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Biotechnology and Bioengineering Sep 2022Lignin is a largely untapped source for the bioproduction of value-added chemicals. Pseudomonas putida KT2440 has emerged as a strong candidate for bioprocessing of...
Lignin is a largely untapped source for the bioproduction of value-added chemicals. Pseudomonas putida KT2440 has emerged as a strong candidate for bioprocessing of lignin feedstocks due to its resistance to several industrial solvents, broad metabolic capabilities, and genetic amenability. Here we demonstrate the engineering of P. putida for the ability to metabolize syringic acid, one of the major products that comes from the breakdown of the syringyl component of lignin. The rational design was first applied for the construction of strain Sy-1 by overexpressing a native vanillate demethylase. Subsequent adaptive laboratory evolution (ALE) led to the generation of mutations that achieved robust growth on syringic acid as a sole carbon source. The best mutant showed a 30% increase in the growth rate over the original engineered strain. Genomic sequencing revealed multiple mutations repeated in separate evolved replicates. Reverse engineering of mutations identified in agmR, gbdR, fleQ, and the intergenic region of gstB and yadG into the parental strain recaptured the improved growth of the evolved strains to varied extent. These findings thus reveal the ability of P. putida to utilize lignin more fully as a feedstock and make it a more economically viable chassis for chemical production.
Topics: Base Sequence; Carbon; Lignin; Metabolic Engineering; Pseudomonas putida
PubMed: 35524438
DOI: 10.1002/bit.28131 -
Communications Biology Dec 2022Despite advances in understanding the metabolism of Pseudomonas putida KT2440, a promising bacterial host for producing valuable chemicals from plant-derived feedstocks,...
Despite advances in understanding the metabolism of Pseudomonas putida KT2440, a promising bacterial host for producing valuable chemicals from plant-derived feedstocks, a strain capable of producing free fatty acid-derived chemicals has not been developed. Guided by functional genomics, we engineered P. putida to produce medium- and long-chain free fatty acids (FFAs) to titers of up to 670 mg/L. Additionally, by taking advantage of the varying substrate preferences of paralogous native fatty acyl-CoA ligases, we employed a strategy to control FFA chain length that resulted in a P. putida strain specialized in producing medium-chain FFAs. Finally, we demonstrate the production of oleochemicals in these strains by synthesizing medium-chain fatty acid methyl esters, compounds useful as biodiesel blending agents, in various media including sorghum hydrolysate at titers greater than 300 mg/L. This work paves the road to produce high-value oleochemicals and biofuels from cheap feedstocks, such as plant biomass, using this host.
Topics: Pseudomonas putida; Fatty Acids, Nonesterified; Biofuels; Biomass; Fatty Acids
PubMed: 36509863
DOI: 10.1038/s42003-022-04336-2 -
BMC Microbiology Jan 2021Pseudomonas putida KT2440 is a metabolically versatile, HV1-certified, genetically accessible, and thus interesting microbial chassis for biotechnological applications....
BACKGROUND
Pseudomonas putida KT2440 is a metabolically versatile, HV1-certified, genetically accessible, and thus interesting microbial chassis for biotechnological applications. However, its obligate aerobic nature hampers production of oxygen sensitive products and drives up costs in large scale fermentation. The inability to perform anaerobic fermentation has been attributed to insufficient ATP production and an inability to produce pyrimidines under these conditions. Addressing these bottlenecks enabled growth under micro-oxic conditions but does not lead to growth or survival under anoxic conditions.
RESULTS
Here, a data-driven approach was used to develop a rational design for a P. putida KT2440 derivative strain capable of anaerobic respiration. To come to the design, data derived from a genome comparison of 1628 Pseudomonas strains was combined with genome-scale metabolic modelling simulations and a transcriptome dataset of 47 samples representing 14 environmental conditions from the facultative anaerobe Pseudomonas aeruginosa.
CONCLUSIONS
The results indicate that the implementation of anaerobic respiration in P. putida KT2440 would require at least 49 additional genes of known function, at least 8 genes encoding proteins of unknown function, and 3 externally added vitamins.
