Fermentation

vignette|Fermentation industrielle. La fermentation est un processus métabolique convertissant généralement des glucides en acides, en gaz ou en alcools pour en extraire une partie de l'énergie chimique tout en ré-oxydant les coenzymes réduites par ces réactions. Il s'agit d'une voie métabolique d'oxydoréduction dans laquelle l'accepteur ultime d'électrons est souvent confondu avec le produit final des réactions. Elle se caractérise par une dégradation partielle de la substance fermentescible et ne permet qu'une production d'énergie limitée. Elle a lieu chez des levures et des bactéries, ainsi que dans les cellules musculaires manquant d'oxygène, c'est-à-dire en conditions anaérobies. Sa caractérisation au contribua à la découverte des enzymes. Louis Pasteur estimait ainsi que des ferments étaient responsables de la fermentation alcoolique chez la levure. Plus précisément, la fermentation est un mode de respiration cellulaire mettant en œuvre un système de transfert d'électrons repo

Influence of pH on dehalorespiring consortia & first steps in the development of a methodology for acid neutralization in enhanced in situ bioremediation

Nicole Munz

Enhanced in situ bioremediation of chlorinated solvents is, especially in source zone remediation, often limited through the accompanied high acidification of groundwater. This acid built-up originating from the dechlorination itself and fermentation products, such as organic acids, presents a draw-back in the efficiency of dehalorespiring bacteria. It is therefore indispensable to develop and implement appropriate buffering strategies to ensure a constant transformation of toxic chloroethenes to the final non-toxic product ethene. In this study pH tolerance of different dehalorespiring bacteria was tested. The limiting acidic pH varies significantly among the consortia. The consortium SL23b4 (PCE to ethene) grows at all the examined pH (pH 5 to pH 7.5) with a pH optimum between 6 and 7 (depending on the dechlorination step), whereas Maroc 6b (PCE to ethene) was only found to grow at incubation pH of 7, and very slowly at pH 6.5. The consortium 2SO (cis-DCE to ethene) seems to be inhibited at acidic pH (no growth at pH 5 and 5.5) having a pH optimum above neutral pH. In most cases incomplete dechlorination was observed in cultures growing at acidic pH or below the pH optima, accumulating toxic intermediate dechlorination products as a consequence. Additionally, high rates of dechlorination were always accompanied by pH drops up to 1.2 pH units, giving evidence of high acid production. pH had also influence on the community composition. In the consortium SL23b4 a shift from a Dehalococcoides- (pH 6-7.5) to a Sulfurospirillum spp.- (pH5 -5.5) dominated community was observed. Consortium Maroc 6b showed significantly different community compositions due to a pH change of only 0.5 pH units. Dehalococcoides spp. and Dehalobacter spp. were dominating at pH 7 and pH 6.5, respectively. Consortium 2SO is dominated by Dehalococcoides spp. at all pH, although at pH 7 a much higher diversity seems to prevail, with the threefold number of OTUs. Tests on the tolerance of the dehalorespiring consortia for elevated PCE concentration showed similar patterns for the different dechlorination steps, as for pH cultures, and confirmed a generally higher sensitivity towards PCE and pH changes for the last steps of dechlorination. Conclusively, Dehalococcoides spp., the only known genera able to perform complete dechlorination, seem to be highly sensitive to pH and PCE changes. Further, abiotic experiments on silicate mineral dissolution, using forsteritic olivine, were performed to simulate the buffer capacity of the minerals for the acid production during microbial reductive dechlorination. In these experiments, the buffer capacity was insufficient to counterbalance the constant acid titration rate, resulting in a final pH between 3 and 4 (initial pH of 7). The formation of precipitates, such as iron oxide, and the re-precipitation of magnesium were observed in some cases, showing a decrease in magnesium concentrations.

2010

Production of a thermostable fumarase in Saccharomyces cerevisiae for the bioconversion of fumarate to L-malate

Coralie Signorell

The gene encoding fumarase (fum) from Thermus thermophilus was expressed in yeast Saccharomyces cerevisiae. The recombinant cells were heated at 70°C to inactivate indigenous enzymes and used for the bioconversion of fumaric acid to L-malic acid. By heating the host cells at 70°C, substrate is able to go across the heat-damaged membrane of the microorganism and a desired product can be formed. This new concept, called Synthetic Metabolic Engineering (SME), has already been applied successfully in Escherichia coli. Unfortunately, E. coli membrane is weakened too much during the heat treatment and enzyme leakage appears. The surface structure of yeast is more rigid than that of E. coli and this might be taken as an advantage for application of SME. When continuous or repeated-batch reaction is carried out, enzyme leakage becomes a major drawback. It is assumed that yeast cells could overcome this problem by retaining more enzymes in the cell during and after the heat treatment. In order to prove this hypothesis, a thermophilic fumarase (FUM) was over-expressed in two hosts, S. cerevisiae as well as E. coli. fum was first modified to be over-expressed in yeast cells and FUM was successfully produced in yeasts. Optimization of SME techniques was carried out for yeast cells. Then, enzyme activity and enzyme leakage was investigated for both strains. E. coli showed high level of FUM expression, though considerable amount of enzyme leaked to supernatant. On the other hand, even though the level of FUM expression in S. cerevisiae was low, yeast cells overcome leakage problem and are re-usable. This study showed the first trial of SME in yeast cells and possibility of utilization of yeast as a host strain for SME.

2011

Influence of pH on dehalorespiring consortia & first steps in the development of a methodology for acid neutralization in enhanced in situ bioremediation

Nicole Munz

2010

Characterization of the autolytic culture Lactobacillus delbrueckii ssp. lactis FAM-10991 during continuous fermentation using immobilized cells

Influence of pH on dehalorespiring consortia & first steps in the development of a methodology for acid neutralization in enhanced in situ bioremediation

Production of a thermostable fumarase in Saccharomyces cerevisiae for the bioconversion of fumarate to L-malate

Characterization of the autolytic culture Lactobacillus delbrueckii ssp. lactis FAM-10991 during continuous fermentation using immobilized cells

Influence of pH on dehalorespiring consortia & first steps in the development of a methodology for acid neutralization in enhanced in situ bioremediation

Production of a thermostable fumarase in Saccharomyces cerevisiae for the bioconversion of fumarate to L-malate