Biodiesel via in situ wet microalgae biotransformation: Zwitter-type ionic liquid supported extraction and transesterification

The production of biodiesel derived from microalgae is among the most forthcoming technologies that provide an ecologic alternative to fossil fuels. Herein, a method was developed that enables the direct extraction and conversion of algal oil to biodiesel without prior isolation. The reaction occurs in aqueous media catalyzed by immobilized Candida antarctica lipase B (Novozyme 435). Zwitter-type ionic liquids were used as cocatalyst to improve the selectivity and reactivity of the enzyme. In a model reaction with sunflower oil, 64% biodiesel was obtained. Applying this method to a slurry of whole-cell Chlorella zof ingiensis in water resulted in 74.8% of lipid extraction, with 27.7% biotransformation products and up to 16% biodiesel. Factors that reduced the lipase activity with whole-cell algae were subsequently probed and discussed. This "in situ" method shows an improvement to existing methods, since it integrates the oil extraction and conversion into an one-pot procedure in aqueous conditions. The extraction is nondisruptive, and is a model for a greener algae to biodiesel process

Experimental and Computational Study of Supercritical Fluid Extraction (SFE) of Omega-3 Components from Fish Oil in Structured Packing

The benefits of polyunsaturated fatty acids and their implications for human health have gained scientific attention to their extraction from biological sources, not being produced by the human body. Most known industrial productions of omega-3 fatty acids often work under operating conditions that may degrade these components and they often use toxic or flammable solvents that can adversely affect human health. In this sense, innovative and interesting prospects are provided by Supercritical Fluid Extraction (SFE). In this work, two parallel studies were carried out: an experimental activity in a laboratory apparatus using supercritical carbon dioxide (scCO2) and preliminary computational fluid dynamics (CFD) simulations, limited to the hydrodynamic aspects of the process. In the experimental apparatus a Sulzer® EX structured packing, made up of corrugated metal gauze sheets, was used as the column filler. The study made it possible to identify the optimal operating conditions leading to an enrichment of the starting mixture in Eicosapentaenoic acid (EPA) and Docosahexaenoic acid (DHA), target products. CFD simulations were based on the Volume of Fluid (VOF) approach, suitable to the present complex multiphase system with two phases in close contact (transesterified fish oil and scCO2). The meatus created by the corrugations of the metal gauze was chosen as the calculation domain representative of the system. The computations were performed by the commercial software Ansys Fluent®, which allowed the prediction of the hydrodynamic evolution of the system through transient simulations. CFD predictions were in qualitative agreement with the experimental result

Kinetic of the Sewage Treatment: The Consumption of Organic Carbon of The Microalga Chlorella sp

As well known, microalgae are eukaryotic or procaryotic microorganisms able to photosynthesize, namely transforming inorganic substrates and sun light into organic compounds and chemical energy. They result very promising in treating civil wastewaters thanks to their ability to employ nitrates and phosphates as nutrients (Lima et al., 2019). Autotrophic microalgae are, anyway, not useful in decreasing the organic carbon content of wastewaters, and for this reason, they cooperate with heterotrophic bacteria. The usefulness of microalgaebacteria consortia in treating wastewaters and the ratio of their inoculum was investigated in a previous work (Lima, 2022a). Contrarily to autotrophic microalgae, mixotrophic microalgae are able to decrease the organic content of the matrix in which they are grown. In this work, we preliminarily investigated the capability of the autochthonous microalga Chlorella sp. CW2 to grow in mixotrophy and decrease the organic content of the artificial wastewater in which they are grown. Several batch cultivations were performed with glucose in different concentrations. Kinetic parameters were obtained and employed to determine the dilution rate (D) ideal for the abatement of glucose from the artificial wastewater

Systematic screening for the biocatalytic hydration of fatty acids from different oily substrates by Elizabethkingia meningoseptica oleate hydratase through a Design-of-experiments approach

