Evaluation of Antibiotic Biodegradation by a Versatile and Highly Active Recombinant Laccase from the Thermoalkaliphilic Bacterium Bacillus sp. FNT.

Laccases are industrially relevant enzymes that have gained great biotechnological importance. To date, most are of fungal and mesophilic origin; however, enzymes from extremophiles possess an even greater potential to withstand industrial conditions. In this study, we evaluate the potential of a recombinant spore-coat laccase from the thermoalkaliphilic bacterium Bacillus sp. FNT (FNTL) to biodegrade antibiotics from the tetracycline, ß-lactams, and fluoroquinolone families. This extremozyme was previously characterized as being thermostable and highly active in a wide range of temperatures (20-90 °C) and very versatile towards several structurally different substrates, including recalcitrant environmental pollutants such as PAHs and synthetic dyes. First, molecular docking analyses were employed for initial ligand affinity screening in the modeled active site of FNTL. Then, the in silico findings were experimentally tested with four highly consumed antibiotics, representatives of each family: tetracycline, oxytetracycline, amoxicillin, and ciprofloxacin. HPLC results indicate that FNTL with help of the natural redox mediator acetosyringone, can efficiently biodegrade 91, 90, and 82% of tetracycline (0.5 mg mL-1) in 24 h at 40, 30, and 20 °C, respectively, with no apparent ecotoxicity of the products on E. coli and B. subtilis. These results complement our previous studies, highlighting the potential of this extremozyme for application in wastewater bioremediation.

New insights into xenobiotic tolerance of Antarctic bacteria: transcriptomic analysis of Pseudomonas sp. TNT3 during 2,4,6-trinitrotoluene biotransformation.

The xenobiotic 2,4,6-trinitrotoluene (TNT) is a highly persistent environmental contaminant, whose biotransformation by microorganisms has attracted renewed attention. In previous research, we reported the discovery of Pseudomonas sp. TNT3, the first described Antarctic bacterium with the ability to biotransform TNT. Furthermore, through genomic analysis, we identified distinctive features in this isolate associated with the biotransformation of TNT and other xenobiotics. However, the metabolic pathways and genes active during TNT exposure in this bacterium remained unexplored. In the present transcriptomic study, we used RNA-sequencing to investigate gene expression changes in Pseudomonas sp. TNT3 exposed to 100 mg/L of TNT. The results showed differential expression of 194 genes (54 upregulated and 140 downregulated), mostly encoding hypothetical proteins. The most highly upregulated gene (> 1000-fold) encoded an azoreductase enzyme not previously described. Other significantly upregulated genes were associated with (nitro)aromatics detoxification, oxidative, thiol-specific, and nitrosative stress responses, and (nitro)aromatic xenobiotic tolerance via efflux pumps. Most of the downregulated genes were involved in the electron transport chain, pyrroloquinoline quinone (PQQ)-related alcohol oxidation, and motility. These findings highlight a complex cellular response to TNT exposure, with the azoreductase enzyme likely playing a crucial role in TNT biotransformation. Our study provides new insights into the molecular mechanisms of TNT biotransformation and aids in developing effective TNT bioremediation strategies. To the best of our knowledge, this report is the first transcriptomic response analysis of an Antarctic bacterium during TNT biotransformation.

Isolation and Identification of Bacteria from Three Geothermal Sites of the Atacama Desert and Their Plant-Beneficial Characteristics.

The Region of Arica and Parinacota (Atacama Desert) offers several unexplored remote sites with unique characteristics that would allow for the formulation of new bioproducts for agriculture. Among them, Jurasi Hot Springs, Polloquere Hot Springs, and Amuyo Lagoons represent a group of open pools fed by thermal water springing from the mountains. Their microbiomes remain unspecified, providing a unique opportunity to characterize the endemic community of these sites and develop new bioproducts for sustainable agriculture. Bacteria were isolated from the sediments of these geothermal sites and characterized by sequencing the 16S rRNA gene, microbiological characterization, and agricultural functional characterization. A total of 57 bacteria were isolated from three geothermal sites north of the Atacama Desert. The sequence analysis showed that the isolates belong to several bacterial genera, including Pantoea, Bacillus, and Pseudomonas, among others. The functional characterization revealed the presence of PGP traits, hydrolytic enzymes, and biocontrol activity against phytopathogenic fungi. These bacteria possess the potential to develop new biobased products for agriculture in arid conditions.

