scholarly journals Identification, Characterization, and Hydrolase Producing Performance of Thermophilic Bacteria: Geothermal Hot Springs in Eastern and Southeastern Anatolia Region of Turkey

2021 ◽  
Author(s):  
Orhan Uluçay ◽  
Arzu Görmez ◽  
Cem Öziç
Keyword(s):  
Hot Springs ◽  
Hot Spring ◽  
Thermophilic Bacteria ◽  
Hydrolytic Enzymes ◽  
Bacillus Coagulans ◽  
Rrna Gene ◽  
Bacterial Isolates ◽  
Thermophilic Bacilli ◽  
Geobacillus Kaustophilus ◽  
The Rich

Abstract In the last twenty years, researchers have increasingly focused on the rich microorganism-based diversity of natural hot spring resources to explore the benefits of thermophiles in industrial and biotechnological fields. For this purpose, a total of 83 thermophilic bacilli were isolated from 7 different geothermal hot springs (at temperatures ranging between 40 and 85 ◦ C) located in the east and Southeastern of Turkey. These isolates were identified by different methods such as physiological, morphological, biochemical, molecular properties. According to 16S rRNA gene sequence analysis, 5 different isolates ( Bacillus coagulans , Bacillus licheniformis , Bacillus subtilis , Bacillus thuringiensis , and Geobacillus kaustophilus ) were identified. It was observed that B. licheniformis and B. subtilis were the most species obtained from the researched hot spring sources. Phylogenetic relationships of isolates were evaluated with the help of a phylogenetic tree. The conditions of bacterial isolates to synthesize various hydrolytic enzymes such as protease, cellulase, lipase, and amylase were investigated. When the potential of isolates to produce hydrolytic enzymes was examined, protease 73 (88%), cellulase 34 (41%), lipase 69 (83%), and amylase 68 (82%) were detected. All isolates have at least one or more extracellular protease, cellulase, amylase, or lipase activity. Besides, 32.8% (27) of bacterial isolates were able to synthesize all of the hydrolytic enzymes.

scholarly journals Molecular identification of amylase-producing thermophilic bacteria isolated from Bukit Gadang Hot Spring, West Sumatra, Indonesia

2020 ◽  
Vol 21 (3) ◽  
Author(s):  
Aulia Ardhi ◽  
Arina Nadenggan Sidauruk ◽  
Nabella Suraya ◽  
Nova Wahyu Pratiwi ◽  
Usman Pato ◽  
...  
Keyword(s):  
High Temperature ◽  
Molecular Identification ◽  
Hot Springs ◽  
Hot Spring ◽  
Thermophilic Bacteria ◽  
Amylase Activity ◽  
Hydrolytic Enzymes ◽  
Molecular Technique ◽  
West Sumatra ◽  
Pharmaceutical Industries

Abstract. Ardhi A, Sidauruk AN, Suraya N, Pratiwi NW, Pato U, Saryono. 2020. Molecular identification of amylase-producing thermophilic bacteria isolated from Bukit Gadang Hot Spring, West Sumatra, Indonesia. Biodiversitas 21: 994-1000.  Amylase is one of the hydrolytic enzymes that is widely used in a wide number of industrial processes such as food, fermentation, textile, paper, detergent, and pharmaceutical industries. Amylase produced by thermophilic bacteria may be thermostable, which is very beneficial in several applications requiring high temperature, for example, the process of gelatinization, liquefaction, and saccharification are performed in high temperature involved in the starch processing. In this study, the amylase-producing ability of thermophilic bacteria isolated from Bukit Gadang hot spring, West Sumatra, Indonesia, was checked and followed by molecular identification. Thirteen isolates that were successfully isolated from the hot springs were microscopically and macroscopically characterized, biochemically tested, and determined their amylase enzyme activity both qualitatively and quantitatively. The isolate that performed the best amylase activity was identified using the molecular technique. The DNA sequencing was carried out in 16S rRNA and continued with BLAST search for species identification. The result of molecular identification showed that the isolate with the best amylase activity was identified as Bacillus licheniformis. The optimum amylase production  (231.33 U/ml)  and the best enzyme-specific activity  (101.79 U/mg) were obtained at the incubation time of 36 hours.

