In this report, we establish proof-of-principle demonstrating for the first time genetic engineering of a photoautotrophic microorganism for bioremediation of naturally occurring cyanotoxins. In model cyanobacterium Synechocystis sp. PCC 6803 we have heterologously expressed Sphingopyxis sp. USTB-05 microcystinase (MlrA) bearing a 23 amino acid N-terminus secretion peptide from native Synechocystis sp. PCC 6803 PilA (sll1694). The resultant whole cell biocatalyst displayed about 3 times higher activity against microcystin-LR compared to a native MlrA host (Sphingomonas sp. ACM 3962), normalized for optical density. In addition, MlrA activity was found to be almost entirely located in the cyanobacterial cytosolic fraction, despite the presence of the secretion tag, with crude cellular extracts showing MlrA activity comparable to extracts from MlrA expressing E. coli. Furthermore, despite approximately 9.4-fold higher initial MlrA activity of a whole cell E. coli biocatalyst, utilization of a photoautotrophic chassis resulted in prolonged stability of MlrA activity when cultured under semi-natural conditions (using lake water), with the heterologous MlrA biocatalytic activity of the E. coli culture disappearing after 4 days, while the cyanobacterial host displayed activity (3% of initial activity) after 9 days. In addition, the cyanobacterial cell density was maintained over the duration of this experiment while the cell density of the E. coli culture rapidly declined. Lastly, failure to establish a stable cyanobacterial isolate expressing native MlrA (without the N-terminus tag) via the strong cpcB560 promoter draws attention to the use of peptide tags to positively modulate expression of potentially toxic proteins.

In this study, a multidisciplinary approach investigated the enzymatic degradation of micropollutants in real, not modified, municipal wastewaters of a plant located in Italy. Stir Bar Sorptive Extraction combined to Gas Chromatography-Mass Spectrometric detection (SBSE-GC-MS) was applied to profile targeted pollutants in wastewaters collected after the primary sedimentation (W1) and the final effluent (W2). Fifteen compounds were detected at ng/L - μg/L, including pesticides, personal care products (PCPs) and drugs. The most abundant micropollutants were bis(2-ethylhexyl) phthalate, diethyl phthalate and ketoprofen. Laccases of Trametes pubescens MUT 2400 were very active against all the target micropollutants: except few cases, their concentration was reduced more than 60%. Chemical analysis and environmental risk do not always come together. To verify whether the treated wastewaters can represent a stressor for the aquatic ecosystem, toxicity was also evaluated. Raphidocelis subcapitata and Lepidium sativum tests showed a clear ecotoxicity reduction, even though they did not evenly respond. Two in vitro tests (E-screen test and MELN assay) were used to evaluate the estrogenic activity. Treatments already operating in the plant (e.g. activated sludge) partially reduced the estradiol equivalent concentration, and it was almost negligible after the laccases treatment. The results of this study suggest that laccases of T. pubescens are promising biocatalysts for the micropollutants transformation in wastewaters and surface waters.

To enhance the dye removal efficiency by natural enzyme, horseradish peroxidase (HRP) was immobilized onto amine-functionalized superparamagnetic iron oxide and used as a biocatalyst for the oxidative degradation of acid black-HC dye. The anchored enzyme was characterized by vibrating sample magnetometry, Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetry, scanning electron microscopy, Brunauer–Emmett–Teller and Barrett–Joyner–Halenda methods, nitrogen adsorption–desorption measurements, Zeta potential, energy dispersive X-ray spectroscopy, and transmission electron microscopy. The Michaelis constant values of free and immobilized HRP were determined to be 4.5 and 5 mM for hydrogen peroxide and 12.5 and 10 mM for guaiacol, respectively. Moreover, the maximum values of free and immobilized HRP were 2.4 and 2 U for H₂O₂, respectively, and 1.25 U for guaiacol. The immobilized enzyme was thermally stable up to 60°C, whereas the free peroxidase was stable only up to 40°C. In the catalytic experiment, the immobilized HRP exhibited superior catalytic activity compared with that of free HRP for the oxidative decolorization and removal of acid black-HC dye. The influence of experimental parameters such as the catalyst dosage, pH, H₂O₂ concentration, and temperature on the removal efficiency was investigated. The reaction followed second-order kinetics, and the thermodynamic activation parameters were determined.

