Steady-state Continuous Cultures Research Articles

Controls on the hydrogen isotope composition of tetraether lipids in an autotrophic ammonia-oxidizing marine archaeon

The stable hydrogen isotope composition of persistent biomolecules is used as a palaeohydrological proxy. While much previous work has focused on plant leaf wax-derived n-alkanes, the potential of prokaryotic lipid biomarkers as carriers of H isotope signatures remains underexplored, particularly in the Archaea. Here we investigated H isotope distributions in the membrane lipids of the ammonia-oxidizing chemoautotroph Nitrosopumilus maritimus strain SCM1. Hydrogen isotope ratios were measured on the cleaved biphytane chains of tetraether membrane lipids extracted from steady-state continuous cultures cultivated at slow, medium, and fast growth rates. In contrast to recent work on bacterial fatty acids, where the direction and magnitude of isotopic fractionation varies widely (ca. 600‰ range) as a function of central C and energy metabolism, archaeal biphytane data in the present work are relatively invariant. The weighted average 2H/1H fractionation values relative to growth water (2εL/W) ranged from –272 to –260‰, despite a three-fold difference in doubling times (30.8–92.5 hr), yielding an average growth-rate effect <0.2‰ hr−1. These 2εL/W values are more negative than most heterotrophic microbial lipid H isotope measurements in the literature, and are on par with those from other autotrophic archaea, as well as with phytol from photoautotrophic algae. N. maritimus values of 2εL/W also varied systematically with the number of internal rings (cyclopentyl + cyclohexyl), increasing for each additional ring by 6.4 ± 2.7‰. Using an isotope flux-balance model in tandem with a comprehensive analysis of the sources of H in archaeal lipid biosynthesis, we use this observation to estimate the kinetic isotope effects (KIEs) of H incorporation from water; from reducing cofactors such as flavins and NADPH, and for the transhydrogenation reaction(s) that convert the electron-donor derived NADH into these cofactors. Consistent with prior studies on bacteria and plants, our results indicate the KIEs of reducing cofactors in archaea are highly fractionating, while those involving exchange of water protons are less so. When combined with the observation of minimal growth-rate sensitivity, our results suggest biphytanes of autotrophic 3HP/4HB utilizing Nitrososphaerota (a.k.a. Thaumarchaeota) may be offset from their growth waters by a nearly constant 2εL/W value. Together with the ring effect, this implies that all biphytanes originating from a common source should have a predictable ordering of their isotope ratios with respect to biphytane ring number, allowing precise reconstruction of the original δ2H value of the environmental water. Collectively, these patterns indicate archaeal biphytanes have potential as paleo-hydrological proxies, either as a complement or an alternative to leaf wax n-alkanes.

Bioinformatics and metabolic flux analysis highlight a new mechanism involved in lactate oxidation in Clostridium tyrobutyricum

Climate change and environmental issues compel us to find alternatives to the production of molecules of interest from petrochemistry. This study aims at understanding the production of butyrate, hydrogen, and CO2 from the oxidation of lactate with acetate in Clostridium tyrobutyricum and thus proposes an alternative carbon source to glucose. This specie is known to produce more butyrate than the other butyrate-producing clostridia species due to a lack of solvent genesis phase. The recent discoveries on flavin-based electron bifurcation and confurcation mechanism as a mode of energy conservation led us to suggest a new metabolic scheme for the formation of butyrate from lactate-acetate co-metabolism. While searching for genes encoding for EtfAB complexes and neighboring genes in the genome of C. tyrobutyricum, we identified a cluster of genes involved in butyrate formation and another cluster involved in lactate oxidation homologous to Acetobacterium woodii. A phylogenetic approach encompassing other butyrate-producing and/or lactate-oxidizing species based on EtfAB complexes confirmed these results. A metabolic scheme on the production of butyrate, hydrogen, and CO2 from the lactate-acetate co-metabolism in C. tyrobutyricum was constructed and then confirmed with data of steady-state continuous culture. This in silico metabolic carbon flux analysis model showed the coherence of the scheme from the carbon recovery, the cofactor ratio, and the ATP yield. This study improves our understanding of the lactate oxidation metabolic pathways and the role of acetate and intracellular redox balance, and paves the way for the production of molecules of interest as butyrate and hydrogen with C. tyrobutyricum.