scholarly journals Microbial Diversity of Terrestrial Geothermal Springs in Armenia and Nagorno-Karabakh: A Review

2021 ◽  
Vol 9 (7) ◽  
pp. 1473
Author(s):  
Ani Saghatelyan ◽  
Armine Margaryan ◽  
Hovik Panosyan ◽  
Nils-Kåre Birkeland
Keyword(s):  
16S Rrna ◽  
Microbial Diversity ◽  
High Altitude ◽  
Hot Springs ◽  
Hot Spring ◽  
Abiotic Factors ◽  
Rrna Gene ◽  
Geothermal Springs ◽  
Bacterial Phyla ◽  
Terrestrial Hot Springs

The microbial diversity of high-altitude geothermal springs has been recently assessed to explore their biotechnological potential. However, little is known regarding the microbiota of similar ecosystems located on the Armenian Highland. This review summarizes the known information on the microbiota of nine high-altitude mineralized geothermal springs (temperature range 25.8–70 °C and pH range 6.0–7.5) in Armenia and Nagorno-Karabakh. All these geothermal springs are at altitudes ranging from 960–2090 m above sea level and are located on the Alpide (Alpine–Himalayan) orogenic belt, a seismically active region. A mixed-cation mixed-anion composition, with total mineralization of 0.5 mg/L, has been identified for these thermal springs. The taxonomic diversity of hot spring microbiomes has been examined using culture-independent approaches, including denaturing gradient gel electrophoresis (DGGE), 16S rRNA gene library construction, 454 pyrosequencing, and Illumina HiSeq. The bacterial phyla Proteobacteria, Bacteroidetes, Cyanobacteria, and Firmicutes are the predominant life forms in the studied springs. Archaea mainly include the phyla Euryarchaeota, Crenarchaeota, and Thaumarchaeota, and comprise less than 1% of the prokaryotic community. Comparison of microbial diversity in springs from Karvachar with that described for other terrestrial hot springs revealed that Proteobacteria, Bacteroidetes, Actinobacteria, and Deinococcus–Thermus are the common bacterial groups in terrestrial hot springs. Contemporaneously, specific bacterial and archaeal taxa were observed in different springs. Evaluation of the carbon, sulfur, and nitrogen metabolism in these hot spring communities has revealed diversity in terms of metabolic activity. Temperature seems to be an important factor in shaping the microbial communities of these springs. Overall, the diversity and richness of the microbiota are negatively affected by increasing temperature. Other abiotic factors, including pH, mineralization, and geological history, also impact the structure and function of the microbial community. More than 130 bacterial and archaeal strains (Bacillus, Geobacillus, Parageobacillus, Anoxybacillus, Paenibacillus, Brevibacillus Aeribacillus, Ureibacillus, Thermoactinomyces, Sporosarcina, Thermus, Rhodobacter, Thiospirillum, Thiocapsa, Rhodopseudomonas, Methylocaldum, Desulfomicrobium, Desulfovibrio, Treponema, Arcobacter, Nitropspira, and Methanoculleus) have been reported, some of which may be representative of novel species (sharing 91–97% sequence identity with their closest matches in GenBank) and producers of thermozymes and biomolecules with potential biotechnological applications. Whole-genome shotgun sequencing of T. scotoductus K1, as well as of the potentially new Treponema sp. J25 and Anoxybacillus sp. K1, were performed. Most of the phyla identified by 16S rRNA were also identified using metagenomic approaches. Detailed characterization of thermophilic isolates indicate the potential of the studied springs as a source of biotechnologically valuable microbes and biomolecules.

scholarly journals Carbon Source Preference in Chemosynthetic Hot Spring Communities

2015 ◽  
Vol 81 (11) ◽  
pp. 3834-3847 ◽  
Author(s):  
Matthew R. Urschel ◽  
Michael D. Kubo ◽  
Tori M. Hoehler ◽  
John W. Peters ◽  
Eric S. Boyd
Keyword(s):  
High Temperature ◽  
National Park ◽  
Hot Springs ◽  
Dissolved Inorganic Carbon ◽  
Hot Spring ◽  
Affinity Constant ◽  
Rrna Gene ◽  
Organic Substrates ◽  
Short Term ◽  
Acetate Assimilation

