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1.
BMC Microbiol ; 24(1): 385, 2024 Oct 02.
Article in English | MEDLINE | ID: mdl-39358715

ABSTRACT

BACKGROUND BURKHOLDERIA: is a phosphorus solubilizing microorganism discovered in recent years, which can dissolve insoluble phosphorus compounds into soluble phosphorus. To investigate the effects of Burkholderia and calcium phosphate on the composting of Torreya grandis branches and leaves, as well as to explain the nutritional and metabolic markers related to the composting process. METHODS: In this study, we employed amplicon sequencing and untargeted metabolomics analysis to examine the interplay among phosphorus (P) components, microbial communities, and metabolites during T. grandis branch and leaf waste composting that underwent treatment with calcium phosphate and phosphate-solubilizing bacteria (Burkholderia). There were four composting treatments, 10% calcium phosphate (CaP) or 5 ml/kg (1 × 108/ml Burkholderia) microbial inoculum (WJP) or both (CaP + WJP), and the control group (CK). RESULTS: The results indicated that Burkholderia inoculation and calcium phosphate treatment affected the phosphorus composition, pH, EC, and nitrogen content. Furthermore, these treatments significantly affected the diversity and structure of bacterial and fungal communities, altering microbial and metabolite interactions. The differential metabolites associated with lipids and organic acids and derivatives treated with calcium phosphate treatment are twice as high as those treated with Burkholderia in both 21d and 42d. The results suggest that calcium phosphate treatment alters the formation of some biological macromolecules. CONCLUSION: Both Burkholderia inoculation and calcium phosphate treatment affected the phosphorus composition, nitrogen content and metabolites of T. grandis branch and leaf waste compost.These results extend our comprehension of the coupling of matter transformation and community succession in composting with the addition of calcium phosphate and phosphate-solubilizing bacteria.


Subject(s)
Burkholderia , Calcium Phosphates , Composting , Phosphorus , Soil Microbiology , Calcium Phosphates/metabolism , Phosphorus/metabolism , Burkholderia/metabolism , Burkholderia/genetics , Burkholderia/drug effects , Bacteria/metabolism , Bacteria/genetics , Bacteria/classification , Bacteria/drug effects , Microbiota/drug effects , Nitrogen/metabolism , Soil/chemistry , Plant Leaves/microbiology , Fungi/metabolism , Fungi/drug effects , Fungi/genetics , Fungi/classification , Hydrogen-Ion Concentration
2.
Microb Cell Fact ; 23(1): 255, 2024 Sep 28.
Article in English | MEDLINE | ID: mdl-39342283

ABSTRACT

Cyclo (Phe-Pro) (cFP), a cyclic dipeptide with notable antifungal, antibacterial, and antiviral properties, shows great promise for biological control of plant diseases. Produced as a byproduct by non-ribosomal peptide synthetases (NRPS), the regulatory mechanism of cFP biosynthesis remains unclear. In a screening test of 997 Tn5 mutants of Burkholderia seminalis strain R456, we identified eight mutants with enhanced antagonistic effects against Fusarium graminearum (Fg). Among these, mutant 88's culture filtrate contained cFP, confirmed through HPLC and LC-MS, which actively inhibited Fg. The gene disrupted in mutant 88 is part of the Dct transport system (Dct-A, -B, -D), responsible for C4-dicarboxylate transport. Knockout mutants of Dct genes exhibited higher cFP levels than the wild type, whereas complementary strains showed no significant difference. Additionally, the presence of exogenous C4-dicarboxylates reduced cFP production in wild type R456, indicating that these substrates negatively regulate cFP synthesis. Given that cFP synthesis is related to NRPS, we previously identified an NRPS cluster in R456, horizontally transferred from algae. Specifically, knocking out gene 2061 within this NRPS cluster significantly reduced cFP production. A Fur box binding site was predicted upstream of gene 2061, and yeast one-hybrid assays confirmed Fur protein binding, which increased with additional C4-dicarboxylates. Knockout of the Fur gene led to up-regulation of gene 2061 and increased cFP production, suggesting that C4-dicarboxylates suppress cFP synthesis by enhancing Fur-mediated repression of gene 2061.


