Characterization of an extremophile bacterial acid phosphatase derived from metagenomics analysis.

Acid phosphatases are enzymes that play a crucial role in the hydrolysis of various organophosphorous molecules. A putative acid phosphatase called FS6 was identified using genetic profiles and sequences from different environments. FS6 showed high sequence similarity to type C acid phosphatases and retained more than 30% of consensus residues in its protein sequence. A histidine-tagged recombinant FS6 produced in Escherichia coli exhibited extremophile properties, functioning effectively in a broad pH range between 3.5 and 8.5. The enzyme demonstrated optimal activity at temperatures between 25 and 50°C, with a melting temperature of 51.6°C. Kinetic parameters were determined using various substrates, and the reaction catalysed by FS6 with physiological substrates was at least 100-fold more efficient than with p-nitrophenyl phosphate. Furthermore, FS6 was found to be a decamer in solution, unlike the dimeric forms of crystallized proteins in its family.

Synthesis of aromatic amino acids from 2G lignocellulosic substrates.

Pseudomonas putida is a highly solvent-resistant microorganism and useful chassis for the production of value-added compounds from lignocellulosic residues, in particular aromatic compounds that are made from phenylalanine. The use of these agricultural residues requires a two-step treatment to release the components of the polysaccharides of cellulose and hemicellulose as monomeric sugars, the most abundant monomers being glucose and xylose. Pan-genomic studies have shown that Pseudomonas putida metabolizes glucose through three convergent pathways to yield 6-phosphogluconate and subsequently metabolizes it through the Entner-Doudoroff pathway, but the strains do not degrade xylose. The valorization of both sugars is critical from the point of view of economic viability of the process. For this reason, a P. putida strain was endowed with the ability to metabolize xylose via the xylose isomerase pathway, by incorporating heterologous catabolic genes that convert this C5 sugar into intermediates of the pentose phosphate cycle. In addition, the open reading frame T1E_2822, encoding glucose dehydrogenase, was knocked-out to avoid the production of the dead-end product xylonate. We generated a set of DOT-T1E-derived strains that metabolized glucose and xylose simultaneously in culture medium and that reached high cell density with generation times of around 100 min with glucose and around 300 min with xylose. The strains grew in 2G hydrolysates from diluted acid and steam explosion pretreated corn stover and sugarcane straw. During growth, the strains metabolized > 98% of glucose, > 96% xylose and > 85% acetic acid. In 2G hydrolysates P. putida 5PL, a DOT-T1E derivative strain that carries up to five independent mutations to avoid phenylalanine metabolism, accumulated this amino acid in the medium. We constructed P. putida 5PLΔgcd (xylABE) that produced up to 250 mg l-1 of phenylalanine when grown in 2G pretreated corn stover or sugarcane straw. These results support as a proof of concept the potential of P. putida as a chassis for 2G processes.

Translation Into Spanish and Proposal to Modify the Orchard Sports Injury Classification System (OSICS) Version 12.

PURPOSE: To publish the Spanish translation of Version 12 of the sports injury classification system called Orchard Sports Injury Classification System and propose a modification to include a numerical code that reflects the impact of the injury on sports functionality. METHODS: The members of the working group on the epidemiology of sports injury, of the Group for the Study of the Muscle-Tendon System (GESMUTE), and of the Spanish Society of Sports Traumatology (SETRADE), carried out a bibliographical review on the epidemiological classification systems of injuries, 3 face-to-face consensus meetings, and various online pieces of work, following the Delphi work methodology. RESULTS: The Spanish translation of Version 12 of the Orchard Sports Injury Classification System is fully accessible and free of charge at https://gesmute.es/traduccion-espanola-osics-12/. The current project proposes to add, at the end of the current coding system of Version 12, a numerical code (0: No Functional impairment; 1: Limits Sports Activity; 2: Prevents Sports Activity; 3: Limits Daily Life Activities), to indicate any functional repercussions caused by the injury. CONCLUSION: We present the Spanish translation of Version 12 of the Orchard Sports Injury Classification System. We propose as an improvement the inclusion of functionality criteria in sports injury classifications; more specifically, our proposal could be an improvement to the Orchard Sports Injury Classification System Version 12.