Study on the antibacterial properties of leathers tanned with natural tannins and their interactions with shoes inhabiting bacteria

Content: Tannins are high molecular weight polyphenols, naturally synthesized by plants to defend themselves against biotic and abiotic stress factors. Their role as antioxidant, antibiotic and antibacterial agent has been known for many years among agriculture, food, pharma and cosmetics industry. If tannins would perform an antibacterial activity in a vegetable tanned leather, the leather itself could be certified as an antibacterial material. This effect could be very interesting for all the applications in which the leather, being in contact with sweat and bacteria, becomes a solution to reduce more or less severe hyperhidrosis and bromhidrosis. The goal of the study was the assessment of the antibacterial activity of vegetable tanned leathers with natural tannins to produce articles in direct contact with human skin and, therefore, their effect on sweat, bacterial growth and metabolite production. Firstly, the antibacterial activity has been evaluated and compared between leathers tanned with Chestnut, Quebracho and Tara extracts, chrome tanned leathers and synthetic materials. The trial was performed in vitro by inoculating gram positive (Staphylococcus aureus) and gram negative (Escherichia coli) bacterial strains. A later step defined the most suitable blend of tannins to obtain, after tanning and/or retanning, an antibacterial natural leather. Furthermore, the vegetable tanned leathers, made with this tannins blend, have been the target of an in vivo trial during which 15 panelists have worn two differently made shoes. The lining and insole inside the right shoe have been made with vegetable tanned leathers with tannins, while the ones inside the left shoe contained only synthetic material. The shoes have been worn for 28 consecutive days, followed by a molecular and bioinformatic analysis of microbiota samples taken from the inner surface of the shoes by using a sterile swab. Lastly, a biochemical analysis of volatile short chain fatty acids has been carried out to investigate the byproducts of the bacteria responsible for the unpleasant odor of shoes. Take-Away: 1. Vegetable tanned leather is a wonderful antibacterial material thanks to the presence of natural tannins, such as chestnut, quebracho and tara. This property is appreciated in the production of insole leather, lining, leather goods and automotive interiors. 2. The problem of bromhidrosis (bad feet odor) can be avoided by using vegetable tanned leather. 3. In particular, vegetable leathers tanned with tannins used to make inside part of the shoes permit to avoid the formation of cheesy and acidic odours thanks to their antibacterial properties and their capacity to absorb sweat

ActS activates peptidoglycan amidases during outer membrane stress in <i>Escherichia coli</i>

The integrity of the cell envelope of E. coli relies on the concerted activity of multi-protein machineries that synthesize the peptidoglycan (PG) and the outer membrane (OM). Our previous work found that the depletion of lipopolysaccharide (LPS) export to the OM induces an essential PG remodeling process involving LD-transpeptidases (LDTs), the glycosyltransferase function of PBP1B and the carboxypeptidase PBP6a. Consequently, cells with defective OM biogenesis lyse if they lack any of these PG enzymes. Here we report that the morphological defects, and lysis associated with a ldtF mutant with impaired LPS transport, are alleviated by the loss of the predicted OM-anchored lipoprotein ActS (formerly YgeR). We show that ActS is an inactive member of LytM-type peptidoglycan endopeptidases due to a degenerated catalytic domain. ActS is capable of activating all three main periplasmic peptidoglycan amidases, AmiA, AmiB, and AmiC, which were previously reported to be activated only by EnvC and/or NlpD. Our data also suggest that in vivo ActS preferentially activates AmiC and that its function is linked to cell envelope stress

Study on the antibacterial properties of leathers tanned with natural tannins and their interactions with shoes inhabiting bacteria

