Purification and characterization of a thermostable glutamate dehydrogenase from a thermophilic microorganism from Deception Island, Antarctica

Glutamate dehydrogenase (GDH) catalyzes the oxidative deamination of glutamate to α-ketoglutarate and ammonium ions. Currently the determination of ammonium and glutamate is carried out using a bovine GDH enzyme, which lacks optimal thermostability for long term storage at room temperature. From samples of Deception Island, Antarctica, we obtained the thermophilic bacteria PID 15 belonging to the Bacillus genera with high GDH specific activity. This new enzyme exhibited NAD+ dependent activity and no activity was observed when NADP+ was used. This enzyme shows a specific activity of 4.7 U∙mg-1 for the oxidative deamination reaction and 15.4 U∙mg-1 for the reduction of α-ketoglutarate. This enzyme has an optimum temperature of 65°C and pH of 8.5 for the oxidative deamination. For the reduction of α-ketoglutarate, the optimum temperature is 60°C, with a pH of 8.0. One of the most important characteristics of this enzyme is its ability to retain more than 60% of its activity when it is incubated for 8 h at 65°C. The enzyme is also able to retain full activity when it is incubated for 48 d at 4°C and over 80% of its activity when it is incubated at 25°C. Characterization of its kinetics suggests that it primarily catalyzes the formation of α-ketoglutarate. This enzyme has an important biological role in the catabolism of glutamate and may have some interesting biotechnological applications based on its thermostable properties

Asociación entre el ambiente volcánico del monte submarino Orca y microorganismos extremófilos

La zona del rift del Bransfield, ubicado entre las islas Shetland del Sur y la península Antártica, es un área geológica única caracterizada por un proceso de extensión cortical (rift) y subducción de tipo roll-back a la cual se asocian procesos volcánicos recientes y actividad hidrotermal subárea y submarina. Estos contrastes hacen oportuno el estudio de las interacciones entre el ambiente geológico-marino con los microorganismos. Unos de los edificios volcánicos submarinos más prominentes del rift del Bransfield es el monte Orca. Durante el crucero ANT-XXV/4 a bordo del buque R/V Polarstern en 2010, se obtuvieron muestras de agua al interior y fuera del cráter del monte Orca. Mediciones oceanográficas y geoquímicas, indicaron que existe una actividad hidrotermal débil. Por otro lado, se encontraron microorganismos termófilos e hipertermofilos en las muestras luego de incubación en varios medios y a temperaturas entre 70 °C y 97 °C, siendo esta la primera observación de estos tipos de organismos en aguas frías (>1 °C) y profundas de la Antártica (> 1000 m). Posteriormente en febrero de 2019, durante el crucero ANTAR XXVI a bordo del buque BAP Carrasco, se obtuvieron muestras de sedimentos en el fondo marino al interior y exterior del cráter del monte Orca, para determinar relaciones entre la composición de los sedimentos con los microorganismos extremófilos, utilizando técnicas similares de análisis. Resultados preliminares muestran que los sedimentos se encuentran enriquecidos en sulfuros polimetálicos y elementos como hierro, cobre y zinc. Además, en los cultivos de las muestras del fondo marino, en todas crecieron microorganismos termófilos (>70 °C), encontrándose una asociación entre la generación de precipitados de sulfuros de hierro y el crecimiento de los termófilos

Alicyclobacillus sp. strain CC2, a thermo-acidophilic bacterium isolated from Deception Island (Antarctica) containing a thermostable superoxide dismutase enzyme

A gram-positive, rod-shaped, aerobic, thermo-acidophilic bacterium CC2 (optimal temperature 55℃ and pH 4.0), belonging to the genus Alicyclobacillus was isolated from geothermal soil collected from “Cerro Caliente”, Deception Island, Antarctica. Owing to the harsh environmental conditions found in this territory, microorganisms are exposed to conditions that trigger the generation of reactive oxygen species (ROS). They must have an effective antioxidant defense system to deal with this oxidative stress. We focused on one of the most important enzymes: superoxide dismutase, which was partially purified and characterized. This study presents the first report of a thermo-acidophilic bacterium isolated from Deception Island with a thermostable superoxide dismutase (SOD)

