Enhancing systematic tolerance in Bermuda grass (Cynodon dactylon L.) through amplified alkB gene expression and bacterial-driven hydrocarbon degradation.

This study aimed to access the impact of soil polluted with petroleum (5, 10 g petroleum kg-1 soil) on Bermuda grass (Cynodon dactylon L.) with and without applied bacterial inoculants (Arthrobacter oxydans ITRH49 and Pseudomonas sp. MixRI75). Both soil and seed were given bacterial inoculation. The evaluated morphological parameters of Bermuda grass were fresh and dry weight. The results demonstrated that applied bacterial inoculants enhanced 5.4%, 20%, 28% and 6.4%, 21%, and 29% shoot and root fresh/dry weights in Bermuda grass under controlled environment. The biochemical analysis of shoot and root was affected deleteriously by the 10 g petroleum kg-1 soil pollution. Microbial inoculants enhanced the activities of enzymatic (catalase, peroxidase, glutathione reductase, ascorbate peroxidase, superoxide dismutase) and non-enzymatic (ɑ-tocopherols, proline, reduced glutathione, ascorbic acid) antioxidant to mitigate the toxic effects of ROS (H2O2) under hydrocarbon stressed condition. The maximum hydrocarbon degradation (75%) was recorded by Bermuda grass at 5 g petroleum kg-1 soil contamination. Moreover, bacterial persistence and alkane hydroxylase gene (alkB) abundance and expression were observed more in the root interior than in the rhizosphere and shoot interior of Bermuda grass. Subsequently, the microbe used a biological tool to propose that the application of plant growth-promoting bacteria would be the most favorable choice in petroleum hydrocarbon polluted soil to conquer the abiotic stress in plants and the effective removal of polyaromatic hydrocarbons in polluted soil.

Heavy Metal Tolerance of Microorganisms Isolated from Coastal Marine Sediments and Their Lead Removal Potential.

In this study, 338 microorganisms, comprising 271 bacteria and 67 fungi, were isolated from sediment samples collected from underexplored Pacific and Caribbean regions of Colombia. Screening trials were conducted on selected strains (n = 276) to assess their tolerance to cadmium (Cd2+), lead (Pb2+), and zinc (Zn2+), leading to the identification of six bacteria capable of withstanding 750 mg·L-1 of each heavy metal ion. Three promising microorganisms, identified as Enterobacter sp. INV PRT213, Pseudomonas sp. INV PRT215, and Stenotrophomonas sp. INV PRT216 were selected for lead removal experiments using LB broth medium supplemented with 400 mg·L-1 Pb2+. Among these, Pseudomonas sp. INV PRT215 exhibited significant potential, removing 49% of initial Pb2+ after 240 min of exposure (16.7 g wet biomass·L-1, pH 5, 30 °C). Infrared spectra of Pb-exposed biomass showed changes in functional groups, including carbonyl groups of amides, carboxylate, phosphate, hydroxyl, and amine groups, compared to the not-exposed control. These changes suggested interactions between the metal and functional groups in the biomass. The findings of this study highlight the potential of microorganisms derived from coastal marine environments as promising candidates for future applications in bioremediation of polluted environments contaminated with heavy metals.

Comparative proteomic analysis of saline tolerant, phosphate solubilizing endophytic Pantoea sp., and Pseudomonas sp. isolated from Eichhornia rhizosphere.