Content: Tannins are high molecular weight polyphenols, naturally synthesized by plants to defend themselves against biotic and abiotic stress factors. Their role as antioxidant, antibiotic and antibacterial agent has been known for many years among agriculture, food, pharma and cosmetics industry. If tannins would perform an antibacterial activity in a vegetable tanned leather, the leather itself could be certified as an antibacterial material. This effect could be very interesting for all the applications in which the leather, being in contact with sweat and bacteria, becomes a solution to reduce more or less severe hyperhidrosis and bromhidrosis. The goal of the study was the assessment of the antibacterial activity of vegetable tanned leathers with natural tannins to produce articles in direct contact with human skin and, therefore, their effect on sweat, bacterial growth and metabolite production. Firstly, the antibacterial activity has been evaluated and compared between leathers tanned with Chestnut, Quebracho and Tara extracts, chrome tanned leathers and synthetic materials. The trial was performed in vitro by inoculating gram positive (Staphylococcus aureus) and gram negative (Escherichia coli) bacterial strains. A later step defined the most suitable blend of tannins to obtain, after tanning and/or retanning, an antibacterial natural leather. Furthermore, the vegetable tanned leathers, made with this tannins blend, have been the target of an in vivo trial during which 15 panelists have worn two differently made shoes. The lining and insole inside the right shoe have been made with vegetable tanned leathers with tannins, while the ones inside the left shoe contained only synthetic material. The shoes have been worn for 28 consecutive days, followed by a molecular and bioinformatic analysis of microbiota samples taken from the inner surface of the shoes by using a sterile swab. Lastly, a biochemical analysis of volatile short chain fatty acids has been carried out to investigate the byproducts of the bacteria responsible for the unpleasant odor of shoes. Take-Away: 1. Vegetable tanned leather is a wonderful antibacterial material thanks to the presence of natural tannins, such as chestnut, quebracho and tara. This property is appreciated in the production of insole leather, lining, leather goods and automotive interiors. 2. The problem of bromhidrosis (bad feet odor) can be avoided by using vegetable tanned leather. 3. In particular, vegetable leathers tanned with tannins used to make inside part of the shoes permit to avoid the formation of cheesy and acidic odours thanks to their antibacterial properties and their capacity to absorb sweat

Study on the antibacterial properties of leathers tanned with natural tannins and their interactions with shoes inhabiting bacteria

Content: Tannins are high molecular weight polyphenols, naturally synthesized by plants to defend themselves against biotic and abiotic stress factors. Their role as antioxidant, antibiotic and antibacterial agent has been known for many years among agriculture, food, pharma and cosmetics industry. If tannins would perform an antibacterial activity in a vegetable tanned leather, the leather itself could be certified as an antibacterial material. This effect could be very interesting for all the applications in which the leather, being in contact with sweat and bacteria, becomes a solution to reduce more or less severe hyperhidrosis and bromhidrosis. The goal of the study was the assessment of the antibacterial activity of vegetable tanned leathers with natural tannins to produce articles in direct contact with human skin and, therefore, their effect on sweat, bacterial growth and metabolite production. Firstly, the antibacterial activity has been evaluated and compared between leathers tanned with Chestnut, Quebracho and Tara extracts, chrome tanned leathers and synthetic materials. The trial was performed in vitro by inoculating gram positive (Staphylococcus aureus) and gram negative (Escherichia coli) bacterial strains. A later step defined the most suitable blend of tannins to obtain, after tanning and/or retanning, an antibacterial natural leather. Furthermore, the vegetable tanned leathers, made with this tannins blend, have been the target of an in vivo trial during which 15 panelists have worn two differently made shoes. The lining and insole inside the right shoe have been made with vegetable tanned leathers with tannins, while the ones inside the left shoe contained only synthetic material. The shoes have been worn for 28 consecutive days, followed by a molecular and bioinformatic analysis of microbiota samples taken from the inner surface of the shoes by using a sterile swab. Lastly, a biochemical analysis of volatile short chain fatty acids has been carried out to investigate the byproducts of the bacteria responsible for the unpleasant odor of shoes. Take-Away: 1. Vegetable tanned leather is a wonderful antibacterial material thanks to the presence of natural tannins, such as chestnut, quebracho and tara. This property is appreciated in the production of insole leather, lining, leather goods and automotive interiors. 2. The problem of bromhidrosis (bad feet odor) can be avoided by using vegetable tanned leather. 3. In particular, vegetable leathers tanned with tannins used to make inside part of the shoes permit to avoid the formation of cheesy and acidic odours thanks to their antibacterial properties and their capacity to absorb sweat

