Rhodococcus aetherivorans BCP1 as cell factory for the production of intracellular tellurium nanorods under aerobic conditions

Background: Tellurite (TeO32-) is recognized as a toxic oxyanion to living organisms. However, mainly anaerobic or facultative-anaerobic microorganisms are able to tolerate and convert TeO32- into the less toxic and available form of elemental Tellurium (Te0), producing Te-deposits or Te-nanostructures. The use of TeO32--reducing bacteria can lead to the decontamination of polluted environments and the development of "green-synthesis" methods for the production of nanomaterials. In this study, the tolerance and the consumption of TeO32- have been investigated, along with the production and characterization of Te-nanorods by Rhodococcus aetherivorans BCP1 grown under aerobic conditions. Results: Aerobically grown BCP1 cells showed high tolerance towards TeO32- with a minimal inhibitory concentration (MIC) of 2800μg/mL (11.2mM). TeO32- consumption has been evaluated exposing the BCP1 strain to either 100 or 500μg/mL of K2TeO3 (unconditioned growth) or after re-inoculation in fresh medium with new addition of K2TeO3 (conditioned growth). A complete consumption of TeO32- at 100μg/mL was observed under both growth conditions, although conditioned cells showed higher consumption rate. Unconditioned and conditioned BCP1 cells partially consumed TeO32- at 500μg/mL. However, a greater TeO32- consumption was observed with conditioned cells. The production of intracellular, not aggregated and rod-shaped Te-nanostructures (TeNRs) was observed as a consequence of TeO32- reduction. Extracted TeNRs appear to be embedded in an organic surrounding material, as suggested by the chemical-physical characterization. Moreover, we observed longer TeNRs depending on either the concentration of precursor (100 or 500μg/mL of K2TeO3) or the growth conditions (unconditioned or conditioned grown cells). Conclusions:Rhodococcus aetherivorans BCP1 is able to tolerate high concentrations of TeO32- during its growth under aerobic conditions. Moreover, compared to unconditioned BCP1 cells, TeO32- conditioned cells showed a higher oxyanion consumption rate (for 100μg/mL of K2TeO3) or to consume greater amount of TeO32- (for 500μg/mL of K2TeO3). TeO32- consumption by BCP1 cells led to the production of intracellular and not aggregated TeNRs embedded in an organic surrounding material. The high resistance of BCP1 to TeO32- along with its ability to produce Te-nanostructures supports the application of this microorganism as a possible eco-friendly nanofactory

Assembly, growth and conductive properties of tellurium nanorods produced by Rhodococcus aetherivorans BCP1

Tellurite (TeO32-) is a hazardous and toxic oxyanion for living organisms. However, several microorganisms can bioconvert TeO32- into the less toxic form of elemental tellurium (Te0). Here, Rhodococcus aetherivorans BCP1 resting (non-growing) cells showed the proficiency to produce tellurium-based nanoparticles (NPs) and nanorods (NRs) through the bioconversion of TeO32-, depending on the oxyanion initial concentration and time of cellular incubation. Te-nanostructures initially appeared in the cytoplasm of BCP1 cells as spherical NPs, which, as the exposure time increased, were converted into NRs. This observation suggested the existence of an intracellular mechanism of TeNRs assembly and growth that resembled the chemical surfactant-assisted process for NRs synthesis. The TeNRs produced by the BCP1 strain showed an average length (>700 nm) almost doubled compared to those observed in other studies. Further, the biogenic TeNRs displayed a regular single-crystalline structure typically obtained for those chemically synthesized. The chemical-physical characterization of the biogenic TeNRs reflected their thermodynamic stability that is likely derived from amphiphilic biomolecules present in the organic layer surrounding the NRs. Finally, the biogenic TeNRs extract showed good electrical conductivity. Thus, these findings support the suitability of this strain as eco-friendly biocatalyst to produce high quality tellurium-based nanomaterials exploitable for technological purposes

Biosynthesis of selenium-nanoparticles and -nanorods as a product of selenite bioconversion by the aerobic bacterium Rhodococcus aetherivorans BCP1

The wide anthropogenic use of selenium compounds represents the major source of selenium pollution worldwide, causing environmental issues and health concerns. Microbe-based strategies for metal removal/recovery have received increasing interest thanks to the association of the microbial ability to detoxify toxic metal/metalloid polluted environments with the production of nanomaterials. This study investigates the tolerance and the bioconversion of selenite (SeO32−) by the aerobically grown Actinomycete Rhodococcus aetherivorans BCP1 in association with its ability to produce selenium nanoparticles and nanorods (SeNPs and SeNRs). The BCP1 strain showed high tolerance towards SeO32− with a Minimal Inhibitory Concentration (MIC) of 500 mM. The bioconversion of SeO32− was evaluated considering two different physiological states of the BCP1 strain, i.e. unconditioned and/or conditioned cells, which correspond to cells exposed for the first time or after re-inoculation in fresh medium to either 0.5 or 2 mM of Na2SeO3, respectively. SeO32− bioconversion was higher for conditioned grown cells compared to the unconditioned ones. Selenium nanostructures appeared polydisperse and not aggregated, as detected by electron microscopy, being embedded in an organic coating likely responsible for their stability, as suggested by the physical-chemical characterization. The production of smaller and/or larger SeNPs was influenced by the initial concentration of provided precursor, which resulted in the growth of longer and/or shorter SeNRs, respectively. The strong ability to tolerate high SeO32− concentrations coupled with SeNP and SeNR biosynthesis highlights promising new applications of Rhodococcus aetherivorans BCP1 as cell factory to produce stable Se-nanostructures, whose suitability might be exploited for biotechnology purposes

Exploitation of the strictly aerobic Rhodococcus aetherivorans BCP1 strain for the production of Selenium- and Tellurium-nanostructures

Tellurite (TeO32-) and selenite (SeO32-) are hazardous and toxic oxyanions for livingorganisms. However, microorganisms that tolerate and bioconvert these oxyanions to the lesstoxic and available form of elemental tellurium (Te0) and selenium (Se0) are seen as ideal candidates not only for bioremediation purposes, but also as novel cell factories to produce valuable metalloid-nanostructures. Here, Rhodococcus aetherivorans BCP1\u2019s tolerance and consumption of both TeO32- and SeO32- have been investigated, along with the production and characterization of metalloid-nanomaterials in the form of Se/Te-nanorods and Se-nanoparticles. BCP1 displayed a Minimal Inhibitory Concentration towards TeO32- and/or SeO32- of 11.2 and 500 mM, respectively, in rich medium under oxic conditions. Generally, pre-adapted BCP1 cells bioconverted a higher amount of oxyanions, regardless the initial concentrations of TeO3 2- (0.4 or 2 mM) and/or SeO32- (0.5 or 2 mM). Transmission Electron Microscopy and Energy-Dispersive X-Ray Spectroscopy showed stable and non-aggregated metalloid-nanomaterials surrounded by an organic coating, whose ability to self-assemble in the nanoscale was assessed through Dynamic Light Scattering. Finally, Zeta potential analyses revealed negative surface charges for both metalloid-nanomaterials and organic coating. We hypothesize that the biogenic Se/T