Subject(s)
Burkholderia , Burkholderia/metabolism , Burkholderia/genetics , Fusarium/metabolism , Fusarium/genetics , Fusarium/drug effects , Peptides, Cyclic/biosynthesis , Peptide Synthases/genetics , Peptide Synthases/metabolism , Dicarboxylic Acids/metabolism , Bacterial Proteins/metabolism , Bacterial Proteins/genetics
3.
Appl Microbiol Biotechnol ; 108(1): 474, 2024 Sep 25.
Article in English | MEDLINE | ID: mdl-39320489

ABSTRACT

(R)-3-Isobutylglutarate monoamide (R-IBM) is a key intermediate in the synthesis of the analgesic drug pregabalin. Recently, the imidase BpIH derived from Burkholderia phytofirmans was identified as a promising catalyst for the industrial production of R-IBM. Notably, this catalyst has the distinct advantage of achieving a 100% theoretical yield from 3-isobutyl glutarimide (IBI). In this study, homology modeling and structure alignment techniques were used to determine the substrate binding pocket of BpIH. Semi-rational design was used to analyze the amino acid residue conservation in the binding pocket region of BpIH. Interestingly, mutations of several low-conserved amino acid located 6-9 Å from the substrate significantly enhanced the catalytic activity of BpIH. Among them, the triple mutant Y37FH133NS226I (YHS-I) showed approximately a fivefold increase in enzyme activity and a significantly improved catalytic efficiency (kcat/Km). Under the same reaction time and conditions, YHS-I successfully converted IBI into R-IBM with a conversion rate of 88.87%, with an enantiomeric excess (ee) of the product exceeding 99.9%. In comparison, wild-type BpIH had a conversion rate of only 38.15%. Molecular dynamics and docking results indicated that YHS-I had higher rigidity around the mutation sites. The synergistic substitutions of Y37F, H133N, and S226I altered the interaction network within the mutation site, enhancing the protein's affinity for the substrate and improving catalytic efficiency. KEY POINTS: • 100% theoretical yield of R-IBM by BpIH compared with 50% by resolution • Semi-rational design of BpIH based on conservativity with homologous enzymes • Mutant with enzyme activity of sixfold and product ee value of 99.9.


Subject(s)
Burkholderia , Burkholderia/enzymology , Burkholderia/genetics , Kinetics , Binding Sites , Substrate Specificity , Models, Molecular , Glutarates/metabolism , Amidohydrolases
4.
Enzymes ; 55: 121-142, 2024.
Article in English | MEDLINE | ID: mdl-39222989

ABSTRACT

Recent research has identified a novel class of carbonic anhydrases (CAs), designated ι-CA, predominantly found in marine diatoms, eukaryotic algae, cyanobacteria, bacteria, and archaea genomes. This class has garnered attention owing to its unique biochemical properties and evolutionary significance. Through bioinformatic analyses, LCIP63, a protein initially annotated with an unknown function, was identified as a potential ι-CA in the marine diatom Thalassiosira pseudonana. Subsequent biochemical characterization revealed that LCIP63 has CA activity and its preference for manganese ions over zinc, indicative of evolutionary adaptation to marine environments. Further exploration of bacterial ι-CAs, exemplified by Burkholderia territorii ι-CA (BteCAι), demonstrated catalytic efficiency and sensitivity to sulfonamide and inorganic anion inhibitors, the classical CA inhibitors (CAIs). The classification of ι-CAs into two variant types based on their sequences, distinguished by the COG4875 and COG4337 domains, marks a significant advancement in our understanding of these enzymes. Structural analyses of COG4337 ι-CAs from eukaryotic microalgae and cyanobacteria thereafter revealed a distinctive structural arrangement and a novel catalytic mechanism involving specific residues facilitating CO2 hydration in the absence of metal ion cofactors, deviating from canonical CA behavior. These findings underscore the biochemical diversity within the ι-CA class and highlight its potential as a target for novel antimicrobial agents. Overall, the elucidation of ι-CA properties and mechanisms advances our knowledge of carbon metabolism in diverse organisms and underscores the complexity of CA evolution and function.


Subject(s)
Carbonic Anhydrases , Carbonic Anhydrases/metabolism , Bacterial Proteins/metabolism , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Bacteria/drug effects , Burkholderia , Diatoms , Cyanobacteria , Carbonic Anhydrase Inhibitors/pharmacology , Carbonic Anhydrase Inhibitors/chemistry
5.
Curr Microbiol ; 81(10): 314, 2024 Aug 20.
Article in English | MEDLINE | ID: mdl-39162848

ABSTRACT

Propanotrophs are a focus of interest because of their ability to degrade numerous environmental contaminants. To explore the phylogeny of microorganisms containing the propane monooxygenase gene cluster (prmABCD), NCBI bacterial genomes and publicly available soil associated metagenomes (from soils, rhizospheres, tree roots) were both examined. Nucleic acid sequences were collected only if all four subunits were located together, were of the expected length and were annotated as propane monooxygenase subunits. In the bacterial genomes, this resulted in data collection only from the phyla Actinomycetota and Pseudomonadota. For the soil associated metagenomes, reads from four studies were subject to quality control, assembly and annotation. Following this, the propane monooxygenase subunit nucleic acid sequences were collected and aligned to the collected bacterial sequences. In total, forty-two propane monooxygenase gene clusters were annotated from the soil associated metagenomes. The majority aligned closely to those from the Actinomycetota, followed by the Alphaproteobacteria, then the Betaproteobacteria. Actinomycetota aligning propane monooxygenase sequences were obtained from all four datasets and most closely aligned to the genera Kribbella and Amycolatopsis. Alphaproteobacteria aligning sequences largely originated from metagenomes associated with miscanthus and switchgrass rhizospheres and primarily aligned with the genera Bradyrhizobium, Acidiphilium and unclassified Rhizobiales. Betaproteobacteria aligning sequences were obtained from only the Red Oak root metagenomes and primarily aligned with the genera Paraburkholderia, Burkholderia and Caballeronia. Interestingly, sequences from the environmental metagenomes were not closely aligned to those from well-studied propanotrophs, such as Mycobacterium and Rhodococcus. Overall, the study highlights the previously unreported diversity of putative propanotrophs in environmental samples. The common occurrence of propane monooxygenase gene clusters has implications for their potential use for contaminant biodegradation.


