Optimization of Lipase Production by Burkholderia sp. Using Response Surface Methodology

Response surface methodology (RSM) was employed to optimize the extracellular lipase production by Burkholderia sp. HL-10. Preliminary tests showed that olive oil, tryptone and Tween-80 exhibited significant effects on the lipase production. The optimum concentrations of these three components were determined using a faced-centered central composite design (FCCCD). The analysis of variance revealed that the established model was significant (p < 0.01). The optimized medium containing 0.65% olive oil (v/v), 2.42% tryptone (w/v) and 0.15% Tween-80 (v/v) resulted in a maximum activity of 122.3 U/mL, about three fold higher than that in basal medium. Approximately 99% of validity of the predicted value was achieved.     

Modification of Low Molecular Weight Polysaccharides from Tremella Fuciformis and Their Antioxidant Activity in Vitro

In this study, sulfated low molecular-weight Tremella fuciformis polysaccharides (SLTP) with different sulfate contents were synthesized and their antioxidant activities, including superoxide anion radical, 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical and hydroxyl radical scavenging activities were investigated. The results indicated that, compared to natural Tremella fuciformis polysaccharide (TP) and low molecular weight Tremella fuciformis polysaccharide (LTP), sulfated LTP (SLTP) exhibited stronger scavenging activity towards superoxide anion, DPPH and hydroxyl radicals. In all the cases the effect was found to be dose dependent. The scavenging activity of SLTP was found to be in parallel with the degree of sulfation of SLTP.     

Biosurfactant-and-Bioemulsifier Produced by a Promising Cunninghamella echinulata Isolated from Caatinga Soil in the Northeast of Brazil

A Mucoralean fungus was isolated from Caatinga soil of Pernambuco, Northeast of Brazil, and was identified as Cunninghamella echinulata by morphological, physiological, and biochemical tests. This strain was evaluated for biosurfactant/bioemulsifier production using soybean oil waste (SOW) and corn steep liquor (CSL) as substrates, added to basic saline solution, by measuring surface tension and emulsifier index and activity. The best results showed the surface water tension was reduced from 72 to 36 mN/m, and an emulsification index (E₂₄) of 80% was obtained using engine oil and burnt engine oil, respectively. A new molecule of biosurfactant showed an anionic charge and a polymeric chemical composition consisting of lipids (40.0% w/w), carbohydrates (35.2% w/w) and protein (20.3% w/w). In addition, the biosurfactant solution (1%) demonstrated its ability for an oil displacement area (ODA) of 37.36 cm², which is quite similar to that for Triton X-100 (38.46 cm²). The stability of the reduction in the surface water tension as well as of the emulsifier index proved to be stable over a wide range of temperatures, in pH, and in salt concentration (4%–6% w/v). The biosurfactant showed an ability to reduce and increase the viscosity of hydrophobic substrates and their molecules, suggesting that it is a suitable candidate for mediated enhanced oil recovery. At the same time, these studies indicate that renewable, relatively inexpensive and easily available resources can be used for important biotechnological processes.     

Electrospinning of chitosan/PVA nanofibrous membrane at ultralow solvent concentration

In this work, chitin flakes were deacetylated with 50% (w/v) sodium hydroxide under nitrogen atmosphere at 120 °C for 80 min to obtain chitosan. The chitosan produced was characterized for degree of deacetylation (DD) and molecular weight. Chitosan with the DD of 78–80% was reproducibly obtained. Molecular weight showed an inverse relationship with concentration of NaOH. Chitosan nanofibrous membrane was prepared via the electrospinning of chitosan/polyvinyl alcohol (CH/PVA) aqueous solutions with varying blend compositions. The characteristics of CH/PVA nanofibrous membranes were studied as a function of viscosity of solution and applied voltage. A uniform nanofibrous membrane of average fibre diameter of 80–300 nm was obtained with blend of 2% (w/v) chitosan solution in 1% (v/v) acetic acid and 5% (w/v) PVA in distilled water in the electric field of 20–25 kV with varying proportion of CH/PVA. With the CH/PVA mass ratios; 40/60 to 10/90, electrospinning of nanofibres could be done. The electrospun nanofibrous membrane was analyzed by scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and Thermo gravimetric analysis (TGA). SEM images showed that the morphology and diameter of the nanofibres were mainly affected by the weight ratio of CH/PVA. XRD and FTIR confirmed the strong intermolecular hydrogen bonding between the molecules of Chitosan and PVA.     