Combination of Genome-Scale Models and Bioreactor Dynamics to Optimize the Production of Commodity Chemicals.

The current production of a number of commodity chemicals relies on the exploitation of fossil fuels and hence has an irreversible impact on the environment. Biotechnological processes offer an attractive alternative by enabling the manufacturing of chemicals by genetically modified microorganisms. However, this alternative approach poses some important technical challenges that must be tackled to make it competitive. On the one hand, the design of biotechnological processes is based on trial-and-error approaches, which are not only costly in terms of time and money, but also result in suboptimal designs. On the other hand, the manufacturing of chemicals by biological processes is almost exclusively carried out by batch or fed-batch cultures. Given that batch cultures are expensive and not easy to scale, technical means must be developed to make continuous cultures feasible and efficient. In order to address these challenges, we have developed a mathematical model able to integrate in a single model both the genome-scale metabolic model for the organism synthesizing the chemical of interest and the dynamics of the bioreactor in which the organism is cultured. Such a model is based on the use of Flexible Nets, a modeling formalism for dynamical systems. The integration of a microscopic (organism) and a macroscopic (bioreactor) model in a single net provides an overall view of the whole system and opens the door to global optimizations. As a case study, the production of citramalate with respect to the substrate consumed by E. coli is modeled, simulated and optimized in order to find the maximum productivity in a steady-state continuous culture. The predicted computational results were consistent with the wet lab experiments.

Production of extracellular α-amylase by single-stage steady-state continuous cultures of Candida wangnamkhiaoensis in an airlift bioreactor.

The kinetics of growth and α-amylase production of a novel Candida wangnamkhiaoensis yeast strain were studied in single-stage steady-state continuous cultures. This was performed in a split-cylinder internal-loop airlift bioreactor, using a variety of carbon sources as fermentation substrates. Results showed that the steady-state yields of cell mass from carbohydrates were practically constant for the range of dilution rates assayed, equaling 0.535 ± 0.030, 0.456 ± 0.033, and 0.491 ± 0.035 g biomass/g carbohydrate, when glucose, maltose, and starch, respectively were used as carbon sources. No α-amylase activity was detected when glucose was used as the carbon source in the influent medium, indicating that α-amylase synthesis of C. wangnamkhiaoensis is catabolically repressed by glucose. Contrastingly, maltose and starch induce synthesis of α-amylase in C. wangnamkhiaoensis, with starch being the best α-amylase inducer. The highest α-amylase volumetric and specific activities (58400 ± 800 U/L and 16900 ± 200 U/g biomass, respectively), and productivities (14000 ± 200 U/L·h and 4050 ± 60 U/g biomass·h, respectively) were achieved at a dilution rate of 0.24 h-1 using starch as the carbon source. In conclusion, single-stage steady-state continuous culture in an airlift bioreactor represents a powerful tool, both for studying the regulatory mechanisms of α-amylase synthesis by C. wangnamkhiaoensis and for α-amylase production. Furthermore, results showed that C. wangnamkhiaoensis represents a potential yeast species for the biotechnological production of α-amylase, which can be used for the saccharification of starch. This offers an attractive renewable resource for the production of biofuels (particularly bioethanol), representing an alternative to fossil fuels with reduced cost of substrates.

