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Biomolecules Nov 2021Ketalpyruvyltransferases belong to a widespread but little investigated class of enzymes, which utilise phosphoenolpyruvate (PEP) for the pyruvylation of saccharides....
Ketalpyruvyltransferases belong to a widespread but little investigated class of enzymes, which utilise phosphoenolpyruvate (PEP) for the pyruvylation of saccharides. Pyruvylated saccharides play pivotal biological roles, ranging from protein binding to virulence. Limiting factors for the characterisation of ketalpyruvyltransferases are the availability of cognate acceptor substrates and a straightforward enzyme assay. We report on a fast ketalpyruvyltransferase assay based on the colorimetric detection of phosphate released during pyruvyltransfer from PEP onto the acceptor via complexation with Malachite Green and molybdate. To optimise the assay for the model 4,6-ketalpyruvyl::ManNAc-transferase CsaB from , a β-d-ManNAc-α-d-GlcNAc-diphosphoryl-11-phenoxyundecyl acceptor mimicking an intermediate of the bacterium's cell wall glycopolymer biosynthesis pathway, upon which CsaB is naturally active, was produced chemo-enzymatically and used together with recombinant CsaB. Optimal assay conditions were 5 min reaction time at 37 °C and pH 7.5, followed by colour development for 1 h at 37 °C and measurement of absorbance at 620 nm. The structure of the generated pyruvylated product was confirmed by NMR spectroscopy. Using the established assay, the first kinetic constants of a 4,6-ketalpyuvyl::ManNAc-transferase could be determined; upon variation of the acceptor and PEP concentrations, a of 19.50 ± 3.50 µM and of 0.21 ± 0.01 s as well as a of 258 ± 38 µM and a of 0.15 ± 0.01 s were revealed. CsaB was inactive on synthetic NP-β-d-ManNAc and β-d-ManNAc-β-d-GlcNAc-1-Me, supporting the necessity of a complex acceptor substrate.
Topics: Catalysis; Hexosamines; Paenibacillus; Phosphates; Phosphoenolpyruvate
PubMed: 34827730
DOI: 10.3390/biom11111732 -
Pathogens (Basel, Switzerland) Apr 2020An emerging need for new classes of antibiotics is, on the one hand, evident as antimicrobial resistance continues to rise. On the other hand, the awareness of the pros...
An emerging need for new classes of antibiotics is, on the one hand, evident as antimicrobial resistance continues to rise. On the other hand, the awareness of the pros and cons of chemically synthesized compounds' extensive use leads to a search for new metabolites in already known reservoirs. Previous research showed that strain ( MP1) recovered from a buckwheat honey sample presented a wide spectrum of antimicrobial activity against both Gram-positive and Gram-negative pathogens. Recent investigation has confirmed that MP1 (deposited at DDBJ/ENA/GenBank under the accession WSQB00000000) produces a proteinaceous, heat-stable compound(s) with the maximum antimicrobial production obtained after 18 hours of MP1 growth in LB medium at 37 °C with continuous shaking at 200 RPM. The highest activity was found in the 40% ammonium sulfate precipitate, with high activity also remaining in the 50% and 60% ammonium sulfate precipitates. Moderate to high antimicrobial activity that is insensitive to proteases or heat treatment, was confirmed against pathogenic bacteria that included FSL - X1-0001 (strain 10403S), L1 - 0030 and O157: H7. Further studies, including de novo sequencing of peptides by mass spectrometry, are in progress.
PubMed: 32344843
DOI: 10.3390/pathogens9050319 -
Plants (Basel, Switzerland) Jan 2021In the last two decades grapevine trunk diseases (GTDs) have emerged as the most significant threat for grapevine sustainability worldwide. The tracheomycotic fungus...
In the last two decades grapevine trunk diseases (GTDs) have emerged as the most significant threat for grapevine sustainability worldwide. The tracheomycotic fungus (Pch) is the predominant GTD-associated species and cannot be controlled with available chemicals. In the present study, we evaluated the effectiveness of two microbial strains ( K165 and F2) against Pch in grapevine. In vitro bioassays, performed in a growth culture medium simulating the xylem environment, indicated that F2 decreased Pch growth and sporulation, whereas K165 did not have any effect on Pch growth. experiments revealed that root-drench and stem-puncture application of K165 and F2 reduced the endophytic relative DNA amount of Pch by 90% and 82%, respectively, compared to controls. However, wood discoloration, the typical symptom of Pch infection, was not reduced in the F2 treated grapevines. Nevertheless, the F2 treated grapevines harbored higher lignin levels compared to mocks, as it was also done by K165. Therefore, F2 and K165 have the potential to be used as biocontrol agents against Pch in grapevines.
PubMed: 33499084
DOI: 10.3390/plants10020207 -
The Pediatric Infectious Disease Journal Mar 2016
Topics: Anti-Bacterial Agents; Fatal Outcome; Female; Gram-Positive Bacterial Infections; Humans; Infant, Newborn; Infant, Premature; Meningoencephalitis; Paenibacillus; Sepsis
PubMed: 26866854
DOI: 10.1097/INF.0000000000001003 -
Journal of Experimental Botany May 2021The biocontrol agent Paenibacillus alvei K165 was previously shown to protect Arabidopsis thaliana plants against Verticillium dahliae. Here we show that K165 also...
