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Molecular Biology Reports Mar 2024A novel lytic bacteriophage (phage) was isolated with Pseudomonas mendocina strain STP12 (P. mendocina) from the untreated site of Sewage Treatment Plant of Lovely...
BACKGROUND
A novel lytic bacteriophage (phage) was isolated with Pseudomonas mendocina strain STP12 (P. mendocina) from the untreated site of Sewage Treatment Plant of Lovely Professional University, India. P. mendocina is a Gram-negative, rod-shaped, aerobic bacterium belonging to the family Pseudomonadaceae and has been reported in fifteen (15) cases of economically important diseases worldwide.
METHODS AND RESULTS
Here, a novel phage specifically infecting and killing P. mendocina strain STP12 was isolated from sewage sample using enrichment, spot test and double agar overlay (DAOL) method and was designated as vB_PmeS_STP12. The phage vB-PmeS-STP12 was viable at wide range of pH and temperature ranging from 4 to10 and - 20 to 70 °C respectively. Host range and efficiency of plating (EOP) analysis indicated that phage vB-PmeS-STP12 was capable of infecting and killing P. mendocina strain STP6 with EOP of 0.34. Phage vB_PmeS_STP12 was found to have a significant bacterial reduction (p < 0.005) at all the doses administered, particularly at optimal MOI of 1 PFU/CFU, compared to the control. Morphological analysis using high resolution transmission electron microscopy (HR-TEM) revealed an icosahedral capsid of ~ 55 nm in diameter on average with a short, non-contractile tail. The genome of vB_PmeS_STP12 is a linear, dsDNA containing 36,212 bp in size with a GC content of 58.87% harbouring 46 open reading frames (ORFs). The 46 predicted ORFs encode proteins with functional information categorized as lysis, replication, packaging, regulation, assembly, infection, immune, and hypothetical. However, the genome of vB_PmeS_STP12 appeared to be devoid of tRNAs, integrase gene, toxins genes, virulence factors, antimicrobial resistance genes (ARGs) and CRISPR arrays. The blast analysis with phylogeny revealed that vB_PmeS_STP12 is genetically similar to Pseudomonas phage PMBT14, Pseudomonas phage Almagne and Serratia phage Serbin with a highest identity of 74.00%, 74.93% and 59.48% respectively.
CONCLUSIONS
Taken together, characterization, morphological analysis and genome-informatics indicated that vB_PmeS_STP12 is podovirus morphotype belonging to the class Caudoviticetes, family Zobellviridae which appeared to be devoid of integrase gene, ARGs, CRISPR arrays, virulence factors and toxins genes, exhibiting stability and infectivity at wide range of pH (4 to10) and temperature (-20 to 70 °C), thereby making vB_PmeS_STP12 suitable for phage therapy or biocontrol. Based on the bibliometric analysis and data availability with respect to sequences deposited in GenBank, this is the first report of a phage infecting Pseudomonas mendocina.
Topics: Humans; Bacteriophages; Pseudomonas; Phage Therapy; Sewage; Genome, Viral; Informatics; Integrases; Virulence Factors; Phylogeny
PubMed: 38483683
DOI: 10.1007/s11033-024-09362-3 -
Colloids and Surfaces. B, Biointerfaces Feb 2014A potential biofilm forming and phenanthrene utilizing marine bacterium Pseudomonas mendocina NR802 was isolated from Rushukulya, Odisha, East Coast of India. The effect...
A potential biofilm forming and phenanthrene utilizing marine bacterium Pseudomonas mendocina NR802 was isolated from Rushukulya, Odisha, East Coast of India. The effect of Ca(2+) and Mg(2+) on biofilm growth and phenanthrene degradation was evaluated. Among the various tested concentrations, 20 mM of Ca(2+) and Mg(2+) showed a significant enhancement in biofilm production by the bacterium. The SEM-EDAX study showed that the elemental composition of the biofilm varied significantly when grown in the presence of Ca(2+) and Mg(2+). The CSLM analysis of biofilms grown in the presence of 20 mM Ca(2+) and Mg(2+) reveal the critical role of these ions on biofilm architectural parameters such as total biomass, biofilm thickness, roughness coefficient and surface to biovolume ratio. Ca(2+) was found to enhance the extracellular polymeric substances (EPS) production and phenanthrene degradation. Ca(2+) enhanced the biofilm growth in a dose dependent manner, whereas Mg(2+) significantly increased the cell growth in biofilm. More than 15% increase in phenanthrene degradation was observed when biofilm was grown in the presence of an additional 20 mM Ca(2+). This study also supports the fundamental role of Ca(2+) in biofilm growth, architecture as well as biofilm-mediated pollutant degradation.
