentation models assume that microorganisms optimise the adenosine triphosphate (ATP) yield on substrate and therefore they do not predict the less ATP efficient lactate production, which limits their application for designing lactate production in mixed-culture fermentations. In this study, a metabolic model taking into account cellular resource allocation and limitation is proposed to predict and analyse under which conditions lactate production from glucose can be beneficial for microorganisms. The model uses a flux balances analysis approach incorporating additional constraints from the resource allocation theory and simulates glucose fermentation in a continuous reactor. This approach predicts lactate production is predicted at high dilution rates, provided that amino acids are in the culture medium. In minimal medium and lower dilution rates, mostly butyrate and no lactate is predicted. Auxotrophy for amino acids of LAB is identified to provide a competitive advantage in rich media because less resources need to be allocated for anabolic machinery and higher specific growth rates can be achieved. The Matlab™ codes required for performing the simulations presented in this study are available at https://doi.org/10.5281/zenodo.4031144., (© 2020 Wiley Periodicals LLC.)

The valorisation of protein-rich residual streams by anaerobic mixed-culture fermentation (MCF) has been barely studied in contrast to carbohydrate-rich wastes. The aim of this work was, therefore, to investigate how protein composition, i.e. the amino acid (AA) profile, affects the individual consumption of amino acids and, consequently, the outcome of the process. Mixed-culture fermentations were performed with two model proteins (casein and gelatin) using continuous and batch reactors at neutral pH values and 25 °C. The acidification was incomplete for both proteins, with casein achieving a higher value than gelatin. Albeit dominated by acetic acid, product spectra were different as well, with n-butyric acid as the second major product for casein and propionic acid for gelatin. The preferential consumption of amino acids was demonstrated, which interestingly depends on protein composition. The previously accepted stoichiometry accurately describes iso and n-butyric acid production, but it fails for propionic, iso and n-valeric acid generation. Overall, this study offers a better understanding of protein fermentation mechanisms, which will help to improve degradation models and to design fermentation processes, based on optimal substrate selection., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

A pH simulator consisting of an efficient numerical solver of a system of nine nonlinear equations was constructed and implemented in the modeling software MATLAB. The pH simulator was integrated in a granular biofilm model and used to simulate the pH profiles within granules performing the nitritation-anammox process for a range of operating points. The simulation results showed that pH profiles were consistently increasing with increasing depth into the granule, since the proton-producing aerobic ammonium-oxidizing bacteria (AOB) were located close to the granule surface. Despite this pH profile, more NH3 was available for AOB than for anaerobic ammonium oxidizers, located in the center of the granules. However, operating at a higher oxygen loading resulted in steeper changes in pH over the depth of the granule and caused the NH3 concentration profile to increase from the granule surface towards the center. The initial value of the background charge and influent bicarbonate concentration were found to greatly influence the simulation result and should be accurately measured. Since the change in pH over the depth of the biofilm was relatively small, the activity potential of the microbial groups affected by the pH did not change more than 5% over the depth of the granules.

A comprehensive and global sensitivity analysis was conducted under a range of operating conditions. The relative importance of mass transfer resistance versus kinetic parameters was studied and found to depend on the operating regime as follows: Operating under the optimal loading ratio of 1.90(gO(2)/m(3)/d)/(gN/m(3)/d), the system was influenced by mass transfer (10% impact on nitrogen removal) and performance was limited by AOB activity (75% impact on nitrogen removal), while operating above, AnAOB activity was limiting (68% impact on nitrogen removal). The negative effect of oxygen mass transfer had an impact of 15% on nitrogen removal. Summarizing such quantitative analyses led to formulation of an optimal operation window, which serves a valuable tool for diagnosis of performance problems and identification of optimal solutions in nitritation/anammox applications., (Copyright © 2012 Elsevier Ltd. All rights reserved.)