Production of (R)-3-Quinuclidinol by E. coli Biocatalysts Possessing NADH-Dependent 3-Quinuclidinone Reductase (QNR or bacC) from Microbacterium luteolum and Leifsonia Alcohol Dehydrogenase (LSADH)

We found two NADH-dependent reductases (QNR and bacC) in Microbacterium luteolum JCM 9174 (M. luteolum JCM 9174) that can reduce 3-quinuclidinone to optically pure (R)-(−)-3-quinuclidinol. Alcohol dehydrogenase from Leifsonia sp. (LSADH) was combined with these reductases to regenerate NAD⁺ to NADH in situ in the presence of 2-propanol as a hydrogen donor. The reductase and LSADH genes were efficiently expressed in E. coli cells. A number of constructed E. coli biocatalysts (intact or immobilized) were applied to the resting cell reaction and optimized. Under the optimized conditions, (R)-(−)-3-quinuclidinol was synthesized from 3-quinuclidinone (15% w/v, 939 mM) giving a conversion yield of 100% for immobilized QNR. The optical purity of the (R)-(−)-3-quinuclidinol produced by the enzymatic reactions was >99.9%. Thus, E. coli biocatalysis should be useful for the practical production of the pharmaceutically important intermediate, (R)-(−)-3-quinuclidinol.     

Does Short-Term Dietary Omega-3 Fatty Acid Supplementation Influence Brain Hippocampus Gene Expression of Zinc Transporter-3?

Dietary omega-3 fatty acids have been recognized to improve brain cognitive function. Deficiency leads to dysfunctional zinc metabolism associated with learning and memory impairment. The objective of this study is to explore the effect of short-term dietary omega-3 fatty acids on hippocampus gene expression at the molecular level in relation to spatial recognition memory in mice. A total of 24 male BALB/c mice were randomly divided into four groups and fed a standard pellet as a control group (CTL, n = 6), standard pellet added with 10% (w/w) fish oil (FO, n = 6), 10% (w/w) soybean oil (SO, n = 6) and 10% (w/w) butter (BT, n = 6). After 3 weeks on the treatment diets, spatial-recognition memory was tested on a Y-maze. The hippocampus gene expression was determined using a real-time PCR. The results showed that 3 weeks of dietary omega-3 fatty acid supplementation improved cognitive performance along with the up-regulation of α-synuclein, calmodulin and transthyretin genes expression. In addition, dietary omega-3 fatty acid deficiency increased the level of ZnT3 gene and subsequently reduced cognitive performance in mice. These results indicate that the increased the ZnT3 levels caused by the deficiency of omega-3 fatty acids produced an abnormal zinc metabolism that in turn impaired the brain cognitive performance in mice.     

Floating Ricobendazole Delivery Systems: A 3D Printing Method by Co-Extrusion of Sodium Alginate and Calcium Chloride

At present, the use of benzimidazole drugs in veterinary medicine is strongly limited by both pharmacokinetics and formulative issues. In this research, the possibility of applying an innovative semi-solid extrusion 3D printing process in a co-axial configuration was speculated, with the aim of producing a new gastro-retentive dosage form loaded with ricobendazole. To obtain the drug delivery system (DDS), the ionotropic gelation of alginate in combination with a divalent cation during the extrusion was exploited. Two feeds were optimized in accordance with the printing requirements and the drug chemical properties: the crosslinking ink, i.e., a water ethanol mixture containing CaCl₂ at two different ratios 0.05 M and 0.1 M, hydroxyethyl cellulose 2% w/v, Tween 85 0.1% v/v and Ricobendazole 5% w/v; and alginate ink, i.e., a sodium alginate solution at 6% w/v. The characterization of the dried DDS obtained from the extrusion of gels containing different amounts of calcium chloride showed a limited effect on the ink extrudability of the crosslinking agent, which on the contrary strongly influenced the final properties of the DDS, with a difference in the polymeric matrix toughness and resulting effects on floating time and drug release.     