Subject(s)
Metagenome , Phylogeny , Soil Microbiology , Multigene Family , Cytochrome P-450 CYP4A/genetics , Cytochrome P-450 CYP4A/metabolism , Burkholderia/genetics , Burkholderia/classification , Burkholderia/enzymology , Bradyrhizobium/genetics , Bradyrhizobium/classification , Bradyrhizobium/enzymology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Genome, Bacterial
6.
Environ Sci Pollut Res Int ; 31(38): 50513-50528, 2024 Aug.
Article in English | MEDLINE | ID: mdl-39096459

ABSTRACT

Cadmium (Cd) is a harmful metal in soil, and reducing Cd accumulation in plants has become a vital prerequisite for maintaining food safety. Phosphate-solubilizing bacteria (PSB) can not only improve plant growth but also inhibit the transportation of metals to roots. However, data on gene expression in PSB Burkholderia sp. strain 'N3' and grafted watermelon plants dealing with Cd remain to be elucidated. In this study, core genes and metabolic pathways of strain 'N3' and grafted plants were analyzed by Illumina sequencing. Results showed that 356 and 2527 genes were upregulated in 'N3' and grafted watermelon plants, respectively, whereas 514 and 1540 genes were downregulated in 'N3' and grafted watermelon plants, respectively. Gene ontology enrichment analysis showed that signal transduction, inorganic ion transport, cell motility, amino acid transport, and metabolism pathways were marked in 'N3'. However, pathways such as secondary metabolite biosynthesis, oxidation-reduction process, electron transfer activity, and channel regulator activity were marked in the grafted plants. Six genes related to pentose phosphate, glycolysis, and gluconeogenesis metabolism were upregulated in the grafted plants. This study paves the way for developing potential strategies to improve plant growth under Cd toxicity.


Subject(s)
Cadmium , Citrullus , Phosphates , Cadmium/toxicity , Citrullus/genetics , Transcriptome/drug effects , Soil Pollutants/toxicity , Gene Expression Profiling , Burkholderia/genetics , Burkholderia/metabolism
7.
Int J Biol Macromol ; 278(Pt 1): 134299, 2024 Oct.
Article in English | MEDLINE | ID: mdl-39097047

ABSTRACT

Quorum sensing (QS) is a cellular communication mechanism in which bacteria secrete and recognize signaling molecules to regulate group behavior. Lipases provide energy for bacterial cell growth but it is unknown whether they influence nutrient-dependent QS by hydrolyzing substrate. A high-yield lipase-producing strain, Burkholderia pyrrocinia WZ10-3, was previously identified in our laboratory, but the composition of its crude enzymes was not elucidated. Here, we identified a key extracellular lipase, Lip1728, in WZ10-3, which accounts for 99 % of the extracellular lipase activity. Lip1728 prefers to hydrolyze triglycerides at sn-1,3 positions, with pNP-C16 being its optimal substrate. Lip1728 exhibited activity at pH 5.0-10.0 and regardless of the presence of metal ions. It had strong resistance to sodium dodecyl sulfate and short-chain alcohols and was activated by phenylmethanesulfonylfluoride (PMSF). Lip1728 knockout significantly affected lipid metabolism and biofilm formation in the presence of olive oil. Finally, oleic acid, a hydrolysate of Lip1728, influenced the production of the signal molecule N-acyl homoserine lactone (AHL) and biofilm formation by downregulating the AHL synthetase gene pyrI. In conclusion, Lip1728, as a key extracellular lipase in B. pyrrocinia WZ10-3, exhibits superior properties that make it suitable for biodiesel production and plays a crucial role in QS.


