UV-Visible spectrum of purified recombinant <i>P</i>. <i>acidilactici</i> CECT 5930 laccase.

<p>Protein was dissolved in 50 mM sodium phosphate, pH 6.5 buffer at a concentration of 14 mg/ml. Only the range from 300 to 800 nm is shown.</p

Amino acid sequence of <i>P</i>. <i>acidilactici</i> CECT 5930 protein homologue of D2EK17.

<p>Shaded and underlined are the motifs corresponding to the four copper ligands which are strictly conserved in MCOs (conserved sequence of these motifs are, successively: HXHG, HXH, HXXHXH and HCHXXXHXXXXM/L/F).</p

Coomassie–blue stained gel after SDS-8% PAGE of different fractions during the purification procedure of recombinant <i>P</i>. <i>acidilactici</i> CECT 5930 laccase.

<p>Lane 1, whole-cell extract from non-induced cells; lane 2, whole-cell extract from post-induction cells; lane 3, protein markers, with the molecular weights indicated on the right (kDa); lanes 4–6, successive fractions of the elution from the metal-chelating chromatography on Ni²⁺-NTA-agarose; lane 7, crude extract corresponding to applied sample on the chromatography column. Arrow marks recombinant protein.</p

pH effect on activity of the recombinant <i>P</i>. <i>acidilactici</i> enzyme toward ABTS (<i>solid line</i>); and tyramine (<i>dashed line</i>) as substrates.

<p>Citrate-phosphate buffer (pH range 2.5–8.0) and carbonate-NaOH buffer (pH range 9.0–10.5) were used at concentration of 50 mM. Enzyme activity is plotted as percentage relative to the maximum value for each substrate (% relative activity). With tyramine as a substrate, the 100% of relative activity means a 43% of tyramine degradation in the reaction mixture, determined by reverse-phase LC-FLD, under the assay conditions described in Material and Methods section. The values are means ± standard deviations of triplicate assays.</p

Ability of Kocuria varians LTH 1540 To Degrade Putrescine: Identification and Characterization of a Novel Amine Oxidase

This work describes the identification and characterization of an amine oxidase from Kocuria varians LTH 1540 (syn. Micrococcus varians) primarily acting on putrescine. Data from MALDI-TOF MS/MS and the identification of Δ¹-pyrroline as degradation product from putrescine indicate that the enzyme is a flavin-dependent putrescine oxidase (PuO). Properties of partially purified enzyme have been determined. The enzyme oxidizes diamines, putrescine and cadaverine, and, to a lesser extent, polyamines, such as spermidine, but not monoamines. The kinetic constants (<i>K</i>_m and <i>V</i>_max) for the two major substrates were 94 ± 10 μM and 2.3 ± 0.1 μmol/min·mg for putrescine and 75 ± 5 μM and 0.15 ± 0.02 μmol/min·mg for cadaverine. Optimal temperature and pH were 45 °C and 8.5, respectively. Enzyme was stable until 50 °C. <i>K. varians</i> PuO is sensitive to human flavin-dependent amine oxidase inhibitors and carboxyl-modifying compounds. The new enzyme has been isolated from a bacterial starter used in the manufacture of fermented meat. One of the problems of fermented foods or beverages is the presence of toxic biogenic amines produced by bacteria. The importance of this works lies in the description of a new enzyme able to degrade two of the most abundant biogenic amines (putrescine and cadaverine), the use of which could be envisaged to diminish biogenic amines content in foods in the future

Effect of potential inhibitors of <i>P</i>. <i>acidilactici</i> laccase.

<p>Liquid assays using ABTS as a substrate were carried out in the presence of 100 μM of each of the indicated compounds. Control was an enzyme assay in the absence of inhibitor and is taken as 100% of activity. Results express the remaining activity as percentage relative to the control. Values are means ± standard deviations of triplicate assays.</p

Function of the ABTS as redox mediator on tyramine oxidation by recombinant <i>P</i>. <i>acidilactici</i> laccase.

<p>Tyramine concentration was quantified by reverse-phase LC-FLD after incubation with recombinant laccase and different concentrations of ABTS for 24 at 28°C. The percentages of tyramine degradation were calculated regarding a control without enzyme. Activity is expressed as percentage relative to the maximum, where a 100% means a 75.2% of tyramine degradation under these assay conditions. The values are means ± standard deviations for triplicate assays.</p

Seasonal variability of lotic macroinvertebrate communities at the habitat scale demonstrates the value of discriminating fine sediment fractions in ecological assessments

Despite lotic systems demonstrating high levels of seasonal and spatial variability, most research and biomonitoring practices do not consider seasonality when interpreting results and are typically focused at the meso‐scale (combined pool/riffle samples) rather than considering habitat patch dynamics. We therefore sought to determine if the sampling season (spring, summer and autumn) influenced observed macroinvertebrate biodiversity, structure and function at the habitat unit scale (determined by substrate composition), and if this in turn influenced the assessment of fine sediment (sand and silt) pressures. We found that biodiversity supported at the habitat level was not seasonally consistent with the contribution of nestedness and turnover in structuring communities varying seasonally. Habitat differences in community composition were evident for taxonomic communities regardless of the season but were not seasonally consistent for functional communities, and, notably, season explained a greater amount of variance in functional community composition than the habitat unit. Macroinvertebrate biodiversity supported by silt habitats demonstrated strong seasonal differences and communities were functionally comparable to sand habitats in spring and to gravel habitats in autumn. Sand communities were impoverished compared to other habitats regardless of the season. Silt habitats demonstrated a strong increase in Ephemeroptera, Plecoptera and Trichoptera (EPT) taxa and functional richness from spring into autumn, while vegetation habitats displayed a peak in EPT abundance in summer. Only silt and sand habitats demonstrated temporal variability in functional evenness suggesting that these habitats are different in terms of their resource partitioning and productivity over time compared to other habitats. Gravel and vegetation habitats appeared to be more stable over time with functional richness and evenness remaining consistent. To accurately evaluate the influence of fine sediment on lotic ecosystems, it is imperative that routine biomonitoring and scientific research discriminate between sand and silt fractions, given they support different biodiversity, particularly during summer and autumn months.</p

Naive realism and the cognitive penetrability of perception

Perceptual experience has representational content. My argument for this claim is an inference to the best explanation. The explanandum is cognitive penetration. In cognitive penetration, perceptual experiences are either causally influenced, or else are partially constituted, by mental states that are representational, including: mental imagery, beliefs, concepts and memories. If perceptual experiences have representational content, then there is a background condition for cognitive penetration that renders the phenomenon prima facie intelligible. Naïve realist or purely relational accounts of perception leave cognitive penetration less well-explained, even when formulated with so-called â��standpointsâ�� or â��third relata.â�

Cyclic voltammetry curves of PANI obtained with 7D5L and SDBS (a) or AOT (b) as templates (after isolation with ethanol) compared with commercial emeraldine salt (c).

<p>Cyclic voltammetry curves of PANI obtained with 7D5L and SDBS (a) or AOT (b) as templates (after isolation with ethanol) compared with commercial emeraldine salt (c).</p