The extraction of disease specific information from Fourier transform infrared (FTIR) spectra human body fluids demands the highest standards accuracy and reproducibility measurements because expected spectral differences between healthy diseased subjects are very small in relation to a large background absorbance whole sample. Here, we demonstrate that with increased sensitivity modern FTIR spectrometers, automatisation sample preparation bioinformatics, it is possible identify validate...

Certain heavy metal ions such as copper and zinc serve essential cofactors of many enzymes, but are toxic at high concentrations. Thus, intracellular levels have to be subtly balanced. P-type ATPases the P(IB)-subclass play a major role in homeostasis. The thermoacidophile Sulfolobus solfataricus possesses two P(IB)-ATPases named CopA CopB. Both enzymes present cells grown copper-depleted medium accumulated upon an increase external concentration. We studied physiological roles both by...

Pseudomonas aeruginosa uses quorum sensing (QS) as a cell-to-cell communication system to orchestrate the expression of virulence determinants. The biosynthesis important quinolone signal (PQS) requires pqsABCDE operon. Here, PqsE acts pathway-specific thioesterase, but it also contributes regulation bacterial via an unknown mechanism. In this manuscript, we report discovery inhibitors tool compounds gain further insights into its different functions. Differential scanning fluorimetry (DSF)...

Pseudomonas aeruginosa is an important nosocomial pathogen that frequently recalcitrant to available antibiotics, underlining the urgent need for alternative therapeutic options against this pathogen. Targeting virulence functions a promising strategy as it expected generate less-selective resistance treatment compared antibiotics. Capitalizing on our nonligand-based benzamide-benzimidazole (BB) core structure compounds reported efficiently block activity of P. multiple factor regulator...

P1 B -ATPases are among the most common resistance factors to metal-induced stress. Belonging superfamily of P-type ATPases, they capable exporting transition metal ions at expense adenosine triphosphate (ATP) hydrolysis. share a conserved structure three cytoplasmic domains linked by transmembrane domain. In addition, possess unique class located N-terminus. bacteria, these primarily associated with binding and either occur individually or as serial copies each other. Within this study,...

Alkyl hydroxyquinoline N-oxides (AQNOs) are antibiotic compounds produced by the opportunistic bacterial pathogen Pseudomonas aeruginosa They products of alkyl quinolone (AQ) biosynthetic pathway, which also generates quorum-sensing molecules 2-heptyl-4(1H)-quinolone (HHQ) and 2-heptyl-3-hydroxy-4(1H)-quinolone (PQS). Although enzymatic synthesis HHQ PQS had been elucidated, route AQNOs synthesized remained elusive. Here, we report on PqsL, key enzyme for AQNO production, structurally...

Synthesis of azetidine-derived natural products by the opportunistic pathogen Pseudomonas aeruginosa is controlled quorum sensing, a process involving production and sensing diffusible signal molecules that decisive for virulence regulation. In this study, we engineered P. titratable expression biosynthetic aze gene cluster, which allowed purification identification two new products, azetidomonamide C diazetidomonapyridone. Diazetidomonapyridone was shown to have highly unusual structure...

Summary Alkyl quinolones (AQs) are multifunctional bacterial secondary metabolites generally known for their antibacterial and algicidal properties. Certain representatives also employed as signalling molecules of Burkholderia strains Pseudomonas aeruginosa . The marine Gammaproteobacterium Microbulbifer sp. HZ11 harbours an AQ biosynthetic gene cluster with unusual topology but does not produce any AQ‐type under laboratory conditions. In this study, we demonstrate the potential strain...

Light-dependent or light-stimulated catalysis provides a multitude of perspectives for implementation in technological biomedical applications. Despite substantial progress made the field photobiocatalysis, number usable light-responsive enzymes is still very limited. Flavoproteins have exceptional potential photocatalytic applications because name-giving cofactor intrinsically features light-dependent reactivity, undergoing photoreduction with variety organic electron donors. However, vast...