Subject(s)
Burkholderia , Lipase , Quorum Sensing , Lipase/metabolism , Lipase/genetics , Quorum Sensing/genetics , Burkholderia/genetics , Burkholderia/enzymology , Burkholderia/physiology , Biofilms/growth & development , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Substrate Specificity , Lipid Metabolism , Nutrients/metabolism , Acyl-Butyrolactones/metabolism
8.
Chemosphere ; 364: 143065, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39128778

ABSTRACT

In this study, a novel strain Burkholderia stabilis TF-2 capable of assimilatory and co-metabolic degradation of chlorobenzenes was obtained. The interaction between chlorobenzene (CB) and target enzymes, as well as the metabolic pathways in TF-2, were elucidated using multi-omics and molecular docking techniques. Results of degradation experiments indicated that TF-2 assimilated CB at a rate of 0.22-0.66 mg·gcell-1·h-1 in concentrations of 20-200 mg L-1. Additionally, TF-2 also used sodium succinate and sodium citrate as substrates to co-metabolize CB, with degradation rates of 0.26-2.00 and 0.31-1.72 mol·gcell-1·h-1, respectively. Whole-genome sequencing revealed over 18 novel genes associated with aromatic hydrocarbon degradation in TF-2. Transcriptomic analysis showed that CB induced the high expression of 119 genes involved in CB metabolism and late mineralization. The significant up-regulation of the bedC1 (encoding a ring-hydroxylated dioxygenase), CatA (chlorocatechol 1,2-dioxygenase), pcaJ (3-oxoadipate CoA-transferase alpha subunit) and fadA (acetyl-CoA acyltransferase) genes facilitated CB metabolism. Based on these findings, a metabolic pathway for CB was constructed, with the key step involving ortho cleavage of the aromatic ring under the action of the catA gene. Furthermore, molecular docking revealed that CB bound to bedC1 with -4.5 kcal mol-1 through hydrophobic bonds, π-stacking, and a halogen bond. These results provide strong support for development of efficient strains to enhance the removal of chlorinated organic compounds.


Subject(s)
Biodegradation, Environmental , Burkholderia , Chlorobenzenes , Molecular Docking Simulation , Chlorobenzenes/metabolism , Burkholderia/metabolism , Bacterial Proteins/metabolism , Bacterial Proteins/genetics , Dioxygenases/metabolism , Dioxygenases/genetics
9.
Mikrochim Acta ; 191(8): 492, 2024 07 27.
Article in English | MEDLINE | ID: mdl-39066907

ABSTRACT

The development and application of an electrochemical sensor is reported for detection of poly(3-hydroxybutyrate) (P3HB) - a bioplastic derived from agro-industrial residues. To overcome the challenges of molecular imprinting of macromolecules such as P3HB, this study employed methanolysis reaction to break down the P3HB biopolymer chains into methyl 3-hydroxybutyrate (M3HB) monomers. Thereafter, M3HB were employed as the target molecules in the construction of molecularly imprinted sensors. The electrochemical device was then prepared by electropolymerizing a molecularly imprinted poly (indole-3-acetic acid) thin film on a glassy carbon electrode surface modified with reduced graphene oxide (GCE/rGO-MIP) in the presence of M3HB. Electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), scanning electron microscopy with field emission gun (SEM-FEG), Raman spectroscopy, attenuated total reflection Fourier-transform infrared (ATR-FTIR) and X-ray Photoelectron Spectroscopy (XPS) were employed to characterize the electrode surface. Under ideal conditions, the MIP sensor exhibited a wide linear working range of 0.1 - 10 nM and a detection limit of 0.3 pM (n = 3). The sensor showed good repeatability, selectivity, and stability over time. For the sensor application, the bioproduction of P3HB was carried out in a bioreactor containing the Burkholderia glumae MA13 strain and sugarcane byproducts as a supplementary carbon source. The analyses were validated through recovery assays, yielding recovery values between 102 and 104%. These results indicate that this MIP sensor can present advantages in the monitoring of P3HB during the bioconversion process.


Subject(s)
Burkholderia , Electrochemical Techniques , Electrodes , Graphite , Hydroxybutyrates , Molecularly Imprinted Polymers , Polyesters , Graphite/chemistry , Polyesters/chemistry , Hydroxybutyrates/chemistry , Burkholderia/chemistry , Burkholderia/metabolism , Electrochemical Techniques/methods , Electrochemical Techniques/instrumentation , Molecularly Imprinted Polymers/chemistry , Limit of Detection , Oxidation-Reduction , Polyhydroxybutyrates
10.
Microb Drug Resist ; 30(9): 354-362, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39029506

