Patrick Jame

A phyloprofile of Frankia genomes was carried out to identify those genes present in symbiotic strains of clusters 1, 1c, 2 and 3 and absent in non-infective strains of cluster 4. At a threshold of 50% AA identity, 108 genes were retrieved. Among these were known symbiosis-associated genes such as nif (nitrogenase), and genes which are not know as symbiosis-associated genes such as can (carbonic anhydrase, CAN). The role of CAN, which supplies carbonate ions necessary for carboxylases and acidifies the cytoplasm, was thus analyzed by staining cells with pH-responsive dyes; assaying for CO₂ levels in N-fixing propionate-fed cells (that require a propionate-CoA carboxylase to yield succinate-CoA), fumarate-fed cells and N-replete propionate-fed cells; conducting proteomics on N-fixing fumarate and propionate-fed cells and direct measurement of organic acids in nodules and in roots. The interiors of both in vitro and nodular vesicles were found to be at a lower pH than that of hyphae. CO₂ levels in N₂-fixing propionate-fed cultures were lower than in N-replete ones. Proteomics of propionate-fed cells showed carbamoyl-phosphate synthase (CPS) as the most overabundant enzyme relative to fumarate-fed cells. CPS combines carbonate and ammonium in the first step of the citrulline pathway, something which would help manage acidity and NH₄⁺. Nodules were found to have sizeable amounts of pyruvate and acetate in addition to TCA intermediates. This points to CAN reducing the vesicles’ pH to prevent the escape of NH₃ and to control ammonium assimilation by GS and GOGAT, two enzymes that work in different ways in vesicles and hyphae. Genes with related functions (carboxylases, biotin operon and citrulline-aspartate ligase) appear to have undergone decay in non-symbiotic lineages.

Saffron is a very high value-added ingredient used in the food supplement market and contains a high level of safranal. Adding synthetic safranal to saffron, which is significantly cheaper, and falsifying the origin of saffron may represent recurrent fraud. Saffron from different countries was analyzed to determine the stable isotope ratios δ¹³C and δ²H from safranal by gas chromatography coupled with isotope-ratio mass spectrometry (GC-C/P-IRMS) and the concentration of saffron metabolites with ultra-high performance liquid chromatography coupled with diode array detector (UHPLC-DAD). The isotopic analysis highlighted a higher ratio of δ²H in synthetic safranal than in natural safranal; the mean values were 36‰ (+/− 40) and −210‰ (+/− 35), respectively. The δ¹³C between Iranian, Spanish and other saffron was significantly different and represents median values of −28.62‰, −30.12‰ and −30.70‰, respectively. Moreover, linear and quadratic discriminant analyses (LDA and QDA) were computed using the two isotope ratios of safranal and the saffron metabolites. A first QDA showed that trans-crocetin and the δ¹³C of safranal, picrocrocin, and crocin C3 concentrations clearly differentiated Iranian saffron from other origins. A second model identified δ¹³C, trans-crocetin, crocin C2, crocin C3, and picrocrocin as good predictors to discriminate saffron samples from Iran, Spain, or other origins, with a total ability score classification matrix of 100% and a prediction matrix of 82.5%. This combined approach may be a useful tool to authenticate the origin of unknown saffron.

Three different types of biomass sourced from forestry waste (eucalyptus residues), agricultural waste (wheat straw), and energy crop (miscanthus) were used as starting materials to produce hemicellulosic sugars, furans (furfural and hydroxymethylfurfural), and oligosaccharides. A two-step hybrid process was implemented; biomass was first autohydrolysed without any additive to extract hemicelluloses and dissolve it in water. Then, the hydrolysate was treated with a solid acid catalyst, TiO₂-WO_x, in order to achieve hydrolysis and produce monomeric sugars and furans. This article investigates the role of the biomass type, autohydrolysis experimental conditions, polymerisation degree and composition of hemicelluloses on the performance of the process coupling autohydrolysis and catalytic hydrolysis. The highest global yields of both oligosaccharides and monomeric sugars were obtained from Eucalyptus (37% and 18%, respectively).

One of the principal theories about the origin of life is based on the abiotic reduction of carbon oxides to various organic molecules in hydrothermal systems. This synthesis is most favored in ultramafic environments undergoing hydrothermal alteration where the serpentinization reaction efficiently produces H2. Nevertheless, decades of hydrothermal experiments have hardly succeeded in producing abundant organic volatiles such as CH4 and short-chain hydrocarbons. On another hand, natural observations have shown the occurrence of other abiotic compounds such as organic acids in fluids and carbonaceous matter (CM) within serpentinized rocks. But organic acids as carbon source and CM as product have not been investigated so far in experiments reproducing hydrothermal peridotite alteration. Here, we explored the effect of formic acid (HCOOH) on the serpentinization reaction and possible feedback effects on carbon speciation in both fluid and solid. We performed reactions at 300°C and 250 bar using peridotite powder (<40 microns) in the presence of  0.1 M formic acid. A temperature of 300°C has been shown to be optimal for olivine serpentinization, while formic acid should partly decomposed into H2, CO, and CO2. After 4 months, H2, CO, CO2, CH4 and short-chain alkanes (mainly ethane) were measured in the fluid, and the powder was completely indurated. The solidified powder displayed a black and white layering perpendicular to fluid diffusion. Its analysis showed the advancement of the serpentinization reaction, and the incorporation of carbon compounds into the solid phase. XRD analysis indicated 70% of serpentinization. SEM-EDX observations showed peculiar texture with large and localized euhedral magnetite grains alternating with larger magnetite grains mixed with C-enriched areas of long chrysotile fibers. FT-IR measurement attested of the widespread formation of carbonaceous material in the solid. Liquid analyses are under progress. Those first results suggest that serpentine formation not only provides additional H2 to the system, but also mineral surfaces that could play a role in the precipitation of carbonaceous material and carbon speciation in natural systems. The nature and formation mechanisms of this latter remain to be addressed but this opens new paths for abiotic organic synthesis under hydrothermal conditions. In addition to their implications as an abiotic carbon source for deep hydrocarbon degraders ecosystems, it could have important implications for the total carbon cycle.