Pleurotus sajor-caju (Fr.) Singer ß-1,3-Glucanoligosaccharide (Ps-GOS) Suppresses RANKL-Induced Osteoclast Differentiation and Function in Pre-Osteoclastic RAW 264.7 Cells by Inhibiting the RANK/NFκB/cFOS/NFATc1 Signalling Pathway.

Edible grey oyster mushroom, Pleurotus sajor-caju, ß (1,3), (1,6) glucan possesses a wide range of biological activities, including anti-inflammation, anti-microorganism and antioxidant. However, its biological activity is limited by low water solubility resulting from its high molecular weight. Our previous study demonstrated that enzymatic hydrolysis of grey oyster mushroom ß-glucan using Hevea ß-1,3-glucanase isozymes obtains a lower molecular weight and higher water solubility, Pleurotus sajor-caju glucanoligosaccharide (Ps-GOS). Additionally, Ps-GOS potentially reduces osteoporosis by enhancing osteoblast-bone formation, whereas its effect on osteoclast-bone resorption remains unknown. Therefore, our study investigated the modulatory activities and underlying mechanism of Ps-GOS on Receptor activator of nuclear factor kappa-Β ligand (RANKL) -induced osteoclastogenesis in pre-osteoclastic RAW 264.7 cells. Cell cytotoxicity of Ps-GOS on RAW 264.7 cells was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and its effect on osteoclast differentiation was determined by tartrate-resistant acid phosphatase (TRAP) staining. Additionally, its effect on osteoclast bone-resorptive ability was detected by pit formation assay. The osteoclastogenic-related factors were assessed by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), Western blot and immunofluorescence. The results revealed that Ps-GOS was non-toxic and significantly suppressed the formation of mature osteoclast multinucleated cells and their resorption activity by reducing the number of TRAP-positive cells and pit formation areas in a dose-dependent manner. Additionally, Ps-GOS attenuated the nuclear factor kappa light chain-enhancer of activated B cells' P65 (NFκB-P65) expression and their subsequent master osteoclast modulators, including nuclear factor of activated T cell c1 (NFATc1) and Fos proto-oncogene (cFOS) via the NF-κB pathway. Furthermore, Ps-GOS markedly inhibited RANK expression, which serves as an initial transmitter of many osteoclastogenesis-related cascades and inhibited proteolytic enzymes, including TRAP, matrix metallopeptidase 9 (MMP-9) and cathepsin K (CTK). These findings indicate that Ps-GOS could potentially be beneficial as an effective natural agent for bone metabolic disease.

Thermosensitive polymer-assisted extraction and purification of fungal laccase from citrus pulp wash effluent.

BACKGROUND: This study explores the use of liquid-liquid extraction with thermosensitive polymers for producing laccase (Lac) from Pleurotus sajor-caju. This process leverages liquid waste from the citrus industry, specifically pulp wash. The research delves into extractive fermentation and thermoseparation, both processes being facilitated by a polymer exhibiting a lower critical solution temperature transition. RESULTS: Key factors considered include the choice of polymer, its concentration, pH, separation temperature, and the behavior of the polymer-rich phase post-extractive fermentation concerning the lower critical solution temperature. Notably, under conditions of 45% by weight of Pluronic L-61 and pH 5.0 at 25 °C, the Lac resulted in an enhancement in the purification factor of 28.4-fold, compared with the Lac obtained directly from the fermentation process on the eighth day. There was an 83.6% recovery of the Lac enzyme in the bottom phase of the system. Additionally, the unique properties of Pluronic L-61, which can induce phase separation and also allow for thermoseparation, led to a secondary fraction (aqueous solution) of Lac with purification factor of 2.1 ± 0.1-fold (at 32 ± 0.9 °C and 30 ± 0.3 min without stirring) from the polymeric phase (top phase). Fourier-transform infrared analysis validated the separation data, particularly highlighting the α-helix content in the amide I region (1600-1700 cm-1 ). CONCLUSION: In summary, the insights from this study pave the way for broader industrial applications of these techniques, underscoring benefits like streamlined process integration, heightened selectivity, and superior separation efficacy. © 2023 Society of Chemical Industry.

Biotransformation of Polyunsaturated Fatty Acids to Trioxilins by Lipoxygenase from Pleurotus sajor-caju.