Existence of a scaling relation in continuous cultures of Scheffersomyces stipitis: the steady states are completely determined by the ratio of carbon and oxygen uptake rates

BackgroundRecently, we showed that steady-state continuous cultures of S. stipitis follow the principles of growth on mixture of two complementary substrates. More precisely, when such cultures are fed with progressively higher concentrations of glucose sf at fixed dilution rate D = 0.1 h−1, oxygen mass-transfer coefficient kla = 50 h−1, and oxygen solubility c_{text{o}}^{*}, they transition from glucose- to oxygen-limited growth through an intermediate dual-limited regime in which both glucose and oxygen are limiting, and ethanol is produced without loss of glucose. It is, therefore, of considerable interest to characterize the dual-limited regime. We found that the dual-limited regime occurs precisely when the operating parameters D, sf, kla, and c_{text{o}}^{*} satisfy the relation Y_{text{os}} < Ds_{text{f}} /left( {k_{text{l}} a cdot c_{text{o}}^{*} } right) < Y_{text{os}}^{prime }, where Yos and Y_{text{os}}^{prime } denote g of glucose consumed per g of oxygen consumed in the carbon- and oxygen-limited regimes. In this work, our goal was to determine if the above characterization of the dual-limited regime holds over a wider range of D, kla, and to understand why the dual-limited regime is determined by the dimensionless ratio Ds_{text{f}} /left( {k_{text{l}} a cdot c_{text{o}}^{*} } right).ResultsTo this end, we performed the foregoing experiments at three additional dilution rates (D = 0.07, 0.15, and 0.20 h−1) and one additional mass-transfer coefficient (kla = 100 h−1). We find that the above characterization of the dual-limited regime is valid for these conditions as well. Furthermore, the boundaries of the dual-limited regime are determined by the dimensionless ratio Ds_{text{f}} /left( {k_{text{l}} a cdot c_{text{o}}^{*} } right), because the steady-state concentrations are completely determined by this ratio. More precisely, if the steady-state concentrations of biomass, glucose, oxygen, and ethanol are suitably scaled, they collapse into a single curve with Ds_{text{f}} /left( {k_{text{l}} a cdot c_{text{o}}^{*} } right) as the independent variable.ConclusionThe dual-limited regime is characterized by the relation Y_{text{os}} < Ds_{text{f}} /left( {k_{text{l}} a cdot c_{text{o}}^{*} } right) < Y_{text{os}}^{{prime }} over the entire range of operating condition 0.07 h−1 ≤ D ≤ 0.20 h−1 and 50 ;{text{h}}^{ - 1} le k_{text{l}} a le 100;{text{h}}^{ - 1}. Since the effect of all operating parameters is embedded in the single parameter Ds_{text{f}} /left( {k_{text{l}} a cdot c_{text{o}}^{*} } right), the dimensionless plot provides a powerful tool to compare, with only a handful of data, various ethanol-producing strains over a wide range of operating conditions.

Kinetics of carbendazim degradation in a horizontal tubular biofilm reactor.

The fungicide carbendazim is an ecotoxic agent affecting aquatic biota. Due to its suspected hormone-disrupting effects, it is considered a "priority hazard substance" by the Water Framework Directive of the European Commission, and its degradation is of major concern. In this work, a horizontal tubular biofilm reactor (HTBR) operating in plug-flow regime was used to study the kinetics of carbendazim removal by an acclimated microbial consortium. The reactor was operated in steady state continuous culture at eight different carbendazim loading rates. The concentrations of the fungicide were determined at several distances of the HTBR. At the loading rates tested, the highest instantaneous removal rates were observed in the first section of the tubular biofilm reactor. No evidence of inhibition of the catabolic activity of the microbial community was found. Strains of the genera Flectobacillus, Klebsiella, Stenotrophomonas, and Flavobacterium were identified in the biofilm; the last three degrade carbendazim in axenic culture.

Simultaneous accumulation of neutral lipids and biomass in Nannochloropsis oceanica IMET1 under high light intensity and nitrogen replete conditions

The difficulty in simultaneously obtaining a high production of neutral lipids and biomass in the oleaginous microalga Nannochloropsis is an attractive challenge that needs to be addressed. To overcome this challenge and unveil the underlying metabolic characteristics, the model oleaginous microalga Nannochloropsis oceanica IMET1, under high light intensity and nitrogen-replete conditions, was prepared via steady-state continuous culture in the present study. The simultaneous accumulation of lipids, particularly neutral lipids, and biomass in N. oceanica IMET1 was triggered at a high light intensity (331.2μmolphotonsm−2s−1) in the light saturation region accompanied by a sufficient nitrogen supply (2000μmolL−1). Moreover, high amounts of valuable fatty acids (arachidonic acid and eicosapentaenoic acid) were also produced. The distribution of fatty acids and intracellular soluble metabolites, the membrane lipid composition and the photosynthetic pathway were associated with the concurrent accumulation of biomass and TAGs in N. oceanica IMET1. The common and different metabolic responses between high-light and nitrogen-depletion strategies for the accumulation of neutral lipids were also discussed. These results provide detailed metabolic clues for the genetic and metabolic engineering of Nannochloropsis for biofuel production.