As olive leaves constitute the main by-product of the olive oil industry with important environmental and economic impact, there is an increasing demand for its valorization. In the present work, we report the development and application of immobilized enzyme batch bioreactors for the chemo-enzymatic treatment of an aqueous Olea europaea leaf extract rich in oleuropein to produce an extract enriched in hydroxytyrosol and other oleuropein hydrolysis products. To this end, a robust biocatalyst was developed through the immobilization of β-glucosidase on chitosan-coated magnetic beads which exhibited high hydrolytic stability after 240 h of incubation at 37 °C. The biocatalyst was successfully used in both a rotating bed-reactor and a stir-tank reactor for the modification of the olive leaf extract leading to high conversion yields of oleuropein (exceeding 90%), while an up to 2.5 times enrichment in hydroxytyrosol was achieved. Over 20 phenolic compounds (from different classes of phytochemicals such as flavonoids, secoiridoids, and their derivatives) were identified, in the extract before and after its modification through various chromatographic and spectroscopic techniques. Finally, the biological activity of both extracts was evaluated. Compared to the non-modified extract, the modified one demonstrated 20% higher antioxidant activity, seven-fold higher antibacterial activity, and enhanced cytotoxicity against leiomyosarcoma cells.

In the present work, pharmaceutical micropollutant degradation by laccase immobilized on silica through an innovative process is proposed. The influence of different parameters on the immobilization conditions was evaluated by a 2³ full factorial design, and parameters leading to the highest activity were identified. Under these conditions, laccase activity reached 14 ± 2 U g⁻¹ of silica with a protein immobilization yield of 35%. The biocatalyst characterization did not show any change in pH and thermal stabilities but enhanced the long-term storage of laccases. Immobilized T. versicolor laccases were then tested to remove four pharmaceutical micropollutants (amoxicillin, ciprofloxacin, carbamazepine, and sulfamethoxazole) in the presence of redox mediators (syringaldehyde, p-coumaric acid, and ABTS). High removal yields (50–100% according to the pollutant) were obtained within 4 h of treatment due to the synergistic effect of laccase-mediator biotransformation and adsorption on the support. Overall, the pharmaceuticals’ removal efficiency was highly influenced by their physicochemical properties; however, the presence of redox mediators impacted not only the oxidation mechanism but also the interactions between the biocatalyst and micropollutants. Finally, the reusability of the biocatalyst was proved during 7 degradation cycles.

Microalgal biomass–based biofuels are a promising alternative to fossil fuels. Microalgal biofuels’ major obstacles are the water and carbon sources for their cultivation and biomass harvest from the liquid medium. To date, an economically viable process is not available for algal based biofuels. The circular bioeconomy is an attractive concept for reuse, reduce, and recycle resources. The recovery of nutrients from waste and effluents by microalgae could significantly impact the escalating demands of energy and nutraceutical source to the growing population. Wastewaters from different sources are enriched with nutrients and carbon, and these resources can be recovered and utilized for the circular bioeconomy approach. However, the utilization of wastewaters and waste seems to be an essential strategy for mass cultivation of microalgae to minimizing freshwater consumption, carbon, nutrients cost, nitrogen, phosphorus removal, and other pollutants loads from wastewater and generating sustainable biomass for value addition for either biofuels or other chemicals. Hence, the amalgamation of wastewater treatment with the mass cultivation of microalgae improved the conventional treatment process and environmental impacts. This review provides complete information on the latest progress and developments of microalgae as potential biocatalyst for the remediation of wastewaters and waste carbon to recover resources through biomass with metabolites for various industrial applications and large-scale cultivation in wastewaters, and future perspectives are discussed.