Topics: Anaerobiosis; Bacterial Proteins; Computer Simulation; Databases, Genetic; Fermentation; Gene Expression Profiling; Metabolic Engineering; Microbial Viability; Pseudomonas putida; Pyrimidines
PubMed: 33407113
DOI: 10.1186/s12866-020-02058-1 -
Journal of Contaminant Hydrology May 2023The clopidogrel bisulfate was degraded under aerobic conditions by two bacterial strains isolated from industrial effluents in El-Harrach, Algeria. The sequencing of...
The clopidogrel bisulfate was degraded under aerobic conditions by two bacterial strains isolated from industrial effluents in El-Harrach, Algeria. The sequencing of their 16S rRNA revealed that these two strains are Pseudomonas aeruginosa and Pseudomonas putida. The experiments showed that this consortium could remove clopidogrel bisulfate at high concentrations (5-1500 mg·L) within 96 h incubation period. The HPLC analysis recorded 75.23% degradation of clopidogrel bisulfate at an initial concentration of 100 mg·L after five days of incubation at pH 7.0 and a temperature of 30 °C. Also, a maximum degradation of 99.08% was carried out at a more basic pH (8.5). While only 41% was degraded at a temperature of 20 °C. Moreover, the presence of supplemental sources of carbon and nitrogen in the mixed culture media effectively improved the biodegradation of clopidogrel bisulfate by the stains. Finally, the morphology of the strains and the properties of the cell's surface were studied using a scanning electron microscope (SEM). This study reports, for the first time, the viability of the aerobic biodegradation of clopidogrel bisulfate in water in a wide range of concentrations.
Topics: Clopidogrel; Pseudomonas aeruginosa; Pseudomonas putida; Wastewater; RNA, Ribosomal, 16S; Biodegradation, Environmental
PubMed: 37167785
DOI: 10.1016/j.jconhyd.2023.104198 -
FEMS Microbiology Letters May 2020Pseudomonas putida and closely-related species such as Pseudomonas fluorescens and Pseudomonas brassicacearum have been reported as potential biocontrol agents and plant...
Pseudomonas putida and closely-related species such as Pseudomonas fluorescens and Pseudomonas brassicacearum have been reported as potential biocontrol agents and plant growth-promoters. Recently, we have described the biocontrol activity of P. putida B2017 against several phytopathogens of agricultural relevance. In this study, its ability to produce potential antibiotic / toxic metabolites was assessed by functional, chromatography-mass spectrometry and genomic analysis. Our results show that B2017 is not able to synthesize surfactants and common antibiotics produced by Pseudomonas spp., i.e. pyrrolnitrin, 2,4-diacetylphloroglucinol, pyoluteorin and pyocyanin, but it produces pyoverdine, a siderophore which is involved in its biocontrol activity. The non-production of other metabolites, such as cyanide, safracin, promysalin and lipopeptides between others, is also discussed. Our data suggest that the mode of action of B2017 is not mainly due to the production of antimicrobial / toxic metabolites. Moreover, these features make P. putida B2017 a promising biocontrol microorganism for plant protection without side effects on environment, non-target organisms and human health.
Topics: Anti-Bacterial Agents; Bacteria; Bacterial Proteins; Biological Control Agents; Fungi; Genome, Bacterial; Oligopeptides; Plant Diseases; Pseudomonas putida; Siderophores; Surface-Active Agents
PubMed: 32347922
DOI: 10.1093/femsle/fnaa075 -
Bulletin of Entomological Research Oct 2020The cabbage aphid, Brevicoryne brassicae L. (Hem: Aphididae), is one of the most serious pests of canola worldwide. In this research, the effects of Pseudomonas putida,...