In Vitro Antioxidant Activities of Sulfated Derivatives of Polysaccharides Extracted from Auricularia auricular

In this research, two types of sulfated polysaccharide derivatives were successfully synthesized. Their antioxidant activities were investigated by employing various established in vitro systems. In addition, the degree of sulfation was evaluated using ion-chromatography and IR spectra. The results verify that, when employing scavenging superoxide radical tests, both the sulfation of acid Auricularia auricular polysaccharides (SAAAP) and the sulfation of neutral Auricularia auricular polysaccharides (SNAAP) derivatives possessed considerable antioxidant activity and had a more powerful antioxidant competence than that of the native non-sulfated polysaccharides (AAAP and NAAP). On the other hand, AAAP and NAAP exhibited stronger activity on scavenging both the hydroxyl radical and lipid peroxidation. Available data obtained with in vitro measurements indicates that the sulfated groups of AAAP and NAAP played an important role on antioxidant activity. In sum, the research demonstrates that the antioxidant activity of sulfated polysaccharide derivatives in vitro has a potential significance for seeking new natural antioxidant protective agents.     

Chemical Composition and Antioxidant Activities of Three Polysaccharide Fractions from Pine Cones

The traditional method of gas chromatography-mass spectrometry for monosaccharide component analysis with pretreatment of acetylation is described with slight modifications and verified in detail in this paper. It was then successfully applied to the quantitative analysis of component monosaccharides in polysaccharides extracted from the pine cones. The results demonstrated that the three pine cone polysaccharides all consisted of ribose, rhamnose, arabinose, xylose, mannose, glucose and galactose in different molar ratios. According to the recovery experiment, the described method was proved accurate and practical for the analysis of pine cone polysaccharides, meeting the need in the field of chemical analysis of Pinus plants. Furthermore; the chemical characteristics, such as neutral sugar, uronic acids, amino acids, molecular weights, and antioxidant activities of the polysaccharides were investigated by chemical and instrumental methods. The results showed that the chemical compositions of the polysaccharides differed from each other, especially in the content of neutral sugar and uronic acid. In the antioxidant assays, the polysaccharide fractions exhibited effective scavenging activities on ABTS radical and hydroxyl radical, with their antioxidant capabilities decreasing in the order of PKP > PAP > PSP. Therefore, although the polysaccharide fractions had little effect on superoxide radical scavenging, they still have potential to be developed as natural antioxidant agents in functional foods or medicine.     

Effect of Glucans from Caripia montagnei Mushroom on TNBS-Induced Colitis

In this study, we evaluated the effect of different doses of polysaccharides extracted from Caripia montagnei mushroom at different intervals of treatment on colonic injury in the model of colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS). The FT-IR analysis and NMR showed that the polysaccharides from this species of mushroom are composed of α- and β-glucans. The colonic damage was evaluated by macroscopic, histological, biochemical and immunologic analyses. The results showed the reduction of colonic lesions in all groups treated with the glucans. Such glucans significantly reduced the levels of IL-6 (50 and 75 mg/kg, p < 0.05), a major inflammatory cytokine. Biochemical analyses showed that the glucans from C. montagnei acted on reducing levels of alkaline phosphatase (75 mg/kg, p < 0.01) and myeloperoxidase (p < 0.001), a result confirmed by the reduction of cellular infiltration observed microscopically. The increase of catalase activity possibly indicates a protective effect of these glucans on colonic tissue, confirming their anti-inflammatory potential.     

Bacterial Molecular Signals in the Sinorhizobium fredii-Soybean Symbiosis

Sinorhizobium (Ensifer) fredii (S. fredii) is a rhizobial species exhibiting a remarkably broad nodulation host-range. Thus, S. fredii is able to effectively nodulate dozens of different legumes, including plants forming determinate nodules, such as the important crops soybean and cowpea, and plants forming indeterminate nodules, such as Glycyrrhiza uralensis and pigeon-pea. This capacity of adaptation to different symbioses makes the study of the molecular signals produced by S. fredii strains of increasing interest since it allows the analysis of their symbiotic role in different types of nodule. In this review, we analyze in depth different S. fredii molecules that act as signals in symbiosis, including nodulation factors, different surface polysaccharides (exopolysaccharides, lipopolysaccharides, cyclic glucans, and K-antigen capsular polysaccharides), and effectors delivered to the interior of the host cells through a symbiotic type 3 secretion system.     