Topics: Biodegradation, Environmental; Biofilms; Calcium; Ions; Magnesium; Molecular Sequence Data; Phenanthrenes; Pseudomonas mendocina; Spectrometry, X-Ray Emission
PubMed: 24216621
DOI: 10.1016/j.colsurfb.2013.10.003 -
International Journal of Biological... Jul 2020In this study, six genes involved in β-oxidation pathway of P. mendocina NK-01 were deleted to construct mutant strains NKU-∆β1 and NKU-∆β5. Compared with the...
In this study, six genes involved in β-oxidation pathway of P. mendocina NK-01 were deleted to construct mutant strains NKU-∆β1 and NKU-∆β5. Compared with the wild strain NKU, the mcl-PHA titers of NKU-∆β5 were respectively increased by 5.58- and 4.85-fold for culturing with sodium octanoate and sodium decanoate. And the mcl-PHA titers of NKU-∆β1 was increased by 10.02-fold for culturing with dodecanoic acid. The contents of dominant monomers 3-hydroxydecanoate (3HD) and 3-hydroxydodecanoate (3HDD) of the mcl-PHA synthesized by NKU-∆β5 were obviously increased to 90.01 and 58.60 mol%, respectively. Further deletion of genes phaG and phaZ, the 3HD and 3HDD contents were further improved to 94.71 and 68.67 mol%, respectively. The highest molecular weight of mcl-PHA obtained in this study was 80.79 × 10 Da, which was higher than the previously reported mcl-PHA. With the increase of dominant monomer contents, the synthesized mcl-PHA showed better thermal properties, mechanical properties and crystallization properties. Interestingly, the cell size of NKU-∆β5 was larger than that of NKU due to the accumulation of more PHA granules. This study indicated that a systematically metabolic engineering approach for P. mendocina NK-01 could significantly improve the mcl-PHA titer, dominant monomer contents and physical properties of mcl-PHA.
Topics: Gene Deletion; Mechanical Phenomena; Metabolic Engineering; Molecular Weight; Polyhydroxyalkanoates; Pseudomonas
PubMed: 31706817
DOI: 10.1016/j.ijbiomac.2019.11.044 -
Carbohydrate Research May 2012Pseudomonas mendocina NK-01 can simultaneously synthesize medium-chain-length polyhydroxyalkanoate (PHA(MCL)) and alginate oligosaccharides (AO) from glucose under...
Pseudomonas mendocina NK-01 can simultaneously synthesize medium-chain-length polyhydroxyalkanoate (PHA(MCL)) and alginate oligosaccharides (AO) from glucose under conditions of limited nitrogen. In this study, the PHA(MCL) synthesis pathway was blocked by a deletion of approximately 57% of the sequence of PHA synthase operon mediated by the suicide plasmid, pEX18TcC1ZC2Amp. Deletion of the PHA synthase operon in P. mendocina NK-01 was confirmed by polymerase chain reaction (PCR) and antibiotic resistance assays to form the gene knockout mutant, P. mendocina C7. Shake-flask and 30 L fermentor cultures of P. mendocina C7 showed a 2.21-fold and 2.64-fold accumulation of AO from glucose, respectively, compared with the wild-type strain. Mass spectrometry and gel permeation chromatography characterization revealed that P. mendocina C7 and P. mendocina NK-01 produced AO were identical in terms of monomer composition and average molecular weight (M(W)). Thus, the mutant P. mendocina C7 has potential use in large scale fermentation of AO. Furthermore, it was demonstrated that the PHA(MCL) and AO synthesis pathways compete for the use of carbon sources in P. mendocina NK-01.