Recombinant expression and characterization of a new laccase, bioinformatically identified, from the Antarctic thermophilic bacterium Geobacillus sp. ID17.

Geobacillus sp. ID17 is a gram-positive thermophilic bacterium isolated from Deception Island, Antarctica, which has shown to exhibit remarkable laccase activity in crude extract at high temperatures. A bioinformatic search using local databases led to the identification of three putative multicopper oxidase sequences in the genome of this microorganism. Sequence analysis revealed that one of those sequences contains the four-essential copper-binding sites present in other well characterized laccases. The gene encoding this sequence was cloned and overexpressed in Escherichia coli, partially purified and preliminary biochemically characterized. The resulting recombinant enzyme was recovered in active and soluble form, exhibiting optimum copper-dependent laccase activity at 55 °C, pH 6.5 with syringaldazine substrate, retaining over 60% of its activity after 1 h at 55 and 60 °C. In addition, this thermophilic enzyme is not affected by common inhibitors SDS, NaCl and L-cysteine. Furthermore, biodecolorization assays revealed that this laccase is capable of degrading 60% of malachite green, 54% of Congo red, and 52% of Remazol Brilliant Blue R, after 6 h at 55 °C with aid of ABTS as redox mediator. The observed properties of this enzyme and the relatively straightforward overexpression and partial purification of it could be of great interest for future biotechnology applications.

Comparative Genomic Analysis of Antarctic Pseudomonas Isolates with 2,4,6-Trinitrotoluene Transformation Capabilities Reveals Their Unique Features for Xenobiotics Degradation.

The nitroaromatic explosive 2,4,6-trinitrotoluene (TNT) is a highly toxic and persistent environmental pollutant. Since physicochemical methods for remediation are poorly effective, the use of microorganisms has gained interest as an alternative to restore TNT-contaminated sites. We previously demonstrated the high TNT-transforming capability of three novel Pseudomonas spp. isolated from Deception Island, Antarctica, which exceeded that of the well-characterized TNT-degrading bacterium Pseudomonas putida KT2440. In this study, a comparative genomic analysis was performed to search for the metabolic functions encoded in the genomes of these isolates that might explain their TNT-transforming phenotype, and also to look for differences with 21 other selected pseudomonads, including xenobiotics-degrading species. Comparative analysis of xenobiotic degradation pathways revealed that our isolates have the highest abundance of key enzymes related to the degradation of fluorobenzoate, TNT, and bisphenol A. Further comparisons considering only TNT-transforming pseudomonads revealed the presence of unique genes in these isolates that would likely participate directly in TNT-transformation, and others involved in the ß-ketoadipate pathway for aromatic compound degradation. Lastly, the phylogenomic analysis suggested that these Antarctic isolates likely represent novel species of the genus Pseudomonas, which emphasizes their relevance as potential agents for the bioremediation of TNT and other xenobiotics.

Redrawing Cities with Children and Adolescents: Development of a Framework and Opportunity Index for Wellbeing-The REDibuja Study Protocol.

Global changes require urgent integration of health and wellbeing into all urban policies. Complex social and environmental factors define wellbeing outcomes and inequities present in cities. Additionally, political decisions are seldom thought and developed considering the needs and participation of children and adolescents. The REDibuja study aims to develop a multidimensional framework of wellbeing for children and adolescents and to validate an index of opportunities for better wellbeing for children and adolescents in the urban context of Temuco, Chile. This child-centered and cross-sectional study will involve mixed methodologies throughout the implementation of five work packages for two years (2022-2023): (1) development of a conceptual framework for child and adolescent wellbeing, (2) integration of available and public data, (3) studies in the local context, (4) data integration using geographic information systems, and (5) validation of the wellbeing opportunity index for children and adolescents. REDibuja will implement methodologies that until now are little used to facilitate political decisions in our regional context. This process and results could be transferred for assessment and decision-making in Latin America and low- and middle-income countries in other regions.

Associations Between Movement Behaviors and Emotional Changes in Toddlers and Preschoolers During Early Stages of the COVID-19 Pandemic in Chile.