A lipoxygenase from Pleurotus sajor-caju (PsLOX) was cloned, expressed in Escherichia coli, and purified as a soluble protein with a specific activity of 629 µmol/min/mg for arachidonic acid (AA). The native PsLOX exhibited a molecular mass of 146 kDa, including a 73-kDa homodimer, as estimated by gel-filtration chromatography. The major products converted from polyunsaturated fatty acids (PUFAs), including AA, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA), were identified as trioxilins (TrXs), namely 13,14,15-TrXB3 , 13,14,15-TrXB4 , and 15,16,17-TrXB5 , respectively, through high-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses. The enzyme displayed its maximum activity at pH 8.0 and 20 °C. Under these conditions, the specific activity and catalytic efficiency of PsLOX for PUFAs exhibited the following order: AA>EPA>DHA. Based on HPLC analysis and substrate specificity, PsLOX was identified as an arachidonate 15-LOX. PsLOX efficiently converted 10 mM of AA, EPA, and DHA to 8.7 mM of 13,14,15-TrXB3 (conversion rate: 87 %), 7.9 mM of 13,14,15-TrXB4 (79 %), and 7.2 mM of 15,16,17-TrXB5 (72 %) in 15, 20, and 20 min, respectively, marking the highest conversion rates reported to date. Collectively, our results demonstrate that PsLOX is an efficient TrXs-producing enzyme.

The optimization of enzymatic oxidation of levofloxacin, a fluoroquinolone antibiotic for wastetwater treatment.

The global presence of antibiotics in the environment has created concerns about the emergence of antibiotic resistance bacteria and potential hazard to humans and the ecosystem. This work aims to study the removal of levofloxacin, a new generation fluoroquinolone antibiotic from aqueous solutions by enzyme mediated oxidation process and optimization of the conditions thereof by response surface methodology (RSM) using Box-Behnken design (BBD). For this study, experiments were conducted to analyze the effect of independent variables namely, pH, temperature, mediator concentration and antibiotic concentration on the degradation percentage of levofloxacin antibiotic using laccase enzyme derived from Trametes versicolor. The residual levofloxacin concentration was determined using high performance liquid chromatography (HPLC). On applying the quadratic regression analysis, among the main parameters, it was found that the percentage degradation was significantly affected by all the four variables. The predicted values for percentage degradation of levofloxacin were close to the experimental values obtained and the R2 (0.95) indicated that the regression was able to give a good prediction of response for the percentage degradation of levofloxacin in the studied range. The optimal conditions for the maximum degradation (99.9%) as predicted by the BBD were: temperature of 37 °C, pH of 4.5, mediator concentration of 0.1 mM and levofloxacin concentration of 5 µg mL-1. The findings of the study were further extended to study the effect of partially purified enzymes isolated from Pleurotus eryngii, Pleurotus florida and Pleurotus sajor caju on the degradation of levofloxacin at concentrations ranging from as low as 0.1 to as high as 50 µg mL-1 in synthetic wastewater utilizing the optimized conditions generated by BBD. A maximum degradation of 88.8% was achieved with the partially purified enzyme isolated from Pleurotus eryngii at 1 µg mL-1 levofloxacin concentration which was at par with the commercial laccase which showed 89% degradation in synthetic wastewater at the optimized conditions. The biodegradation studies were conducted using only 2 units of laccase. Thus, the expensive commercial laccase can be effectively replaced by crude laccase isolated from indigenous macrofungi such as P. eryngii, P. florida and P. sajor caju as a cost effective alternative to degrade levofloxacin present in contaminated wastewater using as low as 2 units of enzyme for a 72 h treatment period.

Nutritional analysis and molecular characterization of hybrid mushrooms developed through intergeneric protoplast fusion between Pleurotus sajor-caju and Calocybe indica with the purpose to achieve improved strains.