Linking dual mode of action of host defense antimicrobial peptide thanatin: structures, lipopolysaccharide and LptAm binding of designed analogs

At present, antibiotics options to cure infections caused by drug resistant Gram-negative pathogens are highly inadequate. LPS outer membrane, proteins involved in LPS transport and biosynthesis pathways are vital targets. Thanatin, an insect derived 21-residue long antimicrobial peptide may be exploited for the development of effective antibiotics against Gram-negative bacteria. As a mode of bacterial cell killing, thanatin disrupts LPS outer membrane and inhibits LPS transport by binding to the periplasmic protein LptAm. Here, we report structure-activity correlation of thanatin and analogs for the purpose of rational design. These analogs of thanatin are investigated, by NMR, ITC and fluorescence, to correlate structure, antibacterial activity and binding with LPS and LptAm, a truncated monomeric variant. Our results demonstrate that an analog thanatin M21F exhibits superior antibacterial activity. In LPS interaction analyses, thanatin M21F demonstrate high affinity binding to outer membrane LPS. The atomic resolution structure of thanatin M21F in LPS micelle reveals four stranded -sheet structure in a dimeric topology whereby the sidechain of aromatic residues Y10, F21 sustained mutual packing at the interface. Strikingly, LptAm binding affinity of thanatin M21F has been significantly increased with an estimated Kd~0.73 nM vs 13 nM for thanatin. Further, atomic resolution structures and interactions of Ala based thanatin analogs define plausible correlations with antibacterial activity and LPS, LptAm interactions. Taken together, the current work provides a frame-work for the designing of thanatin based potent antimicrobial peptides for the treatment of drug resistance Gram-negative bacteria.Ministry of Education (MOE)Submitted/Accepted versionThis work was supported by the grant from Ministry of Education (MOE), Singapore

Thanatin Impairs Lipopolysaccharide Transport Complex Assembly by Targeting LptC–LptA Interaction and Decreasing LptA Stability

International audienceThe outer membrane (OM) of Gram-negative bacteria is a highly selective permeability barrier due to its asymmetric structure with lipopolysaccharide (LPS) in the outer leaflet. In Escherichia coli, LPS is transported to the cell surface by the LPS transport (Lpt) system composed of seven essential proteins forming a transenvelope bridge. Transport is powered by the ABC transporter LptB2FGC, which extracts LPS from the inner membrane (IM) and transfers it, through LptC protein, to the periplasmic protein LptA. Then, LptA delivers LPS to the OM LptDE translocon for final assembly at the cell surface. The Lpt protein machinery operates as a single device, since depletion of any component leads to the accumulation of a modified LPS decorated with repeating units of colanic acid at the IM outer leaflet. Moreover, correct machine assembly is essential for LPS transit and disruption of the Lpt complex results in LptA degradation. Due to its vital role in cell physiology, the Lpt system represents a good target for antimicrobial drugs. Thanatin is a naturally occurring antimicrobial peptide reported to cause defects in membrane assembly and demonstrated in vitro to bind to the N-terminal β-strand of LptA. Since this region is involved in both LptA dimerization and interaction with LptC, we wanted to elucidate the mechanism of inhibition of thanatin and discriminate whether its antibacterial effect is exerted by the disruption of the interaction of LptA with itself or with LptC. For this purpose, we here implemented the Bacterial Adenylate Cyclase Two-Hybrid (BACTH) system to probe in vivo the Lpt interactome in the periplasm. With this system, we found that thanatin targets both LptC-LptA and LptA-LptA interactions, with a greater inhibitory effect on the former. We confirmed in vitro the disruption of LptC-LptA interaction using two different biophysical techniques. Finally, we observed that in cells treated with thanatin, LptA undergoes degradation and LPS decorated with colanic acid accumulates. These data further support inhibition or disruption of Lpt complex assembly as the main killing mechanism of thanatin against Gram-negative bacteria