Background: There is limited evidence about emotional and behavioral responses in toddlers and preschoolers during the novel coronavirus (COVID-19) pandemic, particularly in Latin America. Objective: To assess associations between changes in movement behaviors (physical activity, screen time and sleeping) and emotional changes in toddlers and preschoolers during early stages of the pandemic in Chile. Methods: A cross-sectional study conducted from March 30th to April 27th, 2020. Main caregivers of 1- to 5-year-old children living in Chile answered an online survey that included questions about sociodemographic characteristics, changes in the child's emotions and behaviors, movement behaviors and caregivers' stress during the pandemic. Multiple linear regressions were used to assess the association between different factors and emotional changes in toddlers and preschoolers. Results: In total, 1727 caregivers provided complete data on emotional changes for children aged 2.9 ± 1.36 years old, 47.9% girls. A large proportion of toddlers and preschoolers in Chile experienced emotional and behavioral changes. Most caregivers reported that children "were more affectionate" (78.9%), "more restless" (65.1%), and 'more frustrated' (54.1%) compared with pre-pandemic times. Apart from changes in movement behaviors, factors such as child age, caregivers' age and stress, and residential area (urban/rural) were consistently associated with changes in emotions and behaviors. Conclusion: The pandemic substantially affected the emotions and behaviors of toddlers and preschoolers in Chile. The findings suggest that supportive actions for caregivers may have a positive impact not only on adults but also on children. Mental health promotion programs should consider multilevel approaches in which the promotion of movement behaviors and support for caregivers should be essential pieces for future responses.

Living to the High Extreme: Unraveling the Composition, Structure, and Functional Insights of Bacterial Communities Thriving in the Arsenic-Rich Salar de Huasco Altiplanic Ecosystem.

Microbial communities inhabiting extreme environments such as Salar de Huasco (SH) in northern Chile are adapted to thrive while exposed to several abiotic pressures and the presence of toxic elements such as arsenic (As). Hence, we aimed to uncover the role of As in shaping bacterial composition, structure, and functional potential in five different sites in this altiplanic wetland using a shotgun metagenomic approach. The sites exhibit wide gradients of As (9 to 321 mg/kg), and our results showed highly diverse communities and a clear dominance exerted by the Proteobacteria and Bacteroidetes phyla. Functional potential analyses show broadly convergent patterns, contrasting with their great taxonomic variability. As-related metabolism, as well as other functional categories such as those related to the CH4 and S cycles, differs among the five communities. Particularly, we found that the distribution and abundance of As-related genes increase as the As concentration rises. Approximately 75% of the detected genes for As metabolism belong to expulsion mechanisms; arsJ and arsP pumps are related to sites with higher As concentrations and are present almost exclusively in Proteobacteria. Furthermore, taxonomic diversity and functional potential are reflected in the 12 reconstructed high-quality metagenome assembled genomes (MAGs) belonging to the Bacteroidetes (5), Proteobacteria (5), Cyanobacteria (1), and Gemmatimonadetes (1) phyla. We conclude that SH microbial communities are diverse and possess a broad genetic repertoire to thrive under extreme conditions, including increasing concentrations of highly toxic As. Finally, this environment represents a reservoir of unknown and undescribed microorganisms, with great metabolic versatility, which needs further study. IMPORTANCE As microbial communities inhabiting extreme environments are fundamental for maintaining ecosystems, many studies concerning composition, functionality, and interactions have been carried out. However, much is still unknown. Here, we sampled microbial communities in the Salar de Huasco, an extreme environment subjected to several abiotic stresses (high UV radiation, salinity and arsenic; low pressure and temperatures). We found that although microbes are taxonomically diverse, functional potential seems to have an important degree of convergence, suggesting high levels of adaptation. Particularly, arsenic metabolism showed differences associated with increasing concentrations of the metalloid throughout the area, and it effectively exerts a significant pressure over these organisms. Thus, the significance of this research is that we describe highly specialized communities thriving in little-explored environments subjected to several pressures, considered analogous of early Earth and other planets, that have the potential for unraveling technologies to face the repercussions of climate change in many areas of interest.

Composition and structure of the skin microbiota of rorquals off the Eastern South Pacific.

Recent advances in high-throughput sequencing have enabled the large-scale interrogation of microbiota in the most diverse environments, including host-associated microbiota. This has led to the recognition that the skin microbiota of rorquals is specific and structurally different from that of the ocean. This study reveals the skin microbiome of 85 wild individuals along the Chilean coast belonging to Megaptera novaeangliae, Balaenoptera musculus and Balaenoptera physalus. Alpha diversity analysis revealed significant differences in richness and phylogenetic diversity, particularly among humpback whales from different locations and between blue and humpback whales. Beta diversity was partially explained by host and location but only accounting for up to 17% of microbiota variability (adjusted VPA). Overall, we found that microbiota composition was dominated by bacterial genera such as Cardiobacter, Moraxella, Tenacibaculum, Stenotrophomonas, Flavobacteria and Pseudomonas. We also found that no ASVs were associated with the three rorqual species. Up to four ASVs were specific of a location, indicating a great variability in the microbiota. To the best of our knowledge, this is the first report on the composition and structure of the skin microbiota of whales off the coast of Chile, providing a foundational dataset to understand the microbiota's role in rorquals.