ABSTRACT

Background: Biofilm production in nonfermenting Gram-negative bacteria influences drug resistance. The aim of this work was to evaluate the effect of different antibiotics on biofilm eradication of clinical isolates of Achromobacter, Burkholderia, and Stenotrophomonas maltophilia. Methods: Clinical isolates were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry in a third-level hospital in Monterrey, Mexico. Crystal violet staining was used to determine biofilm production. Drug susceptibility testing was determined by broth microdilution in planktonic cells and biofilm cells. Results: Resistance in planktonic cells was moderate to trimethoprim-sulfamethoxazole, and low to chloramphenicol, minocycline, levofloxacin (S. maltophilia and Burkholderia), ceftazidime, and meropenem (Burkholderia and Achromobacter). Biofilm eradication required higher drug concentrations of ceftazidime, chloramphenicol, levofloxacin, and trimethoprim-sulfamethoxazole than planktonic cells (p < 0.05). Levofloxacin showed biofilm eradication activity in S. maltophilia, minocycline and meropenem in Burkholderia, and meropenem in Achromobacter. Conclusions: Drug resistance increased due to biofilm production for some antibiotics, particularly ceftazidime and trimethoprim-sulfamethoxazole for all three pathogens, chloramphenicol for S. maltophilia and Burkholderia, and levofloxacin for Burkholderia. Some antibiotics could be used for the treatment of biofilm-associated infections in our population, such as levofloxacin for S. maltophilia, minocycline and meropenem for Burkholderia, and meropenem for Achromobacter.


Subject(s)
Achromobacter , Anti-Bacterial Agents , Biofilms , Burkholderia , Gram-Negative Bacterial Infections , Microbial Sensitivity Tests , Stenotrophomonas maltophilia , Biofilms/drug effects , Stenotrophomonas maltophilia/drug effects , Anti-Bacterial Agents/pharmacology , Humans , Burkholderia/drug effects , Achromobacter/drug effects , Gram-Negative Bacterial Infections/microbiology , Gram-Negative Bacterial Infections/drug therapy , Drug Resistance, Bacterial , Trimethoprim, Sulfamethoxazole Drug Combination/pharmacology , Mexico , Ceftazidime/pharmacology , Plankton/drug effects , Drug Resistance, Multiple, Bacterial , Levofloxacin/pharmacology
11.
PLoS Negl Trop Dis ; 18(7): e0012368, 2024 Jul.
Article in English | MEDLINE | ID: mdl-39042701

ABSTRACT

Melioidosis caused by Burkholderia pseudomallei is an infectious disease with a high mortality rate. In acute melioidosis, sepsis is a major cause of death among patients. Once the bacterium enters the bloodstream, immune system dysregulation ensues, leading to cytokine storms. In contrast to B. pseudomallei, a closely related but non-virulent strain B. thailandensis has rarely been reported to cause cytokine storms or death in patients. However, the mechanisms in which the virulent B. pseudomallei causes sepsis are not fully elucidated. It is well-documented that monocytes play an essential role in cytokine production in the bloodstream. The present study, therefore, determined whether there is a difference in the innate immune response to B. pseudomallei and B. thailandensis during infection of primary human monocytes and THP-1 monocytic cells by investigating pyroptosis, an inflammatory death pathway known to play a pivotal role in sepsis. Our results showed that although both bacterial species exhibited a similar ability to invade human monocytes, only B. pseudomallei can significantly increase the release of cytosolic enzyme lactate dehydrogenase (LDH) as well as the increases in caspase-1 and gasdermin D activations in both cell types. The results were consistent with the significant increase in IL-1ß and IL-18 production, key cytokines involved in pyroptosis. Interestingly, there was no significant difference in other cytokine secretion, such as IL-1RA, IL-10, IL-12p70, IL-15, IL-8, and IL-23 in cells infected by both bacterial species. Furthermore, we also demonstrated that ROS production played a crucial role in controlling pyroptosis activation during B. pseudomallei infection in primary human monocytes. These findings suggested that pyroptosis induced by B. pseudomallei in the human monocytes may contribute to the pathogenesis of sepsis in acute melioidosis patients.


Subject(s)
Burkholderia pseudomallei , Burkholderia , Melioidosis , Monocytes , Pyroptosis , Sepsis , Humans , Burkholderia pseudomallei/immunology , Burkholderia pseudomallei/physiology , Monocytes/immunology , Monocytes/microbiology , Melioidosis/microbiology , Melioidosis/immunology , Burkholderia/pathogenicity , Sepsis/microbiology , Sepsis/immunology , Cytokines/metabolism , THP-1 Cells , Immunity, Innate , Cells, Cultured
12.
BMC Genom Data ; 25(1): 73, 2024 Jul 29.
Article in English | MEDLINE | ID: mdl-39075351

ABSTRACT

OBJECTIVES: Rice (Oryza sativa) is the most important food for more than two thirds of the world's population. Bangladesh is the third largest producer and consumer of rice globally. Recently, several symptoms of Bacterial Panicle Blight (BPB) in rice, including seedling blight, sheath rot, floret sterility, and spotted grains, have been detected in the country. In addition, the presence of the most prevalent and virulent causative agent of BPB, Burkholderia glumae, has been confirmed in rice displaying symptoms of the disease. BPB could become one of the next emerging diseases of rice in Bangladesh, and a complete genome of a B. glumae strain from the country will help clarify its origin and devise proper management systems to continue sustainable rice production. DATA DESCRIPTION: We report the first complete genome sequence of a B. glumae strain (BD_21g) isolated from symptomatic rice grains in Bangladesh (Natore District). The genome contains 2 chromosomes (1 and 2, with 3,417,499 and 3,855,283 bp, respectively) and 4 plasmids (1-4, with 123,248, 46,628, 88,744 and 53,064 bp, respectively).


