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Here, we report work on developing an enzymatic process to improve the functionalities of industrial lignin. A kraft lignin sample prepared from marine pine was treated with the high-redox-potential laccase from the basidiomycete fungus Pycnoporus cinnabarinus at three different concentrations and pH conditions, and with and without the chemical mediator 1-hydroxybenzotriazole (HBT). Laccase activity was tested in the presence and absence of kraft lignin. The optimum pH of PciLac was initially 4.0 in the presence and absence of lignin, but at incubation times over 6 h, higher activities were found at pH 4.5 in the presence of lignin. Structural changes in lignin were investigated by Fourier-transform infrared spectroscopy (FTIR) with differential scanning calorimetry (DSC), and solvent-extractable fractions were analyzed using high-performance size-exclusion chromatography (HPSEC) and gas chromatography–mass spectrometry (GC–MS). The FTIR spectral data were analyzed with two successive multivariate series using principal component analysis (PCA) and ANOVA statistical analysis to identify the best conditions for the largest range of chemical modifications. DSC combined with modulated DSC (MDSC) revealed that the greatest effect on glass transition temperature (Tg) was obtained at 130 U g cm−1 and pH 4.5, with the laccase alone or combined with HBT. HPSEC data suggested that the laccase treatments led to concomitant phenomena of oligomerization and depolymerization, and GC–MS revealed that the reactivity of the extractable phenolic monomers depended on the conditions tested. This study demonstrates that P. cinnabarinus laccase can be used to modify marine pine kraft lignin, and that the set of analytical methods implemented here provides a valuable tool for screening enzymatic treatment conditions. [ABSTRACT FROM AUTHOR]

The wastewater from hospitals, pharmaceutical industries and more generally human and animal dejections leads to environmental releases of antibiotics that cause severe problems for all living organisms. The aim of this study was to investigate the capacity of three fungal strains to biotransform the fluoroquinolone levofloxacin. The degradation processes were analyzed in solid and liquid media. Among the three fungal strains tested, Coriolopsis gallica strain CLBE55 (BRFM 3473) showed the highest removal efficiency, with a 15% decrease in antibiogram zone of inhibition for Escherichia coli cultured in solid medium and 25% degradation of the antibiotic in liquid medium based on high-performance liquid chromatography (HPLC). Proteomic analysis suggested that laccases and dye-decolorizing peroxidases such as extracellular enzymes could be involved in levofloxacin degradation, with a putative major role for laccases. Degradation products were proposed based on mass spectrometry analysis, and annotation suggested that the main product of biotransformation of levofloxacin by Coriolopsis gallica is an N-oxidized derivative. [ABSTRACT FROM AUTHOR]

The textile industry generates huge volumes of colored wastewater that require multiple treatments to remove persistent toxic and carcinogenic dyes. Here we studied the decolorization of a recalcitrant azo dye, Reactive Black 5, using laccase-like active cell-free supernatant from Coriolopsis gallica. Decolorization was optimized in a 1 mL reaction mixture using the response surface methodology (RSM) to test the influence of five variables, i.e., laccase-like activity, dye concentration, redox mediator (HBT) concentration, pH, and temperature, on dye decolorization. Statistical tests were used to determine regression coefficients and the quality of the models used, as well as significant factors and/or factor interactions. Maximum decolorization was achieved at 120 min (82 ± 0.6%) with the optimized protocol, i.e., laccase-like activity at 0.5 U mL−1, dye at 25 mg L−1, HBT at 4.5 mM, pH at 4.2 and temperature at 55 °C. The model proved significant (ANOVA test with p < 0.001): coefficient of determination (R²) was 89.78%, adjusted coefficient of determination (R²A) was 87.85%, and root mean square error (RMSE) was 10.48%. The reaction conditions yielding maximum decolorization were tested in a larger volume of 500 mL reaction mixture. Under these conditions, the decolorization rate reached 77.6 ± 0.4%, which was in good agreement with the value found on the 1 mL scale. RB5 decolorization was further evaluated using the UV-visible spectra of the treated and untreated dyes. [ABSTRACT FROM AUTHOR]

