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

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

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

Reproductive timing and fecundity in the Neotropical lizard Enyalius perditus (Squamata: Leiosauridae)

Enyalius perditus is a semi-arboreal lizard species whose reproduction is poorly known. Here, we combine information obtained from preserved and live specimens to describe the reproductive timing (vitellogenesis, gravidity, and egg-laying) and fecundity (clutch size, egg size, and relative clutch mass) in females of E. perditus. Female reproduction is remarkably seasonal and occurs in the warmer and wetter periods of the year. Secondary vitellogenesis occurs from mid to late spring, whereas gravidity and egg-laying occur in early summer. Mating appears to be synchronized with secondary vitellogenesis, indicating an associated reproductive cycle. We suggest that E. perditus females produce only a single clutch per reproductive season. Clutch size ranged from three to 11 eggs and was positively correlated with female body size. Finally, the relative clutch mass was high, a recurrent feature to “sit-and-wait” foragers

Exploitation of Microalgae Biomass Under an Integrated Biorefinery Approach

As known, microalgae are an appealing source of chemicals and high-value compounds which find application in nutraceuticals, cosmetics and pharmaceutics. Fatty acids (FA), in particular, have drawn attention to the possibility of employing them as a source of biodiesel alternatively to fossil fuels. In addition, several lipid derivatives have been found in microalgae and may be employed in several biotechnological applications. Hydroxy fatty acids can be substrates for several industrial applications thanks to their functionalization, which increases their reactivity and, for this reason, can be used as functional building blocks to produce a multitude of bio-based materials. Recently, a promising method for the chemical modification of unsaturated-FAs (U-FA) has appeared. In fact, U-FA may be modified by members of the hydratase enzyme family to produce saturated and unsaturated hydroxy fatty acids with high stereo- and regio-selectivity. These enzymes are able to introduce a water molecule to the double bond present in the free fatty acids (FFA) Oleic Acid (OA), Linoleic Acid (LA), producing 10-hydroxy fatty acids (10-hydroxy-FAs). Furthermore, the carbohydrate component of the microalgal biomass may be converted into furfuryl compounds and, in particular in 5-hydroxyl methyl furfural (5-HMF). This is one of the chemical bio-compound different from petroleum-derived ones with the highest added value and may be obtained through lignocellulosic biomasses or hexoses sugars through acid catalysis. It is defined platform molecule because it is the precursor of several compounds for the chemical industry. In this work, we aimed to optimize a circular bioprocess by performing, starting from the same biomass, two different processes: the biotransformation of microalgal FFAs through the employment of a genetically modified E. coli on one side, and the conversion of the remaining biomass in furfuryl products. The first process allowed the production of very interesting lipid derivatives with biotechnological applications, including 10 hydroxy-stearic acid and 10-hydroxy-octadecenoic acid. The second process was obtained through heterogeneous catalysis based on niobium phosphate. This procedure represents a high-innovative application of microalgal biomass and allows the simultaneous exploitation of FAs and carbohydrates. This may result in an increase in the commercial value of microalgal biomass

Comparative electronic structures of nitrogenase FeMoco and FeVco

An investigation of the active site cofactors of the molybdenum and vanadium nitrogenases (FeMoco and FeVco) was performed using high-resolution X-ray spectroscopy. Synthetic heterometallic iron–sulfur cluster models and density functional theory calculations complement the study of the MoFe and VFe holoproteins using both non-resonant and resonant X-ray emission spectroscopy. Spectroscopic data show the presence of direct iron–heterometal bonds, which are found to be weaker in FeVco. Furthermore, the interstitial carbide is found to perturb the electronic structures of the cofactors through highly covalent Fe–C bonding. The implications of these conclusions are discussed in light of the differential reactivity of the molybdenum and vanadium nitrogenases towards various substrates. Possible functional roles for both the heterometal and the interstitial carbide are detailed.This work was supported by the European Research Council (ERC) under the European Union’s Seventh Framework Programme (FP/2007–2013) ERC Grant Agreement number 615414 (S. D.) and the ERC N-ABLE project (O. E.). Funding was also provided by the Deutsche Forschungsgemeinschaft grants EI-520/7 and RTG1976 (O. E.), the NIH (R01-GM45881 to J. A. K.), and by the Max-Planck–Gesellschaft (S. D., R. B., J. K. K., and F. A. L.). J. A. R. was funded by a graduate study scholarship from the German Academic Exchange Service (DAAD). R. B. acknowledges support from the Icelandic Research Fund, Grant No. 141218051 and the University of Iceland Research Fund. Matthias Gschell and Florian Schneider are thanked for preparing the extracted FeMoco, and Tabea Hamann is thanked for providing samples of the molybdenum cubane. Stefan Hugenbruch, Benjamin Van Kuiken, Rebeca Gómez Castillo, and Anselm Hahn are thanked for assistance with data collection. The ESRF and CHESS are also acknowledged for providing beamtime, and Sara Lafuerza and Pieter Glatzel at beamline ID-26 (ESRF) and Kenneth D. Finkelstein at beamline C-1 (CHESS) are gratefully acknowledged for technical assistance with measurements. CHESS is supported by the National Science Foundation and the National Institutes of Health/National Institute of General Medical Sciences under NSF award DMR-133220. Open Access funding provided by the Max Planck Society.Peer Reviewe