Deglycosylated RBD produced in Pichia pastoris as a low-cost sera COVID-19 diagnosis tool and a vaccine candidate.

During the COVID-19 outbreak, numerous tools including protein-based vaccines have been developed. The methylotrophic yeast Pichia pastoris (synonymous to Komagataella phaffii) is an eukaryotic cost-effective and scalable system for recombinant protein production, with the advantages of an efficient secretion system and the protein folding assistance of the secretory pathway of eukaryotic cells. In a previous work, we compared the expression of SARS-CoV-2 Spike Receptor Binding Domain in P. pastoris with that in human cells. Although the size and glycosylation pattern was different between them, their protein structural and conformational features were indistinguishable. Nevertheless, since high mannose glycan extensions in proteins expressed by yeast may be the cause of a nonspecific immune recognition, we deglycosylated RBD in native conditions. This resulted in a highly pure, homogenous, properly folded and monomeric stable protein. This was confirmed by circular dichroism and tryptophan fluorescence spectra and by SEC-HPLC, which were similar to those of RBD proteins produced in yeast or human cells. Deglycosylated RBD was obtained at high yields in a single step, and it was efficient in distinguishing between SARS-CoV-2-negative and positive sera from patients. Moreover, when the deglycosylated variant was used as an immunogen, it elicited a humoral immune response ten times greater than the glycosylated form, producing antibodies with enhanced neutralizing power and eliciting a more robust cellular response. The proposed approach may be used to produce at a low cost, many antigens that require glycosylation to fold and express, but do not require glycans for recognition purposes.

Microbial associations for bioremediation. What does "microbial consortia" mean?

Microbial associations arise as useful tools in several biotechnological processes. Among them, bioremediation of contaminated environments usually takes advantage of these microbial associations. Despite being frequently used, these associations are indicated using a variety of expressions, showing a lack of consensus by specialists in the field. The main idea of this work is to analyze the variety of microbial associations referred to as "microbial consortia" (MC) in the context of pollutants biodegradation and bioremediation. To do that, we summarize the origin of the term pointing out the features that an MC is expected to meet, according to the opinion of several authors. An analysis of related bibliography was done seeking criteria to rationalize and classify MC in the context of bioremediation. We identify that the microbe's origin and the level of human intervention are usually considered as a category to classify them as natural microbial consortia (NMC), artificial microbial consortia (AMC), and synthetic microbial consortia (SMC). In this sense, NMC are those associations composed by microorganisms obtained from a single source while AMC members come from different sources. SMC are a class of AMC in which microbial composition is defined to accomplish a certain specific task. We propose that the effective or potential existence of the interaction among MC members in the source material should be considered as a category in the classification as well, in combination with the origin of the source and level of intervention. Cross-kingdom MC and new developments were also considered. Finally, the existence of grey zones in the limits between each proposed microbial consortia category is addressed. KEY POINTS: • Microbial consortia for bioremediation can be obtained through different methods. • The use of the term "microbial consortia" is unclear in the specialized literature. • We propose a simplified classification for microbial consortia for bioremediation.

Antarctic wastewater: A local source of microplastic pollution.

Microplastic (MP) particles can be found all around the planet, even in Antarctica where they can be locally originated or transported by marine currents and winds. In this communication, we identify and report for the first time the contribution of a wastewater treatment plant (WWTP) as a local source of MP particles in the region. The analysis of the entire sample using micro-Raman spectroscopy revealed an MP concentration that ranged from 64 to 159 particles per liter of wastewater. >90 % of the identified particles were smaller than 50 µm. Among those analyzed, microplastics were identified as polyethylene, polypropylene, polyvinyl chloride, polytetrafluoroethylene, polyethylene terephthalate, and polystyrene. These findings demonstrate the need for urgent policies and technologies to mitigate this MP contamination source.

Metagenomic strategies identify diverse integron-integrase and antibiotic resistance genes in the Antarctic environment.

The objective of this study is to identify and analyze integrons and antibiotic resistance genes (ARGs) in samples collected from diverse sites in terrestrial Antarctica. Integrons were studied using two independent methods. One involved the construction and analysis of intI gene amplicon libraries. In addition, we sequenced 17 metagenomes of microbial mats and soil by high-throughput sequencing and analyzed these data using the IntegronFinder program. As expected, the metagenomic analysis allowed for the identification of novel predicted intI integrases and gene cassettes (GCs), which mostly encode unknown functions. However, some intI genes are similar to sequences previously identified by amplicon library analysis in soil samples collected from non-Antarctic sites. ARGs were analyzed in the metagenomes using ABRIcate with CARD database and verified if these genes could be classified as GCs by IntegronFinder. We identified 53 ARGs in 15 metagenomes, but only four were classified as GCs, one in MTG12 metagenome (Continental Antarctica), encoding an aminoglycoside-modifying enzyme (AAC(6´)acetyltransferase) and the other three in CS1 metagenome (Maritime Antarctica). One of these genes encodes a class D ß-lactamase (blaOXA-205) and the other two are located in the same contig. One is part of a gene encoding the first 76 amino acids of aminoglycoside adenyltransferase (aadA6), and the other is a qacG2 gene.

Isolation of Papulaspora halima and a new morphological variety of Halosphaeria tubulifera from seawater of potter cove (King George Island, South Shetland Island, Antarctica) / Aislamiento de Papulaspora halima y de una nueva variedad morfológica de Halosphaeria tubulifera desde agua de mar de Potter Cove (Isla Rey Jorge /25 de Mayo, islas Shetland del Sur, Antártida)

Marine fungi ascribed to the ascomycetes and the hyphomycetes are infrequently reported for the Southern Ocean. For this reason, the main objective of the present work was to detect the presence of these fungi seawater of Potter Cove, King George (25 de Mayo) Island, South Shetland Island, Antarctica. For this purpose marine fungi were grown on wood test panels, placed into plastic nets in the tidal zone, exposed to the Antarctic seawater for different periods of time, which ranged between 2 and 12 months.As a result of this survey, we were able to recover and identify two marine fungi, Papulospora halima (which represents the first report for this environment) and a new morphological variety of Halosphaeria tubulifera.

Los ascomicetes e hifomicetes marinos están escasamente documentados para el océano Atlántico Sur. Por este motivo, el principal objetivo del presente trabajo fue detectar la presencia de dichos hongos en las agua marinas de la Potter Cove, en la isla Rey Jorge/25 de Mayo (islas Shetland del Sur, Antártida). Para este propósito, los hongos marinos se desarrollaron en paneles de madera dentro de una red plástica en la zona tidal, expuestos al agua de mar antártica por diferentes períodos de tiempo que oscilaron entre 2 a 12 meses. Como resultado de este estudio, fuimos capaces de recuperar e identificar 2 hongos marinos, Papulospora halima (que representa el primer reporte para este ambiente) y una nueva variedad morfológica de Halosphaeria tubulifera.