The edible plant oils production is associated with the release of different types of by-products. The latter represent cheap and available substrates to produce valuable compounds, such as flavours and fragrances, biologically active compounds and bio-based polymers. Elizabethkingia meningoseptica Oleate hydratases (Em_OhyA) can selectively catalyze the conversion of unsaturated fatty acids, specifically oleic acid, into hydroxy fatty acids, which find different industrial applications. In this study, Design-of-experiment (DoE) strategy was used to screen and identify conditions for reaching high yields in the reaction carried out by Escherichia coli whole-cell carrying the recombinant enzyme Em_OhyA using Waste Cooking Oils (WCO)-derived free fatty acids (FFA) as substrate. The identified reaction conditions for high oleic acid conversion were also tested on untreated triglycerides-containing substrates, such as pomace oil, sunflower oil, olive oil and oil mill wastewater (OMW), combining the triglyceride hydrolysis by the lipase from Candida rugosa and the E. coli whole-cell containing Em_OhyA for the production of hydroxy fatty acids. When WCO, sunflower oil and OMW were used as substrate, the one-pot bioconversion led to an increase of oleic acid conversion compared to the standard reaction. This work highlights the efficiency of the DoE approach to screen and identify conditions for an enzymatic reaction for the production of industrially-relevant products

Identification of a spin-coupled Mo(III) in the nitrogenase iron-molybdenum cofactor

International audienceNitrogenase is a complex enzyme that catalyzes the formation of ammonia utilizing a MoFe7S9C cluster. The presence of a central carbon atom was recently revealed, finally completing the atomic level description of the active site. However, important prerequisites for understanding the mechanism - the total charge, metal oxidation states and electronic structure are unknown. Herein we present high-energy resolution fluorescence detected Mo K-edge X-ray absorption spectroscopy of nitrogenase. Comparison to FeMo model complexes of known oxidation state indicates that the Mo in the FeMo cofactor of nitrogenase is best described as Mo(III), in contrast to the universally accepted Mo(IV) assignment. The oxidation state assignment is supported by theoretical calculations, which reveal the presence of an unusual spin-coupled Mo(III) site. Although so far Mo(III) was not reported to occur in biology the suggestion raises interesting parallels with the known homogenous Mo catalysts for N-2 reduction, where a Mo(III) compound is the N-2-binding species. It also requires a reassignment of the Fe oxidation states in the cofacto

Reliability of a forensic odontology method for age-at-death estimation in adults: a Mexican case study

This study aimed to evaluate the reliability of an age estimation method based on the pulp⁄tooth area ratio by assessing intra- and inter-examiner agreement across five observers at different intervals. Using the same X-ray device and technical parameters, 96 digital periapical X-ray images of upper and lower canines were obtained from 28 deceased people in Central America, whose age at death ranged from 19 to 49 years. Excellent and good agreement of results were achieved, and there were no statistically significant differences. The R2 value for upper teeth (54.0%) was higher than the R2 value for lower teeth (45.7%). The highest intraclass correlation coefficient value was 0.995 (0.993–0.997) and the lowest 0.798 (0.545–0.895). Inter-examiner agreement was high with values of 0.975 (0.965–0.983) and 0.927 (0.879–0.955). This method is adequate for assessing age in missing and unidentified people, including victims of mass disasters

Phylogeny of European Anodontini (Bivalvia: Unionidae) with a redescription of Anodonta exulcerata

