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Long-duration human space missions require considerable amounts of water, oxygen, and nutritious biomass. Additionally, the space vehicles must be well equipped to deal with metabolic human waste. It is therefore important to develop life-support systems which make these missions self-sufficient in terms of water, food, and oxygen production as well as waste management. One such solution is the employment of regenerative life-support systems that use biological and chemical/physical processes to recycle crew waste, revitalize air, and produce water and food. Photosynthetic cyanobacteria Limnospira could play a significant role in meeting these objectives. Limnospira can metabolize CO2 and nitrogen-rich human waste to produce oxygen and edible biomass. So far, life-support system studies have mainly focused on using chemical/physical methods to recycle water, degrade human waste, and recycle CO2 into oxygen. Nowadays, additional microbial processes are considered, such as nitrification of urea–ammonium–rich human waste and then using the nitrate for cyanobacterial cultivation and air vitalization. This cascade of multiple processes tends to increase the complexity of the life-support systems. The possibility of using non-nitrified urine for Limnospira cultivation can partially solve these issues. Our previous studies have shown that it is possible to cultivate Limnospira with urea and ammonium, the prominent nitrogen forms present in non-nitrified urine. In this study, we investigated the possibility of cultivating Limnospira with the different nitrogen forms present in non-nitrified urine and also evaluated their effect on the oxygen production capacity of Limnospira. For this 35-day-long study, we worked on a simplified version of the European Space Agency’s MELiSSA. During this ground demonstration study, we monitored the effect of urea and ammonium (vs. nitrate) on the oxygen production capacity of Limnospira. A deterministic control law, developed and validated on the basis of a stochastic light-transfer model, modulated (increase/decrease) the incident light on the photobioreactor (with Limnospira) to control oxygen levels in the closed loop. The CO2 from the mouse compartment was recycled as a carbon source for Limnospira. We observed that while the system could meet the desired oxygen levels of 20.3% under the nitrate and urea regime, it could only reach a maximum O2 level of 19.5% under the ammonium regime.

The ability of cyanobacterium Arthrospira sp. to assimilate waste nitrogen sources (ammonium and urea) makes it an important candidate for wastewater management. The aim of this work was to evaluate a cultivation approach based on continuous-transitional-feeding regime (nitrate-ammonium-nitrate) in a photobioreactor to assess the effects of ammonium salts on Arthrospira sp. PCC 8005 metabolism. Using a comprehensive biochemical, proteomic and stoichiometric profiling of biomass, this study demonstrated that the proposed cultivation approach could increase the proteins and pigments yields by 20-30%, compared to the respective yields obtained from wild-type Arthrospira sp. strain A light-energy-transfer model was used to predict the biomass and oxygen productivities of Arthrospira sp. cultivated under transitional-feeding regime. 95 ± 2% match was observed between the experimental and simulated productivities. This study thus opened new avenues for use of ammonium rich wastewater for commercial production of high value pigments, biofuel and bioplastics using Arthrospira sp.

The aim of this study was to investigate ammonia load removable by a fixed-bed bioreactor. In this study, high ammonia loads, up to 4.5 kgN/m3 d were applied on a fixed bed reactor colonised by two pures nitrifying bacteria: Nitrosomonas europaea ATCC® 19718 and Nitrobacter winogradskyi ATCC® 25391. The loads were similar to the highest values reported in literature especially with a two bacteria population. The complete ammonia removal efficiency in our installation has reached 2.5 kgN/m3.d for a full nitrification. An N-tank in series model was used to represent the fixed bed reactor and to predict the nitrification in the installation. The developed model was accurate and representative of the nitrification assumed by Nitrosomonas europaea ATCC® 19718 and Nitrobacter winogradskyi ATCC® 25391. This model was also used to predict the biomass and oxygen concentrations during the experiment under various operational conditions. The biofilm fixed on the solid support (polysterene beads 2 mm diameter) was mainly present in the bottom of the reactor where the ammonia load was applied. It is demonstrated that in this region of the column the oxygen concentrations reached the lowest values.

