Insights on the Advancements of In Silico Metabolic Studies of Succinic Acid Producing Microorganisms: A Review with Emphasis on Actinobacillus succinogenes

Succinic acid (SA) is one of the top candidate value-added chemicals that can be produced from biomass via microbial fermentation. A considerable number of cell factories have been proposed in the past two decades as native as well as non-native SA producers. Actinobacillus succinogenes is among the best and earliest known natural SA producers. However, its industrial application has not yet been realized due to various underlying challenges. Previous studies revealed that the optimization of environmental conditions alone could not entirely resolve these critical problems. On the other hand, microbial in silico metabolic modeling approaches have lately been the center of attention and have been applied for the efficient production of valuable commodities including SA. Then again, literature survey results indicated the absence of up-to-date reviews assessing this issue, specifically concerning SA production. Hence, this review was designed to discuss accomplishments and future perspectives of in silico studies on the metabolic capabilities of SA producers. Herein, research progress on SA and A. succinogenes, pathways involved in SA production, metabolic models of SA-producing microorganisms, and status, limitations and prospects on in silico studies of A. succinogenes were elaborated. All in all, this review is believed to provide insights to understand the current scenario and to develop efficient mathematical models for designing robust SA-producing microbial strains.

Influence of the Initial Sugar Concentration and Supplementation with Yeast Extract on Succinic Acid Fermentation in a Lactose-Based Medium

The aim of this study was to investigate the production of succinic acid from lactose concentrate, a by-product of cheese-making, using Actinobacillus succinogenes and Basfia succiniciproducens. Although the ability of these strains to metabolize different sugars is already known, their application in the conversion of lactose bears high potential for optimization. With regard to B. succiniciproducens, this approach is completely novel. In particular, the effect of the medium’s sugar concentration as well as the ability of its supplementation with yeast extract to prevent a lack of essential nutrient proteins and vitamins was examined. Lactose-based media containing sugar concentrations of between 20 and 65 g L−1 and 5 g L−1 of yeast extract were fermented, with both strains showing comparable performances. The best results in terms of succinic acid yield and acid concentration—0.57 g g−1 initial sugar and 23 g L−1—were achieved at an initial sugar concentration of 43 g L−1. The necessity of yeast extract was demonstrated using the sugar-optimized medium without supplementation. As a result, the yield and concentration of succinic acid dropped to 0.34 g g−1 and 13 g L−1 and the sugar consumption decreased from more than 99 to less than 55%. Therefore, the supplementation amount of 5 g L−1 of yeast extract can be regarded as well-balanced.

Identifying Energy Extraction Optimisation Strategies of Actinobacillus succinogenes

A. succinogenes is well known for utilising various catabolic pathways. A multitude of batch fermentation studies confirm flux shifts in the catabolism as time proceeds. It has also been shown that continuous cultures exhibit flux variation as a function of dilution rate. This indicates a direct influence of the external environment on the proteome of the organism. In this work, ATP production efficiency was explored to evaluate the extent of bio-available energy on the production behaviour of A. succinogenes. It was found that the microbe successively utilised its most-to-least efficient energy extraction pathways, providing evidence of an energy optimisation survival strategy. Moreover, data from this study suggest a pyruvate overflow mechanism as a means to throttle acetic and formic acid production, indicating a scenario in which the external concentration of these acids play a role in the energy extraction capabilities of the organism. Data also indicates a fleeting regime where A. succinogenes utilises an oxidised environment to its advantage for ATP production. Here it is postulated that the energy gain and excretion cost of catabolites coupled to the changing environment is a likely mechanism responsible for the proteome alteration and its ensuing carbon flux variation. This offers valuable insights into the microbe’s metabolic logic gates, providing a foundation to understand how to exploit the system.

Well Knowledge of the Physiology of Actinobacillus succinogenes to Improve Succinic Acid Production

The anaerobic fermentation of glucose and fructose was performed by Actinobacillus succinogenes 130Z in batch mode using three different volume of bioreactors (0.25, 1 and 3 L). The strategy used was the addition of MgCO3 and fumaric acid (FA) as mineral carbon and the precursor of succinic acid, respectively, in the culture media. Kinetics and yields of succinic acid (SA) production in the presence of sugars in a relevant synthetic medium were investigated. Work on the bench scale (3 L) showed the best results when compared to the small anaerobic reactor's succinic acid yield and productivity after 96 h of fermentation. For an equal mixture of glucose and fructose used as substrate at 0.4 mol L−1 with the addition of FA as enhancer and under proven optimal conditions (pH 6.8, T = 37 °C, anaerobic condition and 1% v/v of biomass), about 0.5 mol L−1 of SA was obtained, while the theoretical production of succinic acid was 0.74 mol L−1. This concentration corresponded to an experimental yield of 0.88 (mol-C SA/mol-C sugars consumed anaerobically) and a volumetric productivity of 0.48 g-SA L−1 h−1. The succinic acid yield and concentration obtained were significant and in the order of those reported in the literature.