112 ⟨ 111 ⟩ β twins are observed in a superelastic β Ti-24Nb-4Zr-8Sn (wt.%) single crystal after tensile test. A careful Schmid factor analysis shows that these twins are formed in the antitwinning sense regarding the classical 112 ⟨ 111 ⟩ β twinning system of bcc structures. These are then 112 ⟨ 111 ⟩ β antitwins. Moreover, a full stress-induced martensitic (SIM) transformation of β phase into α" martensite is evidenced from in situ synchrotron X-ray diffraction. This transformation is fully accomplished before the onset of plastic deformation and, in turn, the formation of twins. From crystallographic reconstruction, 112 ⟨ 111 ⟩ β antitwins are shown to be passively formed from the reversion, during the reverse SIM transformation when the stress is released, of 110 110 α ″ twins actually formed in α" martensite. The martensitic transformation occurring before twinning plays a key role in the activation of antitwinning systems by reducing both shear and shuffle magnitudes of twinning. Variant selection of stress-induced martensite and Schmid factor analysis show that the classical 112 ⟨ 111 ⟩ β twins can never be activated in β titanium alloys involving SIM transformation, while the non-classical 112 ⟨ 111 ⟩ β twinning system in the antitwinning sense is always favored. [ABSTRACT FROM AUTHOR]

Laccase-treated wood fibres were tested for small-scale wet-process manufacture of hardboards. Two laccases of distinct redox potentials, one from Pycnoporus cinnabarinus and one from Myceliophthora thermophila, were compared in terms of their effect on the physical–chemical properties of the treated fibres and hardboards. Wood fibres were produced from Norway spruce (Picea abies) by thermomechanical pulping. The thermomechanical pulp was treated with each laccase in parallel, in the presence or absence of the synthetic laccase mediator 1-hydroxybenzotriazole (HBT). High-performance size-exclusion chromatography revealed that the ethanolic extractable phenolic compounds in the fibres underwent oligomerization upon enzymatic treatment, and that the extent of oligomerization was dependent on the enzyme source and concentration and on the presence or absence of mediator. Lower lignin oligomerization levels led to higher (up to two-fold) fibre internal bonding, whereas higher lignin oligomerization levels led to higher fibre hydrophobisation. X-ray photoelectron spectroscopy revealed a significant change in surface lignin content. These results demonstrate pre-treatment of spruce fibres with laccase–mediator systems prior to hot processing can improve the mechanical resistance of hardboards while using lower amounts of enzyme. [ABSTRACT FROM AUTHOR]

The ability of Trichoderma reesei, a fungus widely used for the commercial production of hemicellulases and cellulases, to grow and modify technical soda lignin was investigated. By quantifying fungal genomic DNA, T. reesei showed growth and sporulation in solid and liquid cultures containing lignin alone. The analysis of released soluble lignin and residual insoluble lignin was indicative of enzymatic oxidative conversion of phenolic lignin side chains and the modification of lignin structure by cleaving the fi-O-4 linkages. The results also showed that polymerization reactions were taking place. A proteomic analysis conducted to investigate secreted proteins at days 3, 7, and 14 of growth revealed the presence of five auxiliary activity (AA) enzymes in the secretome: AA6, AA9, two AA3 enzymes), and the only copper radical oxidase encoded in the genome of T. reesei. This enzyme was heterologously produced and characterized, and its activity on lignin-derived molecules was investigated. Phylogenetic characterization demonstrated that this enzyme belonged to the AA5_1 family, which includes characterized glyoxal oxidases. However, the enzyme displayed overlapping physicochemical and catalytic properties across the AA5 family. The enzyme was remarkably stable at high pH and oxidized both, alcohols and aldehydes with preference to the alcohol group. It was also active on lignin-derived phenolic molecules as well as simple carbohydrates. HPSEC and LC-MS analyses on the reactions of the produced protein on lignin dimers (SS ββ, SS βO4 and GG β5) uncovered the polymerizing activity of this enzyme, which was accordingly named lignin copper oxidase (TrLOx). Polymers of up 10 units were formed by hydroxy group oxidation and radical formation. The activations of lignin molecules by TrLOx along with the co-secretion of this enzyme with reductases and FAD flavoproteins oxidoreductases during growth on lignin suggest a synergistic mechanism for lignin breakdown. [ABSTRACT FROM AUTHOR]

The functional diversity of the New Caledonian mangrove sediments was examined, observing the distribution of fungal dye-decolorizing peroxidases (DyPs), together with the complete biochemical characterization of the main DyP. Using a functional metabarcoding approach, the diversity of expressed genes encoding fungal DyPs was investigated in surface and deeper sediments, collected beneath either Avicennia marina or Rhizophora stylosa trees, during either the wet or the dry seasons. The highest DyP diversity was observed in surface sediments beneath the R. stylosa area during the wet season, and one particular operational functional unit (OFU1) was detected as the most abundant DyP isoform. This OFU was found in all sediment samples, representing 51–100% of the total DyP-encoding sequences in 70% of the samples. The complete cDNA sequence corresponding to this abundant DyP (OFU 1) was retrieved by gene capture, cloned, and heterologously expressed in Pichia pastoris. The recombinant enzyme, called DyP1, was purified and characterized, leading to the description of its physical–chemical properties, its ability to oxidize diverse phenolic substrates, and its potential to decolorize textile dyes; DyP1 was more active at low pH, though moderately stable over a wide pH range. The enzyme was very stable at temperatures up to 50 °C, retaining 60% activity after 180 min incubation. Its ability to decolorize industrial dyes was also tested on Reactive Blue 19, Acid Black, Disperse Blue 79, and Reactive Black 5. The effect of hydrogen peroxide and sea salt on DyP1 activity was studied and compared to what is reported for previously characterized enzymes from terrestrial and marine-derived fungi. [ABSTRACT FROM AUTHOR]

