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Optimization and characterization of Alginic acid synthesized from a novel strain of Pseudomonas stutzeri.

ABSTRACT: Alginate is a group of water-soluble linear polysaccharides comprising of variable units of ?-l-guluronic and ?-d-mannuronic acid. The alginates are in high demand in biomedical, pharmaceutical and bioengineering applications. In the present study, we have isolated a strain of Pseudomonas stutzeri that has potential alginate synthesis. The biochemical and physiochemical characteristic including Carbazole assay, DSC, FTIR and H NMR were confirmed the alginate synthesis efficacy by P. stutzeri. Evaluation of P. stutzeri alginate for the removal of heavy metals such as Chromium, Cobalt and Lead showed that it effectively adsorbs heavy metals. Further analysis of gelling ability and cytotoxicity evaluation revealed that the alginate can be reconstituted as hydrogel and scaffold. Overall, our findings suggest that the strain P. stutzeri TN_Alg Syn may be used to produce alginate at commercial level that has the potential bioremediation and biomedical applications.

SUBMITTER: S M

PROVIDER: S-EPMC7451856 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

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Optimization and characterization of Alginic acid synthesized from a novel strain of Pseudomonas stutzeri.

S Meena M M Vidya Kalaivani VK Tripathi Abhishek Dutt AD Ts Ramyaa Lakshmi RL

Biotechnology reports (Amsterdam, Netherlands) 20200808

Alginate is a group of water-soluble linear polysaccharides comprising of variable units of α-l-guluronic and β-d-mannuronic acid. The alginates are in high demand in biomedical, pharmaceutical and bioengineering applications. In the present study, we have isolated a strain of Pseudomonas stutzeri that has potential alginate synthesis. The biochemical and physiochemical characteristic including Carbazole assay, DSC, FTIR and H NMR were confirmed the alginate synthesis efficacy by P. st ...[more]

PMID: 32874945

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Project description:UNLABELLED:Cytochrome c oxidases (CcOs), members of the heme-copper containing oxidase (HCO) superfamily, are the terminal enzymes of aerobic respiratory chains. The cbb3-type cytochrome c oxidases (cbb3-CcO) form the C-family and have only the central catalytic subunit in common with the A- and B-family HCOs. In Pseudomonas stutzeri, two cbb3 operons are organized in a tandem repeat. The atomic structure of the first cbb3 isoform (Cbb3-1) was determined at 3.2 Å resolution in 2010 (S. Buschmann, E. Warkentin, H. Xie, J. D. Langer, U. Ermler, and H. Michel, Science 329:327-330, 2010, http://dx.doi.org/10.1126/science.1187303). Unexpectedly, the electron density map of Cbb3-1 revealed the presence of an additional transmembrane helix (TMH) which could not be assigned to any known protein. We now identified this TMH as the previously uncharacterized protein PstZoBell_05036, using a customized matrix-assisted laser desorption ionization (MALDI)-tandem mass spectrometry setup. The amino acid sequence matches the electron density of the unassigned TMH. Consequently, the protein was renamed CcoM. In order to identify the function of this new subunit in the cbb3 complex, we generated and analyzed a CcoM knockout strain. The results of the biochemical and biophysical characterization indicate that CcoM may be involved in CcO complex assembly or stabilization. In addition, we found that CcoM plays a role in anaerobic respiration, as the ?CcoM strain displayed altered growth rates under anaerobic denitrifying conditions. IMPORTANCE:The respiratory chain has recently moved into the focus for drug development against prokaryotic human pathogens, in particular, for multiresistant strains (P. Murima, J. D. McKinney, and K. Pethe, Chem Biol 21:1423-1432, 2014, http://dx.doi.org/10.1016/j.chembiol.2014.08.020). cbb3-CcO is an essential enzyme for many different pathogenic bacterial species, e.g., Helicobacter pylori, Vibrio cholerae, and Pseudomonas aeruginosa, and represents a promising drug target. In order to develop compounds targeting these proteins, a detailed understanding of the molecular architecture and function is required. Here we identified and characterized a novel subunit, CcoM, in the cbb3-CcO complex and thereby completed the crystal structure of the Cbb3 oxidase from Pseudomonas stutzeri, a bacterium closely related to the human pathogen Pseudomonas aeruginosa.

Project description:PurposeSelenium nanoparticles (SeNP) have several applications in the field of biotechnology, including their use as anti-cancer drugs. The purpose of the present study is to analyze the efficacy of green synthesis on the preparation of SeNP and its effect on their anti-cancer properties.MethodsA bacterial strain isolated from a freshwater source was shown to efficiently synthesize SeNP with potential therapeutic properties. The quality and stability of the NP were studied by scanning electron microscopy, X-ray diffraction, zeta-potential and FTIR analysis. A cost-effective medium formulation from biowaste having 6% banana peel extract enriched with 0.25 mM tryptophan was used to synthesize the NP. The NP after optimization was used to analyze their anti-tumor and anti-angiogenic activity. For this purpose, first, the cytotoxicity of the NP against cancer cells was analyzed by MTT assay and then chorioallantoic membrane assay was performed to assess anti-angiogenic activity. Further, cell migration assay and clonogenic inhibition assay were performed to test the anti-tumor properties of SeNP. To assess the cytotoxicity of SeNP on healthy RBC, hemolysis assay was performed.ResultsThe strain identified as Pseudomonas stutzeri (MH191156) produced phenazine carboxylic acid, which aids the conversion of Se oxyanions to reduced NP state, resulting in particles in the size range of 75 nm to 200 nm with improved stability and quality of SeNP, as observed by zeta (ξ) potential of the particles which was found to be -46.2 mV. Cytotoxicity of the SeNP was observed even at low concentrations such as 5 µg/mL against cervical cancer cell line, ie, HeLa cells. Further, neovascularization was inhibited by upto 30 % in CAMs of eggs coinoculated with SeNp when compared with untreated controls, indicating significant anti-angiogenic activity of SeNP. The NP also inhibited the invasiveness of HeLa cells as observed by decreased cell migration and clonogenic proliferation. These observations indicate significant anti-tumor and anti-angiogenic activity of the SeNP in cervical cancer cells.ConclusionP. stutzeri (MH191156) is an efficient source of Se NP production with potential anti-angiogenic and anti-tumor properties, particularly against cervical cancer cells.

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Optimization and characterization of Alginic acid synthesized from a novel strain of Pseudomonas stutzeri.

Publications

Optimization and characterization of Alginic acid synthesized from a novel strain of <i>Pseudomonas stutzeri</i>.

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