Subject(s)
Burkholderia , Genome, Bacterial , Oryza , Plant Diseases , Oryza/microbiology , Burkholderia/genetics , Burkholderia/isolation & purification , Burkholderia/pathogenicity , Bangladesh , Genome, Bacterial/genetics , Plant Diseases/microbiology , Whole Genome Sequencing
13.
Med Microbiol Immunol ; 213(1): 16, 2024 Jul 20.
Article in English | MEDLINE | ID: mdl-39033094

ABSTRACT

Melioidosis is a severe infectious disease caused by Burkholderia pseudomallei, an intracellular pathogen with a high mortality rate and significant antibiotic resistance. The high mortality rate and resistance to antibiotics have drawn considerable attention from researchers studying melioidosis. This study evaluated the effects of various concentrations (75, 50, and 25 µg/mL) of promethazine hydrochloride (PTZ), a potent antihistamine, on biofilm formation and lipase activity after 24 h of exposure to B. thailandensis E264. A concentration-dependent decrease in both biofilm biomass and lipase activity was observed. RT-PCR analysis revealed that PTZ treatment not only made the biofilm structure loose but also reduced the expression of btaR1, btaR2, btaR3, and scmR. Single gene knockouts of quorum sensing (QS) receptor proteins (∆btaR1, ∆btaR2, and ∆btaR3) were successfully constructed. Deletion of btaR1 affected biofilm formation in B. thailandensis, while deletion of btaR2 and btaR3 led to reduced lipase activity. Molecular docking and biological performance results demonstrated that PTZ inhibits biofilm formation and lipase activity by suppressing the expression of QS-regulated genes. This study found that repositioning PTZ reduced biofilm formation in B. thailandensis E264, suggesting a potential new approach for combating melioidosis.


Subject(s)
Biofilms , Burkholderia , Drug Repositioning , Promethazine , Biofilms/drug effects , Biofilms/growth & development , Burkholderia/drug effects , Burkholderia/physiology , Burkholderia/genetics , Promethazine/pharmacology , Molecular Docking Simulation , Anti-Bacterial Agents/pharmacology , Lipase/metabolism , Lipase/genetics , Gene Expression Regulation, Bacterial/drug effects , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Humans , Quorum Sensing/drug effects
14.
NPJ Biofilms Microbiomes ; 10(1): 64, 2024 Jul 30.
Article in English | MEDLINE | ID: mdl-39080326

ABSTRACT

Plant-sucking insects have intricate associations with a diverse array of microorganisms to facilitate their adaptation to specific ecological niches. The midgut of phytophagous true bugs is generally structured into four distinct compartments to accommodate their microbiota. Nevertheless, there is limited understanding regarding the origins of these gut microbiomes, the mechanisms behind microbial community assembly, and the interactions between gut microbiomes and their insect hosts. In this study, we conducted a comprehensive survey of microbial communities within the midgut compartments of a bean bug Riptortus pedestris, soybean plant, and bulk soil across 12 distinct geographical fields in China, utilizing high-throughput sequencing of the 16 S rRNA gene. Our findings illuminated that gut microbiota of the plant-sucking insects predominantly originated from the surrounding soil environment, and plants also play a subordinate role in mediating microbial acquisition for the insects. Furthermore, our investigation suggested that the composition of the insect gut microbiome was probably shaped by host selection and/or microbe-microbe interactions at the gut compartment level, with marginal influence from soil and geographical factors. Additionally, we had unveiled a noteworthy dynamic in the acquisition of core bacterial taxa, particularly Burkholderia, which were initially sourced from the environment and subsequently enriched within the insect midgut compartments. This bacterial enrichment played a significant role in enhancing insect host reproduction. These findings contribute to our evolving understanding of microbiomes within the insect-plant-soil ecosystem, shedding additional light on the intricate interactions between insects and their microbiomes that underpin the ecological significance of microbial partnerships in host adaptation.