The biocontrol agent Paenibacillus alvei K165 was previously shown to protect Arabidopsis thaliana plants against Verticillium dahliae. Here we show that K165 also confers inherited immune resistance to V. dahliae. By performing a histone acetyltransferases mutant screen, ChIP assays, and transcriptomic experiments, we were able to show that histone acetylation significantly contributes to the K165 biocontrol activity and establishment of inheritable resistance to V. dahliae. K165 treatment primed the expression of immune-related marker genes and the cinnamyl alcohol dehydrogenase gene CAD3 through the function of histone acetyltransferases. Our results reveal that offspring of plants treated with K165 have primed immunity and enhanced lignification, both contributing towards the K165-mediated inherited immune resistance. Thus, our study paves the way for the use of biocontrol agents for the establishment of inheritable resistance to agronomically important pathogens.
Topics: Ascomycota; Disease Resistance; Gossypium; Paenibacillus; Plant Diseases; Verticillium
PubMed: 33829257
DOI: 10.1093/jxb/erab154 -
The Plant Pathology Journal Oct 2022This study was performed to reveal phenotypic characters and identity of symbiont bacteria of Nasutitermes as well as investigate their potential as antagonist of plant...
This study was performed to reveal phenotypic characters and identity of symbiont bacteria of Nasutitermes as well as investigate their potential as antagonist of plant pathogenic fungi. Isolation of the symbiont bacteria was carried out from inside the heads and the bodies of soldier and worker termite which were collected from 3 locations of nests. Identification was performed using phenotypic test and sequence of 16S ribosomal DNA (16S rDNA). Antagonistic capability was investigated in the laboratory against 3 phytopathogenic fungi i.e., Phytophthora capsici, Ganoderma boninense, and Rigidoporus microporus. Totally, 39 bacterial isolates were obtained from inside the heads and the bodies of Nasutitermes. All the isolates showed capability to inhibit growth of P. capsici, however, 34 isolates showed capability to inhibit growth of G. boninense and 32 isolates showed capability to inhibit growth of R. microporus. Two bacterial strains (IK3.1P and 1B1.2P) which showed the highest percentage of inhibition were further identified based on their sequence of 16S rDNA. The result showed that 1K3.1P strain was placed in the group of type strain and reference strains of Lysinibacillus fusiformis meanwhile 1B1.2P strain was grouped within type strain and reference strains Paenibacillus alvei. The result of this study supply valuable information on the role of symbiont bacteria of Nasutitermes, which may support the development of the control method of the three above-mentioned phytopathogenic fungi.
PubMed: 36221917
DOI: 10.5423/PPJ.OA.03.2022.0031 -
Glycobiology Jan 2016Flagellin glycosylation impacts, in several documented cases, the functionality of bacterial flagella. The basis of flagellin glycosylation has been studied for various...
Flagellin glycosylation impacts, in several documented cases, the functionality of bacterial flagella. The basis of flagellin glycosylation has been studied for various Gram-negative bacteria, but less is known about flagellin glycans of Gram-positive bacteria including Paenibacillus alvei, a secondary invader of honeybee colonies diseased with European foulbrood. Paenibacillus alvei CCM 2051(T) swarms vigorously on solidified culture medium, with swarming relying on functional flagella as evidenced by abolished biofilm formation of a non-motile P. alvei mutant defective in the flagellin protein Hag. Here, the glycobiology of the polar P. alvei flagella was investigated. Analysis on purified flagellin demonstrated that the 30-kDa Hag protein (PAV_2c01710) is modified with an O-linked trisaccharide comprised of one hexose and two N-acetyl-hexosamine residues, at three sites of glycosylation. Downstream of the hag gene on the bacterial chromosome, two open reading frames (PAV_2c01630, PAV_2c01640) encoding putative glycosyltransferases were shown to constitute a flagellin glycosylation island. Mutants defective in these genes exhibited altered migration in sodium dodecyl sulfate polyacrylamide gel electrophoresis as well as loss of extracellular flagella production and bacterial motility. This study reveals that flagellin glycosylation in P. alvei is pivotal to flagella formation and bacterial motility in vivo, and simultaneously identifies flagella glycosylation as a second protein O-glycosylation system in this bacterium, in addition to the well-investigated S-layer tyrosine O-glycosylation pathway.
Topics: Amino Acid Sequence; Bacterial Proteins; Flagellin; Glycosylation; Glycosyltransferases; Hexoses; Molecular Sequence Data; Mutation; Paenibacillus; Protein Processing, Post-Translational
PubMed: 26405108
DOI: 10.1093/glycob/cwv087 -
BMC Research Notes Jun 2020A Paenibacillus strain isolated in previous research exhibited antimicrobial activity against relevant human pathogens including Staphylococcus aureus and Listeria...