Operational Stability to Changes in Composition of Herbicide Mixtures Fed to a Laboratory-Scale Biobarrier

The main objective of this work was to evaluate the operational stability of a laboratory-scale aerobic biobarrier designed for the treatment of water contaminated by mixtures of three herbicides frequently found in agricultural runoffs, atrazine, simazine and 2,4-dichlorophenoxyacetic acid (2,4-D). The microbial consortium used to degrade the herbicides was composed by six cultivable bacterial strains, identified as members of the genera Variovorax, Sphingopyxis, Hydrocarboniphaga, Methylobacterium, Pseudomonas and Acinetobacter. The effect caused by a seventh member of the microbial consortium, a ciliated protozoa of the genus Colpoda, on the herbicides biodegradation kinetics, was also evaluated. The biodegradation of five combinations of the herbicides 2,4-D, atrazine and simazine was studied in the biobarrier, operated in steady state continuous culture at different volumetric loading rates. In all cases, removal efficiencies determined by chemical oxygen demand (COD) and HPLC were nearly 100%. These results, joined to the null accumulation of aromatic byproducts of atrazine and simazine catabolism, show that after 495days of operation, in the presence of the protozoa, the adaptability of the microbial consortium to changing environmental conditions allowed the complete removal of the mixture of herbicides.

Comparison of Real Time Polymerase Chain Reaction Quantification of Changes in hilA and rpoS Gene Expression of a Salmonella Typhimurium Poultry Isolate Grown at Fast Versus Slow Dilution Rates in an Anaerobic Continuous Culture System

The objective of this study was to determine the genetic responses of a Salmonella enterica Typhimurium poultry isolate during low and high dilution rates (D) in steady state continuous culture (CC) incubations. Samples for genetic analyses were taken from a previous study where S. Typhimurium cells had been grown in two chemostats operated concurrently, which were designated as Trials 1 and 2, with eight dilution rates sampled during steady state. Real-time polymerase chain reaction (PCR) on two target genes (rpoS and hilA) was analyzed as changes in expression of the target gene relative to the reference gene (16S rRNA). At the lowest D (0.0125 h−1) in Trial 2, rpoS expression was more than twofold higher than the second highest relative expression. In Trial 1 hilA expression was 21.8-, 27.8-, and 21-fold higher in D 0.0125 h−1, 0.025 h−1, 0.05 h−1, respectively, compared to D 0.1 h−1, 0.54 h−1, and 1.08 h−1. In Trial 2, D 0.025 h−1 and 0.27 h−1 showed no difference in hilA expression but were significantly higher compared to other D. From these results, low glucose conditions may play an essential role in triggering rpoS induction, as well as contributing to potential virulence.

Photobioreactor design for isotopic non‐stationary 13C‐metabolic flux analysis (INST 13C‐MFA) under photoautotrophic conditions