Subject(s)
Bacteria , Gastrointestinal Microbiome , RNA, Ribosomal, 16S , Soil Microbiology , Animals , RNA, Ribosomal, 16S/genetics , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , China , Glycine max/microbiology , High-Throughput Nucleotide Sequencing , Heteroptera/microbiology , Heteroptera/physiology , Reproduction , Phylogeny , Host Microbial Interactions , Burkholderia/genetics , Burkholderia/physiology , Burkholderia/classification
15.
Anal Bioanal Chem ; 416(25): 5431-5443, 2024 Oct.
Article in English | MEDLINE | ID: mdl-38842688

ABSTRACT

Bacterial quorum sensing is a chemical language allowing bacteria to interact through the excretion of molecules called autoinducers, like N-acyl-homoserine lactones (AHLs) produced by Gram-negative Burkholderia and Paraburkholderia bacteria known as opportunistic pathogens. The AHLs differ in their acyl-chain length and may be modified by a 3-oxo or 3-hydroxy substituent, or C = C double bonds at different positions. As the bacterial signal specificity depends on all of these chemical features, their structural characterization is essential to have a better understanding of the population regulation and virulence phenomenon. This study aimed at enabling the localization of the C = C double bond on such specialized metabolites while using significantly lower amounts of biological material. The approach is based on LC-MS/MS analyses of bacterial extracts after in-solution derivatization by a photochemical Paternò-Büchi reaction, leading to the formation of an oxetane ring and subsequently to specific fragmentations when performing MS/MS experiments. The in-solution derivatization of AHLs was optimized on several standards, and then the matrix effect of bacterial extracts on the derivatization was assessed. As a proof of concept, the optimized conditions were applied to a bacterial extract enabling the localization of C = C bonds on unsaturated AHLs.


Subject(s)
Acyl-Butyrolactones , Quorum Sensing , Tandem Mass Spectrometry , Tandem Mass Spectrometry/methods , Acyl-Butyrolactones/chemistry , Acyl-Butyrolactones/metabolism , Acyl-Butyrolactones/analysis , Chromatography, Liquid/methods , Burkholderia/chemistry , Liquid Chromatography-Mass Spectrometry
16.
J Environ Manage ; 362: 121250, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38833921

ABSTRACT

To investigate the impact and mechanism of Cd-tolerant bacteria in soil on promoting Cd accumulation in Ageratum conyzoides L., we verified the impact of inoculating two strains, B-1 (Burkholderia contaminans HA09) and B-7 (Arthrobacter humicola), on Cd accumulation in A. conyzoides through a pot experiment. Additionally, we investigated the dissolution of CdCO3 and nutrient elements, as well as the release of indoleacetic acid (IAA) by the two strains. The results showed that both strains can significantly improve the dissolution of CdCO3. Strains B-1 and B-7 had obvious effect of dissolving phosphorus, which was 5.63 and 2.76 times higher than that of the control group, respectively. Strain B-7 had significant effect of dissolution potassium, which was 1.79 times higher than that of the control group. Strains B-1 and B-7 had significant nitrogen fixation effect, which was 29.53 and 44.39 times higher than that of the control group, respectively. In addition, inoculating with strain B-1 and B-7 significantly increased the Cd extraction efficiency of A. conyzoides (by 114% and 45% respectively) through enhancing Cd accumulation and the biomass of A. conyzoides. Furthermore, the inoculation of strain B-1 and B-7 led to a significant increase in the activities of CAT and SOD, as well as the content of chlorophyll a and total chlorophyll in the leaves of A. conyzoides. To sum up, strain B-1 and B-7 can promote the phytoremediation efficiency of A. conyzoides on Cd by promoting the biomass and Cd accumulation of A. conyzoides.


Subject(s)
Ageratum , Arthrobacter , Biodegradation, Environmental , Cadmium , Soil Pollutants , Cadmium/metabolism , Arthrobacter/metabolism , Soil Pollutants/metabolism , Ageratum/metabolism , Burkholderia/metabolism , Indoleacetic Acids/metabolism
17.
Int J Mol Sci ; 25(11)2024 May 31.
Article in English | MEDLINE | ID: mdl-38892282

ABSTRACT

The rhizobacterial strain BJ3 showed 16S rDNA sequence similarity to species within the Burkholderia genus. Its complete genome sequence revealed a 97% match with Burkholderia contaminans and uncovered gene clusters essential for plant-growth-promoting traits (PGPTs). These clusters include genes responsible for producing indole acetic acid (IAA), osmolytes, non-ribosomal peptides (NRPS), volatile organic compounds (VOCs), siderophores, lipopolysaccharides, hydrolytic enzymes, and spermidine. Additionally, the genome contains genes for nitrogen fixation and phosphate solubilization, as well as a gene encoding 1-aminocyclopropane-1-carboxylate (ACC) deaminase. The treatment with BJ3 enhanced root architecture, boosted vegetative growth, and accelerated early flowering in Arabidopsis. Treated seedlings also showed increased lignin production and antioxidant capabilities, as well as notably increased tolerance to water deficit and high salinity. An RNA-seq transcriptome analysis indicated that BJ3 treatment significantly activated genes related to immunity induction, hormone signaling, and vegetative growth. It specifically activated genes involved in the production of auxin, ethylene, and salicylic acid (SA), as well as genes involved in the synthesis of defense compounds like glucosinolates, camalexin, and terpenoids. The expression of AP2/ERF transcription factors was markedly increased. These findings highlight BJ3's potential to produce various bioactive metabolites and its ability to activate auxin, ethylene, and SA signaling in Arabidopsis, positioning it as a new Burkholderia strain that could significantly improve plant growth, stress resilience, and immune function.