Comparison between self-reported and accelerometer-derived measurements for classifying children and adolescents as physically active in Chile.

The aim of this study was to compare self-reported with two accelerometer-derived methods to classify Chilean children and adolescents as physically active. In total, 247 students wore an accelerometer on their hips during 7 consecutive days to classify them as physically active based on (1) daily accumulation of ≥ 60 minutes of moderate-to-vigorous physical activity (MVPA) on each of the seven days, and (2) average MVPA ≥ 60 minutes/day. Also, participants were classified as physically active if they reported being active for at least 60 minutes in all seven days. When using the accelerometer data, 0.8% were active in all seven days, while 10.5% recorded ≥ 60 minutes MVPA per day on average. Based on self-report, 7.2% were physically active. The agreement between self-reported and accelerometer estimations were poor. Important differences were observed between the self-reported and device-derived methods for classifying children and adolescents as physically active. When comparing them, some considerations should be taken. The findings suggest that these methods are not interchangeable. Therefore, if possible, they should be used as complementary measurements.

Comparison between self-reported and accelerometer-derived measurements for classifying children and adolescents as physically active in Chile / Comparación entre medidas autoreportadas y provenientes de acelerómetro para clasificar a los niños, niñas y adolescentes como físicamente activos en Chile / Comparação de autorrelato e medidas derivadas de acelerômetro para classificar a atividade física em crianças e adolescentes chilenos

The aim of this study was to compare self-reported with two accelerometer-derived methods to classify Chilean children and adolescents as physically active. In total, 247 students wore an accelerometer on their hips during 7 consecutive days to classify them as physically active based on (1) daily accumulation of ≥ 60 minutes of moderate-to-vigorous physical activity (MVPA) on each of the seven days, and (2) average MVPA ≥ 60 minutes/day. Also, participants were classified as physically active if they reported being active for at least 60 minutes in all seven days. When using the accelerometer data, 0.8% were active in all seven days, while 10.5% recorded ≥ 60 minutes MVPA per day on average. Based on self-report, 7.2% were physically active. The agreement between self-reported and accelerometer estimations were poor. Important differences were observed between the self-reported and device-derived methods for classifying children and adolescents as physically active. When comparing them, some considerations should be taken. The findings suggest that these methods are not interchangeable. Therefore, if possible, they should be used as complementary measurements.

El objetivo fue comparar el autoreporte con dos métodos provenientes de acelerómetría para clasificar a niños, niñas y adolescentes chilenos como físicamente activos. Doscientos cuarenta y siete estudiantes llevaron un acelerómetro en la cintura durante siete días consecutivos y fueron clasificados como físicamente activos, basados en: (1) acumulación diaria de ≥ 60 minutos de actividad física de moderada a vigorosa (AFMV) en cada uno de los siete días, y (2) promedio de AFMV ≥ 60 minutos/día. Asimismo, los participantes fueron clasificados como físicamente activos si reportaron ser activos por al menos 60 minutos en cada uno de los siete días. Al usar los datos del acelerómetro, 0,8% fueron activos los siete días, mientras que un 10,5% registró ≥ 60 minutos AFMV por día promedio. Un 7,2% fue físicamente activos según el autoreporte. El acuerdo entre las estimaciones autoreportadas y el acelerómetro fue pobre. Se observaron importantes diferencias entre los autoreportes y los métodos derivados del dispositivo para clasificar niños, niñas y adolescentes como físicamente activos. Cuando se comparen datos derivados de los acelerómetros y autoreportes esto se debería considerar. Los resultados sugieren que estos métodos no son intercambiables. Por ello, en la medida de lo posible, se deberían usar como medidas complementarias.

