Project description:We report the isolation, whole-genome sequencing, and annotation of Enterobacter sp. strain RIT 637, Pseudomonas sp. strain RIT 778, and <i>Deinococcus</i> sp. strain RIT 780. Disk diffusion assays using spent medium demonstrated that all bacteria produced bactericidal compounds against Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, and Staphylococcus aureus ATCC 25923.
Project description:CONTEXT:Microorganisms produce a variety of pigments and many pigments from bacteria were reported to have therapeutic potential including anticancer effects. AIM:The aim of this study is to evaluate the anticancer potential a yellow pigment from newly isolated Pseudomonas stutzeri JGI 52. MATERIALS AND METHODS:Serial dilution method was adopted for the isolation of pigmented bacteria from soil sources. Pigment extraction was carried out from bacterial isolates using methanol as the solvent and the pigment was purified by thin layer chromatography. Through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, the effect of the pigment fraction on cancer cells was analyzed. Apoptosis induction was evaluated by caspase-3 activity assay, DNA fragmentation analysis, cell morphology observation by AO-EB staining under the fluorescence microscope, and cellular cytotoxicity was analysed by lactate dehydrogenase (LDH) release assay. Characterization of the purified pigment was by high-performance liquid chromatography and electrospray ionization-mass spectrometry analysis. STATISTICAL ANALYSIS:Significance of the results was confirmed by performing one-way analysis of variance. RESULTS:The pigment (PY3) from P. stutzeri inhibited the proliferation of HeLa, HepG2, and Jurkat cells and found to be less toxic to lymphocytes and CHO cells. PY3 exhibited apoptotic potential in the cancer cell lines, as evidenced by cleavage of DNA, LDH release, activation of caspase-3, and decrease in cell count. Results of mass spectra indicated the presence of "fucoxanthinol" which was earlier reported as an anticancer compound from seaweeds. CONCLUSIONS:This study revealed that the pigment PY3 from P. stutzeri has anticancer potential and induced cell death by apoptosis. It was found to have the carotenoid fucoxanthinol, responsible for its observed anticancer activity.
Project description:Pseudomonas sp. strain M1 is a soil isolate with remarkable biotechnological potential. The genome of Pseudomonas sp. M1 was sequenced using both 454 and Illumina technologies. A customized genome assembly pipeline was used to reconstruct its genome sequence to a single scaffold.
Project description:Microbial degradation of lignin and its related aromatic compounds has great potential for the sustainable production of chemicals and bioremediation of contaminated soils. We previously isolated Pseudomonas sp. strain 9.1 from historical waste deposits (forming so-called fiber banks) released from pulp and paper mills along the Baltic Sea coast. The strain accumulated vanillyl alcohol during growth on vanillin, and while reported in other microbes, this phenotype is less common in wild-type pseudomonads. As the reduction of vanillin to vanillyl alcohol is an undesired trait in Pseudomonas strains engineered to accumulate vanillin, connecting the strain 9.1 phenotype with a genotype would increase the fundamental understanding and genetic engineering potential of microbial vanillin metabolism. The genome of Pseudomonas sp. 9.1 was sequenced and assembled. Annotation identified oxidoreductases with homology to Saccharomyces cerevisiae alcohol dehydrogenase ScADH6p, known to reduce vanillin to vanillyl alcohol, in both the 9.1 genome and the model strain Pseudomonas putida KT2440. Recombinant expression of the Pseudomonas sp. 9.1 FEZ21_09870 and P. putida KT2440 PP_2426 (calA) genes in Escherichia coli revealed that these open reading frames encode aldehyde reductases that convert vanillin to vanillyl alcohol, and that P. putida KT2440 PP_3839 encodes a coniferyl alcohol dehydrogenase that oxidizes coniferyl alcohol to coniferyl aldehyde (i.e., the function previously assigned to calA). The deletion of PP_2426 in P. putida GN442 engineered to accumulate vanillin resulted in a decrease in by-product (vanillyl alcohol) yield from 17% to ?1%. Based on these results, we propose the reannotation of PP_2426 and FEZ21_09870 as areA and PP_3839 as calA-II IMPORTANCE Valorization of lignocellulose (nonedible plant matter) is of key interest for the sustainable production of chemicals from renewable resources. Lignin, one of the main constituents of lignocellulose, is a heterogeneous aromatic biopolymer that can be chemically depolymerized into a heterogeneous mixture of aromatic building blocks; those can be further converted by certain microbes into value-added aromatic chemicals, e.g., the flavoring agent vanillin. We previously isolated a Pseudomonas sp. strain with the (for the genus) unusual trait of vanillyl alcohol production during growth on vanillin. Whole-genome sequencing of the isolate led to the identification of a vanillin reductase candidate gene whose deletion in a recombinant vanillin-accumulating P. putida strain almost completely alleviated the undesired vanillyl alcohol by-product yield. These results represent an important step toward biotechnological production of vanillin from lignin using bacterial cell factories.
Project description:Here, we present the complete genome of Pseudomonas sp. UK4. This bacterium was the first Pseudomonas strain shown to produce functional amyloids, and it represents a model organism for studies of functional amyloids in Pseudomonas (Fap).