Purification and Characterization of Aporphine Alkaloids from Leaves of Nelumbo nucifera Gaertn and Their Effects on Glucose Consumption in 3T3-L1 Adipocytes

Aporphine alkaloids from the leaves of Nelumbo nucifera Gaertn are substances of great interest because of their important pharmacological activities, particularly anti-diabetic, anti-obesity, anti-hyperlipidemic, anti-oxidant, and anti-HIV’s activities. In order to produce large amounts of pure alkaloid for research purposes, a novel method using high-speed counter-current chromatography (HSCCC) was developed. Without any initial cleanup steps, four main aporphine alkaloids, including 2-hydroxy-1-methoxyaporphine, pronuciferine, nuciferine and roemerine were successfully purified from the crude extract by HSCCC in one step. The separation was performed with a simple two-phase solvent system composed of n-hexane-ethyl acetate-methanol-acetonitrile-water (5:3:3:2.5:5, v/v/v/v/v). In each operation, 100 mg crude extracts was separated and yielded 6.3 mg of 2-hydroxy-1-methoxyaporphine (95.1% purity), 1.1 mg of pronuciferine (96.8% purity), 8.5 mg of nuciferine (98.9% purity), and 2.7 mg of roemerine (97.4%) respectively. The chemical structure of four aporphine alkaloids are identified by means of electrospray ionization MS (ESI-MS) and nuclear magnetic resonance (NMR) analysis. Moreover, the effects of four separated aporphine alkaloids on insulin-stimulated glucose consumption were examined in 3T3-L1 adipocytes. The results showed that 2-hydroxy-1-methoxyaporphine and pronuciferine increased the glucose consumption significantly as rosiglitazone did.     

Effect of Repetitive Lysine-Tryptophan Motifs on the Eukaryotic Membrane

In a previous study, we synthesized a series of peptides containing simple sequence repeats, (KW)_n–NH₂ (n = 2,3,4 and 5) and determined their antimicrobial and hemolytic activities, as well as their mechanism of antimicrobial action. However, (KW)₅ showed undesirable cytotoxicity against RBC cells. In order to identify the mechanisms behind the hemolytic and cytotoxic activities of (KW)₅, we measured the ability of these peptides to induce aggregation of liposomes. In addition, their binding and permeation activities were assessed by Trp fluorescence, calcein leakage and circular dichrorism using artificial phospholipids that mimic eukaryotic liposomes, including phosphatidylcholine (PC), PC/sphingomyelin (SM) (2:1, w/w) and PC/cholesterol (CH) (2:1, w/w). Experiments confirmed that only (KW)₅ induced aggregation of all liposomes; it formed much larger aggregates with PC:CH (2:1, w/w) than with PC or PC:SM (2:1, w/w). Longer peptide (KW)₅, but not (KW)₃ or (KW)₄, strongly bound and partially inserted into PC:CH compared to PC or PC:SM (2:1, w/w). Calcein release experiments showed that (KW)₅ induced calcein leakage from the eukaryotic membrane. Greater calcein leakage was induced by (KW)₅ from PC:CH than from PC:SM (2:1, w/w) or PC, whereas (KW)₄ did not induce calcein leakage from any of the liposomes. Circular dichroism measurements indicated that (KW)₅ showed higher conformational transition compared to (KW)₄ due to peptide-liposome interactions. Taken together, our results suggest that (KW)₅ reasonably mediates the aggregation and permeabilization of eukaryotic membranes, which could in turn explain why (KW)₅ displays efficient hemolytic activity.     

Tubular multi‐bilayer polysaccharide biofilms on ultra‐thin cellulose fibers

Multiple bilayered polysaccharide biofilms have been assembled by electrostatic layer‐by‐layer (LBL), alternating deposition of cationic chitosan (CS, M_v = 405 kDa) and anionic dextran sulfate (DXS, M_w = 500 kDa) onto ultra‐fine cellulose (CELL) and partially hydrolyzed cellulose acetate fibers with diameters ranging from 350 to 410 nm. While the surfaces of partially hydrolyzed (degrees of substitution of 1.14 or 0.2) and CELL fibers were equally hydrophilic, higher surface charges on the more hydrolyzed fibers afford thicker bilayers. The elestrostatic interactions between CS and DXS were enhanced by the presence of NaCl in the dipping and rinsing solutions to allow uniform deposition of sequential polysaccharide bilayers. At 0.25M NaCl, each CS/DXS bilayer averaged 6.4 to 9.0 nm thick with the total thickness of the five bilayer (CS/DXS)₅ varied from 64 to 77 nm. The CS/DXS bilayers exhibited much reduced BET surface area and pore volume indicating that these polysaccharides were much more densely packed on the fully hydrolyzed CELL fibers. The findings proofed the concept that long chain polysaccharide electrolytes can be self‐assembled as nanometer scale tubular bilayers on ultra‐fine cellulose fibers to afford wholly polysaccharidic fibrous architecture. The electrolytic nature, chemical reactivity, and structural versatility of these ultra‐high specific surface polysaccharides are advantageous and can be further tuned to serve biological functions and for biomedical applications. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Production of Structured Phosphatidylcholine with High Content of DHA/EPA by Immobilized Phospholipase A1-Catalyzed Transesterification

