The yeast Pichia pastoris is a cost-effective and easily scalable system for recombinant protein production. In this work we compared the conformation of the receptor binding domain (RBD) from severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) Spike protein expressed in P. pastoris and in the well established HEK-293T mammalian cell system. RBD obtained from both yeast and mammalian cells was properly folded, as indicated by UV-absorption, circular dichroism and tryptophan fluorescence. They also had similar stability, as indicated by temperature-induced unfolding (observed Tm were 50 °C and 52 °C for RBD produced in P. pastoris and HEK-293T cells, respectively). Moreover, the stability of both variants was similarly reduced when the ionic strength was increased, in agreement with a computational analysis predicting that a set of ionic interactions may stabilize RBD structure. Further characterization by high-performance liquid chromatography, size-exclusion chromatography and mass spectrometry revealed a higher heterogeneity of RBD expressed in P. pastoris relative to that produced in HEK-293T cells, which disappeared after enzymatic removal of glycans. The production of RBD in P. pastoris was scaled-up in a bioreactor, with yields above 45 mg/L of 90% pure protein, thus potentially allowing large scale immunizations to produce neutralizing antibodies, as well as the large scale production of serological tests for SARS-CoV-2.

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Three hydrocarbon-degrading Rhodococcus strains isolated from polluted Antarctic soils proved to be closely related despite their different origins. Strains had a similar hydrocarbon degradation pattern and optimum growth temperature ranged between 25ºC and 30ºC, showing that strains are psychrotolerant but not psychrophiles. Specific growth rate on rich media ranged between 0.12 and 0.21 h−1, higher than those observed on hydrocarbons as carbon source. Results suggest that in Antarctic contaminated soils, closely related Rhodococcus strains are present and could play an important role in decontamination. Microcosm systems showed that, although the natural microflora respond significantly to the pollutants, bioaugmentation with Rhodococcus strain (ADH), improved biodegradation either alone or mixed with a hydrocarbon-degrading Acinetobacter strain. In comparison with microcosm where only ADH was inoculated, a non-significant decrease in hydrocarbon concentration was observed when ADH was inoculated as mixed culture with a previously tested strain. Pollutants dramatically reduced bacterial groups in soils resulting in a dominance of Pseudomonas. Microcosms showed that when natural microflora has no previous history of exposure to the pollutants, bioaugmentation with autochthonous strains improves degradation of the contaminants. The positive response of the native bacteria to the pollutants leaves the question open as to whether bioaugmentation is necessary when soils have a long previous exposure to hydrocarbons.

Complexity involved in the transport of soils and the restrictive legislation for the area makes on‐site bioremediation the strategy of choice to reduce hydrocarbons contamination in Antarctica. The effect of biostimulation (with N and P) and bioaugmentation (with two bacterial consortia and a mix of bacterial strains) was analysed by using microcosms set up on metal trays containing 2·5 kg of contaminated soil from Marambio Station. At the end of the assay (45 days), all biostimulated systems showed significant increases in total heterotrophic aerobic and hydrocarbon‐degrading bacterial counts. However, no differences were detected between bioaugmented and nonbioaugmented systems, except for J13 system which seemed to exert a negative effect on the natural bacterial flora. Hydrocarbons removal efficiencies agreed with changes in bacterial counts reaching 86 and 81% in M10 (bioaugmented) and CC (biostimulated only) systems. Results confirmed the feasibility of the application of bioremediation strategies to reduce hydrocarbon contamination in Antarctic soils and showed that, when soils are chronically contaminated, biostimulation is the best option. Bioaugmentation with hydrocarbon‐degrading bacteria at numbers comparable to the total heterotrophic aerobic counts showed by the natural microflora did not improve the process and showed that they would turn the procedure unnecessarily more complex.