Adaptive metabolic behavior of photoautotrophic microorganisms toward genetic and environmental perturbations can be interpreted in a quantitative depiction of carbon flow through a biochemical reaction network using isotopic non-stationary (13) C-metabolic flux analysis (INST (13) C-MFA). To evaluate (13) C-metabolic flux maps for Chlamydomonas reinhardtii, an original experimental framework was designed allowing rapid, reliable collection of high-quality isotopomer data against time. It involved (i) a short-time (13) C labeling injection device based on mixing control in a torus-shaped photobioreactor with plug-flow hydrodynamics allowing a sudden step-change in the (13) C proportion in the substrate feed and (ii) a rapid sampling procedure using an automatic fast filtration method coupled to a manual rapid liquid nitrogen quenching step. (13) C-substrate labeling enrichment was controlled through the total dissolved inorganic carbon concentration in the pulsed solution. First results were obtained from steady-state continuous culture measurements allowing the characterization of the kinetics of label incorporation into light-limited growing cells cultivated in a photobioreactor operating at the maximal biomass productivity for an incident photon flux density of 200 µmol m(-2) s(-1). (13)C label incorporation was measured for 21 intracellular metabolites using IC-MS/MS in 58 samples collected across a labeling experiment duration of 7 min. The fastest labeling rate was observed for 2/3-phosphoglycerate with an apparent isotopic stationary state reached after 300 s. The labeling rate was consistent with the optimized mixing time of about 4.9 s inside the reactor and the shortest reliable sampling period assessed at 5 s.

Effect of ADH II Deficiency on the Intracellular Redox Homeostasis inZymomonas mobilis

Mutant strain of the facultatively anaerobic, ethanol-producing bacterium Zymomonas mobilis, deficient in the Fe-containing alcohol dehydrogenase isoenzyme (ADH II), showed impaired homeostasis of the intracellular NAD(P)H during transition from anaerobic to aerobic conditions, and also in steady-state continuous cultures at various oxygen supplies. At the same time, ADH II deficiency in aerobically grown cells was accompanied by a threefold increase of catalase activity and by about 50% increase of hydrogen peroxide excretion. It is concluded that ADH II under aerobic conditions functions to maintain intracellular redox homeostasis and to protect the cells from endogenous hydrogen peroxide.

Toxicity evaluation of 2-hydroxybiphenyl and other compounds involved in studies of fossil fuels biodesulphurisation

The acute toxicity of some compounds used in fossil fuels biodesulphurisation studies, on the respiration activity, was evaluated by Gordonia alkanivorans and Rhodococcus erythropolis. Moreover, the effect of 2-hydroxybiphenyl on cell growth of both strains was also determined, using batch (chronic bioassays) and continuous cultures. The IC50 values obtained showed the toxicity of all the compounds tested to both strains, specially the high toxicity of 2-HBP. These results were confirmed by the chronic toxicity data. The toxicity data sets highlight for a higher sensitivity to the toxicant by the strain presenting a lower growth rate, due to a lower cells number in contact with the toxicant. Thus, microorganisms exhibiting faster generation times could be more resistant to 2-HBP accumulation during a BDS process. The physiological response of both strains to 2-HBP pulse in a steady-state continuous culture shows their potential to be used in a future fossil fuel BDS process.

Aerobic Biodegradation of a Sulfonated Phenylazonaphthol Dye by a Bacterial Community Immobilized in a Multistage Packed-Bed BAC Reactor

A microbial community able to aerobically degrade the azo dye Acid Orange 7 was selected from riparian or lacustrine sediments collected at sites receiving textile wastewaters. Three bacterial strains, pertaining to the genera Pseudomonas, Arthrobacter, and Rhizobium, constitute the selected community. The biodegradation of AO7 was carried out in batch-suspended cell culture and in a continuously operated multistage packed-bed BAC reactor. The rapid decolorization observed in batch culture, joined to a delay of about 24 h in COD removal and cell growth, suggests that enzymes involved in biodegradation of the aromatic amines generated after AO7 azo-bond cleavage (1-amino-2-naphthol [1-A2N] and 4-aminobenzenesulfonic acid [4-ABS]), are inducible in this microbial consortium. After this presumptive induction period, the accumulated byproducts, measured through COD, were partially metabolized and transformed in cell mass. At all azo dye loading rates used, complete removal of AO7 and 1-A2N was obtained in the multistage packed-bed BAC reactor (PBR).; however, the overall COD (eta ( COD )) and 4-ABS (eta ( ABS )) removal efficiencies obtained in steady state continuous culture were about 90%. Considering the toxicity of 1-A2N, its complete removal has particular relevance. In the first stages of the packed-bed BAC reactor (Fig. 4a-c), major removal was observed. In the last stage, only a slight removal of COD and 4-ABS was obtained. Comparing to several reported studies, the continuously operated multistage packed-bed BAC reactor showed similar or superior results. In addition, the operation of large-packed-bed BAC reactors could be improved by using several shallow BAC bed stages, because the pressure drop caused by bed compaction of a support material constituted by small and fragile particles can be reduced.

