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1.
BMC Microbiol ; 24(1): 385, 2024 Oct 02.
Artículo en Inglés | MEDLINE | ID: mdl-39358715

RESUMEN

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.


Asunto(s)
Burkholderia , Fosfatos de Calcio , Compostaje , Fósforo , Microbiología del Suelo , Fosfatos de Calcio/metabolismo , Fósforo/metabolismo , Burkholderia/metabolismo , Burkholderia/genética , Burkholderia/efectos de los fármacos , Bacterias/metabolismo , Bacterias/genética , Bacterias/clasificación , Bacterias/efectos de los fármacos , Microbiota/efectos de los fármacos , Nitrógeno/metabolismo , Suelo/química , Hojas de la Planta/microbiología , Hongos/metabolismo , Hongos/efectos de los fármacos , Hongos/genética , Hongos/clasificación , Concentración de Iones de Hidrógeno
2.
PLoS Pathog ; 20(10): e1011862, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39361719

RESUMEN

The plant pathogenic bacterium Burkholderia glumae causes bacterial panicle blight (BPB) in rice-growing areas worldwide. It has been widely accepted that an acyl-homoserine lactone (AHL)-type quorum sensing (QS) system encoded by tofI and tofR genes (TofIR QS) is a key regulatory mechanism underlying the bacterial pathogenesis of B. glumae. In addition, qsmR, which encodes an IclR-family regulatory protein, has been considered an important part of TofIR QS. However, the present study with three strains of B. glumae representing different pathogenic strains revealed that this currently accepted paradigm should be modified. We characterized the regulatory function of TofIR QS and qsmR in three different strains of B. glumae, 336gr-1 (virulent), 411gr-6 (hypervirulent) and 257sh-1 (avirulent). In 336gr-1, both TofIR QS and qsmR were critical for the pathogenesis, being consistent with previous studies. However, in the hypervirulent strain 411gr-6, TofIR QS only partially contributes to the virulence, whereas qsmR was critical for pathogenesis like in 336gr-1. Furthermore, we found that a single nucleotide polymorphism causing T50K substitution in the qsmR coding sequence was the cause of the non-pathogenic trait of the naturally avirulent strain 257sh-1. Subsequent analyses of gene expression and transcriptome revealed that TofIR QS is partially controlled by qsmR at the transcriptional level in both virulent strains. Further genetic tests of additional B. glumae strains showed that 11 out of 20 virulent strains retained the ability to produce toxoflavin even after removing the tofI/tofM/tofR QS gene cluster like 411gr-6. In contrast, all the virulent strains tested lost the ability to produce toxoflavin almost completely upon deletion of the qsmR gene. Taking these results together, qsmR, rather than TofIR QS, is a master regulator that determines the pathogenic trait of B. glumae thus a more appropriate pathogen target for successful management of BPB.


Asunto(s)
Proteínas Bacterianas , Burkholderia , Enfermedades de las Plantas , Factores de Transcripción , Acil-Butirolactonas/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Burkholderia/genética , Burkholderia/patogenicidad , Burkholderia/metabolismo , Regulación Bacteriana de la Expresión Génica , Oryza/microbiología , Enfermedades de las Plantas/microbiología , Percepción de Quorum/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Virulencia
3.
Appl Microbiol Biotechnol ; 108(1): 474, 2024 Sep 25.
Artículo en Inglés | MEDLINE | ID: mdl-39320489

RESUMEN

(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.


Asunto(s)
Burkholderia , Burkholderia/enzimología , Burkholderia/genética , Cinética , Sitios de Unión , Especificidad por Sustrato , Modelos Moleculares , Glutaratos/metabolismo , Amidohidrolasas
4.
Microb Cell Fact ; 23(1): 255, 2024 Sep 28.
Artículo en Inglés | MEDLINE | ID: mdl-39342283

RESUMEN

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.