Soil salinization is a major stress affecting crop production on a global scale. Application of stress tolerant plant growth promoting rhizobacteria (PGPR) in saline soil can be an ideal practice for improving soil fertility. Rhizospheric microbiota of stress tolerant Eichhornia crassipes was screened for saline tolerant phosphate solubilizing bacteria, and the two isolates showing maximum solubilization index at 1 M NaCl were subjected to further analyses. The isolates were identified as Pantoea dispersa and Pseudomonas aeruginosa. Among the two isolates, P. dispersa PSB1 showed better phosphorus (P) solubilization potential under saline stress (335 ± 30 mg/L) than P. aeruginosa PSB5 (200 ± 24 mg/L). The mechanisms of P-solubilization, such as the production of organic acids and phosphatase were found to be influenced negatively by saline stress. The adaptive mechanisms of the isolates to overcome salt stress were analyzed by protein profiling which revealed salt stress induced modulations in protein expression involved in amino acid biosynthesis, carbon metabolisms, chemotaxis, and stress responses. Survival mechanisms such as protein RecA, LexA repressor and iron-sulfur cluster synthesis were upregulated in both the organisms under saline stress. P. dispersa PSB1 showed improved defense mechanisms such as the production of osmotolerants, redox enzymes, and quorum quenchers under saline stress, which may explain its better P solubilization potential than the P. aeruginosa PSB5. This study emphasizes the need for molecular approaches like proteome analysis of PGPR for identifying novel traits like stress tolerance and plant growth promotion before developing them as biofertilizers and biocontrol formulations.

Evaluation of di-rhamnolipid biosurfactants production by a novel Pseudomonas sp. S1WB: Optimization, characterization and effect on petroleum-hydrocarbon degradation.

Rhamnolipid biosurfactants are multifunctional compounds that can play an indispensable role in biotechnological, biomedical, and environmental bioremediation-related fields, and have attracted significant attention in recent years. Herein, a novel strain Pseudomonas sp. S1WB was isolated from an oil-contaminated water sample. The biosurfactants produced by this strain have capabilities to reduce surface tension (SFT) at 32.75 ± 1.63 mN/m and emulsified 50.2 ± 1.13 % in liquid media containing 1 % used engine oil (UEO) as the sole carbon source. However, the lowest SFT reduction (28.25 ± 0.21), highest emulsification index (60.15 ± 0.07), and the maximum yields (900 mg/L) were achieved under optimized conditions; where, the glucose/urea and glycerol/urea combinations were found efficient carbon and nitrogen substrates for improved biosurfactants production. Biosurfactants product was characterized using ultra-high performance liquid chromatography-mass spectrometry (UHPLC- MS) and detected various di- rhamnolipids congeners. In addition, the di-rhamnolipids produced by S1WB strain was found highly stable in terms of surface activity and EI indices at different environmental factors i.e. temperature, pH and various NaCl concentrations, where, emulsifying property was found high stable till 30 days of incubation. Moreover, the stain was capable to degrade hydrocarbon at 42.2 ± 0.04 %, and the Gas chromatography- mass spectrometry (GC-MS) profile showed the majority of peak intensities of hydrocarbons have been completely degraded compared to control.

Nitrogen dependence of rhamnolipid mediated degradation of petroleum crude oil by indigenous Pseudomonas sp. WD23 in seawater.

Effect of oil spills on living forms demands for safe, ecofriendly and cost-effective methods to repair the damage. Pseudomonads have exceptional tolerance to xenobiotics and can grow at varied environmental conditions. This study aims at biosurfactant mediated degradation of petroleum crude oil by an indigenous Pseudomonas sp. WD23 in sea water. Pseudomonas sp. WD23 degraded 34% of petroleum crude oil (1.0% v/v) on supplementation of yeast extract (0.05 g/L) with glucose (1.0 g/L) in seawater. The strain produced a biosurfactant which was confirmed as a rhamnolipid (lipid: rhamnose 1:3.35) by FT-IR, LCMS and quantitative analysis. Produced rhamnolipid had low CMC (20.0 mg/L), emulsified petroleum oils (75-80%) and had high tolreance to varied conditions of pH, temperature and ionic strength. OFAT studies were performed to analyse the effect of petroleum crude oil, glucose, inoculum, yeast extract, pH, agitation speed and incubation time on degradation by Pseudomonas sp. WD23. Petroleum crude oil and glucose had significant effect on biodegradation, rhamnolipid production and growth, further optimized by central composite design. At optimum conditions of 3.414% v/v PCO and 6.53 g/L glucose, maximum degradation of 81.8 ± 0.67% was observed at pH 7.5, 100 RPM, 15.0% v/v inoculum in 28 days, with a 3-fold increase in biodegradation. GCMS analysis revealed degradation (86-100%) of all low and high molecular weight hydrocarbons present in petroleum crude oil. Hence, the strain Pseudomonas sp. WD23 can be effectively developed for management of oil spills in seas and oceans due to its excellent degradation abilities.

