RESUMO
The ability of siderophores to play roles beyond iron acquisition has been recently proven for many of them and evidence continues to grow. An earlier work showed that the siderophore enterobactin is able to increase copper toxicity by reducing Cu2+ to Cu+, a form of copper that is more toxic to cells. Copper toxicity is multifaceted. It involves the formation of reactive oxygen species (ROS), mismetallation of enzymes and possibly other mechanisms. Given that we previously reported on the capacity of enterobactin to alleviate oxidative stress caused by various stressors other than copper, we considered the possibility that the siderophore could play a dual role regarding copper toxicity. In this work, we show a bimodal effect of enterobactin on copper toxicity (protective and harmful) which depends on the siderophore concentration. We found that the absence of enterobactin rendered Escherichia coli cells more sensitive to copper, due to the reduced ability of those cells to cope with the metal-generated ROS. Consistently, addition of low concentrations of the siderophore had a protective effect by reducing ROS levels. We observed that in order to achieve this protection, enterobactin had to enter cells and be hydrolyzed in the cytoplasm. Further supporting the role of enterobactin in oxidative stress protection, we found that both oxygen and copper, induced the expression of the siderophore and also found that copper strongly counteracted the well-known downregulation effect of iron on enterobactin synthesis. Interestingly, when enterobactin was present in high concentrations, cells became particularly sensitive to copper most likely due to the Cu2+ to Cu+ reduction, which increased the metal toxicity leading to cell death.
RESUMO
Trichosporon akiyoshidainum HP-2023 completely discolorised Reactive Black 5 (200 mg/l) in 24 h. Manganese peroxidase and phenoloxidase, but no laccase activities were detected throughout the incubation. Total aromatic amines in media with Reactive Black 5 decreased 83% after 24 h, supporting an oxidative mechanism of azo dye discoloration. To unravel the genetic basis of these activities, the genome of Trichosporon akiyoshidainum HP-2023 was sequenced, assembled and annotated de novo. T. akiyoshidainum HP-2023 genome comprises 30 MB with a G+C content of 60.75% and 9019 gene models. Thirty-three putative carbohydrate-active enzymes with auxiliary activities, probably involved in lignin degradation and dye discoloration, were identified in the annotated genome, including two laccases, four extracellular fungal heme-peroxidases, nineteen hydrogen peroxide-producing enzymes, and four benzoquinone oxidoreductases. This report will facilitate further studies of textile-dye discoloration with this and closely related strains and poses questions about the ligninolytic potential of Trichosporon akiyoshidainum HP-2023 and related species.