Asunto(s)
Burkholderia , Burkholderia/metabolismo , Burkholderia/genética , Fusarium/metabolismo , Fusarium/genética , Fusarium/efectos de los fármacos , Péptidos Cíclicos/biosíntesis , Péptido Sintasas/genética , Péptido Sintasas/metabolismo , Ácidos Dicarboxílicos/metabolismo , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética
5.
J Agric Food Chem ; 72(40): 22132-22143, 2024 Oct 09.
Artículo en Inglés | MEDLINE | ID: mdl-39316703

RESUMEN

Banana anthracnose, caused by Colletotrichum fructicola, significantly reduced the postharvest fruit quality. Employing biocontrol strategies offers a sustainable approach to enhance agricultural practices. The Burkholderia sp. strain BX1 hinders the growth and appressorium formation of C. fructicola, and its sterile filtrate lowers the anthracnose incidence while preserving the fruit quality. Scanning electron microscopy and genomic analyses confirmed BX1 as Burkholderia pyrrocinia. AntiSMASH analysis identified three siderophores with high similarity, and improved MALDI-TOF IMS confirmed the presence of the siderophore pyochelin. Furthermore, the BX1 filtrate suppressed the expression of virulence genes in C. fructicola and induced the expression of disease resistance genes in banana. However, the presence of 80 µM iron ions notably mitigated BX1's inhibitory effects and reversed the changes in related gene expression. These results underscore BX1's robust efficacy as a biocontrol agent in managing banana anthracnose, highlight the effective antifungal compounds, and elucidate the influence of environmental factors on biocontrol effectiveness.


Asunto(s)
Colletotrichum , Frutas , Musa , Enfermedades de las Plantas , Sideróforos , Musa/microbiología , Colletotrichum/fisiología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Frutas/microbiología , Sideróforos/metabolismo , Burkholderia/genética , Burkholderia/metabolismo , Burkholderia/fisiología , Agentes de Control Biológico/farmacología
6.
Int J Biol Macromol ; 278(Pt 1): 134299, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39097047

RESUMEN

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.


Asunto(s)
Burkholderia , Lipasa , Percepción de Quorum , Lipasa/metabolismo , Lipasa/genética , Percepción de Quorum/genética , Burkholderia/genética , Burkholderia/enzimología , Burkholderia/fisiología , Biopelículas/crecimiento & desarrollo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Especificidad por Sustrato , Metabolismo de los Lípidos , Nutrientes/metabolismo , Acil-Butirolactonas/metabolismo
7.
Curr Microbiol ; 81(10): 314, 2024 Aug 20.
Artículo en Inglés | MEDLINE | ID: mdl-39162848

RESUMEN

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.


Asunto(s)
Metagenoma , Filogenia , Microbiología del Suelo , Familia de Multigenes , Citocromo P-450 CYP4A/genética , Citocromo P-450 CYP4A/metabolismo , Burkholderia/genética , Burkholderia/clasificación , Burkholderia/enzimología , Bradyrhizobium/genética , Bradyrhizobium/clasificación , Bradyrhizobium/enzimología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Genoma Bacteriano
8.
Microbiol Spectr ; 12(10): e0366223, 2024 Oct 03.
Artículo en Inglés | MEDLINE | ID: mdl-39136439

RESUMEN

The seed serves as the primary source for establishing microbial populations in plants across subsequent generations, influencing plant growth and overall health. Cropping conditions, especially farming practices, can influence the composition and functionality of the seed microbiome. Very little is known about the differences in seed microbiome between organic and conventional production systems. In this study, we characterized the endophytic microbial populations in seeds of rice grown under organic and conventional management practices through culture-dependent and -independent analyses. The V4 region of 16S rRNA was used for bacterial taxa identification, and the ITS1 region was used for the identification of fungal taxa. Our results revealed significantly higher Shannon and Simpson indices for bacterial diversity in the conventional farming system, whereas the fungal diversity was higher for observed, Shannon, and Simpson indices in the organic farming system. The cultivable endophytic bacteria were isolated and identified using the full-length 16S rRNA gene. There was no difference in culturable endophytic bacterial isolates in rice seeds grown under both conventional and organic farming systems. Among 33 unique isolates tested in vitro, three bacteria-Bacillus sp. ST24, Burkholderia sp. OR5, and Pantoea sp. ST25-showed antagonistic activities against Marasmius graminum, Rhizoctonia solani AG4, and R. solani AG11, the fungal pathogens causing seedling blight in rice. IMPORTANCE: In this paper, we studied the differences in the endophytic microbial composition of rice seeds grown in conventional and organic farming systems. Our results demonstrate a greater bacterial diversity in conventional farming, while organic farming showcases a higher fungal diversity. Additionally, our research reveals the ability of seed bacterial endophytes to inhibit the growth of three fungal pathogens responsible for causing seedling blight in rice. This study provides valuable insights into the potential use of beneficial seed microbial endophytes for developing a novel microbiome-based strategy in the management of rice diseases. Such an approach has the potential to enhance overall plant health and improve crop productivity.