Isolation and characterization of a novel rhamnolipid producer Pseudomonas sp. LGMS7 from a highly contaminated site in Ain El Arbaa region of Ain Temouchent, Algeria.

This study aims to isolate and characterize a novel rhamnolipid producer within the recent bioremediation approaches for treating hydrocarbon-contaminated soils in Algeria. In this context, from a hydrocarbon-contaminated soil, a newly bacterium designated LGMS7 was screened and identified, belonged to the Pseudomonas genus, and was closely related to Pseudomonas mucidolens, with a 16S rRNA sequence similarity of 99.05%. This strain was found to use different hydrocarbons and oils as a sole carbon and energy source for growth. It showed a stable emulsification index E24 (%) of 66.66% ± 3.46 when growing in mineral salts medium (MSM) supplemented with 2% (v/v) glycerol after incubation for 6 days at 30 °C. Interestingly, it was also able to reduce the surface tension of the cell-free supernatant to around 30 ± 0.65 mN m-1 with a critical micelle concentration (CMC) of 800 mg l-1. It was found to be able to produce around 1260 ± 0.57 mg l-1 as the yield of rhamnolipid production. Its biosurfactant has demonstrated excellent stability against pH (pH 2.0-12.0), salinity (0-150 g l-1), and temperature (-20 to 121 °C). Based on various chromatographic and spectroscopic techniques (i.e., TLC, FTIR, 1H-NMR), it was found to belong to the glycolipid class (i.e., rhamnolipids). Taken altogether, the strain LGMS7 and its biosurfactant display interesting biotechnological capabilities for the bioremediation of hydrocarbon-contaminated sites. To the best of our knowledge, this is the first study that described the production of biosurfactants by Pseudomonas mucidolens species. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02751-6.

Investigation on spectral and biomedical characterization of rhamnolipid from a marine associated bacterium Pseudomonas aeruginosa (DKB1).

Bio-surfactants are a principal group of significant molecules obtained from the microbial sources expressed with distinctive characteristics like biodegradation of hydrocarbons and also have different biomedical properties. The present investigation aims to assess the biomedical properties of synthesized bio-surfactant, rhamnolipid from Pseudomonas aeruginosa (DKB1) under in vitro conditions. The candidate bacterium P. aeruginosa (DKB1) was isolated from oil-polluted fishing harbors of Kanyakumari coast. Initially, the bio-surfactant production by this candidate strain was confirmed by oil displacement assay, hemolytic assay, drop collapse assay and emulsification index. Further, the production of bio-surfactant was achieved through submerged fermentation process using Bushnell-Haas mineral salts medium supplemented with 2% olive oil. The yield of the bio-surfactant was attained as 2.4 g/l and confirmed as rhamnolipid through blue agar plate assay; further, the extracted rhamnolipid was purified and characterized through standard procedures. In stability studies, the rhamnolipid could withstand up to pH 12, temperature 100 °C and 15% of NaCl concentration. The biomedical application of rhamnolipid (30 µg ml-1) was determined by antibacterial, antioxidant and cytotoxic studies. It exhibited a maximum growth inhibition against Bacillus subtilis (26 mm) with the MIC value of 8 µg ml-1. In antioxidant test, rhamnolipid expressed significant (P < 0.0001) inhibition of total reducing power (44.11%), DPPH activity (61.60%), hydroxyl radical (83.30%) and nitric oxide (51.86%) scavenging ability at 100 µg ml-1with the respective IC50 values of 130.50, 77.18, 52.08 and 95.43 µg ml-1. The anticancer activity of the rhamnolipid was assessed with the help of MTT test against MCF-7, HT-29 and E-143 cancer cell lines individually, and the viability of the cells was observed, respectively, as 10.24, 17.66 and 13.50% at 250 µg ml-1concentration with the respective IC50 values of 140.2, 81.02 and 138.9 µg ml-1. From the results, it could be concluded that the rhamnolipid produced by P. aeruginosa (DKB1) isolated from oil-polluted area has effective biomedical properties.