The rising global population and worldwide industrialization have led to unprecedented energy demand that is causing fast depletion of fossil reserves. This has led to search for alternative energy sources that are renewable and environment friendly. Use of lignocellulosic biomass for energy generation is considered a promising approach as it does not compete with food supply. However, the lignin component of the biomass acts as a natural barrier that prevents its efficient utilization. In order to remove the lignin and increase the amount of fermentable sugars, the lignocellulosic biomass is pretreated using physical and chemical methods which are costly and hazardous for environment. Moreover, during the traditional pretreatment process, numerous inhibitory compounds are generated that adversely affect the growth of fermentative microbes. Alternatively, biological methods that use microbes and their enzymes disrupt lignin polymers and increase the accessibility of the carbohydrates for the sugar generation. Microbial laccases have been considered as an efficient biocatalyst for delignification and detoxification offering a green initiative for energy generation process. The present review aims to bring together recent studies in bioenergy generation using laccase biocatalyst in the pretreatment processes. The work provides an overview of the sustainable and eco-friendly approach of biological delignification and detoxification through whole-cell and enzymatic methods, use of laccase-mediator system, and immobilized laccases for this purpose. It also summarizes the advantages, associated challenges, and potential prospects to overcome the limitations.

Environmentally friendly ionic solvents such as (a) ionic liquids (ILs) formulated with hydroxyl ammonium cations and various carboxylic acid anions and (b) choline chloride or ethyl ammonium chloride-based deep eutectic solvents (DES) were tested as media for hydrolytic and synthetic reactions catalysed by lipase-inorganic hybrid nanoflowers. The nature of ionic solvents used has a significant effect on the hydrolytic and synthetic activity of the immobilized lipase, as well as on its stability and reusability. In choline chloride-based DES, the activity and especially the operational stability of the biocatalyst are significantly increased compared to those observed in buffer, indicating the potential application of these solvents as green media for various biocatalytic processes of industrial interest.

This manuscript reports on a study of new biocatalysts for the biodegradation of pyrethroid pesticides, such as esfenvalerate. Experiments of esfenvalerate biodegradation by bacteria isolated from Brazilian savannah (Curtobacterium sp. CBMAI 1834, Bacillus sp. 2B, Lysinibacillus sp. CBMAI 1837, and Bacillus sp. 4T), sea (Kocuria sp. CBMAI 135, Kocuria sp. CBMAI 136, Kocuria marina CBMAI 141, and Kocuria sp. CBMAI 145), and a tropical peat usually known as “turfa” soil (Bacillus sp. P5CBNB, Kosakonia sp. CBMAI 1836, Bacillus sp. CBMAI 1833, and Kosakonia sp. CBMAI 1835) were performed. A biodegradation pathway was proposed for a better understanding of the environmental fate of the above mentioned insecticide. Esfenvalerate (S,S-fenvalerate) and its metabolites [3-phenoxybenzaldehyde (PBAld), 3-phenoxybenzoic acid (PBAc), 3-phenoxybenzyl alcohol, and 2-(4-chlorophenyl)-3-methylbutyric acid) (CLAc)] were quantitatively analyzed in triplicate experiments by a validated method. Initially, 100 mg L⁻¹ esfenvalerate (Sumidan 150SC) was added for each experiment. The residual esfenvalerate (104.7–41.6 mg L⁻¹) and formation of PBAc (0.1–8.1 mg L⁻¹), ClAc (1.5–11.0 mg L⁻¹), PBAlc (0.9 mg L⁻¹), and PBAld (completely biotransformed) were quantified. The 12 bacterial strains accelerated (with different efficiencies) the esfenvalerate degradation and increased the metabolites concentrations. A new and more complete biodegradation pathway based on HPLC-time of flight (ToF) and gas chromatography-mass spectrometry (GC-MS) analyses (in which thermal instability products were detected) was proposed. The detected metabolites are smaller and more polar compounds that may be carried by water and contaminate the environment.

Microbial fuel cells (MFC) have been foreseen as a sustainable renewable energy resource to meet future energy demand. In the past, several studies have been executed in both benchtop and pilot scale to produce electrical energy from wastewater. The key role players in this technology that leads to the operation are microbes, mainly bacteria. The dominant among them is termed as “exoelectrogens” that have the capability to produce and transport electron by utilizing waste source. The current review focuses on such electrogenic bacteria’s involvement for enhanced power generation of MFC. The pathway of electron transfer in their cell along and its conduction to the extracellular environment of the MFC system are critically discussed. The interaction of the microbes in various MFC operational conditions, including the role of substrate and solid electron acceptors, i.e., anode, external resistance, temperature, and pH, was also discussed in depth along with biotechnological advancement and future research perspective.