ABSTRACTRates of dissolved inorganic carbon (DIC), formate, and acetate mineralization and/or assimilation were determined in 13 high-temperature (>73°C) hot springs in Yellowstone National Park (YNP), Wyoming, in order to evaluate the relative importance of these substrates in supporting microbial metabolism. While 9 of the hot spring communities exhibited rates of DIC assimilation that were greater than those of formate and acetate assimilation, 2 exhibited rates of formate and/or acetate assimilation that exceeded those of DIC assimilation. Overall rates of DIC, formate, and acetate mineralization and assimilation were positively correlated with spring pH but showed little correlation with temperature. Communities sampled from hot springs with similar geochemistries generally exhibited similar rates of substrate transformation, as well as similar community compositions, as revealed by 16S rRNA gene-tagged sequencing. Amendment of microcosms with small (micromolar) amounts of formate suppressed DIC assimilation in short-term (<45-min) incubations, despite the presence of native DIC concentrations that exceeded those of added formate by 2 to 3 orders of magnitude. The concentration of added formate required to suppress DIC assimilation was similar to the affinity constant (Km) for formate transformation, as determined by community kinetic assays. These results suggest that dominant chemoautotrophs in high-temperature communities are facultatively autotrophic or mixotrophic, are adapted to fluctuating nutrient availabilities, and are capable of taking advantage of energy-rich organic substrates when they become available.

scholarly journals Genomic adaptations enabling Acidithiobacillus distribution across wide-ranging hot spring temperatures and pHs

Microbiome ◽  
2021 ◽  
Vol 9 (1) ◽  
Author(s):  
Chanenath Sriaporn ◽  
Kathleen A. Campbell ◽  
Martin J. Van Kranendonk ◽  
Kim M. Handley
Keyword(s):  
Hot Springs ◽  
Hot Spring ◽  
Thermal Adaptation ◽  
Distinct Species ◽  
Taupo Volcanic Zone ◽  
Rrna Gene ◽  
Ph Homeostasis ◽  
Microbial Composition ◽  
Related Group ◽  
Geothermal Environments

Abstract Background Terrestrial hot spring settings span a broad spectrum of physicochemistries. Physicochemical parameters, such as pH and temperature, are key factors influencing differences in microbial composition across diverse geothermal areas. Nonetheless, analysis of hot spring pools from the Taupo Volcanic Zone (TVZ), New Zealand, revealed that some members of the bacterial genus, Acidithiobacillus, are prevalent across wide ranges of hot spring pHs and temperatures. To determine the genomic attributes of Acidithiobacillus that inhabit such diverse conditions, we assembled the genomes of 19 uncultivated hot spring Acidithiobacillus strains from six geothermal areas and compared these to 37 publicly available Acidithiobacillus genomes from various habitats. Results Analysis of 16S rRNA gene amplicons from 138 samples revealed that Acidithiobacillus comprised on average 11.4 ± 16.8% of hot spring prokaryotic communities, with three Acidithiobacillus amplicon sequence variants (ASVs) (TVZ_G1, TVZ_G2, TVZ_G3) accounting for > 90% of Acidithiobacillus in terms of relative abundance, and occurring in 126 out of 138 samples across wide ranges of temperature (17.5–92.9 °C) and pH (1.0–7.5). We recovered 19 environmental genomes belonging to each of these three ASVs, as well as a fourth related group (TVZ_G4). Based on genome average nucleotide identities, the four groups (TVZ_G1-TVZ_G4) constitute distinct species (ANI < 96.5%) of which three are novel Acidithiobacillus species (TVZ_G2-TVZ_G4) and one belongs to Acidithiobacillus caldus (TVZ_G1). All four TVZ Acidithiobacillus groups were found in hot springs with temperatures above the previously known limit for the genus (up to 40 °C higher), likely due to significantly higher proline and GC contents than other Acidithiobacillus species, which are known to increase thermostability. Results also indicate hot spring-associated Acidithiobacillus have undergone genome streamlining, likely due to thermal adaptation. Moreover, our data suggest that Acidithiobacillus prevalence across varied hot spring pHs is supported by distinct strategies, whereby TVZ_G2-TVZ_G4 regulate pH homeostasis mostly through Na+/H+ antiporters and proton-efflux ATPases, whereas TVZ_G1 mainly relies on amino acid decarboxylases. Conclusions This study provides insights into the distribution of Acidithiobacillus species across diverse hot spring physichochemistries and determines genomic features and adaptations that potentially enable Acidithiobacillus species to colonize a broad range of temperatures and pHs in geothermal environments.