The relevance of enzyme specificity for coenzymes and the presence of 6-phosphogluconate dehydrogenase for polyhydroxyalkanoates production in the metabolism of Pseudomonas sp. LFM046.

Reconstruction of genome-based metabolic model is a useful approach for the assessment of metabolic pathways, genes and proteins involved in the environmental fitness capabilities or pathogenic potential as well as for biotechnological processes development. Pseudomonas sp. LFM046 was selected as a good polyhydroxyalkanoates (PHA) producer from carbohydrates and plant oils. Its complete genome sequence and metabolic model were obtained. Analysis revealed that the gnd gene, encoding 6-phosphogluconate dehydrogenase, is absent in Pseudomonas sp. LFM046 genome. In order to improve the knowledge about LFM046 metabolism, the coenzyme specificities of different enzymes was evaluated. Furthermore, the heterologous expression of gnd genes from Pseudomonas putida KT2440 (NAD+ dependent) and Escherichia coli MG1655 (NADP+ dependent) in LFM046 was carried out and provoke a delay on cell growth and a reduction in PHA yield, respectively. The results indicate that the adjustment in cyclic Entner-Doudoroff pathway may be an interesting strategy for it and other bacteria to simultaneously meet divergent cell needs during cultivation phases of growth and PHA production.

New metabolites from the marine-derived bacterium Pseudomonas sp. ZZ820R.

Six previously undescribed compounds, named monaxanthones A and B, monaphenol A, monathioamide A, monaprenylindole A, and monavalerolactone A, were isolated from the culture of a marine-sourced bacterium Pseudomonas sp. ZZ820R in rice medium. Their structures were elucidated based on the HRESIMS data, NMR and MS-MS spectroscopic analyses, optical rotation and ECD calculations. Monathioamide A is an unprecedented sulfur-contained compound and monavalerolactone A represents the first example of this type of natural products. Monaprenylindole A showed antibacterial activity against methicillin-resistant Staphylococcus aureus.

Chemical screening identifies an extract from marine Pseudomonas sp.-PTR-08 as an anti-aging agent that promotes fission yeast longevity by modulating the Pap1-ctt1+ pathway and the cell cycle.

Aging is a degenerative process characterized by progressive deterioration of cellular components, ultimately resulting in mortality, in which massive accumulation of reactive oxygen species (ROS) and advanced glycation end products (AGEs) are implicated as crucial factors. At the same time, natural products are rich sources from which to isolate and characterize potential anti-aging compounds. The current study was designed to extract compounds from the marine bacterium Pseudomonas sp. and investigate their in vitro antioxidant and anti-glycation activities, as well as their in vivo effects on aging in the model organism Schizosaccharomyces pombe. In vitro assays showed that a Pseudomonas sp. PTR-08 extract exhibited the best antioxidant and anti-glycation activities. Further, direct administration of the extract significantly increased yeast longevity, accompanied by induction of the yeast oxidative stress response. Molecular analyses indicated that selected extract dramatically up-regulated the expression of pap1+, which encodes the transcriptional factor Pap1 and ctt1+, which encodes catalase, following H2O2 treatment. In line with these results, catalase activity significantly increased, leading to a decrease in intracellular ROS. In addition, this extract may delay the G1 phase of the yeast cell cycle, leading to an extended lifespan. Moreover, our findings indicated that the extract contains pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-, which substantially promotes anti-aging activity in yeast. However, further research must be conducted to better understand the role of this compound in our system.

Development of Palm Fatty Acid Distillate-Containing Medium for Biosurfactant Production by Pseudomonas sp. LM19.