Two edible mushrooms Calocybe indica and Pleurotus sajor-caju were chosen as parent strains in this study to approach the concept of hybridization through the protoplast fusion technique. Protoplast fusion in presence of polyethylene glycol (PEG) was conducted between the parent strains and by further double selection screening method, six somatic hybrid lines were developed. Those fruit bodies of the hybrid lines showed phenotypic resemblance with Pleurotus sajor-caju when grown on paddy straw under favorable conditions. The hybridity of the newly developed somatic hybrid strains was established by barrage reaction, morphological traits, fruitbody parameter and, inter single sequence repeat (ISSR) profiling. One-way analysis of variance (ANOVA) was used for the analysis of phenotypic data of hybrid lines and parents. Five ISSR primers were used to generate 51 amplified DNA fragments ranged between 250 and 3000 bp in size in six hybrids and two parents with 90.19% polymorphism. Some of the hybrids contain some non-parental bands which indicate that recombination might happen in the hybrid genome hence confirming the hybridity of newly developed strains. The dendrogram was created using the Average Linkage (Between Groups) method based on ISSR profiling and genetic distance between parent-hybrids and hybrid-hybrid was analyzed by Jaccard's proximity matrix. A definite improvement in nutritional properties and biological activity was observed in the study. Due to ease in their cultivation, it can play a significant role in the rural economic development.

Evaluation of bioremediation strategies for treating recalcitrant halo-organic pollutants in soil environments.

The aim of this study was to investigate the bioremediation potential of polychlorinated biphenyls (PCBs) in soil, mimicking three strategies: (a) mycoaugmentation: by the addition of Trametes sanguinea and Pleurotus sajor-caju co-cultures immobilized on sugarcane bagasse; (b) biostimulation: by supplementation of sugarcane bagasse; and (c) natural attenuation: no amendments. The experiments were done in microcosms using Ultisol soil. Remediation effectiveness was assessed based on pollutants content, soil characteristics, and ecotoxicological tests. Biostimulation and mycoaugmentation demonstrated the highest PCBs-removal (approx. 90%) with a significant toxicity reduction at 90 d. The studied strains were able to survive during the incubation period in non-sterilized soil. Laccase, manganese-peroxidase and endoxylanase activities increased significantly in co-cultures after 60 d. Sugarcane bagasse demonstrated to be not only a suitable support for fungal immobilization but also an efficient substrate for fungal colonization of PCBs-contaminated soils. Mycoaugmentation and biostimulation with sugarcane bagasse improved oxidable organic matter and phosphorous contents as well as dehydrogenase activity in soil. Therefore, biostimulation with sugarcane bagasse and mycoaugmentation applying dual white-rot fungal cultures constitute two efficient bioremediation alternatives to restore PCBs-contaminated soils.

Pleurotus sajor-caju-Mediated Synthesis of Silver and Gold Nanoparticles Active against Colon Cancer Cell Lines: A New Era of Herbonanoceutics.

Herbal medicines are widely used worldwide and much appreciated because of their fewer side effects and the ability to fight diseases at the root cause. Active 'phyto' ingredients require a scientific approach and a mechanism to distribute components at the target site for better therapeutic results. Nanotechnology, on the other hand, has created new hope for cancer treatment but is still far from being proven in clinical settings. This article combines a unique approach to synthesis with the use of Pleurotus sajor-caju, followed by microwave irritation of silver and gold nanoparticles that ensures the capping of the active phyto ingredient and further enhances the effects of nanomedicine to fight colon cancer, thus opening a new era of what we call herbonanoceutics. The article also compares the characteristics and properties of silver (Au) and gold (Ag) nanoparticles synthesized by an in house developed novel microwave-assisted rapid green synthesis method. The as-prepared Ag NPs and Au NPs were compared using ultraviolet-visible spectroscopy (UV-Vis), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM), and energy dispersive spectroscopy (EDS). Our comparative study revealed that both assemblies display face-centred cubic structures (FCCs) and are nanocrystalline in nature. The advantage of the approach was that the sizes of gold and silver were identical in range with a similar distribution pattern. This has helped us to study the activity against colon cancer cell line (HCT-116) without incoherence since size plays a key role in the application. More specifically, morphological changes, cell viability, the production of reactive oxygen species (ROS) and the fragmentation of DNA have been further reported to assess better the results obtained with the two metals. Our results suggest that the newly adopted synthesis method may ensure the dual benefits from phyto ingredients which further enhances the effectiveness of advanced nanomedicine.

Purification and biochemical characterization of a novel thermostable protease from the oyster mushroom Pleurotus sajor-caju strain CTM10057 with industrial interest.

