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Applied Microbiology and Biotechnology Dec 2024Polymyxins are cationic peptide antibiotics and regarded as the "final line of defense" against multidrug-resistant bacterial infections. Meanwhile, some...
Polymyxins are cationic peptide antibiotics and regarded as the "final line of defense" against multidrug-resistant bacterial infections. Meanwhile, some polymyxin-resistant strains and the corresponding resistance mechanisms have also been reported. However, the response of the polymyxin-producing strain Paenibacillus polymyxa to polymyxin stress remains unclear. The purpose of this study was to investigate the stress response of gram-positive P. polymyxa SC2 to polymyxin B and to identify functional genes involved in the stress response process. Polymyxin B treatment upregulated the expression of genes related to basal metabolism, transcriptional regulation, transport, and flagella formation and increased intracellular ROS levels, flagellar motility, and biofilm formation in P. polymyxa SC2. Adding magnesium, calcium, and iron alleviated the stress of polymyxin B on P. polymyxa SC2, furthermore, magnesium and calcium could improve the resistance of P. polymyxa SC2 to polymyxin B by promoting biofilm formation. Meanwhile, functional identification of differentially expressed genes indicated that an ABC superfamily transporter YwjA was involved in the stress response to polymyxin B of P. polymyxa SC2. This study provides an important reference for improving the resistance of P. polymyxa to polymyxins and increasing the yield of polymyxins. KEY POINTS: • Phenotypic responses of P. polymyxa to polymyxin B was performed and indicated by RNA-seq • Forming biofilm was a key strategy of P. polymyxa to alleviate polymyxin stress • ABC transporter YwjA was involved in the stress resistance of P. polymyxa to polymyxin B.
Topics: Paenibacillus polymyxa; Polymyxin B; Paenibacillus; Calcium; Magnesium; Polymyxins
PubMed: 38170316
DOI: 10.1007/s00253-023-12916-3 -
Genomics Jan 2021The legislations on the usage of antibiotics as growth promoters and prophylactic agents have compelled to develop alternative tools to upsurge the animal protection and...
The legislations on the usage of antibiotics as growth promoters and prophylactic agents have compelled to develop alternative tools to upsurge the animal protection and contain antibiotic usage. Probiotics have emerged as an effective antibiotic substitute in animal farming. The present study explores the probiotic perspective of Paenibacillus polymyxa HK4 interlinking the genotypic and phenotypic characteristics. The draft genome of HK4 revealed the presence of ORFs encoding the functions associated with tolerance to gastrointestinal stress and adhesion. The biosynthetic gene clusters encoding non-ribosomally synthesized peptides, polyketides and lanthipeptides such as fusaricidin, tridecaptin, polymyxin, paenilan and paenibacillin were annotated in HK4 genome. The strain harbored the chromosomal gene conferring the resistance to lincosamides. No functional gene encoding virulence or toxins could be identified in the genome of HK4. The genome analysis data was complemented by the in vitro experiments confirming its survival during gastrointestinal transit, antimicrobial potential and antibiotic sensitivity. NUCLEOTIDE SEQUENCE ACCESSION NUMBER: The draft-genome sequence of Paenibacillus polymyxa HK4 has been deposited as whole-genome shotgun project at GenBank under the accession number PRJNA603023.
Topics: Anti-Bacterial Agents; Genome, Bacterial; Paenibacillus polymyxa; Polyketides; Polymyxins; Probiotics
PubMed: 33096257
DOI: 10.1016/j.ygeno.2020.10.017 -
Antonie Van Leeuwenhoek Nov 2020Rapid development of gene sequencing technologies has led to an exponential increase in microbial sequencing data. Genome research of a single organism does not capture...