Acetic acid is one of the major potential inhibitors of cellular growth resulting from acid pretreatment of lignocellulosic biomass. To use lignocellulose hydrolysate in biorefineries it is necessary to assess their effect on the microorganisms likely to be used. An attractive option is to use a part of these hydrolysates for on-site cellulase production. Based on an industrial process that included a fast initial growth phase, the effect on Trichoderma reesei grown on glucose was assessed and modeled at different concentrations and pH. Acetic acid had a strong effect on T. reesei growth rate and yield, which correlated only with the concentration of the undissociated form in solution. The specific growth rate was accurately modeled by two different classic inhibition models as a function of pH and total acetic acid concentration. Co-consumption of glucose and acetic acid was observed, so that the culture medium was gradually detoxified by the cells. With or without glucose, acetic acid was miner...

The production of alcoholic beverages is based on the production of alcohol and aroma by fermentation, in most cases by the yeast Saccharomyces cerevisiae. The overall objective of this chapter is to improve our understanding of the genetic and environmental factors that control the production of the metabolites (ethanol, higher alcohols, and esters) involved in alcoholic beverages. This chapter aims at developing a comprehensive framework for establishing models of yeast growth and the production of metabolites. A general survey of some of the latest studies on yeast metabolism is presented with the aim of getting a better understanding of the production of the main compounds produced during the winemaking and brewery processes.

This study focuses on the use of acetic acid as carbon source for cell mass and lipid production using an oleaginous yeast, Cryptococcus curvatus. To improve this cell mass and lipid production on acetic acid, a numerical model was built to predict the yeast behavior. The model was first calibrated and validated on a fed-batch culture of the oleaginous yeast grown on acetic acid. After the validation step, this model was used as a predictive tool to search for optimal dilution rate and C/N ratios for continuous culture. A continuous culture was then launched using these culture parameters, revealing that the experimental data fitted the model predictions. At steady-state, cell mass concentration was stabilized at 26.7 g/L containing 48–53% of lipid content. A maximal lipid productivity of 0.54 g L−1 h−1 was obtained, which was superior to those reported in literature. This model could be qualified as a mechanistic one since it allows understanding reactions underlying the lipid production process. The experimental data fitted the model predictions, which confirmed that the use of a model is a powerful and predictive tool which could help the operator to process parameters to improve the culture.

International audience; Cereus triangularis (Cactaceae) is a cactus used in food decoction as a traditional medicine in the North region of Madagascar to reduce stomach ache and intestinal diseases. Hydrocolloids were sequentially extracted from its cladodes with a yield of 24% (240 mg/g based on dried cladodes powder). Structural analyses has revealed that this polysaccharide with a molecular mass of 8430,000 g/mol was mainly composed of a galactan backbone of a (1 → 4) linked β-d-Galp residues probably substituted at position 3 by l-arabinofuranosyl residues. In vitro antioxidant activity of this arabinogalactan-rich fraction was detected and quantified by radical DPPH scavenging, hydroxyl radical scavenging, radical anion superoxide scavenging and reducing power method.

Cell walls of yeasts and bacteria are able to complex with mycotoxins and limit their bioavailability in the digestive tract when these yeasts and bacteria are given as feed additives to animals. To identify the component(s) of the yeast cell wall and the chemical interaction(s) involved in complex formation with zearalenone, four strains of Saccharomyces cerevisiae differing in their cell wall glucan and mannan content were tested. Laboratory strains wt292, fks1, and mnn9 were compared with industrial S. cerevisiae strain sc1026. The complex-forming capacity of the yeast cell walls was determined in vitro by modelling the plots of amount of toxin bound versus amount of toxin added using Hill's model. A cooperative relationship between toxin and adsorbent was shown, and a correlation between the amount of beta-D-glucans in cell walls and complex-forming efficacy was revealed (R2 = 0.889). Cell walls of strains wt292 and mnn9, which have higher levels of beta-D-glucans, were able to complex larger amounts of zearalenone, with higher association constants and higher affinity rates than those of the fks1 and sc1026 strains. The high chitin content in strains mnn9 and fks1 increased the alkali insolubility of beta-D-glucans from isolated cell walls and decreased the flexibility of these cell walls, which restricted access of zearalenone to the chemical sites of the beta-D-glucans involved in complex formation. The strains with high chitin content thus had a lower complex-forming capacity than expected based on their beta-D-glucans content. Cooperativity and the three-dimensional structure of beta-D-glucans indicate that weak noncovalent bonds are involved in the complex-forming mechanisms associated with zearalenone. The chemical interactions between beta-D-glucans and zearalenone are therefore more of an adsorption type than a binding type.