Thermoanaerobaculum aquaticum gen. nov., sp. nov., the first cultivated member of Acidobacteria subdivision 23, isolated from a hot spring

2013 ◽  
Vol 63 (Pt_11) ◽  
pp. 4149-4157 ◽  
Author(s):  
Nathaniel A. Losey ◽  
Bradley S. Stevenson ◽  
Hans-Jürgen Busse ◽  
Jaap S. Sinninghe Damsté ◽  
W. Irene C. Rijpstra ◽  
...  
Keyword(s):  
Gene Sequence ◽  
Hot Springs ◽  
Hot Spring ◽  
Rrna Gene ◽  
Strictly Anaerobic ◽  
Content Type ◽  
Novel Bacterium ◽  
Fermentative Growth ◽  
Novel Genus And Species ◽  
The 16S Rrna Gene

A novel bacterium was isolated from a freshwater hot spring, the Hale House Spring, located at Hot Springs National Park, Hot Springs, AR, USA. Cells of strain MP-01T stained Gram-negative, were rod-shaped, non-motile, strictly anaerobic and chemo-organotrophic and did not form spores. Growth occurred at 50–65 °C, with an optimum at 60 °C, at pH 6.0–8.0, with an optimum at pH 6.5–7.0, and at NaCl concentrations up to 0.5 % (w/v), with optimum growth in the absence of NaCl. Strain MP-01T was capable of fermentative growth on pyruvate or proteinaceous substrates as well as reducing Fe(III) and Mn(IV). Major fatty acids were iso-C15 : 0, iso-C16 : 0, anteiso-C17 : 0 and iso-C17 : 0. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine and the major isoprenoid quinone was MK-10. In the polyamine pattern, sym-homospermidine was the predominant compound. The DNA G+C content was 62.7 mol%. Analysis of the 16S rRNA gene sequence of the isolate indicated that strain MP-01T represents the first reported cultivated member of subdivision 23 of the Acidobacteria . It is proposed that strain MP-01T represents a novel genus and species, for which the name Thermoanaerobaculum aquaticum gen. nov., sp. nov. is proposed. The type strain of Thermoanaerobaculum aquaticum is MP-01T ( = DSM 24856T = JCM 18256T).

Anoxybacillus sp. AH1, an α-amylase-producing thermophilic bacterium isolated from Dargeçit hot spring

Biologia ◽  
2015 ◽  
Vol 70 (7) ◽  
Author(s):  
Ömer Acer ◽  
Hemşe Pırınççiğlu ◽  
Fatma Matpan Bekler ◽  
Reyhan Gül-Güven
Keyword(s):  
Hot Springs ◽  
Hot Spring ◽  
Thermophilic Bacteria ◽  
Potato Starch ◽  
Nitrogen Sources ◽  
Soluble Starch ◽  
Thermophilic Bacterium ◽  
Carbon And Nitrogen Sources ◽  
Carbon And Nitrogen ◽  
Optimum Growth Temperature

AbstractThe present study was conducted to isolate α-amylase-producing thermophilic bacteria from Darge¸cit hot springs in Turkey. The morphological, biochemical and physiological characterisation, as well as genetic analysis by 16S rRNA sequences indicated that the isolated strain AH1 was a member of Anoxybacillus genus. The strain was aerobe, Gram-positive and spore-forming rod, exhibiting optimum growth temperature and pH of 60ºC and 7.0-7.5, respectively. Optimization of growth medium and enzyme assay conditions for extracellular α-amylase production by the novel thermophilic Anoxybacillus sp. AH1 were carried out in many different media containing a variety of carbon and nitrogen sources. Among various carbon and nitrogen sources, peptone (2054.1 U/mL) at 1% and maltose (1862.9 U/mL) at 0.5% increased α-amylase activity, compared to controls. Moreover, a high enzyme production was observed with potato starch at 0.5% and 1% (2668.4 U/mL and 3627 U/mL, respectively), as well as with 1% soluble starch (2051.9 U/mL). The enzyme activity was found to be rather high in the presence of CaCl