BACKGROUND: Proteases are hydrolytic enzymes that catalyze peptide linkage cleavage reactions at the level of proteins and peptides with different degrees of specificity. This group draws the attention of industry. More than one protease in three is a serine protease. Classically, they are active at neutral to alkaline pH. The serine proteases are researched for industrial uses, especially detergents. They are the most commercially available enzyme group in the world market. Overall, fungi produced extracellular proteases, easily separated from mycelium by filtration. RESULTS: A new basidiomycete fungus CTM10057, a hyperproducer of a novel protease (10,500 U/mL), was identified as Pleurotus sajor-caju (oyster mushroom). The enzyme, called SPPS, was purified to homogeneity by heat-treatment (80 °C for 20 min) followed by ammonium sulfate precipitation (35-55%)-dialysis, then UNO Q-6 FPLC ion-exchange chromatography and finally HPLC-ZORBAX PSM 300 HPSEC gel filtration chromatography, and submitted to biochemical characterization assays. The molecular mass was estimated to be 65 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Native-PAGE, casein-zymography, and size exclusion by HPLC. A high homology with mushroom proteases was displayed by the first 26 amino-acid residues of the NH2-terminal aminoacid sequence. Phenylmethanesulfonyl fluoride (PMSF) and diiodopropyl fluorophosphates (DFP) strongly inhibit SPPS, revealing that it is a member of the serine-proteases family. The pH and temperature optima were 9.5 and 70 °C, respectively. Interestingly, SPPS possesses the most elevated hydrolysis level and catalytic efficiency in comparison with SPTC, Flavourzyme® 500 L, and Thermolysin type X proteases. More remarkably, a high tolerance towards organic solvent tolerance was exhibited by SPPS, together with considerable detergent stability compared to the commercial proteases Thermolysin type X and Flavourzyme® 500 L, respectively. CONCLUSIONS: This proves the excellent proprieties characterizing SPPS, making it a potential candidate for industrial applications especially detergent formulations.

Phenol removal by laccases and other phenol oxidases of Pleurotus sajor-caju PS-2001 in submerged cultivations and aqueous mixtures.

In this work, phenol removal from aqueous solutions by Pleurotus sajor-caju PS-2001 phenol oxidases was assessed under different conditions. In stirred-tank reactor (STR), 77, 82, 92 and 36% of removal were attained when initial concentrations of 1.0, 2.0, 3.0 and 4.0 mmol L-1 phenol, respectively, were used. Among the different enzymes produced by this fungus, phenol removal seems to be related to the activity of laccases that attained maximum values between 33 and 91 U mL-1 in STR. With an internal-loop airlift reactor (ILAR), phenol concentrations of 1.0, 2.0, 3.0, 4.0 and 5.0 mmol L-1 were evaluated, and removal of 70, 76, 82, 77 and 82%, respectively, were observed. In ILAR, however, superior maximum titres of laccases were quantified (80-285 U mL-1). Crude enzyme broths have also been tested for phenol removal from 3.0 mmol L-1 aqueous solutions, the best result (55% of removal) being obtained at pH 3.2 and 30 °C, without agitation, using 60 U mL-1 laccases. According to the data presented, phenol can be efficiently removed from liquid media in submerged cultures of P. sajor-caju PS-2001 even when carried out in a simple pneumatic reactor, whereas significantly less amount of the pollutant is degraded when a crude enzyme broth is used.

Pleurotus sajor-caju can be used to synthesize silver nanoparticles with antifungal activity against Candida albicans.

BACKGROUND: Green synthesis of silver nanoparticles (AgNPs) has become widely practiced worldwide. In this study, AgNPs were synthesized using a hot-water extract of the edible mushroom Pleurotus sajor-caju. The product, PSC-AgNPs, was characterized by using UV-visible spectra, dynamic light scattering analysis, transmission electron microscopy (TEM), X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectrometry. To assess its antifungal activity against Candida albicans, gene transcription and protein expression analyses were conducted for CaICL1 and its product, ICL, using real-time quantitative polymerase chain reaction and western blot, respectively. RESULTS: PSC-AgNPs with an average particle size of 11.68 nm inhibited the growth of the pathogenic yeast C. albicans. Values for minimum inhibitory concentration and minimum fungicidal concentration were 250 and 500 mg L-1 , respectively. TEM images revealed that the average particle size of PSC-AgNPs was 16.8 nm, with the values for zeta potential and the polydispersity index being -8.54 mV and 0.137, respectively. XRD and FTIR spectra showed PSC-AgNPs to have a face-centered cubic crystalline structure. The polysaccharides and amino acid residues present in P. sajor-caju extract were found to be involved in reducing Ag+ to AgNP. Both CaICL1 transcription and ICL protein expression were found to be suppressed in the cells treated with PSC-AgNPs as compared with the control. CONCLUSION: Our PSC-AgNP preparation makes for a promising antifungal agent that can downregulate isocitrate lyase. © 2017 Society of Chemical Industry.