O objetivo foi comparar o autorrelato com dois métodos derivados de acelerômetro para classificar crianças e adolescentes chilenos como fisicamente ativas ou inativas. Um total de 247 alunos usaram um acelerômetro no quadril durante sete dias consecutivos e foram classificados como fisicamente ativos com base em: (1) acúmulo diário de ≥ 60 minutos de atividade física de moderada a vigorosa intensidade (AFMV) em cada um dos sete dias e (2) AFMV média por dia de ≥ 60 minutos. Além disso, os participantes foram classificados como fisicamente ativos com base no autorrelato de ser ativo por pelo menos 60 minutos em todos os sete dias. Com o uso dos dados de acelerômetro, 0,8% foram classificados como ativos em todos os sete dias, enquanto 10,5% registravam ≥ 60 minutos de AFMV por dia em média e 7,2% eram fisicamente ativos com base no autorrelato. A concordância foi baixa entre o autorrelato e as estimativas por acelerômetro. Foram observadas diferenças importantes entre o autorrelato e os métodos baseados em dispositivos para classificar as crianças e adolescentes como fisicamente ativos. Algumas considerações são relevantes ao comparar estimativas baseadas em acelerômetro e o autorrelato de atividade física. Os achados sugerem que esses métodos não são intercambiáveis. Portanto, quando possível, devem ser usados como medidas complementares.

Genomic Variation and Arsenic Tolerance Emerged as Niche Specific Adaptations by Different Exiguobacterium Strains Isolated From the Extreme Salar de Huasco Environment in Chilean - Altiplano.

Polyextremophilic bacteria can thrive in environments with multiple stressors such as the Salar de Huasco (SH). Microbial communities in SH are exposed to low atmospheric pressure, high UV radiation, wide temperature ranges, salinity gradient and the presence of toxic compounds such as arsenic (As). In this work we focus on arsenic stress as one of the main adverse factors in SH and bacteria that belong to the Exiguobacterium genus due to their plasticity and ubiquity. Therefore, our aim was to shed light on the effect of niche conditions pressure (particularly arsenic), on the adaptation and divergence (at genotypic and phenotypic levels) of Exiguobacterium strains from five different SH sites. Also, to capture greater diversity in this genus, we use as outgroup five As(III) sensitive strains isolated from Easter Island (Chile) and The Great Salt Lake (United States). For this, samples were obtained from five different SH sites under an arsenic gradient (9 to 321 mg/kg: sediment) and isolated and sequenced the genomes of 14 Exiguobacterium strains, which had different arsenic tolerance levels. Then, we used comparative genomic analysis to assess the genomic divergence of these strains and their association with phenotypic differences such as arsenic tolerance levels and the ability to resist poly-stress. Phylogenetic analysis showed that SH strains share a common ancestor. Consequently, populations were separated and structured in different SH microenvironments, giving rise to multiple coexisting lineages. Hence, this genotypic variability is also evidenced by the COG (Clusters of Orthologous Groups) composition and the size of their accessory genomes. Interestingly, these observations correlate with physiological traits such as growth patterns, gene expression, and enzyme activity related to arsenic response and/or tolerance. Therefore, Exiguobacterium strains from SH are adapted to physiologically overcome the contrasting environmental conditions, like the arsenic present in their habitat.

Sequences of Endophytic Fungal and Bacterial Communities from Araucaria araucana [(Molina) K. Koch, 1869] in the Coastal and Andes Mountain Ranges, Chile.

Here, we report the results from PCR and sequencing of bacterial 16S rRNA and fungal internal transcribed spacer 1 (ITS1) genes from needle, branch, trunk, and root samples of Araucaria araucana, plus soil and associated insects, collected along the entirety of its geographic distribution in Chile (January 2017 and 2018).

2018 Chilean Physical Activity Report Card for Children and Adolescents: Full Report and International Comparisons.

BACKGROUND: The study summarizes the findings of the 2018 Chilean Report Card (RC) on Physical Activity (PA) for Children and Adolescents and compares the results with the first Chilean RC and with other countries from the Global Matrix 3.0. METHODS: A Research Work Group using a standardized methodology from the Global Matrix 3.0 awarded grades for 13 PA-related indicators based on the percentage of compliance for defined benchmarks. Different public data sets, government reports, and papers informed the indicators. RESULTS: The grades assigned were for (1) "behaviors that contribute to overall PA levels": overall PA, D-; organized sport participation, D-; active play, INC; and active transportation, F; (2) "factors associated with cardiometabolic risk": sedentary behavior, C-; overweight and obesity, F; fitness, D; sleep, INC; and (3) "factors that influence PA": family and peers, F; school, D; inclusion, INC; community and built environment, B; government strategies and investments, B-. CONCLUSIONS: Chile's grades remained low compared with the first RC. On the positive side, Chile is advancing in environmental and policy aspects. Our findings indicate that the implementation of new strategies should be developed through collaboration between different sectors to maximize effective investments for increasing PA and decreasing sedentary time among children and adolescents in Chile.