High production costs of biosurfactants are mainly caused by the usage of the expensive substrate and long fermentation period which undermines their potential in bioremediation processes, food, and cosmetic industries even though they, owing to the biodegradability, lower toxicity, and raise specificity traits. One way to circumvent this is to improvise the formulation of biosurfactant-production medium by using cheaper substrate. A culture medium utilizing palm fatty acid distillate (PFAD), a palm oil refinery by-product, was first developed through one-factor-at-a-time (OFAT) technique and further refined by means of the statistical design method of factorial and response surface modeling to enhance the biosurfactant production from Pseudomonas sp. LM19. The results shows that, the optimized culture medium containing: 1.148% (v/v) PFAD; 4.054 g/L KH2PO4; 1.30 g/L yeast extract; 0.023 g/L sodium-EDTA; 1.057 g/L MgSO4·7H2O; 0.75 g/L K2HPO4; 0.20 g/L CaCl2·2H2O; 0.080 g/L FeCl3·6H2O gave the maximum biosurfactant productivity. This study demonstrated that the cell concentration and biosurfactant productivity could reach up to 8.5 × 109 CFU/mL and 0.346 g/L/day, respectively after seven days of growth, which were comparable to the values predicted by an RSM regression model, i.e., 8.4 × 109 CFU/mL and 0.347 g/L/day, respectively. Eleven rhamnolipid congeners were detected, in which dirhamnolipid accounted for 58% and monorhamnolipid was 42%. All in all, manipulation of palm oil by-products proved to be a feasible substrate for increasing the biosurfactant production about 3.55-fold as shown in this study.

Development of plant-microbe phytoremediation system for petroleum hydrocarbon degradation: An insight from alkb gene expression and phytotoxicity analysis.

Aim of present work was to assess in-planta association potential of isolated endophytic bacterial strain Pseudomonas sp. (J10) (KY608252) with two cultivars of Lolium perenne L. (small & jumbo) and Arabidopsis thaliana L. for total petroleum hydrocarbon (TPH) degradation, alkane monooxygenase (alkb) gene expression and phytotoxicity analysis. A plant-microbe phytoremediation system was established to investigate the bacteria's ability to colonize the plant body and quantification of alkb gene to help withstand TPH stress in soil as well as in hydroponics. A real-time PCR method was developed to analyze bacterial colonization and survival within the plant body. Analysis revealed that J10 efficiently colonized all the tested plant species and expressed alkb gene under hydrocarbon stress ranging between 3.7 × 102-3.9 × 106 in A. thaliana and L. perenne (small), respectively. The colonization was more pronounced in soil as compared to hydroponic system. J10 inoculation reduced phytotoxicity and suggested that inoculation had a positive effect on plant growth under stress conditions as compared to control. L. perenne (small) showed significant TPH removal efficiency (45.6%) followed by L. perenne jumbo (24.5%) and A. thaliana (6.2%). In hydroponics, L. perenne (small) degraded about 28.2% TPH followed by L. perenne (jumbo) as 24.4%. Potential of the indigenously isolated plant endophytes may be exploited further for phytoremediation efficiency and industrial applications.

Phosphate-solubilizing bacteria and silicon synergistically augment phosphorus (P) uptake by wheat (Triticum aestivum L.) plant fertilized with soluble or insoluble P source.