Solid State NMR Studies of Intact Lipopolysaccharide Endotoxin

International audienceLipopolysaccharides (LPS) are complex glycolipids forming the outside layer of Gram-negative bacteria. Their hydrophobic and heterogeneous nature greatly hampers their structural study in an environment similar to the bacterial surface. We have studied LPS purified from E. coli and pathogenic P. aeruginosa with long O-antigen polysaccharides assembled in solution as vesicles or elongated micelles. Solid-state NMR with magic-angle spinning permitted the identification of NMR signals arising from regions with different flexibilities in the LPS, from the lipid components to the O-antigen polysaccharides. Atomic scale data on the LPS enabled the study of the interaction of gentamicin antibiotic bound to P. aeruginosa LPS, for which we could confirm that a specific oligosaccharide is involved in the antibiotic binding. The possibility to study LPS alone and bound to a ligand when it is assembled in membrane-like structures opens great prospects for the investigation of proteins and antibiotics that specifically target such an important molecule at the surface of Gram-negative bacteria

Large-Scale Identification of Virulence Genes from Streptococcus pneumoniae

Streptococcus pneumoniae is the major cause of bacterial pneumonia, and it is also responsible for otitis media and meningitis in children. Apart from the capsule, the virulence factors of this pathogen are not completely understood. Recent technical advances in the field of bacterial pathogenesis (in vivo expression technology and signature-tagged mutagenesis [STM]) have allowed a large-scale identification of virulence genes. We have adapted to S. pneumoniae the STM technique, originally used for the discovery of Salmonella genes involved in pathogenicity. A library of pneumococcal chromosomal fragments (400 to 600 bp) was constructed in a suicide plasmid vector carrying unique DNA sequence tags and a chloramphenicol resistance marker. The recent clinical isolate G54 was transformed with this library. Chloramphenicol-resistant mutants were obtained by homologous recombination, resulting in genes inactivated by insertion of the suicide vector carrying a unique tag. In a mouse pneumonia model, 1.250 candidate clones were screened; 200 of these were not recovered from the lungs were therefore considered virulence-attenuated mutants. The regions flanking the chloramphenicol gene of the attenuated mutants were amplified by inverse PCR and sequenced. The sequence analysis showed that the 200 mutants had insertions in 126 different genes that could be grouped in six classes: (i) known pneumococcal virulence genes; (ii) genes involved in metabolic pathways; (iii) genes encoding proteases; (iv) genes coding for ATP binding cassette transporters; (v) genes encoding proteins involved in DNA recombination/repair; and (vi) DNA sequences that showed similarity to hypothetical genes with unknown function. To evaluate the virulence attenuation for each mutant, all 126 clones were individually analyzed in a mouse septicemia model. Not all mutants selected in the pneumonia model were confirmed in septicemia, thus indicating the existence of virulence factors specific for pneumonia

Tungsten Trioxide and Its TiO2 Mixed Composites for the Photocatalytic Degradation of NOx and Bacteria (Escherichia coli) Inactivation

The increased air pollution and its impact on the environment and human health in several countries have caused global concerns. Nitrogen oxides (NO2 and NO) are principally emitted from industrial activities that strongly contribute to poor air quality. Among bacteria emanated from the fecal droppings of livestock, wildlife, and humans, Escherichia coli is the most abundant, and is often associated with the health risk of water. TiO2/WO3 heterostructures represent emerging systems for photocatalytic environmental remediation. However, the results reported in the literature are conflicting, depending on several parameters. In this work, WO3 and a series of TiO2/WO3 composites were properly synthesized by an easy and fast method, abundantly characterized by several techniques, and used for NOx degradation and E. coli inactivation under visible light irradiation. We demonstrated that the photoactivity of TiO2/WO3 composites towards NO2 degradation under visible light is strongly related to the WO3 content. The best performance was obtained by a WO3 load of 20% that guarantees limited e&minus;/h+ recombination. On the contrary, we showed that E. coli could not be degraded under visible irradiation of the TiO2/WO3 composites