Subject(s)
Arabidopsis , Burkholderia , Stress, Physiological , Burkholderia/genetics , Burkholderia/metabolism , Burkholderia/growth & development , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis/microbiology , Stress, Physiological/genetics , Plant Development/genetics , Indoleacetic Acids/metabolism , Gene Expression Regulation, Plant , Genomics/methods , Plant Growth Regulators/metabolism , Plant Roots/microbiology , Plant Roots/growth & development , Plant Roots/genetics , Plant Roots/metabolism , Ethylenes/metabolism
18.
Sci Rep ; 14(1): 13350, 2024 06 10.
Article in English | MEDLINE | ID: mdl-38858437

ABSTRACT

Lignin, a heterogeneous aromatic polymer present in plant biomass, is intertwined with cellulose and hemicellulose fibrils, posing challenges to its effective utilization due to its phenolic nature and recalcitrance to degradation. In this study, three lignin utilizing bacteria, Klebsiella sp. LEA1, Pseudomonas sp. LEA2, and Burkholderia sp. LEA3, were isolated from deciduous forest soil samples in Nan province, Thailand. These isolates were capable of growing on alkali lignin and various lignin-associated monomers at 40 °C under microaerobic conditions. The presence of Cu2+ significantly enhanced guaiacol oxidation in Klebsiella sp. LEA1 and Pseudomonas sp. LEA2. Lignin-related monomers and intermediates such as 2,6-dimethoxyphenol, 4-vinyl guaiacol, 4-hydroxybenzoic acid, benzoic acid, catechol, and succinic acid were detected mostly during the late stage of incubation of Klebsiella sp. LEA1 and Pseudomonas sp. LEA2 in lignin minimal salt media via GC-MS analysis. The intermediates identified from Klebsiella sp. LEA1 degradation suggested that conversion and utilization occurred through the ß-ketoadipate (ortho-cleavage) pathway under limited oxygen conditions. The ability of these bacteria to thrive on alkaline lignin and produce various lignin-related intermediates under limited oxygen conditions suggests their potential utility in oxygen-limited processes and the production of renewable chemicals from plant biomass.


Subject(s)
Forests , Klebsiella , Lignin , Oxygen , Pseudomonas , Soil Microbiology , Lignin/metabolism , Pseudomonas/metabolism , Pseudomonas/isolation & purification , Oxygen/metabolism , Klebsiella/metabolism , Klebsiella/isolation & purification , Burkholderia/metabolism , Burkholderia/isolation & purification , Biodegradation, Environmental
19.
Am J Trop Med Hyg ; 111(2): 281-286, 2024 Aug 07.
Article in English | MEDLINE | ID: mdl-38917784

ABSTRACT

Burkholderia spp. are opportunistic pathogens that cause infection in patients with disrupted immunity. The study intended to demonstrate the epidemiology and clinical features associated with Burkholderia spp. bacteremia. This retrospective study was performed to assess the clinical and laboratory characteristics of patients whose blood cultures were growing Burkholderia spp. and, based on their underlying comorbidities, were subjected to survival analysis from January 2022 to December 2022 at a university hospital in northern India. Three hundred patients with Burkholderia spp. bacteremia were included in this study conducted over 1 year. The mean age of the patients was 33.86 years with a male predominance of 56.67% (170/300, 56.67%). Underlying malignancies (207/300, 69.0%) were the most common clinical diagnosis, and catheter in situ (300/300, 100.0%) was the most common risk factor. Burkholderia cenocepacia (244/300, 81.33%) was the most common Burkholderia spp. isolated. All isolates were highly susceptible to minocycline. Kidney disease (P = 0.029), hypertension (P = 0.005), type 2 diabetes mellitus (P = 0.039), and respiratory disease (P <0.001) in patients were significantly associated with death owing to Burkholderia spp. bacteremia, whereas patients with malignancies (P <0.001) and undergoing treatment were significantly associated with a better outcome when the microorganism was susceptible to empirical antibiotics. The presence of indwelling devices, mechanical ventilation (P <0.001), and a hemodialysis catheter (P = 0.026) were statistically significant risk factors associated with poor outcomes.


Subject(s)
Bacteremia , Burkholderia Infections , Burkholderia , Humans , India/epidemiology , Male , Female , Bacteremia/epidemiology , Bacteremia/microbiology , Burkholderia Infections/epidemiology , Burkholderia Infections/microbiology , Adult , Retrospective Studies , Burkholderia/isolation & purification , Middle Aged , Young Adult , Risk Factors , Adolescent , Anti-Bacterial Agents/therapeutic use , Child , Aged , Neoplasms/complications , Neoplasms/epidemiology
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