Rapid development of gene sequencing technologies has led to an exponential increase in microbial sequencing data. Genome research of a single organism does not capture the changes in the characteristics of genetic information within a species. Pan-genome analysis gives us a broader perspective to study the complete genetic information of a species. Paenibacillus polymyxa is a Gram-positive bacterium and an important plant growth-promoting rhizobacterium with the ability to produce multiple antibiotics, such as fusaricidin, lantibiotic, paenilan, and polymyxin. Our study explores the pan-genome of 14 representative P. polymyxa strains isolated from around the world. Heap's law model and curve fitting confirmed an open pan-genome of P. polymyxa. The phylogenetic and collinearity analyses reflected that the evolutionary classification of P. polymyxa strains are not associated with geographical area and ecological niches. Few genes related to phytohormone synthesis and phosphate solubilization were conserved; however, the nif cluster gene associated with nitrogen fixation exists only in some strains. This finding is indicative of nitrogen fixing ability is not stable in P. polymyxa. Analysis of antibiotic gene clusters in P. polymyxa revealed the presence of these genes in both core and accessory genomes. This observation indicates that the difference in living environment led to loss of ability to synthesize antibiotics in some strains. The current pan-genomic analysis of P. polymyxa will help us understand the mechanisms of biological control and plant growth promotion. It will also promote the use of P. polymyxa in agriculture.
Topics: Genome, Plant; Nitrogen Fixation; Paenibacillus polymyxa; Phylogeny; Plant Development; Plant Growth Regulators; Rhizosphere
PubMed: 32816227
DOI: 10.1007/s10482-020-01461-y -
Methods in Molecular Biology (Clifton,... 2020The skin contains three primary layers: epidermis, dermis, and hypodermis. Separation of epidermal components from dermis (dermal-epidermal separation) is an important...
The skin contains three primary layers: epidermis, dermis, and hypodermis. Separation of epidermal components from dermis (dermal-epidermal separation) is an important basic investigation technique for pharmacology, toxicology, and biology. There are different systems of epidermal separation, including typical methods of chemical, enzyme, heat, etc. Each approach has advantages versus disadvantages, and thus the appropriate method should be chosen for a given research question. Here we described the method of enzyme separation.
Topics: Bacterial Proteins; Cell Separation; Dermis; Endopeptidases; Epidermal Cells; Humans; Paenibacillus polymyxa; Skin; Trypsin
PubMed: 31792753
DOI: 10.1007/7651_2019_267 -
Frontiers in Bioengineering and... 2022Production of some antimicrobial peptides by bacterial producers is a resource-intensive process, thus, using inexpensive growth media and simplifying antimicrobial...
Production of some antimicrobial peptides by bacterial producers is a resource-intensive process, thus, using inexpensive growth media and simplifying antimicrobial extraction and down-stream processing are highly desirable. Acid whey, a dairy industry waste, is explored as a medium for production of broad-spectrum antimicrobials from selected bacteriocinogenic bacteria. Neutralized and yeast extract-supplemented acid whey was suitable for production of antimicrobials by four tested strains, but OSY-EC was the most prolific antimicrobial producer. Concentrating synthesized antimicrobials during culture incubation using beads of polymeric adsorbent resin, followed by solvent extraction and freeze-drying, resulted in antimicrobials-rich powder (AMRP). Under these conditions, OSY-EC produced paenibacillin, polymyxin E, and fusaricidin, which are active against Gram-positive and Gram-negative bacteria and fungi, respectively. When media containing 2x and 4x minimum inhibitory concentrations of AMRP were inoculated with and , microbial populations decreased by ≥4-log CFU ml in tryptic soy broth and ≥3.5-log CFU ml in milk. The antimicrobial mechanism of action of AMRP solutions was attributed to the disruption of cytoplasmic membrane of indicator strains, and . These findings exemplify promising strategies for valorization of acid whey microbial bioreactions to yield potent antimicrobials.
PubMed: 35646844
DOI: 10.3389/fbioe.2022.869778 -
Enzyme and Microbial Technology Mar 2023Recently, there has been increased interest in the synthesis of nanoparticles by using natural polysaccharides. These polysaccharides are eco-friendly, nontoxic, and...