International audience; The isolated cell wall of Saccharomyces cerevisiae has some capacity to adsorb zearalenone (affinity near 30%) and reduce the bioavailability of toxins in the digestive tract. The adsorption process was quantified in vitro, and the data obtained when plotted with Hill's equation indicated a cooperative process. The model showed that the adsorption capacity was related to the yeast cell wall composition. This work focused on the role of various beta-D-glucan types in the efficacy of zearalenone adsorption by yeast cell wall and sought to elucidate some of the adsorption mechanisms. Zearalenone was mixed at 37 degreesC with a constant quantity of alkali-soluble or alkali-insoluble beta-D-glucans isolated from yeast cell walls, and the amount of adsorbed zearalenone was measured. Given that the alkali solubility of beta-D-glucans is a determining factor for their three-dimensional conformation and that the alkali-insoluble fraction had a greater affinity (up to 50%) than the alkali-soluble fraction (similar to16%), it was concluded that the three-dimensional structure strongly influences the adsorption process. The alkali insolubility of beta-D-glucans led to the formation of single and/or triple helices, which have been identified as the most favorable structures for zearalenone adsorption efficacy. The beta(1,3)-D-glucan and beta(1,6)-D-glucan compositions of the two alkali-extracted fractions and their involvement in the adsorption process are discussed.

Three models based on sigmoidal plotting were tested for their ability to describe zearalenone adsorption on Saccharomyces cerevisiae cell walls in vitro. All three models closely fitted the experimental data, but Hill's equation gave the most accurate parameters, and provided information on the physical and chemical mechanisms involved in the adsorption of mycotoxin on yeast cell walls.

International audience; Background: On-site cellulase production using locally available lignocellulosic biomass (LCB) is essential for cost-effective production of 2nd-generation biofuels. Cellulolytic enzymes (cellulases and hemicellulases) must be produced in fed-batch mode in order to obtain high productivity and yield. To date, the impact of the sugar composition of LCB hydrolysates on cellulolytic enzyme secretion has not been thoroughly investigated in industrial conditions. Results: The effect of sugar mixtures (glucose, xylose, inducer) on the secretion of cellulolytic enzymes by a glucose-derepressed and cellulase-hyperproducing mutant strain of Trichoderma reesei (strain CL847) was studied using a small-scale protocol representative of the industrial conditions. Since production of cellulolytic enzymes is inducible by either lactose or cellobiose, two parallel mixture designs were performed separately. No significant difference between inducers was observed on cellulase secretion performance, probably because a common induction mechanism occurred under carbon flux limitation. The characteristics of the enzymatic cocktails did not correlate with productivity, but instead were rather dependent on the substrate composition. Increasing xylose content in the feed had the strongest impact. It decreased by 2-fold cellulase, endoglucanase, and cellobiohydrolase activities and by 4-fold β-glucosidase activity. In contrast, xylanase activity was increased 6-fold. Accordingly, simultaneous high β-glucosidase and xylanase activities in the enzymatic cocktails seemed to be incompatible. The variations in enzymatic activity were modelled and validated with four fed-batch cultures performed in bioreactors. The overall enzyme production was maintained at its highest level when substituting up to 75% of the inducer with non-inducing sugars. Conclusions: The sugar substrate composition strongly influenced the composition of the cellulolytic cocktail secreted by T. reesei in fed-batch mode. Modelling can be used to predict cellulolytic activity based on the sugar composition of the culture-feeding solution, or to fine tune the substrate composition in order to produce a desired enzymatic cocktail.