Important nutritional constituents, flavour components, antioxidant and antibacterial properties of Pleurotus sajor-caju.

Oyster mushroom (Pleurotus sajor-caju) cultivated in the laboratory was studied for nutritional constituents, flavor components, antioxidant and antibacterial properties. Nutritional constituents estimated per 100 g dry weight (d.w.) include protein (29.3 g), carbohydrate (62.97 g), crude fat (0.91 g), ash (6.82 g) and crude fiber (12.3 g). Energy value of this mushroom was about 297.5 kcal/100 g d.w. Major mineral components estimated include Ca, Fe, and Mg with highest level of 505.0, 109.5 and 108.7 mg/100 g respectively. Methanolic extract containing significant amounts of phenols and flavonoids showed free radical scavenging potential and antibacterial activities against various spp. of Gram positive and Gram negative bacteria. Compounds responsible for antibacterial activities analyzed by GC-MS include ß- Sistosterol, Cholestanol, 1,5-Dibenzoylnaphthalene and 1,2-Benzenedicarboxylic acid. Flavor components extracted by hot extraction method were found to be higher in number and concentration than the cold extraction method. The characteristic flavor component of mushroom i.e. 1-Octen-3-ol was better extracted by hot than the cold.

Exploring fungal-mediated solutions and its molecular mechanistic insights for textile dye decolorization.

Decolorization of textile dyes and study of their intermediate compounds is necessary to comprehend the mechanism of dye degradation. In the present study, different fungal mediated solutions were explored to provide an alternative to treat the reactive dyes. Growing biomass of Pleurotus sajor caju showed 83% decolorization (249.99 mg L-1 removal) of Reactive Blue 13 (RB 13) and 63% decolorization (188.83 mg L-1) of Reactive Black 5 (RB 5) at 300 mg L-1 initial concentration on 8 d. Higher laccase activity was positively correlated with increase in decolorization. However, increasing dye concentration has inhibitory effect on fungal biomass due to increase in toxicity. In laccase mediated decolorization, laccase produced from P. sajor caju using carbon rich waste material as substrate showed 89% decolorization (276.36 mg L-1 removal) of RB 13 and 33% decolorization (105.37 mg L-1 removal) of RB 5 at 300 mg L-1 initial dye concentration in 100 min at 30 °C and pH 3.0'. Comparing the two methods, laccase-mediated decolorization shows better decolorization in less time and does not produce sludge. Further, the present work also attempted to study the dye degradation pathway for Reactive blue 13 via laccase mediated process. Fourier-transform infrared spectroscopy (FTIR), high-performance liquid chromatography (HPLC), and gas chromatography-mass spectrometry (GC-MS) were utilized to identify the degraded products. The GC-MS analysis showed the formation of naphthalene, naphthalene 2-ol, benzene,1-2, dicarboxylic acid, 4, amino, 6,chloro, 1-3-5, triazin-2-ol as the final degraded products after enzymatic degradation of RB 13. These findings provide in-depth study of laccase-mediated textile dye degradation mechanism.

Sustainable production of Pleurotus sajor-caju mushrooms and biocomposites using brewer's spent and agro-industrial residues.

Brazil is one of the world's largest beer producers and also a major food producer. These activities generate a large amount of residues which, if disposed of inappropriately, can have adverse effects on the environment. The objective of this research was to evaluate the potential of using these residues for both mushroom cultivation (traditional use) and the production of mycelium-based composites (innovative use). Mushroom production (Pleurotus sajor-caju) was conducted using only brewer's spent grains (fresh and dried) and also mixed with banana leaves (1:1) or peach palm leaves (1:1), which are residues widely available in the northern region of Santa Catarina, Brazil. The productivity of mushrooms cultivated using fresh and dried brewer's spent grains did not exhibit a statistically significant difference, indicating that this residue can be utilized shortly after its generation in the industrial process, thereby reducing costs associated with production. Combining brewer's spent grains with banana or peach palm leaves resulted in enhanced mushroom production (0.41 and 0.38 g day-1, respectively) compared to using the leaves as a sole substrate. The mushrooms produced contain sugars and a minimal sodium content, and are considered a source of phosphorus. In addition, no toxic elements (Hg and Pb) were present. The mycelium-based composites produced using the residual substrate (after the mushroom harvest) exhibited better mechanical properties (compressive strength = 0.04 MPa, density = 242 kg m-3, and low humidity sorption) than those produced using fresh substrate. The results demonstrate the synergistic effect of combining the two approaches under investigation. The use of brewer´s spent enhance the mushroom productivity and the residual substrate enhance the mechanical properties of mycelium-based composites. The compressive strength, density, and air humidity sorption properties are essential for determining the potential applications of mycelium-based composites. The use of brewer's spent grains mixed with banana leaves demonstrated significant promise for mushroom production and subsequent application in the development of mycelium-based composites. These sequential approaches contribute to waste valorization and the rational utilization of natural resources, as the mycelium-based composites are considered for substitution of synthetic materials, thereby promoting sustainability for future generations.