Freshwater bivalves are highly threatened and globally declining due to multiple anthropogenic impacts, making them important conservation targets. Because conservation policies and actions generally occur at the species level, accurate species identification and delimitation is critical. A recent phylogenetic study of Italian mussel populations revalidated an Anodonta species bringing the number of known European Anodontini from three to four species. The current study contributes to the clarification of the taxonomy and systematics of European Anodontini, using a combination of molecular, morphological and anatomical data, and constructs phylogenies based on complete mitogenomes. A redescription of A. exulcerata and a comparative analysis of morphological and anatomical characters with respect to the other two species of Anodonta present in the area are provided. No reliable diagnostic character has emerged from comparative analysis of the morphometric characters of 109 specimens from 16 sites across the Italian peninsula. In fact, the discriminant analysis resulted in a greater probability of correct assignment to the site of origin than to the species. This confirms the difficulties of an uncritical application of visual characters for the delimitation of species, especially for Anodontinae.This research was developed under ConBiomics: the missing approach for the conservation of freshwater bivalves project Nº NORTE-01-0145-FEDER-030286, cofinanced by COMPETE 2020, Portugal 2020 and the European Union through the ERDF - European Regional Development Fund and by FCT - Fundação para a Ciência e a Tecnologia, through national funds (UID/Multi/04423/2019). FCT also supported Manuel Lopes-Lima (SFRH/BD/115728/2016).info:eu-repo/semantics/publishedVersio

Kinetic of the Sewage Treatment: The Consumption of Organic Carbon of The Microalga Chlorella sp.

As well known, microalgae are eukaryotic or procaryotic microorganisms able to photosynthesize, namely transforming inorganic substrates and sun light into organic compounds and chemical energy. They result very promising in treating civil wastewaters thanks to their ability to employ nitrates and phosphates as nutrients (Lima et al., 2019). Autotrophic microalgae are, anyway, not useful in decreasing the organic carbon content of wastewaters, and for this reason, they cooperate with heterotrophic bacteria. The usefulness of microalgae-bacteria consortia in treating wastewaters and the ratio of their inoculum was investigated in a previous work (Lima, 2022a). Contrarily to autotrophic microalgae, mixotrophic microalgae are able to decrease the organic content of the matrix in which they are grown. In this work, we preliminarily investigated the capability of the autochthonous microalga Chlorella sp. CW2 to grow in mixotrophy and decrease the organic content of the artificial wastewater in which they are grown. Several batch cultivations were performed with glucose in different concentrations. Kinetic parameters were obtained and employed to determine the dilution rate (D) ideal for the abatement of glucose from the artificial wastewater

Human Intrinsic Factor Expression for Bioavailable Vitamin B12 Enrichment in Microalgae

Dietary supplements and functional foods are becoming increasingly popular complements to regular diets. A recurring ingredient is the essential cofactor vitamin B12 (B12). Microalgae are making their way into the dietary supplement and functional food market but do not produce B12, and their B12 content is very variable. In this study, the suitability of using the human B12-binding protein intrinsic factor (IF) to enrich bioavailable B12 using microalgae was tested. The IF protein was successfully expressed from the nuclear genome of the model microalga Chlamydomonas reinhardtii and the addition of an N-terminal ARS2 signal peptide resulted in efficient IF secretion to the medium. Co-abundance of B12 and the secreted IF suggests the algal produced IF protein is functional and B12-binding. Utilizing IF expression could be an efficient tool to generate B12-enriched microalgae in a controlled manner that is suitable for vegetarians and, potentially, more bioavailable for humans

Experimental and theoretical correlations between vanadium K-edge X-ray absorption and Kβ emission spectra

A series of vanadium compounds was studied by K-edge X-ray absorption (XAS) and K[Formula: see text] X-ray emission spectroscopies (XES). Qualitative trends within the datasets, as well as comparisons between the XAS and XES data, illustrate the information content of both methods. The complementary nature of the chemical insight highlights the success of this dual-technique approach in characterizing both the structural and electronic properties of vanadium sites. In particular, and in contrast to XAS or extended X-ray absorption fine structure (EXAFS), we demonstrate that valence-to-core XES is capable of differentiating between ligating atoms with the same identity but different bonding character. Finally, density functional theory (DFT) and time-dependent DFT calculations enable a more detailed, quantitative interpretation of the data. We also establish correction factors for the computational protocols through calibration to experiment. These hard X-ray methods can probe vanadium ions in any oxidation or spin state, and can readily be applied to sample environments ranging from solid-phase catalysts to biological samples in frozen solution. Thus, the combined XAS and XES approach, coupled with DFT calculations, provides a robust tool for the study of vanadium atoms in bioinorganic chemistry