Asunto(s)
Bacterias , Endófitos , Hongos , Microbiota , Agricultura Orgánica , Oryza , ARN Ribosómico 16S , Semillas , Oryza/microbiología , Endófitos/aislamiento & purificación , Endófitos/clasificación , Endófitos/genética , Semillas/microbiología , Microbiota/genética , ARN Ribosómico 16S/genética , Bacterias/clasificación , Bacterias/aislamiento & purificación , Bacterias/genética , Hongos/aislamiento & purificación , Hongos/clasificación , Hongos/genética , Burkholderia/genética , Burkholderia/aislamiento & purificación , Burkholderia/clasificación , Rhizoctonia/aislamiento & purificación , Rhizoctonia/genética , Rhizoctonia/crecimiento & desarrollo , Bacillus/aislamiento & purificación , Bacillus/genética , Bacillus/clasificación , Pantoea/aislamiento & purificación , Pantoea/genética , Pantoea/clasificación , Enfermedades de las Plantas/microbiología , Agricultura/métodos
9.
Environ Sci Pollut Res Int ; 31(38): 50513-50528, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-39096459

RESUMEN

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.


Asunto(s)
Cadmio , Citrullus , Fosfatos , Cadmio/toxicidad , Citrullus/genética , Transcriptoma/efectos de los fármacos , Contaminantes del Suelo/toxicidad , Perfilación de la Expresión Génica , Burkholderia/genética , Burkholderia/metabolismo
10.
NPJ Biofilms Microbiomes ; 10(1): 64, 2024 Jul 30.
Artículo en Inglés | MEDLINE | ID: mdl-39080326

RESUMEN

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.


Asunto(s)
Bacterias , Microbioma Gastrointestinal , ARN Ribosómico 16S , Microbiología del Suelo , Animales , ARN Ribosómico 16S/genética , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , China , Glycine max/microbiología , Secuenciación de Nucleótidos de Alto Rendimiento , Heterópteros/microbiología , Heterópteros/fisiología , Reproducción , Filogenia , Interacciones Microbiota-Huesped , Burkholderia/genética , Burkholderia/fisiología , Burkholderia/clasificación
11.
BMC Genom Data ; 25(1): 73, 2024 Jul 29.
Artículo en Inglés | MEDLINE | ID: mdl-39075351

RESUMEN

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).


Asunto(s)
Burkholderia , Genoma Bacteriano , Oryza , Enfermedades de las Plantas , Oryza/microbiología , Burkholderia/genética , Burkholderia/aislamiento & purificación , Burkholderia/patogenicidad , Bangladesh , Genoma Bacteriano/genética , Enfermedades de las Plantas/microbiología , Secuenciación Completa del Genoma
12.
Med Microbiol Immunol ; 213(1): 16, 2024 Jul 20.
Artículo en Inglés | MEDLINE | ID: mdl-39033094

RESUMEN

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.


Asunto(s)
Biopelículas , Burkholderia , Reposicionamiento de Medicamentos , Prometazina , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Burkholderia/efectos de los fármacos , Burkholderia/fisiología , Burkholderia/genética , Prometazina/farmacología , Simulación del Acoplamiento Molecular , Antibacterianos/farmacología , Lipasa/metabolismo , Lipasa/genética , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Humanos , Percepción de Quorum/efectos de los fármacos
13.
J Hazard Mater ; 475: 134936, 2024 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-38889456

