d-Glucan from Fruiting Body and Mycelium of Cerrena unicolor (Bull.) Murrill: Structural Characterization and Use as a Novel Inducer of Mutanase

Water-insoluble, alkali-soluble polysaccharide (marked as ASP) was extracted from the vegetative mycelium and fruiting body of Cerrena unicolor strain. Monosaccharide examination of ASP demonstrated that the isolated biopolymer was composed mainly of glucose, xylose, and mannose monomers. The methylation investigation of studied polymers indicated that (1→3)-linked -d-Glcp is the major chain constituent (92.2% for glucans isolated from fruiting body and 90.1% from mycelium). 1 H NMR, FT-IR, and immunofluorescent labelling determinations confirmed that the polysaccharides isolated from both fruiting body and mycelium of . unicolor are (1→3)--d-glucans. The obtained (1→3)--d-glucans showed differences in viscosity and similar characteristics in optical rotations. (1→3)--d-Glucans extracted from mycelium and fruiting body of C. unicolor were also used as potential and specific inducers of mutanase synthesis by Trichoderma harzianum. The highest mutanase activity (0.38 U/mL) was obtained after induction of enzyme by (1→3)--d-glucan isolated from the mycelium of C. unicolor, and this biopolymer has been suggested as a new alternative to streptococcal mutan for the mutanase induction in T. harzianum. (1→3)--d-Glucan-induced mutanase showed high hydrolysis potential in reaction with dextranase-pretreated mutan, where maximal degree of saccharification and solubilization of this bacterial homoglucan (83.1% and 78.4%, resp.) was reached in 3 h at 45 ∘ C

Female within-nest spawning-site preference in a paternal brooding blenny and its effect on the female mate choice

Female spawning-site preference within a nest and its effect on the female mate choice in a paternal brooding blenny Rhabdoblennius ellipes, were examined in rocky intertidal pools using artificial nests. The number of eggs deposited at the nest entrance site was lower than the number deposited at the center and at deep sites. Moreover, the within-nest difference in the egg developmental stage indicated that eggs at the nest entrance site were deposited after those at the center and deep sites. These results indicated that females prefer to spawn eggs in the center and deep sites rather than at the entrance site. Owing to the higher egg mortality rate at the entrance site, females may avoid spawning at such sites. An analysis of the nests in the study area suggested that the within-nest site preference affects female mate choice that is females avoided nests where only the entrance site was available for spawning and instead spawned in nests where the center and deep sites were available

Perspektywy zastosowania biotechnologii w produkcji lotnych zwiazkow smakowo-zapachowych

Praca prezentuje obecny stan badań nad mikrobiologiczną produkcją naturalnych związków smakowo-zapachowych, zwłaszcza laktonów, związków aromatycznych (waniliny i aldehydu benzoesowego) oraz terpenów. Szczególny nacisk położono na przedstawienie zalet, ograniczeń oraz perspektyw wykorzystania w tym celu procesów biotransformacji.This rewiev presents the current state of the art of microbiological production of natural flavours, particulary lactones, aromatic compounds (vanillin, benzaldehyde) and terpenes. Special emphasis is placed on advantages, disadvantages and prospects for application to this end biotransformation processes

Optimization of mutanase production by Trichoderma harzianum

The present paper describes optimization of fermentation conditions in shaken flasks and scale-up of fermentor production up to 115 L. The response surface methodology (RSM) has been successfully applied in standardization of mutanase production by Trichoderma harzianum CCM F-340. The model was very well fitted to the experimental data and explained more than 96% of the whole variation of the response (adjusted R2 = 0.962). In order to confirm the adequacy of the regression model based on the experimental data, validation cultures were grown in conditions created through optimization. The highest enzyme activity (0.747 U/mL) was reached in shaken flask cultures on Mandels’ medium in a volume of 140 mL modified in terms of carbon (cell wall preparation from the polypore fungus Laetiporus sulphureus 8.08 g/L) and nitrogen (soybean peptone 1.38 g/L) sources, under culture conditions 30°C, pH 5.3, agitation 270 rpm. The scale-up of the culture in the bioreactors with a working volume of 5 and 115 L resulted in a slight decrease in the mutanase activity (0.734 and 0.682 U/mL, respectively). The validation experiment showed a 70.6% increase in the production of mutanase compared with the culture before optimization. The results proved that the cultures could be scaled-up successfully from shaken flasks to the bioreactor scale. Our results indicate that in optimal conditions, T. harzianum could be a highly effective extracellular mutanase source. This report is the first to deal with optimization of mutanase biosynthesis using a mathematical model and scale-up of enzyme production in controlled fermentors with a view to facilitate application thereof in industry.Keywords: Mutanase, Trichoderma harzianum, response surface methodology (RSM), bioreactors, submerged culture.African Journal of Biotechnology, Vol 13(25) 2538-254