Structural Characterization of a Pleurotus sajor-caju Polysaccharide and Its Neuroprotection Related to the Inhibition of Oxidative Stress.

A novel polysaccharide PSP2-1 was isolated and purified from Pleurotus sajor-caju. The structural characterization data displayed that the molecular weight of PSP2-1 was 44.9 kDa, and PSP2-1 consisted of fucose, galactose, glucose, and mannose. The methylation results showed that the glycosidic bonds of PSP2-1 included T-Fuc, 1,6-Gal, T-Glc, 1,6-Glc, 1,3,6-Glc, 1,3-Man, 1,2,6-Man, and T-Man. Neuroprotective studies indicated that PSP2-1 significantly improved the cell viability of the H2O2-induced oxidatively damaged neuronal cell HT22, reduced the release of LDH, inhibited apoptosis and release of cytochrome c, and alleviated the decline of mitochondrial membrane potential and ROS accumulation. Furthermore, PSP2-1 decreased the phosphorylation levels of cleaved PARP and cleaved caspase-3, and increased the ratio of bcl-2/bax. Additionally, PSP2-1 could inhibit the phosphorylation of MAPK family members including JNK, p38, and Erk. Finally, animal experiments showed that PSP2-1 could improve the oxidative stress injury and the learning and memory ability of mice with aging induced by D-galactose. Our results confirmed that PSP2-1 significantly ameliorated the oxidative stress injury, inhibited the apoptosis in H2O2-induced neuronal cells via MAPK pathway, and also improved cognition in mice with aging induced by D-galactose. Our research gives the foundation for the functional food application of P. sajor-caju polysaccharides in the future.

Evaluation of Volatile Profile and In Vitro Antioxidant Activity of Fermented Green Tea Infusion With Pleurotus sajor-caju (Oyster Mushroom).

Green tea has distinct astringency, bitter taste, and typical green flavor because of its post-harvest treatment without withering and enzymatic oxidation. Microbial fermentation has been identified as a promising strategy that could give green tea infusion a special taste flavor. This might be linked to the metabolic transformation ability of microorganisms. In this study, starter culture of edible mushroom Pleurotus sajor-caju (oyster mushroom) was used for submerged fermentation of green tea infusion in order to improve its flavor and taste quality. The volatile profile determined by headspace solid-phase microextraction, coupled with gas chromatography mass spectrometry, showed that the contents of (Z)-2-penten-1-ol and methyl heptadienone in green tea infusion were decreased significantly by the fermentation with the basidiomycete P. sajor-caju (p < 0.01), which would alleviate the herbal and grass flavor of green tea infusion to a certain extent. Meanwhile, the contents of linalool and geraniol were increased 9.3 and 11.3 times, respectively, whereas methyl salicylate was newly produced after fermentation by P. sajor-caju, endowing the fermented tea infusion with a pleasant flower and fruit aroma. In addition, the polyphenol profile was determined using high-performance liquid chromatography equipped with ion trap mass spectrometry, and the results indicated that the contents of most polyphenols in green tea infusion decreased significantly after fermentation by P. sajor-caju. The reduction of catechins and anthocyanins in fermented green tea infusion alleviated the astringency and bitterness. Moreover, the antioxidant activity of fermented green tea infusion was obviously decreased, especially the DPPH-free radical-scavenging ability and the ferric-reducing power. However, it is noteworthy that the ABTS-free radical scavenging ability was improved compared with the unfermented one, indicating that the increased tea pigments and volatile metabolites (such as linalool and geraniol) after fermentation with P. sajor-caju may also contribute to the antioxidant capacity of fermented green tea infusion. Overall, the innovative approach driven by P. sajor-caju fermentation has achieved promising potential to manipulate the green tea flavor.