RESUMEN

Biotic-abiotic hybrid systems have recently emerged as a potential technique for stable and efficient removal of persistent contaminants due to coupling of microbial catabolic with abiotic adsorption/redox processes. In this study, Burkholderia vietnamensis C09V (B.V.C09V) was successfully integrated with a Zeolitic Imidazolate Framework-8 (ZIF-8) to construct a state-of-art biotic-abiotic system using polyvinyl alcohol/ sodium alginate (PVA/SA) as media. The biotic-abiotic system (PVA/SA-ZIF-8 @B.V.C09V) was able to remove 99.0 % of 2,4-DCP within 168 h, which was much higher than either PVA/SA, PVA/SA-ZIF-8 or PVA/SA@B.V.C09V (53.8 %, 72.6 % and 67.2 %, respectively). Electrochemical techniques demonstrated that the carrier effect of PVA/SA and the driving effect of ZIF-8 collectively accelerated electron transfer processes associated with enzymatic reactions. In addition, quantitative-PCR (Q-PCR) revealed that ZIF-8 stimulated B.V.C09V to up-regulate expression of tfdB, tfdC, catA, and catC genes (2.40-, 1.68-, 1.58-, and 1.23-fold, respectively), which encoded the metabolism of related enzymes. Furthermore, the effect of key physical, chemical, and biological properties of PVA/SA-ZIF-8 @B.V.C09V on 2,4-DCP removal were statistically investigated by Spearman correlation analysis to identify the key factors that promoted synergistic removal of 2,4-DCP. Overall, this study has created an innovative new strategy for the sustainable remediation of 2,4-DCP in aquatic environments.


Asunto(s)
Clorofenoles , Alcohol Polivinílico , Contaminantes Químicos del Agua , Zeolitas , Clorofenoles/química , Contaminantes Químicos del Agua/química , Alcohol Polivinílico/química , Zeolitas/química , Alginatos/química , Burkholderia/metabolismo , Burkholderia/genética , Adsorción , Imidazoles/química , Biodegradación Ambiental , Estructuras Metalorgánicas/química
14.
Int J Mol Sci ; 25(11)2024 May 31.
Artículo en Inglés | MEDLINE | ID: mdl-38892282

RESUMEN

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.


Asunto(s)
Arabidopsis , Burkholderia , Estrés Fisiológico , Burkholderia/genética , Burkholderia/metabolismo , Burkholderia/crecimiento & desarrollo , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Arabidopsis/microbiología , Estrés Fisiológico/genética , Desarrollo de la Planta/genética , Ácidos Indolacéticos/metabolismo , Regulación de la Expresión Génica de las Plantas , Genómica/métodos , Reguladores del Crecimiento de las Plantas/metabolismo , Raíces de Plantas/microbiología , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Etilenos/metabolismo
15.
Microb Genom ; 10(6)2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38860878

RESUMEN

Endofungal Mycetohabitans (formerly Burkholderia) spp. rely on a type III secretion system to deliver mostly unidentified effector proteins when colonizing their host fungus, Rhizopus microsporus. The one known secreted effector family from Mycetohabitans consists of homologues of transcription activator-like (TAL) effectors, which are used by plant pathogenic Xanthomonas and Ralstonia spp. to activate host genes that promote disease. These 'Burkholderia TAL-like (Btl)' proteins bind corresponding specific DNA sequences in a predictable manner, but their genomic target(s) and impact on transcription in the fungus are unknown. Recent phenotyping of Btl mutants of two Mycetohabitans strains revealed that the single Btl in one Mycetohabitans endofungorum strain enhances fungal membrane stress tolerance, while others in a Mycetohabitans rhizoxinica strain promote bacterial colonization of the fungus. The phenotypic diversity underscores the need to assess the sequence diversity and, given that sequence diversity translates to DNA targeting specificity, the functional diversity of Btl proteins. Using a dual approach to maximize capture of Btl protein sequences for our analysis, we sequenced and assembled nine Mycetohabitans spp. genomes using long-read PacBio technology and also mined available short-read Illumina fungal-bacterial metagenomes. We show that btl genes are present across diverse Mycetohabitans strains from Mucoromycota fungal hosts yet vary in sequences and predicted DNA binding specificity. Phylogenetic analysis revealed distinct clades of Btl proteins and suggested that Mycetohabitans might contain more species than previously recognized. Within our data set, Btl proteins were more conserved across M. rhizoxinica strains than across M. endofungorum, but there was also evidence of greater overall strain diversity within the latter clade. Overall, the results suggest that Btl proteins contribute to bacterial-fungal symbioses in myriad ways.