Recently, there has been increased interest in the synthesis of nanoparticles by using natural polysaccharides. These polysaccharides are eco-friendly, nontoxic, and cheap to prepare. On the other hand, the attention in hydrocolloids and films has significantly enhanced, and their application is very promising in the food, pharmaceutical, perfumery and cosmetics, oil, paper, and textile industries. In this context, the present study is aimed to prepare silver nanoparticles by using viscous and superviscous exopolysaccharides of the rhizobacterium Paenibacillus polymyxa strains, CCM 1465 and 88A, and examined the properties of the resultant nanoparticles. We examined the synthesis and properties of silver nanoparticles under variable synthetic conditions by using exopolysaccharides of the rhizobacteria Paenibacillus polymyxa CCM 1465 and 88A. To prepare nanoparticles, we used different combinations of exopolysaccharide and silver nitrate concentrations: 1-10 mg/mL and 1-40 mM, respectively. The resulting solutions were alkalinized from pH 7.5-12 and heated for 15, 30, and 60 min to determine the optimal synthetic conditions. We found that the exopolysaccharides of strains CCM 1465 and 88A reduced silver ions and acted as nanoparticle stabilizers. The prepared spherical, oval, and triangular particles were stable and ranged in size from 2 to 40 nm, depending on the strain and on the experimental conditions. The nanoparticles showed antibacterial and antifungal activity against Escherichia coli K-12, Pseudomonas aeruginosa 50.3, Bacillus subtilis 26-D, and Fusarium oxysporum. In addition, the nanoparticles were active against SK-MEL-2 human melanoma cells. This finding shows the promise of further research on the exopolysaccharides of P. polymyxa 1465 and 88А in different fields of science, including medicine.
Topics: Humans; Paenibacillus polymyxa; Metal Nanoparticles; Escherichia coli K12; Silver; Polysaccharides; Anti-Bacterial Agents; Escherichia coli
PubMed: 36508942
DOI: 10.1016/j.enzmictec.2022.110174 -
Scientific Reports Sep 2022Pectin is one of the main structural components in fruits and an indigestible fiber made of D-galacturonic acid units with α (1-4) linkage. This study investigates the...
Pectin is one of the main structural components in fruits and an indigestible fiber made of D-galacturonic acid units with α (1-4) linkage. This study investigates the microbial degradation of pectin in apple waste and the production of bioactive compounds. Firstly, pectin-degrading bacteria were isolated and identified, then pectinolytic activity was assessed by DNS. The products were evaluated by TLC and LC-MS-ESI. The antioxidative effects were investigated using DPPH and anti-cancer effects and cytotoxicity were analyzed by MTT and flow cytometry. In this study two new bacterial isolates, Alcaligenes faecalis AGS3 and Paenibacillus polymyxa S4 with the pectinolytic enzyme were introduced. Structure analysis showed that the products of enzymatic degradation include unsaturated mono, di, tri, and penta galacturonic acids with 74% and 69% RSA at 40 mg/mL for A. faecalis and P. polymyxa S4, respectively. The results of anti-tumor properties on MCF-7 cells by MTT assay, for products of AGS3 and S4 at 40 mg/mL after 48 h, showed 7% and 9% survival, respectively. In the flow cytometric assessment, the compounds of AGS3 at 40 mg/mL were 100% lethal in 48 h and regarding S4 isolate caused 98% death. Cytotoxicity evaluation on L-929 cells showed no significant toxicity on living cells.
Topics: Alcaligenes faecalis; Hexuronic Acids; Malus; Paenibacillus; Paenibacillus polymyxa; Pectins; Polygalacturonase
PubMed: 36138114
DOI: 10.1038/s41598-022-20011-2 -
Microbiological Research Jul 2021The diseases caused by phytopathogens account for huge economic losses in the agricultural sector. Paenibacillus polymyxa is one of the agriculturally important...
The diseases caused by phytopathogens account for huge economic losses in the agricultural sector. Paenibacillus polymyxa is one of the agriculturally important biocontrol agents and plant growth promoting bacterium. This study describes the antifungal potential of P. polymyxa HK4 against an array of fungal phytopathogens and its ability to stimulate seed germination of cumin and groundnut under in vitro conditions. The cumin and groundnut seeds bacterized with HK4 exhibited enhanced germination efficiency in comparison to controls. The use of HK4 as a soil inoculant significantly promoted the shoot length and fresh weight of groundnut plants in pot studies. The draft genome analysis of HK4 revealed the genetic attributes for motility, root colonization, antagonism, phosphate solubilization, siderophore production and production of volatile organic compounds. The bacterium HK4 harnessed several hydrolytic enzymes that may assist its competence in the rhizosphere. The PCR amplification and sequence analysis of the conserved region of the fusA gene amplicon revealed the ability of HK4 to produce fusaricidin. Furthermore, the LC-ESI-MS/MS of crude cell pellet extract of HK4 confirmed the presence of fusaricidin as a major antifungal metabolite. This study demonstrated the potential of HK4 as a biocontrol agent and a plant growth promoter.