Topics: Acyltransferases; Alginates; Fermentation; Glucuronic Acid; Hexuronic Acids; Mutation; Oligosaccharides; Operon; Plasmids; Polyhydroxyalkanoates; Pseudomonas mendocina; Sequence Deletion
PubMed: 22429775
DOI: 10.1016/j.carres.2012.02.024 -
Scientific Reports Feb 2019Polyhydroxyalkanoate (PHA) can be produced by microorganisms from renewable resources and is regarded as a promising bioplastic to replace petroleum-based plastics....
Polyhydroxyalkanoate (PHA) can be produced by microorganisms from renewable resources and is regarded as a promising bioplastic to replace petroleum-based plastics. Pseudomonas mendocina NK-01 is a medium-chain-length PHA (mcl-PHA)-producing strain and its whole-genome sequence is currently available. The yield of mcl-PHA in P. mendocina NK-01 is expected to be improved by applying a promoter engineering strategy. However, a limited number of well-characterized promoters has greatly restricted the application of promoter engineering for increasing the yield of mcl-PHA in P. mendocina NK-01. In this work, 10 endogenous promoters from P. mendocina NK-01 were identified based on RNA-seq and promoter prediction results. Subsequently, 10 putative promoters were characterized for their strength through the expression of a reporter gene gfp. As a result, five strong promoters designated as P4, P6, P9, P16 and P25 were identified based on transcriptional level and GFP fluorescence intensity measurements. To evaluate whether the screened promoters can be used to enhance transcription of PHA synthase gene (phaC), the three promoters P4, P6 and P16 were separately integrated into upstream of the phaC operon in the genome of P. mendocina NK-01, resulting in the recombinant strains NKU-4C1, NKU-6C1 and NKU-16C1. As expected, the transcriptional levels of phaC1 and phaC2 in the recombinant strains were increased as shown by real-time quantitative RT-PCR. The phaZ gene encoding PHA depolymerase was further deleted to construct the recombinant strains NKU-∆phaZ-4C1, NKU-∆phaZ-6C1 and NKU-∆phaZ-16C1. The results from shake-flask fermentation indicated that the mcl-PHA titer of recombinant strain NKU-∆phaZ-16C1 was increased from 17 to 23 wt% compared with strain NKU-∆phaZ. This work provides a feasible method to discover strong promoters in P. mendocina NK-01 and highlights the potential of the screened endogenous strong promoters for metabolic engineering of P. mendocina NK-01 to increase the yield of mcl-PHA.
Topics: Acyltransferases; Bacterial Proteins; Metabolic Engineering; Polyhydroxyalkanoates; Promoter Regions, Genetic; Pseudomonas mendocina; Real-Time Polymerase Chain Reaction
PubMed: 30755729
DOI: 10.1038/s41598-019-39321-z -
Journal of Bacteriology Jul 2011Pseudomonas mendocina NK-01 can synthesize medium-chain-length polyhydroxyalkanoate (PHA(MCL)) and alginate oligosaccharides (AO) simultaneously from glucose under...
Pseudomonas mendocina NK-01 can synthesize medium-chain-length polyhydroxyalkanoate (PHA(MCL)) and alginate oligosaccharides (AO) simultaneously from glucose under conditions of limited nitrogen. Here, we report the complete sequence of the 5.4-Mbp genome of Pseudomonas mendocina NK-01, which was isolated from farmland soil in Tianjin, China.
Topics: Alginates; China; DNA, Bacterial; Genome, Bacterial; Glucose; Glucuronic Acid; Hexuronic Acids; Molecular Sequence Data; Oligosaccharides; Polyhydroxyalkanoates; Pseudomonas mendocina; Sequence Analysis, DNA; Soil Microbiology
PubMed: 21551299
DOI: 10.1128/JB.05068-11 -
Bioresource Technology Jan 2024Bio-mitigation of plastics by microorganisms generates carbon dioxide (CO) that can be utilized for algal biomass generation. Pseudomonas mendocina ABF786, reportedly...
