Novel Bifunctional Acylase from Actinoplanes utahensis: A Versatile Enzyme to Synthesize Antimicrobial Compounds and Use in Quorum Quenching Processes

Many intercellular communication processes, known as quorum sensing (QS), are regulated by the autoinducers N-acyl-L-homoserine lactones (AHLs) in Gram-negative bacteria. The inactivation of these QS processes using different quorum quenching (QQ) strategies, such as enzymatic degradation of the autoinducers or the receptor blocking with non-active analogs, could be the basis for the development of new antimicrobials. This study details the heterologous expression, purification, and characterization of a novel N-acylhomoserine lactone acylase from Actinoplanes utahensis NRRL 12052 (AuAHLA), which can hydrolyze different natural penicillins and N-acyl-homoserine lactones (with or without 3-oxo substitution), as well as synthesize them. Kinetic parameters for the hydrolysis of a broad range of substrates have shown that AuAHLA prefers penicillin V, followed by C12-HSL. In addition, AuAHLA inhibits the production of violacein by Chromobacterium violaceum CV026, confirming its potential use as a QQ agent. Noteworthy, AuAHLA is also able to efficiently synthesize penicillin V, besides natural AHLs and phenoxyacetyl-homoserine lactone (POHL), a nonnatural analog of AHLs that could be used to block QS receptors and inhibit signal of autoinducers, being the first reported AHL acylase capable of synthesizing AHLs

A Novel Lipase from Streptomyces exfoliatus DSMZ 41693 for Biotechnological Applications

Genome mining of Streptomyces exfoliatus DSMZ 41693 has allowed us to identify four different lipase-encoding sequences, and one of them (SeLipC) has been successfully cloned and extracellularly expressed using Rhodococcus sp. T104 as a host. SeLipC was purified by one-step hydrophobic interaction chromatography. The enzyme is a monomeric protein of 27.6 kDa, which belongs to subfamily I.7 of lipolytic enzymes according to its phylogenetic analysis and biochemical characterization. The purified enzyme shows the highest activity at 60 °C and an optimum pH of 8.5, whereas thermal stability is significantly improved when protein concentration is increased, as confirmed by thermal deactivation kinetics, circular dichroism, and differential scanning calorimetry. Enzyme hydrolytic activity using p-nitrophenyl palmitate (pNPP) as substrate can be modulated by different water-miscible organic cosolvents, detergents, and metal ions. Likewise, kinetic parameters for pNPP are: KM = 49.6 µM, kcat = 57 s−1, and kcat/KM = 1.15 × 106 s−1·M−1. SeLipC is also able to hydrolyze olive oil and degrade several polyester-type polymers such as poly(butylene succinate) (PBS), poly(butylene succinate)-co-(butylene adipate) (PBSA), and poly(ε-caprolactone) (PCL). Moreover, SeLipC can catalyze the synthesis of different sugar fatty acid esters by transesterification using vinyl laurate as an acyl donor, demonstrating its interest in different biotechnological applications.Ministerio de Ciencia, Innovación y Universidades (España)Sección Deptal. de Bioquímica y Biología Molecular (Biológicas)Depto. de Química en Ciencias FarmacéuticasFac. de Ciencias BiológicasFac. de FarmaciaTRUEpu

Development of active learning strategies in Biochemistry Laboratory BBM1

Con objeto de fomentar el aprendizaje activo en el laboratorio de Bioquímica BBM1, se diseñaron, emplearon y evaluaron una serie de actividades y herramientas: (i) test pre-laboratorio, realizados mediante la aplicación Socrative; (ii) herramientas de apoyo al aprendizaje autónomo del estudiante, relacionadas con la elaboración del cuaderno de laboratorio y los procesos de evaluación; (iii) actividades formativas complementarias, tales como el desarrollo de un mini-congreso y una visita a las instalaciones del Departamento. El objetivo final del proyecto es que el estudiante entienda de verdad lo que hace en el laboratorio y sea capaz de aplicar lo que ha aprendido a nuevas situaciones.In order to promote active learning in the BBM1 Biochemistry laboratory, several activities and tools were designed, used and evaluated: (i) pre-laboratory tests, carried out using Socrative application; (ii) tools to support the student's autonomous learning, related to preparation of the laboratory notebook and evaluation processes; (iii) supplementary training activities, such as development of a mini-congress and a visit to the Department's facilities. The ultimate goal of the project is that the student really understands what he does in the laboratory and is able to apply what he has learned to new situations.Depto. de Bioquímica y Biología MolecularFac. de Ciencias QuímicasFALSEsubmitte

Applied Biocatalysis: effective catalysis for sustainable development

Proyecto de aprendizaje basado en investigación grupal, tutorizada, modular e integrable en diversos títulos y asignaturas de ciencias experimentales e ingeniería química y bioquímicaUniversidad Complutense de Madrid. Vicerrectorado de Calidad.Depto. de Ingeniería Química y de MaterialesFac. de Ciencias QuímicasFALSEsubmitte

Laboratorio virtual en Biotecnología: aplicación y evaluación

La implementación de prácticas virtuales mediante plataformas de simulación experimental en los estudios de máster en el área de la Biotecnología y otras áreas afines de grado, con el fin de avanzar en la digitalización e internacionalización de las asignaturas, evaluando diferentes recursos de varias plataformas comerciales y abierta