OBJECTIVE
A Paenibacillus strain isolated in previous research exhibited antimicrobial activity against relevant human pathogens including Staphylococcus aureus and Listeria monocytogenes. In this study, the genome of the aforementioned strain, designated as MP1, was shotgun sequenced. The draft genome of strain MP1 was subject to multiple genomic analyses to taxonomically characterize it and identify the genes potentially responsible for its antimicrobial activity.
RESULTS
Here we report the draft genome sequence of an antimicrobial producing Paenibacillus strain, MP1. Average Nucleotide Identity (ANI) analysis established strain MP1 as a new strain of the previously characterized Paenibacillus alvei. The genomic analysis identified several putative secondary metabolite clusters including seven Nonribosomal Peptide Synthetase clusters (NRPS) (> 10,000 nt), one bacteriocin or other unspecified Ribosomally Synthesized and Post-Translationally modified Peptide Product (RiPP), one lanthipeptide, and six hybrid clusters (NRPS-Type I Polyketide synthase (T1PKS) and NRPS-trans Amino Transferase Polyketide Synthase (AT-PKS)).
Topics: Anti-Infective Agents; Genome, Bacterial; Paenibacillus; Whole Genome Sequencing
PubMed: 32517793
DOI: 10.1186/s13104-020-05124-z -
Current Microbiology Apr 2021The increased prevalence of multidrug-resistant pathogens poses a significant clinical threat, and hence, the discovery of novel antibiotics is the need of the hour....
The increased prevalence of multidrug-resistant pathogens poses a significant clinical threat, and hence, the discovery of novel antibiotics is the need of the hour. Several attempts are being made worldwide to screen and identify newer antibiotics from various microbial sources. The genus Paenibacillus is known for its biosynthetic potential and metabolic versatility in producing several secondary metabolites. In this study, we isolated Paenibacillus alvei strain JR949 from the soil, which exhibited antimicrobial activity against Enteropathogenic Escherichia coli (EPEC), Pseudomonas aeruginosa (PAO1), and methicillin-resistant Staphylococcus aureus (MRSA). The whole genome of this strain was sequenced using the Illumina platform. The genome mining of the draft genome sequence revealed a total of 31 biological gene clusters (BGCs) responsible for the synthesis of secondary metabolites. The construction of the similarity network of the BGCs and the comparative analysis with the genetically related strains aided the identification of metabolites produced by this strain. We identified BGCs coding for paenibactin, paenibacterin, anabaenopeptin NZ857, icosalide A/B, polymyxin, and bicornutinA1/A2 with 100% similarity. The BGCs with lower sequence similarity to paenibacterin, polymyxin B, colistin A/B, pellasoren, tridecaptin, pelgipeptin, and marthiapeptide were also identified. Furthermore, 13 putative NRPS BGCs, 3 NRPS-T1PKS hybrid clusters, a T1PKS, and a bacteriocin BGC were identified with very low similarity (≤ 25%) or no similarity with known antibiotics. Further experimental investigations may result in the discovery of novel antimicrobial drugs.
Topics: Anti-Infective Agents; Methicillin-Resistant Staphylococcus aureus; Multigene Family; Paenibacillus; Sequence Analysis
PubMed: 33616690
DOI: 10.1007/s00284-021-02393-0 -
International Journal of Systematic... Jul 1996The name "Bacillus apiarius" Katznelson 1955 was not included on the Approved Lists of Bacterial Names and thus lost standing in bacterial nomenclature. The genetic...
The name "Bacillus apiarius" Katznelson 1955 was not included on the Approved Lists of Bacterial Names and thus lost standing in bacterial nomenclature. The genetic homogeneity of "B. apiarius" strains was assessed by determining their G+C contents by the buoyant density method and by measuring the levels of DNA relatedness by spectrophotometric reassociation procedures. The G+C contents of the 15 strains examined, ranged from 52 to 54 mol%. DNA reassociation revealed the presence of two clusters, each with high levels of intragroup relatedness (60 to 100%). One cluster consisted of six strains highly related to Bacillus thiaminolyticus, and the other consisted of nine strains related to the designated type strain of "B. apiarius." The strains in the second cluster were not closely related genetically to the type strains of organisms frequently associated with honey bees (namely, Paenibacillus alvei, Paenibacillus larvae, Bacillus laterosporus, and Paenibacillus pulvifaciens). The "B. apiarius" strains in the second cluster were also phenotypically homogeneous and distinguishable from the previously described species. Comparative analyses of the 16S rRNA gene DNA sequence showed that the proper phylogenetic position of the second cluster was in the genus Paenibacillus. These findings justify the proposal of a new species with the name Paenibacillus apiarius. The type strain is NRRL NRS-1438.
Topics: Bacillus; Base Sequence; DNA, Bacterial; Fatty Acids; Molecular Sequence Data; Phenotype; Phylogeny; RNA, Bacterial; RNA, Ribosomal, 16S
PubMed: 8782677
DOI: 10.1099/00207713-46-3-688