Comparison of cell wall polysaccharides in Schizophyllum commune after changing phenotype by mutation.

The Agaricomycetes fungi produce various compounds with pharmaceutical, medicinal, cosmetic, environmental and biotechnological properties. In addition, some polysaccharides extracted from the fungal cell wall have antitumor and immunomodulatory actions. The aim of this study was to use genetic modification to transform Schizophyllum commune and identify if the phenotype observed (different from the wild type) resulted in changes of the cell wall polysaccharides. The plasmid pUCHYG-GPDGLS, which contains the Pleurotus ostreatus glucan synthase gene, was used in S. commune transformations. Polysaccharides from cell wall of wild (ScW) and mutants were compared in this study. Polysaccharides from the biomass and culture broth were extracted with hot water. One of the mutants (ScT4) was selected for further studies and, after hydrolysis/acetylation, the GLC analysis showed galactose as the major component in polysaccharide fraction from the mutant and glucose as the major monomer in the wild type. Differences were also found in the elution profiles from HPSEC and NMR analyses. From the monosaccharide composition it was proposed that mannogalactans are components of S. commune cell wall for both, wild and mutant, but in different proportions. To our knowledge, this is the first time that mannogalactans are isolated from S. commune liquid culture.

Production of Pleurotus sajor-caju crude enzyme broth and its applicability for the removal of bisphenol A.

Bisphenol A is an endocrine interfering compound, produced and used on a large scale worldwide. Chemical and biologic methods can be used to remove it from the environment. Biological methods are considered less costly, safer and, according to green chemistry definitions, an environmentally correct method. Considering the use of a crude enzyme broth, without any downstream process, the costs could be mostly reduced. Thus, the removal of bisphenol A by Pleurotus sajor-caju crude enzyme broth was investigated. Initially, the agro-industrial wastes were characterized and, the composition of the culture medium and the bioreactor culture conditions were defined. The enzyme produced in the highest concentration was characterized and the crude broth used in the bisphenol A removal assays. The OXI45 culture medium presented the highest laccase activity (1,850.7 U L-1, 350 rpm). Greater laccase stability was observed at 20 - 40 oC and pHs 5 - 7. Vanillin and ferulic acid (considered mediator compounds) were identified in the crude broth, probably helping on the obtention of the high value of removal effectiveness (0.052 mg U-1 h-1). The results indicate the potential use of the Pleurotus sajor-caju crude enzyme broth to obtain an enzymatic formulation for application in the environmental area.

A polysaccharide fraction extracted from Pleurotus ostreatus mycelial biomass inhibit Sarcoma 180 tumor.

Fungi of Pleurotus genus have attracted a great interest due to their medicinal properties such as anti-inflammatory, antimicrobial and antitumor. These properties are attributed mainly to polysaccharides synthesized by Pleurotus. This work aimed to study the mycelial growth of P. ostreatus in submerged culture, evaluating the influence of the initial concentration of substrate (20 and 40 g/L of glucose) and the pH (4 and 6) on kinetic parameters of production of biomass. The effectiveness of different doses (10, 30 and 50 mg/kg) of a mycelium polysaccharide fraction extracted from P. ostreatus in reducing Sarcoma 180 development in mice was also verified. In the range of this study, maximum concentration of mycelial biomass (about 12.8 g/L) was obtained using 40.0 g/L of glucose, at pH 4.0. The total biomass productivity (Px) was not significantly affected by substrate concentration and pH, reaching values of 0.034 g/L.h. Sarcoma 180 tumor weight was reduced in 74.1, 75.5 and 53.7% when 10, 30 and 50 mg/kg were administered, respectively. These results show the high antitumor potential of intracellular polysaccharide fraction of mycelial biomass of P. ostreatus, particularly at lower doses of 10 and 30 mg/kg.

A polysaccharide fraction extracted from Pleurotus ostreatus mycelial biomass inhibit Sarcoma 180 tumor

ABSTRACT Fungi of Pleurotus genus have attracted a great interest due to their medicinal properties such as anti-inflammatory, antimicrobial and antitumor. These properties are attributed mainly to polysaccharides synthesized by Pleurotus. This work aimed to study the mycelial growth of P. ostreatus in submerged culture, evaluating the influence of the initial concentration of substrate (20 and 40 g/L of glucose) and the pH (4 and 6) on kinetic parameters of production of biomass. The effectiveness of different doses (10, 30 and 50 mg/kg) of a mycelium polysaccharide fraction extracted from P. ostreatus in reducing Sarcoma 180 development in mice was also verified. In the range of this study, maximum concentration of mycelial biomass (about 12.8 g/L) was obtained using 40.0 g/L of glucose, at pH 4.0. The total biomass productivity (Px) was not significantly affected by substrate concentration and pH, reaching values of 0.034 g/L.h. Sarcoma 180 tumor weight was reduced in 74.1, 75.5 and 53.7% when 10, 30 and 50 mg/kg were administered, respectively. These results show the high antitumor potential of intracellular polysaccharide fraction of mycelial biomass of P. ostreatus, particularly at lower doses of 10 and 30 mg/kg.

Exopolysaccharide produced by Pleurotus sajor-caju: its chemical structure and anti-inflammatory activity.

Edible mushrooms are high nutritional value foods, which contain proteins, fibers, minerals, vitamins, and carbohydrates. Among their carbohydrates are some polysaccharides with recognized therapeutic effects. It was reported in this manuscript the structural characterization and antinociceptive and anti-inflammatory activities of an exopolysaccharide (EPS) produced by Pleurotus sajor-caju. The purified EPS was a mannogalactan (PEIsR), which was composed by mannose (37.0%), galactose (39.7%), and 3-O-methyl-galactose (23.3%). The polysaccharide was purified by freeze-thawing and dialysis, and it was characterized by GC-MS analysis and NMR spectroscopy. The mannogalactan is constituted by a main chain of (1 → 6)-linked α-D-Galp and 3-O-methyl-α-D-Galp units. Some of the α-D-Galp units were substituted at O-2 by non-reducing end units of ß-D-Manp. According to the literature review conducted, this is the first time that a methylated polysaccharide was observed on EPS of P. sajor-caju. The mannogalactan was able to reduce the nociception, in vivo, in the writhing and formalin tests and also reduced the carrageenan-induced paw edema, which indicates that it could be an effective antinociceptive and anti-inflammatory agent.

Structural characterization and anti-inflammatory activity of a linear ß-D-glucan isolated from Pleurotus sajor-caju.

Glucans comprise an important class of polysaccharides present in basidiomycetes with potential biological activities. A (1 → 3)-ß-D-glucan was isolated from Pleurotus sajor-caju via extraction with hot water followed by fractionation by freeze-thawing and finally by dimethyl sulfoxide extraction. The purified polysaccharide showed a (13)C-NMR spectrum with six signals consisting of a linear glucan with a ß-anomeric signal at 102.8 ppm and a signal at 86.1 ppm relative to O-3 substitution. The other signals at 76.2, 72.9, 68.3, and 60.8 ppm were attributed to C5, C2, C4, and C6, respectively. This structure was confirmed by methylation analysis, and HSQC studies. The ß-d-glucan from P. sajor-caju presented an immunomodulatory activity on THP-1 macrophages, inhibited the inflammatory phase of nociception induced by formalin in mice, and reduced the number of total leukocytes and myeloperoxidase levels induced by LPS. Taken together, these results demonstrate that this ß-d-glucan exhibits a significant anti-inflammatory activity.