The helium and carbon isotope characteristics of the Andean Convergent Margin

© The Author(s), 2022. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Barry, P. H., De Moor, J. M., Chiodi, A., Aguilera, F., Hudak, M. R., Bekaert, D. V., Turner, S. J., Curtice, J., Seltzer, A. M., Jessen, G. L., Osses, E., Blamey, J. M., Amenabar, M. J., Selci, M., Cascone, M., Bastianoni, A., Nakagawa, M., Filipovich, R., Bustos, E., Schrenk, M. O. , Buongiorno, J., Ramírez, C. J., Rogers, T. J., Lloyd, K. G. & Giovannelli, D. The helium and carbon isotope characteristics of the Andean Convergent Margin. Frontiers in Earth Science, 10, (2022): 897267, https://doi.org/10.3389/feart.2022.897267.Subduction zones represent the interface between Earth’s interior (crust and mantle) and exterior (atmosphere and oceans), where carbon and other volatile elements are actively cycled between Earth reservoirs by plate tectonics. Helium is a sensitive tracer of volatile sources and can be used to deconvolute mantle and crustal sources in arcs; however it is not thought to be recycled into the mantle by subduction processes. In contrast, carbon is readily recycled, mostly in the form of carbon-rich sediments, and can thus be used to understand volatile delivery via subduction. Further, carbon is chemically-reactive and isotope fractionation can be used to determine the main processes controlling volatile movements within arc systems. Here, we report helium isotope and abundance data for 42 deeply-sourced fluid and gas samples from the Central Volcanic Zone (CVZ) and Southern Volcanic Zone (SVZ) of the Andean Convergent Margin (ACM). Data are used to assess the influence of subduction parameters (e.g., crustal thickness, subduction inputs, and convergence rate) on the composition of volatiles in surface volcanic fluid and gas emissions. He isotopes from the CVZ backarc range from 0.1 to 2.6 RA (n = 23), with the highest values in the Puna and the lowest in the Sub-Andean foreland fold-and-thrust belt. Atmosphere-corrected He isotopes from the SVZ range from 0.7 to 5.0 RA (n = 19). Taken together, these data reveal a clear southeastward increase in 3He/4He, with the highest values (in the SVZ) falling below the nominal range associated with pure upper mantle helium (8 ± 1 RA), approaching the mean He isotope value for arc gases of (5.4 ± 1.9 RA). Notably, the lowest values are found in the CVZ, suggesting more significant crustal inputs (i.e., assimilation of 4He) to the helium budget. The crustal thickness in the CVZ (up to 70 km) is significantly larger than in the SVZ, where it is just ∼40 km. We suggest that crustal thickness exerts a primary control on the extent of fluid-crust interaction, as helium and other volatiles rise through the upper plate in the ACM. We also report carbon isotopes from (n = 11) sites in the CVZ, where δ13C varies between −15.3‰ and −1.2‰ [vs. Vienna Pee Dee Belemnite (VPDB)] and CO2/3He values that vary by over two orders of magnitude (6.9 × 108–1.7 × 1011). In the SVZ, carbon isotope ratios are also reported from (n = 13) sites and vary between −17.2‰ and −4.1‰. CO2/3He values vary by over four orders of magnitude (4.7 × 107–1.7 × 1012). Low δ13C and CO2/3He values are consistent with CO2 removal (e.g., calcite precipitation and gas dissolution) in shallow hydrothermal systems. Carbon isotope fractionation modeling suggests that calcite precipitation occurs at temperatures coincident with the upper temperature limit for life (122°C), suggesting that biology may play a role in C-He systematics of arc-related volcanic fluid and gas emissions.This work was principally supported by the NSF-FRES award 2121637 to PB, KL, and JM. Field work was also supported by award G-2016-7206 from the Alfred P. Sloan Foundation and the Deep Carbon Observatory to PB, KL, DG, and JM. Additional support came from The National Fund for Scientific and Technological Development of Chile (FONDECYT) Grant 11191138 (The National Research and Development Agency of Chile, ANID Chile), and COPAS COASTAL ANID FB210021 to GJ. DG was partially supported by funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program Grant Agreement No. 948972—COEVOLVE—ERC-2020-STG

Convalescent plasma in patients admitted to hospital with COVID-19 (RECOVERY): a randomised controlled, open-label, platform trial

SummaryBackground Azithromycin has been proposed as a treatment for COVID-19 on the basis of its immunomodulatoryactions. We aimed to evaluate the safety and efficacy of azithromycin in patients admitted to hospital with COVID-19.Methods In this randomised, controlled, open-label, adaptive platform trial (Randomised Evaluation of COVID-19Therapy [RECOVERY]), several possible treatments were compared with usual care in patients admitted to hospitalwith COVID-19 in the UK. The trial is underway at 176 hospitals in the UK. Eligible and consenting patients wererandomly allocated to either usual standard of care alone or usual standard of care plus azithromycin 500 mg once perday by mouth or intravenously for 10 days or until discharge (or allocation to one of the other RECOVERY treatmentgroups). Patients were assigned via web-based simple (unstratified) randomisation with allocation concealment andwere twice as likely to be randomly assigned to usual care than to any of the active treatment groups. Participants andlocal study staff were not masked to the allocated treatment, but all others involved in the trial were masked to theoutcome data during the trial. The primary outcome was 28-day all-cause mortality, assessed in the intention-to-treatpopulation. The trial is registered with ISRCTN, 50189673, and ClinicalTrials.gov, NCT04381936.Findings Between April 7 and Nov 27, 2020, of 16 442 patients enrolled in the RECOVERY trial, 9433 (57%) wereeligible and 7763 were included in the assessment of azithromycin. The mean age of these study participants was65·3 years (SD 15·7) and approximately a third were women (2944 [38%] of 7763). 2582 patients were randomlyallocated to receive azithromycin and 5181 patients were randomly allocated to usual care alone. Overall,561 (22%) patients allocated to azithromycin and 1162 (22%) patients allocated to usual care died within 28 days(rate ratio 0·97, 95% CI 0·87–1·07; p=0·50). No significant difference was seen in duration of hospital stay (median10 days [IQR 5 to >28] vs 11 days [5 to >28]) or the proportion of patients discharged from hospital alive within 28 days(rate ratio 1·04, 95% CI 0·98–1·10; p=0·19). Among those not on invasive mechanical ventilation at baseline, nosignificant difference was seen in the proportion meeting the composite endpoint of invasive mechanical ventilationor death (risk ratio 0·95, 95% CI 0·87–1·03; p=0·24).Interpretation In patients admitted to hospital with COVID-19, azithromycin did not improve survival or otherprespecified clinical outcomes. Azithromycin use in patients admitted to hospital with COVID-19 should be restrictedto patients in whom there is a clear antimicrobial indication