Bile salts such as cholate are surface-active steroid compounds with functions for digestion and signaling in vertebrates. Upon excretion into soil water bile an electron- carbon-rich growth substrate environmental bacteria. Degradation of proceeds via intermediates a 3-keto-Δ1,4 -diene structure the skeleton shown e.g. Pseudomonas spp. Recently, we isolated bacteria degrading 3-keto-7-deoxy-Δ4,6 -structure suggesting existence second pathway degradation. This potential new was investigated...

The nosocomial pathogen Pseudomonas aeruginosa regulates its virulence via a complex quorum sensing network, which, besides N -acylhomoserine lactones, includes the alkylquinolone signal molecules 2-heptyl-3-hydroxy-4(1 H )-quinolone ( quinolone [PQS]) and 2-heptyl-4(1 (HHQ). Mycobacteroides abscessus subsp. , an emerging pathogen, is capable of degrading PQS also HHQ. Here, we show that although M.

Summary Programmed ribosomal frameshifting (PRF) is a translational anomaly causing the ribosome to shift into an alternative reading frame. PRFs are common in viral genomes, using single nucleotide sequence code for two proteins overlapping frames. In bacteria and eukaryota, less frequent. We report on PRF copper detoxification system of Escherichia coli where metallochaperone generated out first 69 amino acids C‐terminal out‐of‐frame glycine gene copA . besides codes P 1B ‐ATPase CopA,...

In contrast to many steroid hormones and cholesterol, mammalian bile salts are 5β-steroids, which leads a bent structure of the core. Bile surface-active steroids excreted into environment in large amounts, where they subject bacterial degradation. Bacterial degradation is initiated by oxidation A-ring leading canonical Δ 4 -3-keto with double bond A-ring. For 5β-bile salts, this -double introduced 3-keto-bile 5β-Δ -ketosteroid dehydrogenase (5β-Δ -KSTD). With Nov2c019 protein from bile-salt...

The opportunistic pathogen Pseudomonas aeruginosa uses quorum sensing to control its virulence. One of major signal molecules, the quinolone PQS, has high affinity membranes and is known be trafficked mainly via outer membrane vesicles (OMVs). We previously reported that several 3-hydroxy-4(1H)-quinolone 2,4-dioxygenases (HQDs) catalyze cleavage PQS thus act as quenching enzymes. Further analysis showed that, in contrast other HQDs, activity HQD from Streptomyces bingchenggensis (HQDS.b.)...

Many natural products comprise N ‐ O containing functional groups with crucial roles for biological activity. Their enzymatic formation is predominantly achieved by oxidation of an amine to form a hydroxylamine, which enables further functionalization. N‐ hydroxylation flavin‐dependent enzymes has so far been attributed distinct group flavoprotein monooxygenases (FPMOs) two dinucleotide binding domains. Here, we present three hydroxylases that exhibit glutathione reductase 2 (GR2)‐type...

Heavy metals can cause damage to biomolecules such as proteins and DNA in multiple ways. Cells therefore strive for keeping intracellular (heavy) metal ions bound specific that are capable of handling detoxification, export or integration cofactors. Metal binding usually provide coordination sites bind certain with ultrahigh affinity, the thermodynamic driving force being stability organometallic complexes. However, properties these be highly variable. Therefore transfer between separate...

The multiple biological activities of 2-alkylquinolones (AQs) are crucial for virulence Pseudomonas aeruginosa, conferring advantages during infection and in polymicrobial communities. Whereas 2-heptyl-3-hydroxyquinolin-4(1H)-one (the "Pseudomonas quinolone signal" [PQS]) is an important quorum sensing signal molecule, 2-alkyl-1-hydroxyquinolin-4(1H)-ones (also known as 2-alkyl-4-hydroxyquinoline N-oxides [AQNOs]) antibiotics inhibiting respiration. Hydroxylation the PQS precursor...

Pseudomonas aeruginosa is a major threat to public health. Its ability thrive in clinical settings, water distribution systems, or even jet fuel tanks linked detoxification and degradation of diverse hydrophobic substrates that are metabolized via alcohol intermediates.