Asunto(s)
Burkholderia , Rhizopus , Simbiosis , Rhizopus/genética , Rhizopus/metabolismo , Burkholderia/genética , Burkholderia/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Filogenia , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Variación Genética
16.
World J Microbiol Biotechnol ; 40(8): 242, 2024 Jun 13.
Artículo en Inglés | MEDLINE | ID: mdl-38869634

RESUMEN

Lignocellulosic biomass is a valuable, renewable substrate for the synthesis of polyhydroxybutyrate (PHB), an ecofriendly biopolymer. In this study, bacterial strain E5-3 was isolated from soil in Japan; it was identified as Burkholderia ambifaria strain E5-3 by 16 S rRNA gene sequencing. The strain showed optimal growth at 37 °C with an initial pH of 9. It demonstrated diverse metabolic ability, processing a broad range of carbon substrates, including xylose, glucose, sucrose, glycerol, cellobiose, and, notably, palm oil. Palm oil induced the highest cellular growth, with a PHB content of 65% wt. The strain exhibited inherent tolerance to potential fermentation inhibitors derived from lignocellulosic hydrolysate, withstanding 3 g/L 5-hydroxymethylfurfural and 1.25 g/L acetic acid. Employing a fed-batch fermentation strategy with a combination of glucose, xylose, and cellobiose resulted in PHB production 2.7-times that in traditional batch fermentation. The use of oil palm trunk hydrolysate, without inhibitor pretreatment, in a fed-batch fermentation setup led to significant cell growth with a PHB content of 45% wt, equivalent to 10 g/L. The physicochemical attributes of xylose-derived PHB produced by strain E5-3 included a molecular weight of 722 kDa, a number-average molecular weight of 191 kDa, and a polydispersity index of 3.78. The amorphous structure of this PHB displayed a glass transition temperature of 4.59 °C, while its crystalline counterpart had a melting point of 171.03 °C. This research highlights the potential of lignocellulosic feedstocks, especially oil palm trunk hydrolysate, for PHB production through fed-batch fermentation by B. ambifaria strain E5-3, which has high inhibitor tolerance.


Asunto(s)
Biomasa , Burkholderia , Fermentación , Hidroxibutiratos , Lignina , Aceite de Palma , ARN Ribosómico 16S , Xilosa , Lignina/metabolismo , Aceite de Palma/metabolismo , Hidroxibutiratos/metabolismo , Burkholderia/metabolismo , Burkholderia/genética , Burkholderia/crecimiento & desarrollo , Xilosa/metabolismo , ARN Ribosómico 16S/genética , Microbiología del Suelo , Glucosa/metabolismo , Poliésteres/metabolismo , Concentración de Iones de Hidrógeno , Furaldehído/metabolismo , Furaldehído/análogos & derivados , Celobiosa/metabolismo
17.
Enzyme Microb Technol ; 179: 110469, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38878426

RESUMEN

Esomeprazole is the most popular proton pump inhibitor for treating gastroesophageal reflux disease. Previously, a phenylacetone monooxygenase mutant LnPAMOmu15 (LM15) was obtained by protein engineering for asymmetric synthesis of esomeprazole using pyrmetazole as substrate. To scale up the whole cell asymmetric synthesis of esomeprazole and reduce the cost, in this work, an Escherichia coli whole-cell catalyst harboring LM15 and formate dehydrogenase from Burkholderia stabilis 15516 (BstFDH) were constructed through optimized gene assembly patterns. CRISPR/Cas9 mediated insertion of Ptrc promoter in genome was done to enhance the expression of key genes to increase the cellular NADP supply in the whole cell catalyst, by which the amount of externally added NADP+ for the asymmetric synthesis of esomeprazole decreased to 0.05 mM from 0.3 mM for reducing the cost. After the optimization of reaction conditions in the reactor, the scalable synthesis of esomeprazole was performed using the efficient LM15-BstFDH whole-cell as catalyst, which showed the highest reported space-time yield of 3.28 g/L/h with 50 mM of pyrmetazole loading. Isolation procedure was conducted to obtain esomeprazole sodium of 99.55 % purity and > 99.9 % ee with 90.1 % isolation yield. This work provides the basis for production of enantio-pure esomeprazole via cost-effective whole cell biocatalysis.


