Alachlor is a herbicide that is widely used worldwide to protect plant crops against broadleaf weeds and annual grasses. However, due to its endocrine-disrupting activity, its application has been banned in the European Union. As described in our earlier work, Paecilomyces marquandii is a microscopic fungus capable of alachlor removal by N-acetyl oxidation. Our current work employs proteomics and metabolomics to gain a better understanding of alachlor biodegradation by the microscopic fungus P. marquandii. The data revealed that the addition of alachlor reduced culture growth and glucose consumption rate. At the same time, the rates of glycolysis and the tricarboxylic acid (TCA) cycle increased during the initial stage of growth, and there was a shift toward the formation of supplementary materials (UDP-glucose/galactose) and reactive oxygen species (ROS) scavengers (ascorbate). Proteomic analysis revealed that the xenobiotic presence resulted in a strong upregulation of enzymes related to energy, sugar metabolism and ROS production. However, the unique overexpression of cyanide hydratase in alachlor-containing cultures may implicate this enzyme as the key protein involved in the alachlor biodegradation pathway. The characterization of P. marquandii-mediated alachlor removal in terms of cell structure and function resulted in a deeper insight into microorganism strategy toward xenobiotic biodegradation.

Abstract4-n-nonylphenol (4-n-NP) is an endocrine disrupting compound (EDC); pollutants that cause serious disturbances in the environment. This study shows the degradation pathway and initial proteome analysis in cultures of a fungus that actively degrades 4-n-NP, Metarhizium robertsii. The research revealed the presence of 14 4-n-NP metabolites formed as a result of the oxidation of the alkyl chain and benzene ring, which leads to the complete decomposition of the compound. Based on the trend and quantitative analysis of the formation of 4-n-NP derivatives, the best conditions for proteome analysis were established. The data collected allowed the formulation of an explanation of the microorganism's strategy towards the removal of 4-n-NP. The main groups of proteins engaged in the removal of the xenobiotic are: oxidation-reduction systems related to nitroreductaselike proteins, ROS defense systems (peroxiredoxin and superoxide dismutase), the TCA cycle and energyrelated systems. Principal components analysis was applied to unidentified proteins, resulting in the formulation of three subgroups and initial classification of these proteins.

The aim of this study was to overproduce, identify and apply novel laccase-like multicopper oxidases (LMCOs) from Myrothecium roridum in a dye removal process. LMCOs' production was enhanced by modifying the medium and adding copper ions. After purification, two proteins, LMCO1 and LMCO2, with molecular masses of 46.7 and 66.3 kDa were discovered. Peptide analysis by mass spectrometry revealed that they belong to the cupredoxin superfamily. Characteristic peptide sequences were obtained for MCOs and bilirubin oxidases. Crude enzymes were applied in a dye decolorization process. Supplementation with 1 mM of vanillin allowed an almost complete elimination of the Indigo carmine within 3 hours. The dye was removed from a solution containing metals, surfactants and organic solvents. The in-gel assessment of the activity and decolorization ability of MCOs, followed by protein extraction and SDS-PAGE, confirmed that only LMCO2 was responsible for the dye removal. MCOs produced by Myrothecium sp. have been poorly studied before. The obtained results broaden knowledge on this subject and may contribute to the development of an eco-friendly method of dye elimination.

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