The cabbage aphid, Brevicoryne brassicae L. (Hem: Aphididae), is one of the most serious pests of canola worldwide. In this research, the effects of Pseudomonas putida, salicylic acid (SA), and integrated application of both inducers were studied on the resistance of canola to B. brassicae. In free-choice situation, the number of B. brassicae attracted on canola plants under treatments containing P. putida and SA was significantly lower compared to control plants. In the life table study, pre-adult survival, longevity, reproductive period, and fecundity of this aphid were lowest on plants treated with P. putida + SA. The net reproductive rate (R0), intrinsic rate of population increase (r), and finite rate of increase (λ) of B. brassicae decreased significantly in the following order: control (47.19 offspring, 0.293 and 1.340 day-1), P. putida (16.7 offspring, 0.238 and 1.269 day-1), SA (6.37 offspring, 0.163 and 1.178 day-1), and P. putida + SA (3.24 offspring, 0.112 and 1.119 day-1). Moreover, the beneficial effect of the integrated application of P. putida and SA on plant growth parameters was significantly evident in our study. The highest values of glucosinolates, total phenol, and flavonoids were recorded in P. putida + SA treatment. We concluded that canola plants treated with P. putida + SA are more resistant to the cabbage aphid. These findings demonstrated that SA integrated with P. putida on canola plants act effectively for reducing the population of B. brassicae and can be used in integrated management programs of this pest.
Topics: Animals; Antibiosis; Aphids; Brassica napus; Female; Fertility; Pseudomonas putida; Salicylic Acid
PubMed: 32252840
DOI: 10.1017/S0007485320000097 -
Current Microbiology Feb 2021Brassica napus L. is a main oilseed crop cultivated around the world. Plant growth-promoting rhizobacteria (PGPR) are generally applied to a wide range of agricultural...
Brassica napus L. is a main oilseed crop cultivated around the world. Plant growth-promoting rhizobacteria (PGPR) are generally applied to a wide range of agricultural crops for the growth enhancement. In this study, an I-plate technique was used to investigate the plant growth-promoting activity of Pseudomonas putida (strain ATCC12633) on B. napus plants. The volatile organic compounds (VOCs) produced by P. putida were determined by gas chromatography-mass spectrometric (GC-MS) analysis. Furthermore, P. putida were evaluated for its efficacy to induce resistance-related enzymes like peroxidase (POD), phenylalanine ammonia-lyase (PAL), catalase (CAT), and other biochemical compounds such as proline (Pro) and hydrogen peroxide (HO) in B. napus plants. According to the results, P. putida significantly increased the growth of B. napus compared to control. The major VOCs released by P. putida were 2-Butynedioic acid, dimethyl ester, Dimethyl ester of 4,7-dimethylnaphthalene-1,2-dicarboxylic acid, N-[3-Methylaminopropyl]aziridine, Cyclododecane, and Hexadecanoic acid. B. napus seeds treatment with P. putida caused enhanced activities of POD, PAL, CAT, Pro, and HO compared to control. So, the results of the present study showed that inoculation of B. napus with P. putida could serve as a useful tool for promoting the plant growth and inducing systemic resistance.
Topics: Brassica napus; Hydrogen Peroxide; Pseudomonas putida; Seeds; Volatile Organic Compounds
PubMed: 33403487
DOI: 10.1007/s00284-020-02335-2 -
Medicine Nov 2019The aim of this study was to analyze the risk factors, clinical features, and antimicrobial resistance of Pseudomonas putida (P putida) isolated from Tongji Hospital in... (Observational Study)
Observational Study
The aim of this study was to analyze the risk factors, clinical features, and antimicrobial resistance of Pseudomonas putida (P putida) isolated from Tongji Hospital in Wuhan, China.The data of 44 patients with P putida infections were retrospectively reviewed in this study. All cases of P putida strains were detected by the clinical laboratory of Tongji Hospital in the period of January 2010 to December 2017. Antimicrobial susceptibility testing was conducted using Kirby-Bauer method.Forty-four effective strains of P putida were isolated, including 32 inpatients and 12 outpatients. The 32 inpatients cases were obtained from various departments, which were urosurgery wards (n = 5, 15.6%), pediatrics wards (n = 4, 12.5%), hepatic surgery wards (n = 4, 12.5%), among others. The isolates had been discovered from urine specimens (28.2%), blood specimens (21.9%), sputum specimens (12.5%), and so on. Twenty-five patients had histories of catheterization before the isolation of P putida. Twenty-four patients were in immunocompromised states, 5 patients had undergone surgery, catheterization and were taking immunosuppressive therapy simultaneously. Polymicrobial infections were found in some P putida cases, especially Stenotrophomonas maltophilia, Pseudomonas aeruginosa, and Escherichia coli. All the patients had treated by antimicrobial before culture. Multi-drug-resistant strains were detected in 75% of P putida isolates. The P putida strains were resistant to trimethoprim/sulfamethoxazole (97.7%), aztreonam (88.6%), minocyline (74.3%), ticarcillin/clavulanic acid (72.7%), and sensitive to amikacin (86.4%), imipenem (62.8%), gentamicin (56.8%).Catheterization or other invasive procedures, immunocompromised states, and underlying diseases increased the risks of P putida infections. Moreover, the P putida strains were highly resistant to trimethoprim/sulfamethoxazole, aztreonam, minocyline, ticarcillin/clavulanic acid.