Asunto(s)
Biocatálisis , Burkholderia , Escherichia coli , Esomeprazol , Esomeprazol/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Burkholderia/genética , Burkholderia/enzimología , Burkholderia/metabolismo , Coenzimas/metabolismo , Vías Biosintéticas , Ingeniería Metabólica , Formiato Deshidrogenasas/metabolismo , Formiato Deshidrogenasas/genética , Sistemas CRISPR-Cas , Oxigenasas de Función Mixta/metabolismo , Oxigenasas de Función Mixta/genética
18.
Int J Biol Macromol ; 272(Pt 1): 132630, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38810853

RESUMEN

Biocatalyst catalyzing the synthesis of esters under aqueous phase is an alternative with green and sustainable characteristics. Here, a biocatalyst esterase Bur01 was identified through genome sequencing and gene library construction from a Burkholderia ambifaria BJQ0010 with efficient ester synthesis property under aqueous phase for the first time. Bur01 was soluble expressed and the purified enzyme showed the highest activity at pH 4.0 and 40 °C. It had a broad substrate spectrum, especially for ethyl esters. The structure of Bur01 was categorized as a member of α/ß fold hydrolase superfamily. The easier opening of lid under aqueous phase and the hydrophobicity of substrate channel contribute to easier access to the active center for substrate. Molecular docking and site-directed mutation demonstrated that the oxyanion hole Ala22, Met112 and π-bond stacking between His24 and Phe217 played essential roles in catalytic function. The mutants V149A, V149I, L159I and F137I enhanced enzyme activity to 1.42, 1.14, 1.32 and 2.19 folds due to reduced spatial resistance and increased hydrophobicity of channel and ethyl octanoate with the highest conversion ratio of 68.28 % was obtained for F137I. These results provided new ideas for developing green catalysts and catalytic basis of mechanistic studies for ester synthetase under aqueous phase.


Asunto(s)
Biocatálisis , Burkholderia , Esterasas , Ésteres , Simulación del Acoplamiento Molecular , Esterasas/metabolismo , Esterasas/genética , Esterasas/química , Ésteres/metabolismo , Ésteres/química , Burkholderia/enzimología , Burkholderia/genética , Especificidad por Sustrato , Concentración de Iones de Hidrógeno , Agua/química , Dominio Catalítico , Mutagénesis Sitio-Dirigida , Cinética
19.
Emerg Infect Dis ; 30(6): 1249-1252, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38782141

RESUMEN

Burkholderia semiarida was previously identified solely as a plant pathogen within the Burkholderia cepacia complex. We present a case in China involving recurrent pneumonia attributed to B. semiarida infection. Of note, the infection manifested in an immunocompetent patient with no associated primary diseases and endured for >3 years.


Asunto(s)
Infecciones por Burkholderia , Burkholderia , Recurrencia , Humanos , Infecciones por Burkholderia/diagnóstico , Infecciones por Burkholderia/microbiología , Infecciones por Burkholderia/tratamiento farmacológico , China , Burkholderia/aislamiento & purificación , Burkholderia/genética , Masculino , Inmunocompetencia , Antibacterianos/uso terapéutico , Persona de Mediana Edad , Neumonía Bacteriana/microbiología , Neumonía Bacteriana/diagnóstico , Neumonía Bacteriana/tratamiento farmacológico
20.
Emerg Infect Dis ; 30(5): 1055-1057, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38666739

RESUMEN

We report a clinical isolate of Burkholderia thailandensis 2022DZh obtained from a patient with an infected wound in southwest China. Genomic analysis indicates that this isolate clusters with B. thailandensis BPM, a human isolate from Chongqing, China. We recommend enhancing monitoring and surveillance for B. thailandensis infection in both humans and livestock.


Asunto(s)
Infecciones por Burkholderia , Burkholderia , Filogenia , Infección de Heridas , Humanos , Masculino , Burkholderia/genética , Burkholderia/aislamiento & purificación , Burkholderia/clasificación , Infecciones por Burkholderia/microbiología , Infecciones por Burkholderia/diagnóstico , China/epidemiología , Genoma Bacteriano , Infección de Heridas/microbiología , Persona de Mediana Edad
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