This paper presents the synthesis of structured phosphatidylcholine (PC) enriched with docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) by transesterification of DHA/EPA-rich ethyl esters with PC using immobilized phospholipsase A₁ (PLA₁) in solvent-free medium. Firstly, liquid PLA₁ was immobilized on resin D380, and it was found that a pH of 5 and a support/PLA₁ ratio (w/v) of 1:3 were the best conditions for the adsorption. Secondly, the immobilized PLA₁ was used to catalyze transesterification of PC and DHA/EPA-rich ethyl esters. The maximal incorporation of DHA and EPA achieved was 30.7% for 24 h of reaction at 55 °C using a substrate mass ratio (PC/ethyl esters) of 1:6, an immobilized PLA₁ loading of 15% and water dosage of 1.25%. Then the reaction mixture was analyzed by ³¹P nuclear magnetic resonance (NMR). The composition of reaction product included 16.5% PC, 26.3% 2-diacyl-sn-glycero-3-lysophosphatidylcholine (1-LPC), 31.4% 1-diacyl-sn-glycero-3-lysophosphatidylcholine (2-LPC), and 25.8% sn-glycerol-3-phosphatidylcholine (GPC).     

Fracture behaviour of polyethylene naphthalate (PEN)

Fracture toughness of a semi-crystalline polyethylene naphthalate (PEN) film of thicknesses 0.050, 0.075 and 0.125 mm was measured as a function of temperature and loading rate using both double edge notched tension (DENT) and single edge notched tension (SENT) specimens. The specific essential work of fracture (EWF) and the multi-specimen J-integral methods were used to evaluate fracture toughness. The variation of the specific total work of fracture (w_f) with ligament length (L) was linear for ligament lengths between 5 and 15 mm. Within this range, w_f versus L was independent of thickness at all temperatures but was dependent on both temperature and loading rate. The specific EWF (w_e) was found to be independent of thickness and loading rate but showed three regions of varying temperature dependence. Between 23 and 80°C (region I) w_e was essentially independent of temperature but increased with temperature between 80 and 120°C (region II) and decreased with temperature thereafter (region III). At glass transition temperature (i.e. 120°C), w_e reached a maximum value of 75 kJ/m². The specific non-EWF (βw_p) increased with both loading rate and temperature. The greatest change in βw_p value with respect to temperature was obtained in region II.The plot of J-integral versus crack extension (Δa) was independent of thickness but was dependent upon temperature. w_e was found to be equivalent to both J_0.2 and J₀.     

Effects of Ration Levels on Growth and Reproduction from Larvae to First-Time Spawning in the Female Gambusia affinis

Somatic growth and reproduction were examined in individual laboratory-grown female Gambusia affinis fed with high (H), medium (M) and low (L) ration levels from birth to the first-time spawning. Results showed that the body length and weight, condition factor (CF), wet weight gain (WG_w), specific growth rate in wet weight (SGR_w) and ration levels in terms of energy (RL_e) decreased significantly (p < 0.05) with decreasing ration levels from birth to first-time spawning. On the contrary, the food conversion efficiency in terms of energy (FCE_e) increased significantly (p < 0.05) with the decreasing ration levels from birth to first-time sexual maturity. Furthermore, higher percentages of energy intake from food were allocated to somatic and gonad growth in M and L groups compared to the H group before sexual maturity; In addition, the time for first-time spawning in groups M and L was longer than that of the H group. As a result, the gonad-somatic index (GSI) and oocytes/embryos weight in M and L groups were similar to that of the H group, although the ovary weight and oocytes/embryos numbers were all lower than that of the H group. Also, similar growth performances were observed in second-generation offspring, which were produced by female parents fed with different ration levels. These findings suggest that the female G. affinis could produce a number of healthy offspring under conditions of food restriction, and that this could be achieved by increasing the energy allocated to gonad development, reducing fecundity and delaying spawning time. These life strategies ensured that G. affinis could survive and thrive in adverse environmental conditions and exhibit characteristics of invasive fish species.     