Background: Environmental pollution is one of the major problems that the world is facing today. Several approaches have been taken, from physical and chemical methods to biotechnological strategies (e.g. the use of oxidoreductases). Oxidative enzymes from microorganisms offer eco-friendly, cost–effective processes amenable to biotechnological applications, such as in industrial dye decolorization. The aim of this study was to screen marine-derived fungal strains isolated from three coastal areas in Tunisia to identify laccase-like activities, and to produce and characterize active cell-free supernatants of interest for dye decolorization. Results: Following the screening of 20 fungal strains isolated from the harbors of Sfax and Monastir (Tunisia), five strains were identified that displayed laccase-like activities. Molecular-based taxonomic approaches identified these strains as belonging to the species Trichoderma asperellum, Stemphylium lucomagnoense and Aspergillus nidulans. Among these five isolates, one T. asperellum strain (T. asperellum 1) gave the highest level of secreted oxidative activities, and so was chosen for further studies. Optimization of the growth medium for liquid cultures was first undertaken to improve the level of laccase-like activity in culture supernatants. Finally, the culture supernatant of T. asperellum 1 decolorized different synthetic dyes belonging to diverse dye families, in the presence or absence of 1-hydroxybenzotriazole (HBT) as a mediator. Conclusions: The optimal growth conditions to produce laccase-like active cell-free supernatants from T. asperellum 1 were 1.8 mM CuSO4 as an inducer, 1% NaCl to mimic a seawater environment and 3% sucrose as a carbon source. The culture supernatant of T. asperellum 1 effectively decolorized different synthetic dyes belonging to diverse chemical classes, and the presence of HBT as a mediator improved the decolorization process. [ABSTRACT FROM AUTHOR]

Microalgae are versatile sources of bioproducts, a solution for many environmental problems. However, and despite its importance, one of the main problems in large-scale cultures—the presence of contaminants—is rarely systematically approached. Contamination, or the presence of undesirable organisms in a culture, is deleterious for the culture and frequently leads to culture crashes. To avoid contamination, closed systems can be used; however, for very large-scale open systems, contamination is unavoidable and remediation procedures are necessary—ranging from physicochemical treatment to addition of biocidal substances. In all cases, early detection and culture monitoring are paramount. This article describes the biological contaminants, contamination mechanisms, and control systems used in open and closed cultures, discussing the latest advances and techniques in the area. It also discusses the complex interactions of algae with other microorganisms that can be expected in cultivation systems. [ABSTRACT FROM AUTHOR]

Main conclusion: Gibberellic acid is a plant growth hormone that promotes cell expansion and division. Studies have aimed at optimizing and reducing production costs, which could make its application economically viable for different cultivars.Gibberellins consist of a large family of plant growth hormones discovered in the 1930s, which are synthesized via the terpenes route from the geranylgeranyl diphosphate and feature a basic structure formed by an ent-gibberellane tetracyclic skeleton. Among them, only four have biological activity, including gibberellic acid (GA3), which acts as a natural plant growth regulator, especially for stem elongation, seed germination, and increased fruit size. It can be obtained from plants, fungi, and bacteria. There are also some reports about microalgae GA3 producers. Fungi, especially Gibberella fujikuroi, are preferred for GA3 production via submerged fermentation or solid-state fermentation. Many factors may affect its production, some of which are related to the control and scale-up of fermentation parameters. Different GA3 products are available on the market. They can be found in liquid or solid formulations containing only GA3 or a mixture of other biological active gibberellins, which can be applied on a wide variety of cultivars, including crops and fruits. However, the product’s cost still limits its large and continuous application. New low-cost and efficient GA3 production alternatives are surely welcome. This review deals with the latest scientific and technological advances on production, recovery, formulation, and applications of this important plant growth hormone. [ABSTRACT FROM AUTHOR]