scholarly journals Diversity analysis and metagenomic insights into the Antibiotic Resistance and Metal Resistances among Hot Spring Bacteriobiome-insinuating inherent environmental baseline levels of Antibiotic and Metal tolerance

2019 ◽  
Author(s):  
Ishfaq Nabi Najar ◽  
Mingma Thundu Sherpa ◽  
Sayak Das ◽  
Nagendra Thakur
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Hot Springs ◽  
Metal Tolerance ◽  
Hot Spring ◽  
Thermophilic Bacteria ◽  
Antibiotic Resistance Genes ◽  
Metal Resistance ◽  
Culture Independent ◽  
Maximum Similarity

AbstractMechanisms of occurrence and expressions of antibiotic resistance genes (ARGs) in thermophilic bacteria are still unknown owing to limited research and data. The evolution and proliferation of ARGs in the thermophilic bacteria is unclear and needs a comprehensive study. In this research, comparative profiling of antibiotic resistance genes and metal tolerance genes among the thermophilic bacteria has been done by culture-independent functional metagenomic methods. Metagenomic analysis showed the dominance of Proteobacteria, Actinobacteria. Firmicutes and Bacteroidetes in these hot springs. ARG analysis through shotgun gene sequencing was found to be negative in case of thermophilic bacteria. However, few of genes were detected but they were showing maximum similarity with mesophilic bacteria. Concurrently, metal resistance genes were also detected in the metagenome sequence of hot springs. Detection of metal resistance gene and absence of ARG’s investigated by whole genome sequencing, in the reference genome sequence of thermophilic Geobacillus also conveyed the same message. This evolutionary selection of metal resistance over antibiotic genes may have been necessary to survive in the geological craters which are full of different metals from earth sediments rather than antibiotics. Furthermore, the selection could be environment driven depending on the susceptibility of ARG’s in thermophilic environment as it reduces the chances of horizontal gene transfer. With these findings this article highlights many theories and culminates different scopes to study these aspects in thermophiles.

scholarly journals Distinct microbial composition and functions in an underground high-temperature hot spring at different depths

2019 ◽  
Author(s):  
Shijie Bai ◽  
Xiaotong Peng
Keyword(s):  
High Temperature ◽  
Microbial Communities ◽  
Hot Springs ◽  
Hot Spring ◽  
Hydrogen Oxidation ◽  
Archaeal Community ◽  
Limited Area ◽  
Rrna Gene ◽  
Microbial Composition ◽  
Different Depths

Abstract. The microbial diversity and functions of three high-temperature neutral hot springs water samples at different depths (0 m, 19 m and 58 m) were investigated based on 16S rRNA gene sequencing and a functional gene array (GeoChip 5.0). The results revealed that the bacterial communities were distinct at different depths in the hot springs. Additionally, in response to the depths, bacterial/archaeal community compositions exhibited shifts over the depth profiles. Aquificae, Alpha-proteobacteria, and Deinococcus-Thermus were the dominating phyla at 0 m, 19 m, and 58 m, respectively. Hydrogenobacter, Sphingobium, and Thermus were the most abundant genera at 0 m, 19 m, and 58 m, respectively. The phylum Thaumarchaeota was the most abundant member of the archaeal community in the samples at different hot spring depths. Functional results of the microbial communities indicated that microbial metabolic functions were mainly related to sulfur, nitrogen cycling, and hydrogen oxidation. In summary, our results demonstrated that distinct microbial communities and functions were found at different depths of hot springs in a very limited area. These findings will provide new insights into the deep-subsurface biosphere associated with terrestrial hot springs.

scholarly journals Characterizing and Evaluating the Diverse Microbial Communities and Their Mercury Resistance Potential from Hot Spring Sites Representing Gradients in Temperature and pH in Yellowstone National Park