Extraction,Preliminary Characterization and Evaluation of in Vitro Antitumor and Antioxidant Activities of Polysaccharides from Mentha piperita

This study describes the extraction, preliminary characterization and evaluation of the in vitro antitumor and antioxidant activities of polysaccharides extracted from Mentha piperita (MPP). The optimal parameters for the extraction of MPP were obtained by Box-Behnken experimental design and response surface methodology (RSM) at the ratio of water to raw material of 20, extraction time of 1.5 h and extraction temperature at 80 °C. Chemical composition analysis showed that MPP was mainly composed of glucuronic acid, galacturonic acid, glucose, galactose and arabinose, and the molecular weight of its two major fractions were estimated to be about 2.843 and 1.139 kDa, respectively. In vitro bioactivity experiments showed that MPP not only inhibited the growth of A549 cells but possessed potent inhibitory action against DNA topoisomerase I (topo I), and an appreciative antioxidant action as well. These results indicate that MPP may be useful for developing safe natural health products.     

The hepatoprotective activity against ethanol-induced cytotoxicity by aqueous extract of Antrodia cinnamomea

The objective of this study was to evaluate the hepatoprotective activity against the ethanol-induced cytotoxicity by Antrodia cinnamomea which were produced from a large-scale fermentation. The production of biomass, extracellular polysaccharide (EPS), intracellular polysaccharide (IPS), and triterpenoids was 27.1, 1.2, 0.7, and 2.0 g/L, respectively, using a 5000-L agitated bioreactor with 3500 L of A. cinnamomea medium (ACM). The hepatoprotective effects of the water extract from the mycelia of A. cinnamomea (WAC) were evaluated in vitro using ethanol-induced cytotoxicity on AML12 hepatocytes. The cytotoxicity and the apoptosis-associated phosphatidyl serine redistribution of plasma membrane to AML12 cells induced by 300 mM ethanol were effectively reduced by adding 500 mg/L of WAC. From the compositional analysis, the major component in WAC was polysaccharides that showed a high galactose content (70.9 mg/g crude polysaccharide). Hence, the large-scale fermentation was efficient in producing cell mass and its metabolites which were able to protect hepatocytes from the damage by ethanol.     

A High Redox Potential Laccase from Pycnoporus sanguineus RP15: Potential Application for Dye Decolorization

Laccase production by Pycnoporus sanguineus RP15 grown in wheat bran and corncob under solid-state fermentation was optimized by response surface methodology using a Central Composite Rotational Design. A laccase (Lacps1) was purified and characterized and the potential of the pure Lacps1 and the crude culture extract for synthetic dye decolorization was evaluated. At optimal conditions (eight days, 26 °C, 18% (w/w) milled corncob, 0.8% (w/w) NH₄Cl and 50 mmol·L⁻¹ CuSO₄, initial moisture 4.1 mL·g⁻¹), the laccase activity reached 138.6 ± 13.2 U·g⁻¹. Lacps1 was a monomeric glycoprotein (67 kDa, 24% carbohydrate). Optimum pH and temperature for the oxidation of 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonate) (ABTS) were 4.4 and 74.4 °C, respectively. Lacps1 was stable at pH 3.0–8.0, and after two hours at 55–60 °C, presenting high redox potential (0.747 V vs. NHE). ABTS was oxidized with an apparent affinity constant of 147.0 ± 6.4 μmol·L⁻¹, maximum velocity of 413.4 ± 21.2 U·mg⁻¹ and catalytic efficiency of 3140.1 ± 149.6 L·mmol⁻¹·s⁻¹. The maximum decolorization percentages of bromophenol blue (BPB), remazol brilliant blue R and reactive blue 4 (RB4), at 25 or 40 °C without redox mediators, reached 90%, 80% and 60%, respectively, using either pure Lacps1 or the crude extract. This is the first study of the decolorization of BPB and RB4 by a P. sanguineus laccase. The data suggested good potential for treatment of industrial dye-containing effluents.