Enzymatic synthesis of mono- and trifluorinated alanine enantiomers expands the scope of fluorine biocatalysis.

Fluorinated amino acids serve as an entry point for establishing new-to-Nature chemistries in biological systems, and novel methods are needed for the selective synthesis of these building blocks. In this study, we focused on the enzymatic synthesis of fluorinated alanine enantiomers to expand fluorine biocatalysis. The alanine dehydrogenase from Vibrio proteolyticus and the diaminopimelate dehydrogenase from Symbiobacterium thermophilum were selected for in vitro production of (R)-3-fluoroalanine and (S)-3-fluoroalanine, respectively, using 3-fluoropyruvate as the substrate. Additionally, we discovered that an alanine racemase from Streptomyces lavendulae, originally selected for setting an alternative enzymatic cascade leading to the production of these non-canonical amino acids, had an unprecedented catalytic efficiency in ß-elimination of fluorine from the monosubstituted fluoroalanine. The in vitro enzymatic cascade based on the dehydrogenases of V. proteolyticus and S. thermophilum included a cofactor recycling system, whereby a formate dehydrogenase from Pseudomonas sp. 101 (either native or engineered) coupled formate oxidation to NAD(P)H formation. Under these conditions, the reaction yields for (R)-3-fluoroalanine and (S)-3-fluoroalanine reached >85% on the fluorinated substrate and proceeded with complete enantiomeric excess. The selected dehydrogenases also catalyzed the conversion of trifluoropyruvate into trifluorinated alanine as a first-case example of fluorine biocatalysis with amino acids carrying a trifluoromethyl group.

Hydrogeochemical characterization and natural background level determination of selected inorganic substances in groundwater from a semi-confined aquifer in Midwestern Burkina Faso, West Africa.

Hydrogeochemical processes that govern selected inorganic substances distribution in a semi-confined aquifer were characterized using traditional hydrogeochemical approaches and natural background levels (NBLs). Saturation indices and bivariate plots were used to investigate the effects of water-rock interactions on natural evolution of the groundwater chemistry, whereas Q-mode hierarchical cluster analysis and one-way analysis of variance classified the groundwater samples into three distinct groups. To highlight the groundwater status, NBLs and threshold values (TVs) of the substances were calculated using pre-selection method. Piper's diagram showed that the Ca-Mg-HCO3 water type was the only hydrochemical facies of the groundwaters. Although all samples, except a borewell with a high NO3- concentration, had major ion and transition metal concentrations within the World Health Organization's recommended guideline values for drinking water, Cl-, NO3- and PO43- exhibited scattered distribution patterns, reflecting their nonpoint anthropogenic sources in the groundwater system. The bivariate and saturation indices revealed that silicate weathering and possible gypsum and anhydrite dissolution contributed to the groundwater chemistry. In contrast, NH4+, FeT and Mn abundance appeared to be influenced by redox conditions. Strong positive spatial correlations between pH, FeT, Mn and Zn suggested that mobility of these metals was controlled by pH. The relative high F- concentrations in lowland areas may imply the impact of evaporation on this ion's abundance. Contrary to TVs of HCO3-, those of Cl-, NO3-, SO42-, F- and NH4+ were below the guideline values, confirming the influence of chemical weathering on the groundwater chemistry. Based on the present findings, further studies that take into account more inorganic substances are required for NBLs and TVs determination in the area, thereby setting up a robust sustainable management plan for the regional groundwater resources.

Altering the fatty acid profile of Yarrowia lipolytica to mimic cocoa butter by genetic engineering of desaturases.

BACKGROUND: Demand for Cocoa butter is steadily increasing, but the supply of cocoa beans is naturally limited and under threat from global warming. One route to meeting the future demand for cocoa butter equivalent (CBE) could be to utilize microbial cell factories such as the oleaginous yeast Yarrowia lipolytica. RESULTS: The main goal was to achieve triacyl-glycerol (TAG) storage lipids in Y. lipolytica mimicking cocoa butter. This was accomplished by replacing the native Δ9 fatty acid desaturase (Ole1p) with homologs from other species and changing the expression of both Ole1p and the Δ12 fatty acid desaturase (Fad2p). We thereby abolished the palmitoleic acid and reduced the linoleic acid content in TAG, while the oleic acid content was reduced to approximately 40 percent of the total fatty acids. The proportion of fatty acids in TAG changed dramatically over time during growth, and the fatty acid composition of TAG, free fatty acids and phospholipids was found to be very different. CONCLUSIONS: We show that the fatty acid profile in the TAG of Y. lipolytica can be altered to mimic cocoa butter. We also demonstrate that a wide range of fatty acid profiles can be achieved while maintaining good growth and high lipid accumulation, which, together with the ability of Y. lipolytica to utilize a wide variety of carbon sources, opens up the path toward sustainable production of CBE and other food oils.

Occurrence, behaviour and environmental risk assessment of trace metals in stream sediments from southwestern Burkina Faso, West Africa.

In this study, the influence of source area weathering and provenance on the stream sediment geochemistry as well as the environmental impacts of selected potentially toxic trace elements (PTE) were evaluated. Four components derived from the R-mode factor analysis of additive logratio-transformed data pointed to the combined influence of weathering of granitoids, sedimentary rocks and greenstone belts and gold mineralisation on the stream geochemistry. Although 13 PTE were enriched in the majority of the samples, only five elements (As, Cr, Cu, Pb and Ni) were likely to have adverse biological effects. Based on the geochemical maps, the areas susceptible to produce adverse biological effects due to As enrichments are solely drained by the greenstone belts, whereas enrichment of Pb was pronounced in areas of high population densities. The linear regression between enrichment factor (EF) and adverse effect indices (AEI) indicated that in order for Pb to pose threats to the benthic organisms, EF should be 4.5, whereas EF for sediment toxicity are 1 for Cu, Cr and Ni and 2 for As. Consequently, Cu, Cr, Ni, As and, to a lesser degree, Pb pose serious environmental threats to benthic organisms in southwestern Burkina Faso. The stream sediment geochemical data of this study constitute a baseline for assessing future environmental risks.