The artificial ecosystem MELiSSA, supported by the European Space Agency is a closed loop system consisting of 5 compartments in which food, water and oxygen are produced out of organic waste. The first compartment is conceived as a thermophilic anaerobic membrane bioreactor liquefying organic waste into VFAs, ammonium and CO 2 without methane. A 20 L reactor was assembled to demonstrate the selected design and process at prototype scale. We characterized system performance from start-up to steady state and evaluated process efficiencies with special attention drawn to the mass balances. An overall efficiency for organic matter biodegradation of 50% was achieved. The dry matter content was stabilized around 40–50 g L −1 and VFA production around 5–6 g L −1 . The results were consistent for the considered substrate mixture and can also be considered relevant in a broader context, as a first processing step to produce building blocks for synthesis of primary energy vectors.

As a part of a natural biological N-cycle, nitrification is one of the steps included in the conception of artificial ecosystems designed for extraterrestrial life support systems (LSS) such as Micro-Ecological Life Support System Alternative (MELiSSA) project, which is the LSS project of the European Space Agency. Nitrification in aerobic environments is carried out by two groups of bacteria in a two-step process. The ammonia-oxidizing bacteria (Nitrosomonas europaea) realize the oxidation of ammonia to nitrite, and the nitrite-oxidizing bacteria (Nitrobacter winogradskyi), the oxidation of nitrite to nitrate. In both cases, the bacteria achieve these oxidations to obtain an energy and reductant source for their growth and maintenance. Furthermore, both groups also use CO2 predominantly as their carbon source. They are typically found together in ecosystems, and consequently, nitrite accumulation is rare. Due to the necessity of modeling accurately conversion yields and transformation rates to achieve a complete modeling of MELiSSA, the present study focuses on the experimental determination of nitrogen to biomass conversion yields. Kinetic and mass balance studies for axenic cultures of Nitrosomonas europaea and Nitrobacter winogradskyi in autotrophic conditions are performed. The follow-up of these cultures is done using flow cytometry for assessing biomass concentrations and ionic chromatography for ammonium, nitrite, and nitrate concentrations. A linear correlation is observed between cell count and optical density (OD) measurement (within a 10 % accuracy) validating OD measurements for an on-line estimation of biomass quantity even at very low biomass concentrations. The conversion between cell count and biomass concentration has been determined: 7.1 × 1012 cells g dry matter (DM)−1 for Nitrobacter and 6.3 × 1012 cells g DM−1 for Nitrosomonas. Nitrogen substrates and products are assessed redundantly showing excellent agreement for mass balance purposes and conversion yields determination. Although the dominant phenomena are the oxidation of NH 4 + into nitrite (0.95 mol mol N−1 for Nitrosomonas europaea within an accuracy of 3 %) and nitrite into nitrate (0.975 mol mol N−1 for Nitrobacter winogradskyi within an accuracy of 2 %), the Nitrosomonas europaea conversion yield is estimated to be 0.42 g DM mol N−1, and Nitrobacter winogradskyi conversion yield is estimated to be 0.27 g DM mol N−1. The growth rates of both strains appear to be dominated by the oxygen transfer into the experimental setups.

In a previous paper we reported that beta-D-glucans isolated from Saccharomyces cerevisiae could adsorb zearalenone, reduce its bioavailability in the digestive tract, and protect animals against its adverse effects. We have now investigated, in vitro, the kinetics of the interaction between other mycotoxins and beta-D-glucans from several sources at three pH values found along the digestive tract (3.0, 6.0, and 8.0). Acid and neutral conditions gave the highest affinity rates for aflatoxins B1 > deoxynivalenol > ochratoxin A and involved both the (1 --> 3)-beta-D-glucans and the (1 --> 6)-beta-D-glucans. Alkaline conditions, owing to their destructuring action on glucans, were favorable only for the adsorption of patulin. Using molecular mechanics, we found that hydroxyl, ketone, and lactone groups are involved in the formation of both hydrogen bonds and van der Waals interactions between aflatoxins B1, deoxynivalenol and patulin, and beta-D-glucans. Differences in the binding capacity of the mycotoxins are due to their specific physical and chemical characteristics.