Antiviral, Cytotoxic, and Antioxidant Activities of Three Edible Agaricomycetes Mushrooms: Pleurotus columbinus, Pleurotus sajor-caju, and Agaricus bisporus.

In this study, we investigated aqueous extracts of three edible mushrooms: Agaricus bisporus (white button mushroom), Pleurotus columbinus (oyster mushroom), and Pleurotus sajor-caju (grey oyster mushroom). The extracts were biochemically characterized for total carbohydrate, phenolic, flavonoid, vitamin, and protein contents besides amino acid analysis. Triple TOF proteome analysis showed 30.1% similarity between proteomes of the two Pleurotus spp. All three extracts showed promising antiviral activities. While Pleurotus columbinus extract showed potent activity against adenovirus (Ad7, selectivity index (SI) = 4.2), Agaricus bisporus showed strong activity against herpes simplex II (HSV-2; SI = 3.7). The extracts showed low cytotoxicity against normal human peripheral blood mononuclear cells (PBMCs) and moderate cytotoxicity against prostate (PC3, DU-145); colorectal (Colo-205); cecum carcinoma (LS-513); liver carcinoma (HepG2); cervical cancer (HeLa); breast adenocarcinoma (MDA-MB-231 and MCF-7) as well as leukemia (CCRF-CEM); acute monocytic leukemia (THP1); acute promyelocytic leukemia (NB4); and lymphoma (U937) cell lines. Antioxidant activity was evaluated using 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radical scavenging, 2,2'-Azinobis-(3-Ethylbenzthiazolin-6-Sulfonic Acid) ABTS radical cation scavenging, and oxygen radical absorbance capacity (ORAC) assays. The three extracts showed potential antioxidant activities with the maximum activity recorded for Pleurotus columbinus (IC50 µg/mL) = 35.13 ± 3.27 for DPPH, 13.97 ± 4.91 for ABTS, and 29.42 ± 3.21 for ORAC assays.

Optimization of growth conditions for enhancing the production of microbial laccase and its application in treating antibiotic contamination in wastewater.

In this work, seven indigenous macrofungal isolates were selected to screen for their laccase production capability. Among them, isolates viz., Pleurotus eryngii, Pleurotus florida, Pleurotus sajor caju and Gandoderma lucidum were found to exhibit high laccase activity in the preliminary studies and were thus selected for the optimization studies with an aim to enhance laccase production. The pH optimization studies were carried out between pH range of 4-6. The laccase activity and biomass were found to be optimum at pH 4, 4.5, 4.5 and 5 for P. eryngii, P. florida, P. sajor caju and G. lucidum, respectively. Optimization studies with chemical inducers namely, tannic acid, 2,6 dimethoxyphenol and copper sulphate at three different concentration levels were conducted and tannic acid at 2 mM concentration was found to increase the laccase activity to about 45% followed by 2,6 dimethoxyphenol (2 mM) with an increase of about 43% and copper sulphate (0.1 mM) showing 21% increase in the yield. Biodegradation studies utilizing laccase isolated from P. eryngii, P. florida and P. sajor caju was carried out for a commonly detected fluoroquinolone antibiotic, levofloxacin, in water and pharmaceutical wastewater. The results indicated that the degradation efficiency of levofloxacin using laccase isolated from P. eryngii (88.9%) was comparable to commercial laccase (89%). When the cost economics of using crude laccase was evaluated against commercial laccase it was evident that the total cost of the treatment could be reduced by 71.7% if commercial grade laccase was replaced by crude enzyme extracted from indigenous macrofungi such Pleurotus eryngii, Pleurotus florida, and Pleurotus sajor caju indicating a promising and cost-effective alternative for wastewater treatment.

Pro-Health and Anti-Cancer Activity of Fungal Fractions Isolated from Milk-Supplemented Cultures of Lentinus (Pleurotus) Sajor-caju.

