Pre-treatment of a sugarcane bagasse-based substrate prior to saccharification: Effect of coffee pulp and urea on laccase and cellulase activities of Pycnoporus sanguineus.

Production of second-generation bioethanol uses lignocellulose from agricultural by-products such as sugarcane bagasse (SCB). A lignocellulose pre-treatment is required to degrade lignin, ensuring further efficient saccharification. Two experimental designs were set up to define culture conditions of Pycnoporus sanguineus in mesocosms to increase laccase activities and thus delignification. The first experimental design tested the effect of phenolic complementation (via coffee pulp) and the use of urea as a simple nitrogen source and the second defined more precisely the percentages of coffee pulp and urea to enhance delignification. The responses measured were: lignocellulolytic activities, laccase isoform profiles by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the chemical transformation of the substrate using solid-state NMR of 13C. Adding 10% of coffee pulp increased laccase activities and fungal biomass (32.5% and 16% respectively), enhanced two constitutive isoforms (Rf 0.23 and 0.27), induced a new isoform (Rf 0.19) and led to a decrease in total aromatics. However, higher concentrations of coffee pulp (25%) decreased laccase and cellulase activities but no decrease in aromaticity was observed, potentially due to the toxic effect of phenols from coffee pulp. Moreover, laccase production was still inhibited even for lower concentrations of urea (0-5%). Our findings revealed that an agricultural by-product like coffee pulp can enhance laccase activity -though to a threshold- and that urea limited this process, indicating that other N-sources should be tested for the biological delignification of SCB.

Proximal Composition, Nutraceutical Properties, and Acute Toxicity Study of Culinary-Medicinal Oyster Mushroom Powder, Pleurotus ostreatus (Agaricomycetes).

A compositional study was performed on fruiting-body powder of the culinary-medicinal oyster mushroom Pleurotus ostreatus for applications as a nutraceutical/functional food. Carbohydrates (55 g/100 g dry weight [dw]) and proteins (27.45 g/100 g dw, with an in vitro digestibility of 75%) appear to be the major components, but fat content was low (4 g/100 g dw). Pleurotus powder has important micronutrients such as minerals (Fe, Cu, Zn, Mn, Mg, and Co) and ascorbic acid, as well as nonnutrients (i.e., phenolics) with antioxidant potential. A powder-derived aqueous extract had a phenolic compound content of 138 mg/100 g that showed 2,2-diphenyl-1-picrylhydrazyl radical scavenging and inhibition of membrane-lipid peroxidation activities of 58.3% and 61.4%, respectively. The presence of ß-1,3-1,6-D-glucans was also demonstrated (1.54 g/100 g). An acute toxicity test proved that Pleurotus powder was safe after oral administration to both male and female mice at a dose of 2000 mg/kg. The combination of rich nutritional composition, bioactivity, and safety in P. ostreatus fruiting-body powder highlights its potential as a nutraceutical agent promoting health and life quality.

Do spawn storage conditions influence the colonization capacity of a wheat-straw-based substrate by Agaricus subrufescens?

Storage conditions of the spawn of edible fungi are of major importance to facilitate the production of mushrooms. Here, standard storage conditions at 10°C or 15°C were used and the potential of colonization of standard European compost by the tropical species Agaricus subrufescens was assessed during the spawn running phase. Two lignocellulolytic activities, laccase and CMC-cellulase, were enhanced after storage compared to control as well as substrate transformation, as described by the aromaticity ratio and a humification ratio calculated from NMR data. This result indicates that mycelium growth probably occurred during storage at 10 or 15°C, leading to a larger amount of biomass in the inoculum. Moreover, the microbial functional diversity of the substrate was favored, showing that the electivity of the substrate was maintained. Thus, these findings indicate that recommendations for the mushroom producers can be established for A. subrufescens cultivation under European standard conditions.

Microcultures of lactic acid bacteria: characterization and selection of strains, optimization of nutrients and gallic acid concentration.

Eighteen lactic acid bacteria (LAB) strains, isolated from coffee pulp silages were characterized according to both growth and gallic acid (GA) consumption. Prussian blue method was adapted to 96-well microplates to quantify GA in LAB microcultures. Normalized data of growth and GA consumption were used to characterize strains into four phenotypes. A number of 5 LAB strains showed more than 60% of tolerance to GA at 2 g/l; whereas at 10 g/l GA growth inhibition was detected to a different extent depending on each strain, although GA consumption was observed in seven studied strains (>60%). Lactobacillus plantarum L-08 was selected for further studies based on its capacity to degrade GA at 10 g/l (97%). MRS broth and GA concentrations were varied to study the effect on growth of LAB. Cell density and growth rate were optimized by response surface methodology and kinetic analysis. Maximum growth was attained after 7.5 h of cultivation, with a dilution factor of 1-1/2 and a GA concentration between 0.625 and 2.5 g/l. Results indicated that the main factor affecting LAB growth was GA concentration. The main contribution of this study was to propose a novel adaptation of a methodology to characterize and select LAB strains with detoxifying potential of simple phenolics based on GA consumption and tolerance. In addition, the methodology presented in this study integrated the well-known RSM with an experimental design based on successive dilutions.