Phosphorus (P) deficiency is one of the major problems in agricultural soils for crop production around the world. Use of silicon (Si) and phosphate-solubilizing bacteria (PSB) is known as one of the most effective and economical ways for increasing P availability and improving P use efficiency under low P conditions. However, little is known about the alleviative role of Si and PSB together in mitigating P-deficiency stress and in improving P use efficiency in Triticum aestivum L. (wheat), as one of the most important crop plants worldwide. Consequently, aim of the research was to study the combined and single effects of Si (0, 150, 300, and 600 mg kg-1 added as silicic acid) and PSB (B0, Bacillus simplex UT1, and Pseudomonas sp. FA1) on P uptake by wheat plant fertilized with soluble or insoluble P (Esfordi rock phosphate, RP) in a completely randomized design with factorial arrangement through a perlite potted experiment. In addition, the effects of various treatments on wheat shoot and root dry weight, activity of catalase, superoxide dismutase, and peroxidase enzymes, and the uptake of Si and potassium (K) by this plant were also investigated. Both shoot and root biomass of wheat plants were markedly reduced when grown in RP-fertilized medium compared with those grown in soluble P-fertilized medium. The PSB strains and Si levels independently improved all the aforementioned parameters. Application of Si to wheat plants grown in soluble P or insoluble P medium markedly enhanced P use efficiency. According to the results of this study, Si not only increased the uptake of P from sparingly soluble-P source (RP), but also enhanced uptake of P from water-soluble P source. Both Pseudomonas sp. FA1 and B. simplex UT1 showed a considerable role in improvement of root and shoot biomass and uptake of P (and K and Si) under both soluble and insoluble P fertilization conditions with Pseudomonas sp. FA1 being more effective than B. simplex UT1. However, the combined application of the PSB with Si resulted in the greatest enhancement in wheat plant P uptake and other measured parameters. Addition of 600 mg Si kg-1 and Pseudomonas sp. FA1 significantly increased the P shoot concentration of wheat plant fertilized with RP to an adequate level (>0.3%) in the range of P-fertilized plants. Therefore, in addition to PSB application, Si should be considered as soil amendment in agricultural soils deficient in plant available Si as a means of sustainable agriculture with respect to possible savings of scarce P resources (P-fertilizers). The information on the availability of P following PSB and Si addition to plant growth medium may help in better management of P fertilization.

Co-culture development and bioformulation efficacy of psychrotrophic PGPRs to promote growth and development of Pea (Pisum sativum) plant.

Numerous microbes reside in the rhizosphere having plant growth promoting activity, and enhancing the property by increasing plant yield. Plant growth promoting rhizobacteria (PGPR) has gradually increased in agriculture and offers an attractive way to replace chemical fertilizers, pesticides and supplements. Soil was collected from the rhizosphere of an agricultural farm and the psychrotrophic bacterial strains STA3 (KY888133) and RM2 (KY888134) were successfully isolated, and screened on the basis of phosphate solubilization. Further characterization was carried out by morphological, biochemical, and 16S rDNA characterization methods. The unique nature of psychrotrophic Pentoea ananatis and a suitable combination with Pseudomonas fluorescens regarding plant growth promotion activity has not been studied before to our knowledge. An assessment of various parameters of plant growth promoting activity, such as IAA, phosphate solubilization, bio-control activity, HCN and siderophore production, has been carried out. Both strains were found to be positive in various parameters except HCN and Biocontrol activity, which were positive only for the strain RM2. Also, shelf life and their efficacy was determined before and after formulation. A great consistency was observed in all the cultures, even after 70 days of storage under bio-formulation at room temperature, while in the case of the co-culture CPP-2, the cfu ml-1 was greater, followed by RM2 and STA3. Moreover, the growth indices of the pea plant were found to be better in the co-culture CPP-2 compared with individual strains, followed by RM2 and STA3. Thus, the study suggests that the co-culture CPP-2 has a great potential for plant growth promotion as compared with individual strains followed by RM2 and STA3.

Pseudomonas sp. BUP6, a novel isolate from Malabari goat produces an efficient rhamnolipid type biosurfactant.

This study describes the characteristics of a biosurfactant produced by Pseudomonas sp. BUP6, a rumen bacterium, and optimization of parameters required for its production. Initial screening of five parameters (pH, temperature, agitation, incubation, and substrate concentration) was carried out employing Plackett-Burman design, which reduced the number of parameters to 3 (pH, temperature, and incubation) according to their significance on the yield of biosurfactant. A suitable statistical model for the production of biosurfactant by Pseudomonas sp. BUP6 was established according to Box-Behnken design, which resulted in 11% increase (at pH 7, 35 °C, incubation 75 h) in the yield (2070 mg L-1 ) of biosurfactant. The biosurfactant was found stable at a wide range of pH (3-9) with 48 mg L-1 critical micelle concentration; and maintained over 90% of its emulsification ability even after boiling and in presence of sodium chloride (0.5%). The highest cell hydrophobicity (37%) and emulsification (69%) indices were determined with groundnut oil and kerosene, respectively. The biosurfactant was found to inhibit the growth and adhesion of E. coli and S. aureus significantly. From the phytotoxicity studies, the biosurfactant did not show any adverse effect on the germinating seeds of rice and green gram. The structural characterization of biosurfactant employing orcinol method, thin layer chromatography and FT-IR indicated that it is a rhamnolipid (glycolipid). Thus, Pseudomonas sp. BUP6, a novel isolate from Malabari goat is demonstrated as a producer of an efficient rhamnolipid type biosurfactant suitable for application in various industries.

