However, little is known about their biological role or functional interactions with other fungal biomolecules. Noteworthy, AuAHLA is also able to efficiently synthesize penicillin V, besides natural AHLs and phenoxyacetyl-homoserine lactone (POHL), a non-natural 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.Īntifungal proteins (AFPs) from ascomycete fungi could help the development of antimycotics. In addition, AuAHLA inhibits the production of violacein by Chromobacterium violaceum CV026, confirming its potential use as a QQ agent. Kinetic parameters for the hydrolysis of a broad range of substrates have shown that AuAHLA prefers penicillin V, followed by C12-HSL. 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. 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. Many intercellular communication processes, known as quorum sensing (QS), are regulated by the autoinducers N-acyl-l-homoserine lactones (AHLs) in Gram-negative bacteria. Keywords: Proteins Protein Identification Protein Structural Analysis protein purification Ĭhromatography ELISA Protein Quantitation Western blotting This chapter is intended to give a rudimentary understanding of the methodsĪnd technologies behind proteins and their application. Measures for protein content has lit up comparative and functional biology of Furthermore, the application of physicochemical Understanding the protein structures and functions. Sanger, proteomics, which is the large-scale study of proteins, became an essential tool in Since the first protein sequencing in 1953 by Frederic Protein study leading to a deeper understanding of their role in disease biochemical pathwayĪnd also in drug metabolism. A majorīreakthrough in protein purification technologies has greatly changed the landscape of Interaction is critical to understand cellular processes and rational drug design. Unraveling the importance of protein folding, protein�protein Being composed of aminoĪcids despite their similar structure, proteins are surrounded by several other molecules ofĭifferent chemical structures creating an overcrowded environment that makes them different
Being one of the impressing groups of macromolecules, proteins and their applicationĪre crucial to understand the molecular logic of living cells.
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