Topics: Adult; Aged; Aged, 80 and over; Anti-Bacterial Agents; Child; Child, Preschool; China; Drug Resistance, Bacterial; Female; Humans; Male; Microbial Sensitivity Tests; Middle Aged; Pseudomonas Infections; Pseudomonas putida; Retrospective Studies; Risk Factors; Young Adult
PubMed: 31689866
DOI: 10.1097/MD.0000000000017812 -
Biomacromolecules Sep 2023The bioconversion of homogeneous linear catechyl lignin (C-lignin) to polyhydroxyalkanoates (PHA) was examined for the first time in this study. C-lignins from vanilla,...
The bioconversion of homogeneous linear catechyl lignin (C-lignin) to polyhydroxyalkanoates (PHA) was examined for the first time in this study. C-lignins from vanilla, euphorbia, and candlenut seed coats (denoted as C1, C2, and C3, respectively) varied in their molecular structures, which showed different molecular weight distributions, etherification degrees, and contents of hydroxyl groups. A notable amount of nonetherified catechol units existed within C1 and C2 lignins, and these catechol units were consumed during fermentation. These results suggested that the nonetherified catechol structure was readily converted by KT2440. Since the weight-average molecular weight of C2 raw lignin was 26.7% lower than that of C1, the bioconversion performance of C2 lignin was more outstanding. The KT2440 cell amount reached the maximum of 9.3 × 10 CFU/mL in the C2 medium, which was 37.9 and 82.4% higher than that in the C1 and C3 medium, respectively. Accordingly, PHA concentration reached 137 mg/L within the C2 medium, which was 41.2 and 149.1% higher than the C1 and C3 medium, respectively. Overall, C-lignin, with a nonetherified catechol structure and low molecular weight, benefits its microbial conversion significantly.
Topics: Lignin; Polyhydroxyalkanoates; Fermentation; Pseudomonas putida
PubMed: 37555845
DOI: 10.1021/acs.biomac.3c00288 -
Advances in Applied Microbiology 2020This article addresses the lifestyle of Pseudomonas and focuses on how Pseudomonas putida can be used as a model system for biotechnological processes in agriculture,...
This article addresses the lifestyle of Pseudomonas and focuses on how Pseudomonas putida can be used as a model system for biotechnological processes in agriculture, and in the removal of pollutants from soils. In this chapter we aim to show how a deep analysis using genetic information and experimental tests has helped to reveal insights into the lifestyle of Pseudomonads. Pseudomonas putida is a Plant Growth Promoting Rhizobacteria (PGPR) that establishes commensal relationships with plants. The interaction involves a series of functions encoded by core genes which favor nutrient mobilization, prevention of pathogen development and efficient niche colonization. Certain Pseudomonas putida strains harbor accessory genes that confer specific biodegradative properties and because these microorganisms can thrive on the roots of plants they can be exploited to remove pollutants via rhizoremediation, making the consortium plant/Pseudomonas a useful tool to combat pollution.
Topics: Bacterial Proteins; Biodegradation, Environmental; Biofilms; Chemotaxis; Plant Development; Plants; Pseudomonas putida; Rhizosphere; Soil Microbiology; Symbiosis
PubMed: 32386604
DOI: 10.1016/bs.aambs.2019.12.002