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Published on: May 4, 2018
Genome features of Pseudomonas putida LS46, a novel polyhydroxyalkanoate producer and its comparison with other P.
Parveen K Sharma1, Jilagamazhi Fu1, Xiangli Zhang2
1Department of Biosystems Engineering, University of Manitoba, Winnipeg R3T 2N2, MB, Canada.
Abstract:
A novel strain of Pseudomonas putida LS46 was isolated from wastewater on the basis of its ability to synthesize medium chain-length polyhydroxyalkanoates (mcl-PHAs). P.putida LS46 was differentiated from other P.putida strains on the basis of cpn60 (UT). The complete genome of P.putida LS46 was sequenced and annotated. Its chromosome is 5,86,2556 bp in size with GC ratio of 61.69. It is encoding 5316 genes, including 7 rRNA genes and 76 tRNA genes. Nucleotide sequence data of the complete P. putida LS46 genome was compared with nine other P. putida strains (KT2440, F1, BIRD-1, S16, ND6, DOT-T1E, UW4, W619 and GB-1) identified either as biocontrol agents or as bioremediation agents and isolated from different geographical region and different environment. BLASTn analysis of whole genome sequences of the ten P. putida strains revealed nucleotide sequence identities of 86.54 to 97.52%. P.putida genome arrangement was LS46 highly similar to P.putida BIRD1 and P.putida ND6 but was markedly different than P.putida DOT-T1E, P.putida UW4 and P.putida W619. Fatty acid biosynthesis (fab), fatty acid degradation (fad) and PHA synthesis genes were highly conserved among biocontrol and bioremediation P.putida strains. Six genes in pha operon of P. putida LS46 showed >98% homology at gene and proteins level. It appears that polyhydroxyalkanoate (PHA) synthesis is an intrinsic property of P. putida and was not affected by its geographic origin. However, all strains, including P. putida LS46, were different from one another on the basis of house keeping genes, and presence of plasmid, prophages, insertion sequence elements and genomic islands. While P. putida LS46 was not selected for plant growth promotion or bioremediation capacity, its genome also encoded genes for root colonization, pyoverdine synthesis, oxidative stress (present in other soil isolates), degradation of aromatic compounds, heavy metal resistance and nicotinic acid degradation, manganese (Mn II) oxidation. Genes for toluene or naphthalene degradation found in the genomes of P. putida F1, DOT-T1E, and ND6 were absent in the P. putida LS46 genome. Heavy metal resistant genes encoded by the P. putida W619 genome were also not present in the P. putida LS46 genome. Despite the overall similarity among genome of P.putida strains isolated for different applications and from different geographical location a number of differences were observed in genome arrangement, occurrence of transposon, genomic islands and prophage. It appears that P.putida strains had a common ancestor and by acquiring some specific genes by horizontal gene transfer it differed from other related strains.
Insights
The genome of Pseudomonas putida LS46, isolated for polyhydroxyalkanoates (PHAs) synthesis, was sequenced and compared to other strains. PHA synthesis is common in P. putida, but genomic variations exist due to horizontal gene transfer.
Area of Science:
- Microbiology
- Genomics
- Biotechnology
Background:
- Pseudomonas putida strains are known for diverse metabolic capabilities, including biocontrol and bioremediation.
- Medium chain-length polyhydroxyalkanoates (mcl-PHAs) are biodegradable polymers with industrial applications.
- Understanding the genomic basis of P. putida's traits is crucial for optimizing its use.
Purpose of the Study:
- To sequence and annotate the complete genome of a novel PHA-producing P. putida strain, LS46.
- To compare the LS46 genome with other P. putida strains to identify conserved and unique genetic features.
- To investigate the genetic basis of PHA synthesis and other metabolic potentials in P. putida.
Main Methods:
- Isolation and characterization of P. putida LS46 from wastewater.
- Whole-genome sequencing and annotation of P. putida LS46.
- Comparative genomic analysis using BLASTn against nine other P. putida strains.
Main Results:
- The P. putida LS46 genome is 5.86 Mb with a GC content of 61.69%, encoding 5316 genes.
- PHA synthesis genes (pha operon) are highly conserved (>98% homology) across P. putida strains, suggesting it's an intrinsic property.
- LS46 genome lacks genes for toluene/naphthalene degradation and heavy metal resistance found in other strains, but possesses genes for root colonization and Mn oxidation.
Conclusions:
- PHA synthesis is a conserved trait in P. putida, irrespective of geographic origin.
- Genomic variations, including plasmids, prophages, and genomic islands, contribute to strain-specific differences.
- P. putida strains share a common ancestor, with divergence driven by horizontal gene transfer and acquisition of specific genes.
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