The present study aimed to demonstrate Lentinus (formerly Pleurotus) sajor-caju (PSC) as a good source of pro-health substances. It has also shown that supplementation of its culture medium with cow milk may further improve its beneficial properties. Intracellular fractions from fungi grown on a medium supplemented with cow milk were analyzed using various biochemical methods for determination of the nutrient composition. Furthermore, anti-cancer properties of selected extracts were investigated on colorectal cancer cell lines (HT-29, LS 180, and SW948) in vitro. Biochemical analysis showed enrichment in health-enhancing compounds, such as proteins or polysaccharides (about 3.5- and 4.5-fold increase in concentration of proteins and carbohydratesin extracts of mycelia cultured on whole milk (PSC2-I), respectively), with a decrease in the level of free radicals (10-fold decrease in extract grown on milk and medium mixture (1:1) (PSC3-II)), which was related to increased catalase and superoxide dismutase activity (7.5-fold increase in catalase activity and 5-fold in SOD activity in PSC3-II compared to the control). Moreover, the viability of the cancer cells was diminished (to 60.0 ± 6.8% and 40.0 ± 8.6% of the control, on HT-29 and SW948 cells, respectively), along with pro-apoptotic (to 18.8 ± 11.8 and 14.7 ± 8.0% towards LS 180 and SW948 cells, respectively) and NO-secreting effects (about 2-fold increase) of the extracts. This study suggests that PSC has multiple nutritional and anti-cancer properties and can be used as a source of healthy biomolecules in modern medicine or functional foods.

Enhancing Activity of Pleurotus sajor-caju (Fr.) Sing ß-1,3-Glucanoligosaccharide (Ps-GOS) on Proliferation, Differentiation, and Mineralization of MC3T3-E1 Cells through the Involvement of BMP-2/Runx2/MAPK/Wnt/ß-Catenin Signaling Pathway.

Osteoporosis is a leading world health problem that results from an imbalance between bone formation and bone resorption. ß-glucans has been extensively reported to exhibit a wide range of biological activities, including antiosteoporosis both in vitro and in vivo. However, the molecular mechanisms responsible for ß-glucan-mediated bone formation in osteoblasts have not yet been investigated. The oyster mushroom Pleurotus sajor-caju produces abundant amounts of an insoluble ß-glucan, which is rendered soluble by enzymatic degradation using Hevea glucanase to generate low-molecular-weight glucanoligosaccharide (Ps-GOS). This study aimed to investigate the osteogenic enhancing activity and underlining molecular mechanism of Ps-GOS on osteoblastogenesis of pre-osteoblastic MC3T3-E1 cells. In this study, it was demonstrated for the first time that low concentrations of Ps-GOS could promote cell proliferation and division after 48 h of treatment. In addition, Ps-GOS upregulated the mRNA and protein expression level of bone morphogenetic protein-2 (BMP-2) and runt-related transcription factor-2 (Runx2), which are both involved in BMP signaling pathway, accompanied by increased alkaline phosphatase (ALP) activity and mineralization. Ps-GOS also upregulated the expression of osteogenesis related genes including ALP, collagen type 1 (COL1), and osteocalcin (OCN). Moreover, our novel findings suggest that Ps-GOS may exert its effects through the mitogen-activated protein kinase (MAPK) and wingless-type MMTV integration site (Wnt)/ß-catenin signaling pathways.

Fungal biosynthesis of lignin-modifying enzymes from pulp wash and Luffa cylindrica for azo dye RB5 biodecolorization using modeling by response surface methodology and artificial neural network.

This study demonstrates the evaluation between the artificial neural network technique coupled to the genetic algorithm (ANN-GA) and the response surface methodology (RSM) for prediction of Reactive Black 5 (RB5) decolorization by crude enzyme from Pleurotus. sajor-caju. Fungal lignin-modifying enzymes (FLME) were synthesized using pulp wash (PW) as an inducing substrate, and L. cylindrica (L.C) for cell immobilization. When grown in PW, the fungus showed higher Lac activity (126.5 IU. mL-1), whereas when immobilized a higher MnP activity was achieved (22.79 IU. mL-1), but both methods were capable of decolorizing the dye in about 89.4 % and 75 %, respectively. This indicates applicability of PW as an alternative substrate for FLME induction and viability of immobilization for MnP synthesis. For RB5 decolorization, the action of the crude enzyme extract was considered as a function of pH, dye concentration, temperature, and reaction time. The models are well adjusted to predict the efficiency of biodecolorization, with no statistical difference between ANN-GA and RSM, which indicates potential for green enzymes prospecting application in bioprocess industry.