Polymer Film-Based Screening and Isolation of Polylactic Acid (PLA)-Degrading Microorganisms.

Polylactic acid (PLA) has been highlighted as an alternative renewable polymer for the replacement of petroleum-based plastic materials, and is considered to be biodegradable. On the other hand, the biodegradation of PLA by terminal degraders, such as microorganisms, requires a lengthy period in the natural environment, and its mechanism is not completely understood. PLA biodegradation studies have been conducted using mainly undefined mixed cultures, but only a few bacterial strains have been isolated and examined. For further characterization of PLA biodegradation, in this study, the PLA-degrading bacteria from digester sludge were isolated and identified using a polymer film-based screening method. The enrichment of sludge on PLA granules was conducted with the serial transference of a subculture into fresh media for 40 days, and the attached biofilm was inoculated on a PLA film on an agar plate. 3D optical microscopy showed that the isolates physically degraded the PLA film due to bacterial degradation. 16S rRNA gene sequencing identified the microbial colonies to be Pseudomonas sp. MYK1 and Bacillus sp. MYK2. The two isolates exhibited significantly higher specific gas production rates from PLA biodegradation compared with that of the initial sludge inoculum.

Screening and characterizing a denitrifying phosphorus-accumulating bacterium isolated from a circular plug-flow reactor.

Denitrifying phosphorus-accumulating organisms (DNPAO) are viewed as one of the most effective means to solve the removal contradiction of nitrogen and phosphorus in wastewater treatment. In this study, we isolated a DNPAO (C-17, accession number: KU745702) from activated sludge in a patented circular plug-flow reactor, physiologically to Pseudomonas sp. based on 16S rRNA sequence and phenotypic characteristics. The results of denitrifying phosphorus-accumulating experiment showed that Pseudomonas C-17 has high removal efficiencies for [Formula: see text] and NO3-N, 75% and 87%, respectively. The ratio of phosphorus release was 25.0 mg [Formula: see text] (with anabolism) and 26.8 mg [Formula: see text] (without anabolism), respectively. Our results indicated that Pseudomonas C-17 had strong capacity of phosphorus release, and its uptake is often imprecisely evaluated by ignoring the part of metabolic consumption. Pseudomonas C-17 is capable of utilizing oxygen, nitrate and nitrite as electron acceptors under experimental conditions.

Heterologous production of Pseudomonas aeruginosa rhamnolipid under anaerobic conditions for microbial enhanced oil recovery.

AIMS: The ex situ application of rhamnolipid to enhance oil recovery is costly and complex in terms of rhamnolipid production and transportation, while in situ production of rhamnolipid is restricted by the oxygen-deficient environments of oil reservoirs. To overcome the oxygen-limiting conditions and to circumvent the complex regulation of rhamnolipid biosynthesis in Pseudomonas aeruginosa, an engineered strain Pseudomonas stutzeri Rhl was constructed for heterologous production of rhamnolipid under anaerobic conditions. METHODS AND RESULTS: The rhlABRI genes for rhamnolipid biosynthesis were cloned into a facultative anaerobic strain Ps. stutzeri DQ1 to construct the engineered strain Rhl. Anaerobic production of rhamnolipid was confirmed by thin layer chromatography and Fourier transform infrared analysis. Rhamnolipid product reduced the air-water surface tension to 30.3 mN m(-1) and the oil-water interfacial tension to 0.169 mN m(-1). Rhl produced rhamnolipid of 1.61 g l(-1) using glycerol as the carbon source. Rhl anaerobic culture emulsified crude oil up to EI24 ≈ 74. An extra 9.8% of original crude oil was displaced by Rhl in the core flooding test. CONCLUSIONS: Strain Rhl achieved anaerobic production of rhamnolipid and worked well for enhanced oil recovery in the core flooding model. The rhamnolipid produced by Rhl was similar to that of the donor strain SQ6. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to achieve anaerobic and heterologous production of rhamnolipid. Results demonstrated the potential feasibility of Rhl as a promising strain to enhance oil recovery through anaerobic production of rhamnolipid.

