Putative 3-nitrotyrosine detoxifying genes identified in the yeast Debaryomyces hansenii: in silico search of regulatory sequences responsive to salt and nitrogen stress

Background: During salt stress, the yeast Debaryomyces hansenii synthesizes tyrosine as a strategy to avoid the oxidation of proteins. Tyrosine reacts with nitrogen radicals to form 3-nitrotyrosine. 3-nitrotyrosine prevents the effects of associated oxidative stress and thus contributes to the high halotolerace of the yeast. However, the mechanism of how D. hansenii counteracts the presence of this toxic compound is unclear. In this work, we evaluated D. hansenii's capacity to assimilate 3-nitrotyrosine as a unique nitrogen source and measured its denitrase activity under salt stress. To identify putative genes related to the assimilation of 3-nitrotyrosine, we performed an in silico search in the promoter regions of D. hansenii genome. Results: We identified 15 genes whose promoters had binding site sequences for transcriptional factors of sodium, nitrogen, and oxidative stress with oxidoreductase and monooxygenase GO annotations. Two of these genes, DEHA2E24178g and DEHA2C00286g, coding for putative denitrases and having GATA sequences, were evaluated by RT-PCR and showed high expression under salt and nitrogen stress. Conclusions: D. hansenii can grow in the presence of 3-nitrotyrosine as the only nitrogen source and has a high specific denitrase activity to degrade 3-nitrotyrosine in 1 and 2 M NaCl stress conditions. The results suggest that given the lack of information on transcriptional factors in D. hansenii, the genes identified in our in silico analysis may help explain 3-nitrotyrosine assimilation mechanisms.

Structure and Functional Relationship of Halotolerant Proteins- A Review.

With increasing need for various industries for stable enzymes which can outperform under harsh conditions of pH, temperature and saline environments evoke the need for extensive research on extremophilic microorganisms which found to be a suitable source for enzymes with novel properties. Especially, enzymes capable of functioning over a wide range of salt concentrations are in great demand for industries which mainly include proteases, xylanases, mannases, lipases, amylases and cellulases. In case of halophilic, extremely halophilic or halotolerant organisms it has been scientifically proven that there is a correlation between the salt requirement and bacterial growth and its enzymatic activities. When compared to halophilic organism, halotolerant bacteria can survive from low to moderate level of salinity, which gives them unique property over halophilic organisms. The halotolerant organisms synthesize enzymes with greater conformational stability and they can be potentially employed in many industrial processes even under harsher conditions over a range of salinity conditions where enzymes from non-halophilic and halophilic organisms found to be unfit. Less research has been carried out on halotolerant organisms when compared to thermophilic, alkaliphilic and halophilic organisms and so yet to explore the biotechnological applications of these halotolerant enzymes to use them in industrial applications.

Halotolerance, ligninase production and herbicide degradation ability of basidiomycetes strains

Fungi have been recently recognized as organisms able to grow in presence of high salt concentration with halophilic and halotolerance properties and their ligninolytic enzyme complex have an unspecific action enabling their use to degradation of a number of xenobiotic compounds. In this work, both the effect of salt and polyols on growth of the basidiomycetes strains, on their ability to produce ligninolytic enzyme and diuron degradation were evaluated. Results showed that the presence of NaCl in the culture medium affected fungal specimens in different ways. Seven out of ten tested strains had growth inhibited by salt while Dacryopinax elegans SXS323, Polyporus sp MCA128 and Datronia stereoides MCA167 fungi exhibited higher biomass production in medium containing 0.5 and 0.6 mol.L-1 of NaCl, suggesting to be halotolerant. Polyols such as glycerol and mannitol added into the culture media improved the biomass and ligninases production by D. elegans but the fungus did not reveal consumption of these polyols from media. This fungus degraded diuron in medium control, in presence of NaCl as well as polyols, produced MnP, LiP and laccase.

The effect of organic osmoprotectors on Aeromonas trota and A. hydrophila grown under high sodium chloride concentrations

The effect of organic solutes on the growth of Aeromonas trota and A. hydrophila was evaluated. Proline and glutamic acid were not effective as osmoprotectors, but betaine exerts osmoprotection allowing the growth of both strains in inhibitory concentrations of NaCl. Growth kinetics suggests that the halotolerance difference between the strains is associated with the synthesis of osmolytes rather than betaine uptake.

O efeito de solutos orgânicos no crescimento de Aeromonas trota e A. hydrophila foi avaliado. A prolina e o ácido glutâmico não apresentaram efeito osmoprotetor, mas a betaina exerceu osmoproteção permitindo o crescimento de ambas bactérias em concentrações inibitórias de NaCl. A cinética do crescimento sugere que a diferença em halotolerância entre as linhagens está associada à síntese de osmolitos, mais do que à capacidade de acúmulo de betaina.

The effect of organic osmoprotectors on Aeromonas trota and A. hydrophila grown under high sodium chloride concentrations

The effect of organic solutes on the growth of Aeromonas trota and A. hydrophila was evaluated. Proline and glutamic acid were not effective as osmoprotectors, but betaine exerts osmoprotection allowing the growth of both strains in inhibitory concentrations of NaCl. Growth kinetics suggests that the halotolerance difference between the strains is associated with the synthesis of osmolytes rather than betaine uptake.

O efeito de solutos orgânicos no crescimento de Aeromonas trota e A. hydrophila foi avaliado. A prolina e o ácido glutâmico não apresentaram efeito osmoprotetor, mas a betaina exerceu osmoproteção permitindo o crescimento de ambas bactérias em concentrações inibitórias de NaCl. A cinética do crescimento sugere que a diferença em halotolerância entre as linhagens está associada à síntese de osmolitos, mais do que à capacidade de acúmulo de betaina.