Tissue engineering (TE) is the use of a combination of biological cells, engineering and materials methods, and of suitable biochemical and physicochemical factors, in order to improve or replace biological functions. It has brought the advent of entirely new classes of hierarchically organized, multiporous materials, consisting of both chemically and biologically produced parts. Here, we aim at contributing to the unsettled question of the mechanical functioning of bone tissue-engineering scaffolds with tissue-engineered bone—from a theoretical and applied mechanics viewpoint. Therefore, we build on recently developed microelasticity models for vertebrate bone and hydroxyapatite biomaterials, respectively. Tissue engineering scaffolds with tissue-engineered bone are micromechanically represented as tissue-engineered bone-coated macropores in a matrix built up by microporous hydroxyapatite polycrystals, based on an extension toward anisotropy, of Herve–Zaoui’s n-layered inclusion problem. The stiffness of macroporous hydroxyapatite-based TE scaffolds with newly ingrown bone is mainly governed by their porosities [vascular (macro) porosity defined through initial scaffold design and volume fraction of ingrown bone; and intercrystalline (micro) porosity between the hydroxyapatite crystals of the scaffold matrix material], while being less influenced by the type of bone growing inside the macropores. For a given degeneration kinetics of the scaffold, the microelastic models suggest apposition rates of bone needed to maintain the stiffness characteristics of the overall biomaterial-bone construct. This can be seen as a first step toward computer-aided engineering design of tissue-engineering scaffolds for large bone defect regeneration. [ABSTRACT FROM AUTHOR]

A 3.7-GHz 700-kW klystron in continuous-wave (CW) operation has been developed to upgrade the lower hybrid RF plasma heating power in a tokamak up to 10 MW. The klystron is equipped with a diode gun, a five-cavity RF structure, two BeO-disk RF windows, and a large-size X-ray-shielded hypervapotron collector. The output power is recombined in a four-port junction which we also developed. The tube is designed to deliver 620-kW CW RF power with a mismatched load (VSWR = 1.4) and 700-kW CW with a matched load. Several prototypes have been built with successive design improvements. The major improvement was to change one single RF output into two RF outputs. The most recently built prototype meets all design specifications at 73.1 kV and 20.7 A, with an efficiency of 47% on a matched load and 40% with a 1.4 : 1 VSWR load, worst case phase. The power losses dissipated in the body have been measured as low as 17 kW, which corresponds to the RF heating and implies low beam interception. The measured temperatures of the output cavity noses and collector wall have been kept below 130 °C and 200 °C, respectively, which results in large thermal margin. [ABSTRACT FROM AUTHOR]

Mangrove sediments from New Caledonia were screened for xylanase sequences. One enzyme was selected and characterized both biochemically and for its industrial potential. Using a specific cDNA amplification method coupled with a MiSeq sequencing approach, the diversity of expressed genes encoding GH11 xylanases was investigated beneath Avicenia marina and Rhizophora stylosa trees during the wet and dry seasons and at two different sediment depths. GH11 xylanase diversity varied more according to tree species and season, than with respect to depth. One complete cDNA was selected (OFU29) and expressed in Pichia pastoris. The corresponding enzyme (called Xyn11-29) was biochemically characterized, revealing an optimal activity at 40–50 °C and at a pH of 5.5. Xyn11-29 was stable for 48 h at 35 °C, with a half-life of 1 h at 40 °C and in the pH range of 5.5–6. Xyn11-29 exhibited a high hydrolysis capacity on destarched wheat bran, with 40% and 16% of xylose and arabinose released after 24 h hydrolysis. Its activity on wheat straw was lower, with a release of 2.8% and 6.9% of xylose and arabinose, respectively. As the protein was isolated from mangrove sediments, the effect of sea salt on its activity was studied and discussed. [ABSTRACT FROM AUTHOR]

Only a few studies have examined how marine-derived fungi and their enzymes adapt to salinity and plant biomass degradation. This work concerns the production and characterisation of an oxidative enzyme identified from the transcriptome of marine-derived fungus Stemphylium lucomagnoense. The laccase-encoding gene SlLac2 from S. lucomagnoense was cloned for heterologous expression in Aspergillus niger D15#26 for protein production in the extracellular medium of around 30 mg L−1. The extracellular recombinant enzyme SlLac2 was successfully produced and purified in three steps protocol: ultrafiltration, anion-exchange chromatography, and size exclusion chromatography, with a final recovery yield of 24%. SlLac2 was characterised by physicochemical properties, kinetic parameters, and ability to oxidise diverse phenolic substrates. We also studied its activity in the presence and absence of sea salt. The molecular mass of SlLac2 was about 75 kDa, consistent with that of most ascomycete fungal laccases. With syringaldazine as substrate, SlLac2 showed an optimal activity at pH 6 and retained nearly 100% of its activity when incubated at 50°C for 180 min. SlLac2 exhibited more than 50% of its activity with 5% wt/vol of sea salt. [ABSTRACT FROM AUTHOR]