Assessing biological activity of carboxymethylated derivatives of alpha-(1→3)-glucans isolated from fruiting bodies of cultivated pleurotus species

α-(1→3)-Glukany stanowią najmniej poznaną grupę polisacharydów budujących ścianę komórkową grzybów. Celem pracy była ocena aktywności biologicznej karboksymetylowanych pochodnych α-(1→3)-glukanów uzyskanych z owocników uprawnych gatunków rodzaju Pleurotus (P. citrinopileatus, P. djamor, P. erynii i P. precoce). Polisacharydy wyizolowane z owocników poddano analizom strukturalnym (FT-IR i ¹H NMR), w wyniku czego wykazano, że są one liniowymi α-(1→3)-glukanami. Po zastosowaniu metody karboksymetylacji nierozpuszczalne w wodzie α-(1→3)-glukany przeprowadzono w formy rozpuszczalne, tj. karboksymetyl-α-(1→3)-glukany (KM-α-(1→3)-glukany). Następnie analizowano ich wpływ na żywotność komórek prawidłowych CCD 841 CoTr (ludzkie komórki nabłonka jelita grubego) i CCD-18Co (miofibroblasty jelitowe) oraz komórek nowotworowych HeLa (ludzki rak szyjki macicy). Wykonano analizy aktywności metabolicznej i toksyczności (MTT i NR) oraz zdolności do redukcji wolnych rodników (DPPH). Wykazano, że żywotność komórek nowotworowych była najsilniej redukowana przez KM-α-(1→3)-glukan otrzymany z owocników P. citrinopileatus. Z kolei metodą NR wykazano, że wszystkie badane KM-α-glukany w całym zakresie stężeń powodowały statystycznie istotny wzrost żywotności komórek prawidłowych (CCD 841 CoTr i CCD-18Co). Badane KM-α-glukany nie wykazywały aktywności zmierzającej do redukcji wolnych rodników tlenowych. Na podstawie analizy wyników różnicowego barwienia fluorescencyjnego stwierdzono, że KM-α-glukan z owocników P. citrinopileatus zaburza integralność błony komórek nowotworowych. Z kolei barwienie fluorescencyjne filamentów F-aktynowych (F-aktyny) wykazało, że nie wpływał on destrukcyjnie na cytoszkielet badanych komórek.α-(1→3)-Glucans constitute the least known group of polysaccharides that make up the fungal cell wall. The objective of the study was to assess the biological activity of carboxymethylated derivatives of α-(1→3)-glucans extracted from fruiting bodies of the cultivated species of Pleurotus (P. citrinopileatus, P. djamor, P. erynii, and P. precoce) genus. The polysaccharides extracted from the fruiting bodies were structurally analyzed (using FT-IR and ¹H NMR) and, based on the analysis results, it was proved that they were linear α-(1→3)-glucans. With the use of a carboxymethylation method, the water-insoluble α-(1→3)-glucans were converted into soluble forms, i.e. carboxymethyl-α-(1→3)-glucans (KM-α-(1→3)-glucans). Next, their effect was investigated on the viability of normal CCD 841 CoTr cells (normal human colonic epithelial cells), CCD-18Co (intestinal myofibroblasts), and HeLa tumour cells (human cervical cancer). The metabolic activity, toxicity (MTT and NR), and free radical scavenging ability (DPPH) were analyzed. It was shown that the effect of KM-α-(1→3)-glucan derived from the P. citrinopileatus fruiting bodies was the strongest as regards the reduction of the cancer cell viability. Then, the NR method used showed that all the tested KM-α-(1→3)-glucans within the entire concentration range caused the viability of normal cells (CCD 841 CoTr and CCD-18Co) to increase statistically significantly. The KM-α-glucans analyzed did not exhibit any free oxygen radical reduction activity. Based on the results of the differential fluorescent staining analysis, it was confirmed that the KM-α-glucan from fruiting bodies of P. citrinopileatus disturbed the membrane integrity of tumour cells. On the other hand, the fluorescent staining of the F-actin filaments proved that this KM-α-glucan did not have any destructive effect on the cytoskeleton of the cells studied