Bio-mitigation of plastics by microorganisms generates carbon dioxide (CO) that can be utilized for algal biomass generation. Pseudomonas mendocina ABF786, reportedly the most efficient plastic-degrading bacteria, was screened using the modified most probable number technique. This study highlights the use of an integrative prototype for the production of microalgal biomass (Chlorella vulgaris) in combination with bio-mitigation of plastics, which serves a dual purpose: (i) increased plastic-degradation capability by microorganisms (53%-85% increase in plastic weight loss) due to removal of CO feedback inhibition and (ii) increased algal biomass generation (200%-237%) due to supply of extra CO from plastic degradation to the algal cultivation flask. Whole-genome sequencing and functional annotation confirmed that all the genes involved in the mineralization of plastic to CO are present within the genome of P. mendocina ABF786. Using two or more microbial cultures for remediation may increase the process efficiency.
Topics: Chlorella vulgaris; Biofuels; Carbon Dioxide; Pseudomonas mendocina; Microalgae; Biomass
PubMed: 37925087
DOI: 10.1016/j.biortech.2023.129952 -
Journal of Basic Microbiology 2003The regulation of aspartate transcarbamoylase activity in cell extracts of Pseudomonas alcaligenes ATCC 14909 and Pseudomonas mendocina ATCC 25411 was compared. Under... (Comparative Study)
Comparative Study
The regulation of aspartate transcarbamoylase activity in cell extracts of Pseudomonas alcaligenes ATCC 14909 and Pseudomonas mendocina ATCC 25411 was compared. Under saturating substrate concentrations, pyrophosphate, CTP, UDP and ADP were highly inhibitory of the P. alcaligenes transcarbamoylase activity while pyrophosphate, UDP, ADP, ATP and GTP were the most effective inhibitors of the P. mendocina transcarbamoylase. By examining transcarbamoylase inhibition by ribonucleotide triphosphates, it was possible to differentiate these species assigned to different DNA homology groups and such an analysis might prove useful in the reclassification of Pseudomonas species.
Topics: Aspartate Carbamoyltransferase; Diphosphates; Nucleotides; Phosphates; Pseudomonas
PubMed: 12596245
DOI: 10.1002/jobm.200390008 -
Journal of Global Antimicrobial... Sep 2018
Topics: Adult; Burns; Colistin; Fatal Outcome; Humans; Male; Middle Aged; Multiple Organ Failure; Pseudomonas Infections; Pseudomonas mendocina; Skin Transplantation; Treatment Outcome; beta-Lactamases
PubMed: 30118765
DOI: 10.1016/j.jgar.2018.08.002 -
International Journal of Environmental... Jan 2019To remove nitrate in wastewater treatment plant effluent, an aerobic denitrifier was newly isolated from the surface flow constructed wetland and identified as strain...
To remove nitrate in wastewater treatment plant effluent, an aerobic denitrifier was newly isolated from the surface flow constructed wetland and identified as strain GL6. It exhibited efficient aerobic denitrification ability, with the nitrate removal rate of 6.61 mg (N)·L·h. Sequence amplification indicated that the denitrification genes , , , and were present in strain GL6. Nitrogen balance analysis revealed that approximately 74.5% of the initial nitrogen was removed as gas products. In addition, the response surface methodology experiments showed that the maximum removal of total nitrogen occurred at pH 7.76, C/N ratio of 11.2, temperature of 27.8 °C, and with shaking at 133 rpm. Furthermore, under the optimized cultivation condition, strain GL6 was added into wastewater treatment plant effluent and the removal rates of nitrate nitrogen and total nitrogen reached 95.6% and 73.6%, respectively. Thus, strain GL6 has high denitrification potential for deep improvement of effluent quality.
Topics: Aerobiosis; Biodegradation, Environmental; China; Denitrification; Nitrates; Pseudomonas mendocina; Wastewater; Water Purification
PubMed: 30696062
DOI: 10.3390/ijerph16030364