Biotransformation of 2,4,6-Trinitrotoluene by Pseudomonas sp. TNT3 isolated from Deception Island, Antarctica.

2,4,6-Trinitrotoluene (TNT) is a nitroaromatic explosive, highly toxic and mutagenic for organisms. In this study, we report for the first time the screening and isolation of TNT-degrading bacteria from Antarctic environmental samples with potential use as bioremediation agents. Ten TNT-degrading bacterial strains were isolated from Deception Island. Among them, Pseudomonas sp. TNT3 was selected as the best candidate since it showed the highest tolerance, growth, and TNT biotransformation capabilities. Our results showed that TNT biotransformation involves the reduction of the nitro groups. Additionally, Pseudomonas sp. TNT3 was capable of transforming 100 mg/L TNT within 48 h at 28 °C, showing higher biotransformation capability than Pseudomonas putida KT2440, a known TNT-degrading bacterium. Functional annotation of Pseudomonas sp. TNT3 genome revealed a versatile set of molecular functions involved in xenobiotic degradation pathways. Two putative xenobiotic reductases (XenA_TNT3 and XenB_TNT3) were identified by means of homology searches and phylogenetic relationships. These enzymes were also characterized at molecular level using homology modelling and molecular dynamics simulations. Both enzymes share different levels of sequence similarity with other previously described TNT-degrading enzymes and with their closest potential homologues in databases.

Sociodemographic Predictors of Changes in Physical Activity, Screen Time, and Sleep among Toddlers and Preschoolers in Chile during the COVID-19 Pandemic.

The aim was to examine the sociodemographic predictors associated with changes in movement behaviors (physical activity, screen time, and sleep) among toddlers and preschoolers during the early stages of the coronavirus disease 2019 pandemic in Chile. Caregivers of 1- to 5-year-old children completed an online survey between 30 March and 27 April 2020. Information about the child's movement behaviors before (retrospectively) and during the pandemic, as well as family characteristics were reported. In total, 3157 participants provided complete data (mean children age: 3.1 ± 1.38 years). During early stages of the pandemic, time spent in physical activity decreased, recreational screen time and sleep duration increased, and sleep quality declined. Toddlers and preschoolers with space to play at home and living in rural areas experienced an attenuated impact of the pandemic restrictions on their physical activity levels, screen time, and sleep quality. Older children, those whose caregivers were aged ≥35-<45 years and had a higher educational level, and those living in apartments had greater changes, mainly a decrease in total physical activity and increase in screen time. This study has shown the significant impact of the pandemic restrictions on movement behaviors in toddlers and preschoolers in Chile.

Characterization of a novel thermostable (S)-amine-transaminase from an Antarctic moderately-thermophilic bacterium Albidovulum sp. SLM16.

Amine-transaminases (ATAs) are enzymes that catalyze the reversible transfer of an amino group between primary amines and carbonyl compounds. They have been widely studied in the last decades for their application in stereoselective synthesis of chiral amines, which are one of the most valuable building blocks in pharmaceuticals manufacturing. Their excellent enantioselectivity, use of low-cost substrates and no need for external cofactors has turned these enzymes into a promising alternative to the chemical synthesis of chiral amines. Nevertheless, its application at industrial scale remains limited mainly because most of the available ATAs are scarcely tolerant to harsh reaction conditions such as high temperatures and presence of organic solvents. In this work, a novel (S)-ATA was discovered in a thermophilic bacterium, Albidovulum sp. SLM16, isolated from a geothermal Antarctic environmental sample, more specifically from a shoreline fumarole in Deception Island. The transaminase-coding gene was identified in the genome of the microorganism, cloned and overexpressed in Escherichia coli for biochemical characterization. The activity of the recombinant ATA was optimal at 65 °C and pH 9.5. Molecular mass estimates suggest a 75 kDa homodimeric structure. The enzyme turned out to be highly thermostable, maintaining 80% of its specific activity after 5 days of incubation at 50 °C. These results indicate that ATA_SLM16 is an excellent candidate for potential applications in biocatalytic synthesis. To the best of our knowledge, this would be the first report of the characterization of a thermostable (S)-ATA discovered by means of in vivo screening of thermophilic microorganisms.

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

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.

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

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.