Cell yields and fermentation responses of a Salmonella Typhimurium poultry isolate at different dilution rates in an anaerobic steady state continuous culture

The objectives of these studies were to determine cell yield and fermentation responses of a Salmonella enterica serovar Typhimurium poultry isolate using various dilution rates in steady state continuous culture incubations. S. enterica Typhimurium cells were propagated in continuous cultures with a total volume of 0.50 l of Luria Bertani medium containing 0.1% glucose. Dilution rates from 0.0125 to 1.44/h were used. Cell protein concentration generally increased linearly with increased dilution rate up to a rate of 0.54/h and declined at the higher dilution rates. Glucose consumption gave a similar pattern to cell protein concentration by declining at the three highest dilution rates. Short chain fatty acid production was inconsistently influenced by dilution rate. Acetate, the most predominant fatty acid produced, declined at the higher dilution rates, as did propionate. Ammonia production remained stable at the lowest dilution rates, but increased significantly at a dilution rates above 0.27/h.

Coordination of Growth Rate, Cell Cycle, Stress Response, and Metabolic Activity in Yeast

We studied the relationship between growth rate and genome-wide gene expression, cell cycle progression, and glucose metabolism in 36 steady-state continuous cultures limited by one of six different nutrients (glucose, ammonium, sulfate, phosphate, uracil, or leucine). The expression of more than one quarter of all yeast genes is linearly correlated with growth rate, independent of the limiting nutrient. The subset of negatively growth-correlated genes is most enriched for peroxisomal functions, whereas positively correlated genes mainly encode ribosomal functions. Many (not all) genes associated with stress response are strongly correlated with growth rate, as are genes that are periodically expressed under conditions of metabolic cycling. We confirmed a linear relationship between growth rate and the fraction of the cell population in the G0/G1 cell cycle phase, independent of limiting nutrient. Cultures limited by auxotrophic requirements wasted excess glucose, whereas those limited on phosphate, sulfate, or ammonia did not; this phenomenon (reminiscent of the "Warburg effect" in cancer cells) was confirmed in batch cultures. Using an aggregate of gene expression values, we predict (in both continuous and batch cultures) an "instantaneous growth rate." This concept is useful in interpreting the system-level connections among growth rate, metabolism, stress, and the cell cycle.

Continuous biodegradation of single and mixed chlorophenols by a mixed microbial culture constituted by Burkholderia sp., Microbacterium phyllosphaerae, and Candida tropicalis

In this work, the biodegradation of mixed chlorophenols by a microbial consortium was studied in steady state continuous culture. Amplified ribosomal DNA restriction analysis (ARDRA) was used to evaluate the consortium's predominating bacterial species when chemostat was fed with pure or mixed chlorophenols. Removal efficiencies, cell growth yields, volumetric and specific biodegradation rates of phenol and chlorophenols were compared to those obtained with pure strains in the same culture conditions. Using phenol as the primary carbon and energy source, the microbial consortium cometabolically degraded mono-, di- and trichlorophenols, with overall removal efficiencies ranging from 95 to 99.8% and COD removal efficiencies from 85 to 97.8%. In all cultures, the microbial consortium showed better overall removal efficiencies than single strains. Binary mixtures of chlorophenols produced a remarkably increase in specific biodegradation rates of substrates q c, beyond those obtained when the consortium grew on single compounds. The results suggest that chlorophenols collapsed cell's proton motive force; thus, energy spilling, reflected by the q c increase, could be the cell's response to compensate the short-circuit in membrane electrochemical proton gradient.

