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- New bacterial strains for ibuprofen biodegradation: drug removal, transformation, and potential catabolic genesPublication . Lara-Moreno, Alba; Costa, Maria Clara; Vargas‐Villagomez, Ayleen; Carlier, JorgeIbuprofen (IBU) is a significant contaminant frequently found in wastewater treatment plants due to its widespread use and limited removal during treatment processes. This leads to its discharge into the environment, causing considerable environmental concerns. The use of microorganisms has recently been recognized as a sustainable method for mitigating IBU contamination in wastewater. In this study, new bacteria capable of growing in a solid medium with IBU as the only carbon source and removing IBU from a liquid medium were isolated from environmental samples, including soil, marine, mine, and olive mill wastewater. Four bacterial strains, namely Klebsiella pneumoniae TIBU2.1, Klebsiella variicola LOIBU1.1, Pseudomonas aeruginosa LOIBU1.2, and Mycolicibacterium aubagnense HPB1.1, were identified through 16S rRNA gene sequencing. These strains demonstrated significant IBU removal efficiencies, ranging from 60 to 100% within 14 days, starting from an initial IBU concentration of 5 mg per litre. These bacteria have not been previously reported in the literature as IBU degraders, making this work a valuable contribution to further studies in the field of bioremediation in environments contaminated by IBU. Based on the IBU removal results, the most promising bacteria, K. pneumoniae TIBU2.1 and M. aubagnense HPB1.1, were selected for an in silico analysis to identify genes potentially involved in IBU biodegradation. Interestingly, in the tests with TIBU2.1, a peak of IBU transformation product(s) was detected by high-performance liquid chromatography, while in the tests with HPB1.1, it was not detected. The emerging peak was analysed by liquid chromatography-mass spectrometry, indicating the presence of possible conjugates between intermediates of IBU biodegradation. The proteins encoded on their whole-genome sequences were aligned with proteins involved in an IBU-degrading pathway reported in bacteria with respective catabolic genes. The analysis indicated that strain HPB1.1 possesses genes encoding proteins similar to most enzymes reported associated with the IBU metabolic pathways used as reference bacteria, while strain TIBU2.1 has genes encoding proteins similar to enzymes involved in both the upper and the lower part of that pathway. Notably, in the tests with the strain having more candidate genes encoding IBU-catabolic enzymes, no IBU transformation products were detected, while in the tests with the strain having fewer of these genes, detection occurred. This study explores the use of bacteria to mitigate ibuprofen (IBU) contamination in wastewater. Four new bacterial strains were identified that can grow with IBU as the only carbon source and demonstrated significant IBU removal efficiencies. In-depth analysis revealed that two strains, Klebsiella pneumoniae TIBU2.1 and Mycolicibacterium aubagnense HPB1.1, possess genes encoding proteins similar to enzymes involved in IBU degradation, providing a promising avenue for bioremediation in IBU-contaminated environments. image
- Batch studies on the biodegradation of paracetamol and 1,4-hydroquinone by novel bacterial strains isolated from extreme environmental samples and the identification of candidate catabolic genesPublication . Lara-Moreno, Alba; Fatma El-Sayed; Cox, Cymon; Costa, Maria Clara; Carlier, JorgeThe emerging pollutant paracetamol (APAP) is one of the most prescribed drugs worldwide. In addition, APAP and its main metabolites, namely, 4-aminophenol (4-AP), hydroquinone (H2Q), benzoquinone (BQ), and 2,5-dihydroxy-1,4-benzoquinone (2,5-OH-BQ), among others, are frequently detected in wastewater treatment plants (WWTPs) influents, effluents, and the environment. Thus, continuous release into the environment, especially aquatic environments, is a source of general concern. Six APAP-degrading bacterial strains were isolated from two mine samples from the Iberian Pyrite Belt (Lousal and Poderosa mines). Mycolicibacterium aubagnense HPB1.1, which was isolated using enrichment cultures from the Poderosa mine sample in the presence of H2Q as the sole carbon source, also showed APAP biodegrading capabilities. Pure cultures of this strain degraded 34.3 mg L-1 of APAP in 5 days and 9.4 mg L-1 of H2Q in 4 days. Interestingly, BQ and 2,5-OH-BQ were detected as metabolites resulting from H2Q abiotic degradation, but these compounds were removed in the strain's cultures. Furthermore, M. aubagnense HPB1.1 whole-genome was sequenced, and its encoded proteins were aligned with enzymes of APAP-degrading bacteria recovered from databases and literature aiming to identify candidate catabolic genes. Putative amidases, deaminases, hydroxylases, and dioxygenases, responsible for the degradation of APAP by the HPB1.1 strain, were identified by similarity, corroborating its ability to transform APAP and its intermediate metabolite H2Q into less toxic metabolic compounds due to their capacity to break the aromatic ring of these molecules.