Objective of this diploma thesis was to select the most appropriate composition and thermal treatment of a substrate and the optimal temperature for planting and development of the sporocarps of mushrooms Pleurotus eryngii and Pleurotus nebrodensis. Within the objective of this thesis there was a hypothesis that different levels of the thermal treatment of a substrate have an impact on the colonization of such substrate by mycelium and subsequent crop of sporocarps Pleurotus eryngii and Pleurotus nebrodensis and that within those selected strains there can be found a strain which tolerates temperatures in a grow room above 15 °C. Straw pellets, beech sawdust, spruce sawdust and Douglas tree sawdust were selected as substrates for the growth of sporocarps P. eryngii and P. nebrodensis. Some substrates were enhanced by a wheat bran or coniferous sawdust was fermented without and with 5% fugate. Obtained data were statistically processed using the Statistica (StatSoft) programme by the ANOVA method. Statistical assessment of results is presented graphically in the thesis. Results of the experiment proved that Pleurotus eryngii and Pleurotus nebrodensis have the highest growth of their sporocarps on the substrate from fermented coniferous sawdust with added fugate. The results further shown that the mushrooms had higher growth when they were on the substrate without a wheat bran compared to the substrate with a wheat bran, which was even more prone to its contamination. Optimal temperature for planting and development of the sporocarps Pleurotus eryngii was determined to be 12 - 15 °C. There were significant differences in the fructification of P. eryngii and P. nebrodensis which produced only minimal crop. Duration of a growing cycle of Pleurotus eryngii was 72 - 134 days. Development of sporocarps lasted 11 - 21 days.

Enzymatic oxidation of 5-hydroxymethylfurfural (HMF) and its oxidized derivatives was studied using three fungal enzymes: wild-type aryl alcohol oxidase (AAO) from three fungal species, wild-type peroxygenase from Agrocybe aegerita (AaeUPO), and recombinant galactose oxidase (GAO). The effect of pH on different reaction steps was evaluated and apparent kinetic data (Michaelis-Menten constants, turnover numbers, specific constants) were calculated for different enzyme-substrate ratios and enzyme combinations. Finally, the target product, 2,5-furandicarboxylic acid (FDCA), was prepared in a multi-enzyme cascade reaction combining three fungal oxidoreductases at micro-scale. Furthermore, an oxidase-like reaction is proposed for heme-containing peroxidases, such as UPO, horseradish peroxidase, or catalase, causing the conversion of 5-formyl-2-furancarboxylic acid into FDCA in the absence of exogenous hydrogen peroxide.

Enzymatic oxidation of 5-hydroxymethylfurfural (HMF) and its oxidized derivatives was studied using three fungal enzymes: wild-type aryl alcohol oxidase (AAO) from three fungal species, wild-type peroxygenase from Agrocybe aegerita (AaeUPO), and recombinant galactose oxidase (GAO). The effect of pH on different reaction steps was evaluated and apparent kinetic data (Michaelis-Menten constants, turnover numbers, specific constants) were calculated for different enzyme-substrate ratios and enzyme combinations. Finally, the target product, 2,5-furandicarboxylic acid (FDCA), was prepared in a multi-enzyme cascade reaction combining three fungal oxidoreductases at micro-scale. Furthermore, an oxidase-like reaction is proposed for heme-containing peroxidases, such as UPO, horseradish peroxidase, or catalase, causing the conversion of 5-formyl-2-furancarboxylic acid into FDCA in the absence of exogenous hydrogen peroxide.