The use of multi‐parameter flow cytometry to study the impact of limiting substrate, agitation intensity, and dilution rate on cell aggregation during Bacillus licheniformis CCMI 1034 aerobic continuous culture fermentations

The main objective of this work was to establish those factors either physical (power input) or chemical (limiting substrate or dilution rate) that enhance cell aggregation (biofilm or floc formation) and cell physiological state during aerobic continuous cultures of Bacillus licheniformis. Glucose-limited steady-state continuous cultures growing at a dilution rate between 0.64 and 0.87/h and 1,000 rpm (mean specific energy dissipation rate (epsilonT) = 6.5 W/kg), led to the formation of a thin biofilm on the vessel wall characterized by the presence of a high proportion of healthy cells in the broth (after aggregate disruption by sonication) defined as having intact polarized cytoplasmic membranes. An increased epsilonT (from 6.5 W/kg to 38 W/kg) was found to hinder cell aggregation under carbon limitation. The carbon recovery calculated from glucose indicated that additional extracellular polymer was being produced at dilution rates >0.87/h. B. licheniformis growth under nitrogen limitation led to floc formation which increased in size with dilution rate. Counter-intuitively the flocs became more substantial with an increase in epsilonT from 6.5 W/kg to 38 W/kg under nitrogen limitation. Indeed the best culture conditions for enhanced metabolically active cell aggregate formation was under nitrogen limitation at epsilonT = 6.5 W/kg (leading to floc formation), and under carbon limitation at a dilution rate of between 0.64 and 0.87/h, at epsilonT = 6.5 W/kg (leading to vessel wall biofilm formation). This information could be used to optimize culture conditions for improved cell aggregation and hence biomass separation, during thermophilic aerobic bioremediation processes.

Sulfur Formation by Steady-state Continuous Cultures of a Sulfoxidizing Consortium And Thiobacillus thioparus ATCC 23645

The elemental sulfur formation by the partial oxidation of thiosulfate by both a sulfoxidizing consortium and by Thiobacillus thioparus ATCC 23645 was studied under aerobic conditions in chemostat. Steady state was attained with essentially total conversion to sulfate when the dissolved oxygen concentration was 5 mgO2 l-1 and below a dilution rate (D) of 3.0 d-1 for the consortium and 0.9 d-1 for T thioparus. The consortium formed elemental sulfur in steady state under oxygen limitation. Fifty percent of the theoretical elemental sulfur yield was obtained with a dissolved oxygen concentration of 0.2 mgO2 l-1. Growth of T thioparus was negatively affected with a concentration below 1.9 mgO2 l-1. Consortium yield from batch cultures was 2.1 g-1 proteinmol-1 thiosulfate, which was comparable with the values obtained in the chemostat at dilution rates of 0.4 d-1 and 1.2 d-1. The consortium showed a maximum degradation rate of 0.105 gthiosulfate g-1 proteinmin-1 and a saturation rate for S2O3 2-of 1.9 mM.

Sulfur Formation by Steady-state Continuous Cultures of a Sulfoxidizing Consortium And Thiobacillus thioparus ATCC 23645

Sulfur Formation by Steady-state Continuous Cultures of a Sulfoxidizing Consortium And Thiobacillus thioparus ATCC 23645

Metabolic changes in Clostridium absonum ATCC 27555 accompanying induction of epimerization of a primary bile acid.

Some parameters of fermentation have been determined for Clostridium absonum in a chemostat by using a chemically defined medium with glucose as the sole source of carbon and energy. Steady-state continuous cultures were achieved at two dilution rates (D). Trends of the carbon flow were determined by comparison of ratios between the specific rates of formation of the three products of metabolism (lactate, acetate, butyrate). Chenodeoxycholate induced the 7alpha- and 7beta-hydroxysteroid dehydrogenases of C. absonum. In the presence of this inducer, the growth yield and the carbon recovery decreased, the carbon flow distribution was altered favoring acetate production, and a deficit in the reoxydation of nucleotidic cofactors was observed. In the presence of chenodeoxycholate, C. absonum would favor the production of energy at the expense of the reoxidation of nucleotidic cofactors so as to ensure its growth, and the epimerization of chenodeoxycholate to ursodeoxycholate.