Isolation and partial characterization of a new moderate thermophilic Albidovulum sp. SLM16 with transaminase activity from Deception Island, Antarctica

Abstract Background A moderately thermophilic, slightly halophilic, aerobic, Gram-stain negative, bacterial strain, SLM16, was isolated from a mixed of seawater–sand-sediment sample collected from a coastal fumarole located in Whalers Bay, Deception Island, Antarctica. The aim was to screen for thermophilic microorganisms able to degrade primary amines and search for amine transaminase activity for potential industrial application. Results Identification and partial characterization of the microorganism SLM16 were carried out by means of morphological, physiological and biochemical tests along with molecular methods. Cells of strain SLM16 were non-motile irregular rods of 1.5–2.5 μm long and 0.3–0.45 μm wide. Growth occurred in the presence of 0.5–5.5% NaCl within temperature range of 35–55 °C and pH range of 5.5–9.5, respectively. The DNA G+C composition, estimated from ftsY gene, was 66% mol. Phylogenetic analysis using de 16S rRNA gene sequence showed that strain SLM16 belongs to the marine bacterial genus Albidovulum. Conclusion Strain SLM16 is a moderate thermophilic Gram negative microorganisms which belongs to the marine bacterial genus Albidovulum and is closely related to Albidovulum inexpectatum species based on phylogenetic analysis. Additionally, amine-transaminase activity towards the arylaliphatic amine α-methylbenzylamine was detected

Biotechnological applications of archaeal enzymes from extreme environments

Abstract To date, many industrial processes are performed using chemical compounds, which are harmful to nature. An alternative to overcome this problem is biocatalysis, which uses whole cells or enzymes to carry out chemical reactions in an environmentally friendly manner. Enzymes can be used as biocatalyst in food and feed, pharmaceutical, textile, detergent and beverage industries, among others. Since industrial processes require harsh reaction conditions to be performed, these enzymes must possess several characteristics that make them suitable for this purpose. Currently the best option is to use enzymes from extremophilic microorganisms, particularly archaea because of their special characteristics, such as stability to elevated temperatures, extremes of pH, organic solvents, and high ionic strength. Extremozymes, are being used in biotechnological industry and improved through modern technologies, such as protein engineering for best performance. Despite the wide distribution of archaea, exist only few reports about these microorganisms isolated from Antarctica and very little is known about thermophilic or hyperthermophilic archaeal enzymes particularly from Antarctica. This review summarizes current knowledge of archaeal enzymes with biotechnological applications, including two extremozymes from Antarctic archaea with potential industrial use, which are being studied in our laboratory. Both enzymes have been discovered through conventional screening and genome sequencing, respectively

Purification and characterization of a thermostable glutamate dehydrogenase from a thermophilic bacterium isolated from a sterilization drying oven

Glutamate dehydrogenase from axenic bacterial cultures of anew microorganism, called GWE1, isolated from the interior ofa sterilization drying oven, was purified by anion-exchange andmolecular-exclusion liquid chromatography. The apparent molecularmass of the native enzyme was 250.5 kDa and wasshown to be an hexamer with similar subunits of molecularmass 40.5 kDa. For glutamate oxidation, the enzyme showedan optimal pH and temperature of 8.0 and 70oC, respectively.In contrast to other glutamate dehydrogenases isolated frombacteria, the enzyme isolated in this study can use both NAD+and NADP+ as electron acceptors, displaying more affinity forNADP+ than for NAD+. No activity was detected with NADHor NADPH, 2-oxoglutarate and ammonia. The enzyme was exceptionallythermostable, maintaining more than 70% of activityafter incubating at 100oC for more than five hours suggestingbeing one of the most thermoestable enzymes reported inthe family of dehydrogenases. [BMB reports 2012; 45(2): 91-95