Topics: Antifungal Agents; Bacterial Proteins; Crop Protection; Cuminum; Depsipeptides; Fungi; Genome, Bacterial; Genomics; Mass Spectrometry; Paenibacillus polymyxa; Phylogeny; Plant Diseases; Plant Roots; Plant Shoots
PubMed: 33690069
DOI: 10.1016/j.micres.2021.126734 -
Microbiological Research May 2024Soil salinity negatively affects microbial communities, soil fertility, and agricultural productivity and has become a major agricultural problem worldwide. Plant...
Soil salinity negatively affects microbial communities, soil fertility, and agricultural productivity and has become a major agricultural problem worldwide. Plant growth-promoting rhizobacteria (PGPR) with salt tolerance can benefit plant growth under saline conditions and diminish the negative effects of salt stress on plants. In this study, we aimed to understand the salt-tolerance mechanism of Paenibacillus polymyxa at the genetic and metabolic levels and elucidate the mechanism of strain SC2 in promoting maize growth under saline conditions. Under salt stress, we found that strain SC2 promoted maize seedling growth, which was accompanied by a significant upregulation of genes encoding for the biosynthesis of peptidoglycan, polysaccharide, and fatty acid, the metabolism of purine and pyrimidine, and the transport of osmoprotectants such as trehalose, glycine betaine, and K in strain SC2. To further enhance the salt resistance of strain SC2, three mutants (SC2-11, SC2-13, and SC2-14) with higher capacities for salt resistance and exopolysaccharide synthesis were obtained via atmospheric and room-temperature plasma mutagenesis. In saline-alkaline soil, the mutants showed better promoting effect on maize seedlings than wild-type SC2. The fresh weight of maize seedlings was increased by 68.10% after treatment with SC2-11 compared with that of the control group. The transcriptome analysis of maize roots demonstrated that SC2 and SC2-11 could induce the upregulation of genes related to the plant hormone signal transduction, starch and sucrose metabolism, reactive oxygen species scavenging, and auxin and ethylene signaling under saline-alkaline stress. In addition, various transcription factors, such as zinc finger proteins, ethylene-responsive-element-binding protein, WRKY, myeloblastosis proteins, basic helix-loop-helix proteins, and NAC proteins, were up-regulated in response to abiotic stress. Moreover, the microbial community composition of maize rhizosphere soil after inoculating with strain SC2 was varied from the one after inoculating with mutant SC2-11. Our results provide new insights into the various genes involved in the salt resistance of strain SC2 and a theoretical basis for utilizing P. polymyxa in saline-alkaline environments.
Topics: Seedlings; Paenibacillus polymyxa; Zea mays; Soil; Ethylenes
PubMed: 38354626
DOI: 10.1016/j.micres.2024.127639 -
Microorganisms Mar 2023Polymyxin-producing bacteria within the complex have broad-spectrum activities against fungi and bacteria. Their antibacterial activities against soft rot and...
Polymyxin-producing bacteria within the complex have broad-spectrum activities against fungi and bacteria. Their antibacterial activities against soft rot and phytopathogens containing multiple polymyxin-resistant genes were not clear. Here, we selected nine strains within the complex having broad-spectrum antagonistic activities against phytopathogenic fungi and a polymyxin-resistant strain causing stem and root rot disease of sweet potato and did antagonistic assays on nutrient agar and sweet potato tuber slices. These strains within the complex showed clear antagonistic activities against in vitro and in vivo. The most effective antagonistic strain ShX301 showed broad-spectrum antagonistic activities against all the test and strains, completely eliminated from sweet potato seed tubers, and promoted the growth of sweet potato seedlings. Cell-free culture filtrate of ShX301 inhibited growth, swimming motility, and biofilm formation and disrupted plasma membranes, releasing nucleic acids and proteins. Multiple lipopeptides produced by ShX301 may play a major role in the bactericidal and bacteriostatic actions. This study clarifies that the antimicrobial spectrum of polymyxin-producing bacteria within the complex includes the polymyxin-resistant and phytopathogens and strengthens the fact that bacteria within the complex have high probability of being effective biocontrol agents and plant growth promoters.
PubMed: 37110240
DOI: 10.3390/microorganisms11040817