Antagonistic Effect of Pseudomonas sp. CMI-1 onFoodborne Pathogenic Listeria monocytogenes.

Bacterial isolates derived from food or raw food materials of animal origin were screened for potential antagonistic activity against foodborne pathogenic Listeria monocytogenes. Using the agar spot method, ten out of the 94 tested bacteria showed antilisterial activity. All of the antagonistic isolates identified by sequence analysis as strains of the genus Pseudomonas were able to inhibit the growth of all the examined Listeria species including the ruminal pathogenic L. ivanovii and the opportunistic human pathogenic L. innocua. Pseudomonas sp. CMI-1 had the highest inhibitory effect on the growth of different Listeria strains. Co-culturing studies revealed that the inhibition of L. monocytogenes could not be achieved efficiently. Although the population of the Pseudomonas sp. CMI-1 strain increased by up to 10 orders of magnitude during 2 days of culturing period at 20 °C in the presence of L. monocytogenes, the cell count of the pathogen also increased by approx. 6 orders of magnitude. At the same time, appropriate inhibition of cell-free supernatants generated from 6-day-old cultures of Pseudomonas sp. CMI-1 was observed. The inhibitory compound of this antagonistic strain is presumably a chromopeptide siderophore, whose activity and production can be affected by iron supplementation, and which had an absorption maximum typical of siderophores of fluorescent Pseudomonas species. Production of the antilisterial substance was influenced by the oxygen concentration, as in static cultures the concentration of the siderophore was higher than in shake flask cultures.

Quantitative approach to track lipase producing Pseudomonas sp. S1 in nonsterilized solid state fermentation.

UNLABELLED: Proliferation of the inoculated Pseudomonas sp. S1 is quantitatively evaluated using ERIC-PCR during the production of lipase in nonsterile solid state fermentation an approach to reduce the cost of enzyme production. Under nonsterile solid state fermentation with olive oil cake, Pseudomonas sp. S1 produced 57·9 IU g(-1) of lipase. DNA fingerprints of unknown bacterial isolates obtained on Bushnell Haas agar (BHA) + tributyrin exactly matched with that of Pseudomonas sp. S1. Using PCR-based enumeration, population of Pseudomonas sp. S1 was proliferated from 7·6 × 10(4) CFU g(-1) after 24 h to 4·6 × 10(8) CFU g(-1) after 96 h, which tallied with the maximum lipase activity as compared to control. Under submerged fermentation (SmF), Pseudomonas sp. S1 produced maximum lipase (49 IU ml(-1) ) using olive oil as substrate, while lipase production was 9·754 IU ml(-1) when Pseudomonas sp. S1 was grown on tributyrin. Optimum pH and temperature of the crude lipase was 7·0 and 50°C. Crude enzyme activity was 71·2% stable at 50°C for 360 min. Pseudomonas sp. S1 lipase was also stable in methanol showing 91·6% activity in the presence of 15% methanol, whereas 75·5 and 51·1% of activity were retained in the presence of 20 and 30% methanol, respectively. Thus, lipase produced by Pseudomonas sp. S1 is suitable for the production of biodiesel as well as treatment of oily waste water. SIGNIFICANCE AND IMPACT OF STUDY: This study presents the first report on the production of thermophilic organic solvent tolerant lipase using agro-industry waste in nonsterile solid state fermentation. Positive correlation between survival of Pseudomonas sp. S1 and lipase production under nonsterile solid state fermentation was established, which may emphasize the need to combine molecular tools and solid state fermentation in future studies. Our study brings new insights into the lipase production in cost-effective manner, which is an industrially relevant approach.