2020 ◽  
Vol 1 (1) ◽  
Author(s):  
Yelizaveta Rassadkina ◽  
Spencer Roth ◽  
Tamar Barkay
Keyword(s):  
Microbial Communities ◽  
Yellowstone National Park ◽  
National Park ◽  
Hot Springs ◽  
Hot Spring ◽  
Thermophilic Bacteria ◽  
Sequence Data ◽  
Mercury Resistance ◽  
Sample Site ◽  
Microorganism Community

Yellowstone National Park is home to many different hot springs, lakes, geysers, pools, and basins that range in pH, chemical composition, and temperature. These different environmental variations provide a broad range of conditions that select and grow diverse communities of microorganisms. In this study, we collected samples from geochemically diverse lakes and springs to characterize the microbial communities present through 16S rRNA metagenomic analysis. This information was then used to observe how various microorganisms survive in high mercury environments. The results show the presence of microorganisms that have been studied in previous literature. The results also depict gradients of microorganisms including thermophilic bacteria and archaea that exist in these extreme environments. In addition, beta diversity analyses of the sequence data showed site clustering based primarily on temperature instead of pH or sample site, suggesting that while pH, temperature, and sample site were all shown to be significant, temperature is the strongest factor driving microorganism community development. While it is important to characterize the microorganism community present, it is also important to understand how this community functions as a result of its selection. Along with looking at community composition, genomic material was tested to see if it contained mercury methylating (hgcA) or mercury reducing (merA) genes. Out of 22 samples, three of them were observed to have merA genes, while no samples had hgcA genes. These results indicate that microorganisms in Mustard and Nymph Springs may use mercury reduction. Understanding how microorganisms survive in environments with high concentrations of toxic pollutants is crucial because it can be used as a model to better understand mechanisms of resistance and the biogeochemical cycle, as well as for bioremediation and other solutions to anthropogenic problems.

scholarly journals ISOLATION OF THERMOPHILIC LIPASE PRODUCING BACTERIUM FROM HOT SPRINGS AT THE EAST COAST OF PENINSULAR MALAYSIA

2021 ◽  
Vol 11 (1) ◽  
pp. 1-10
Author(s):  
Tengku Hamid ◽  
◽  
Nur Abidin ◽  
Nurmusfirah Hasan ◽  
◽  
...  
Keyword(s):  
Hot Springs ◽  
East Coast ◽  
Hot Spring ◽  
Thermophilic Bacteria ◽  
Natural Habitat ◽  
Primary Source ◽  
Industrial Applications ◽  
Peninsular Malaysia ◽  
Thermostable Enzymes ◽  
Thermophilic Microorganisms

Hot spring is a natural habitat for thermophilic bacteria and the primary source of thermostable enzymes useful in industrial applications. In Malaysia, the search for thermophilic organisms has been focused on hot springs, especially on the peninsular West coast. In this work, lipase or esterase producing thermophilic microorganisms were isolated from East coast hot springs in Pahang and Terengganu's states. Morphological and biochemical analysis were carried out on Isolates LH1, LH2, LH3, LH4, LH5, B2B2 and S1B4, which showed that they are gram positive, aerobic, spore forming, and motile organisms. All of the seven isolates showed the ability to grow at 45°C and formed hydrolysis zones on tributyrin agar plates. However, only isolate B2B2 and S1B4 were able to thrive at higher temperatures of up to 65°C. The genotypic characterisation was carried out using 16S rRNA sequencing. Bacillus and Geobacillus species were found to be the dominant bacteria isolated from these hot springs. From La hot spring, isolate LH1 (MT 645486), Isolates LH2 (MT645483), LH3 (MT645484), LH4 (MT 645485) and LH5 (MT 645487) were all closely related to Bacillus sp. (at 97.3-97.9%). Meanwhile, from Bentong and Sungai Lembing hot springs, isolates B2B2 (MT668631) and S1B4 (MT668632) were near related to either Geobacillus kaustophilus or Geobacillus thermoleovorans; each at 98.5% and 97.9% similarity, respectively. These strains from Geobacillus sp. were able to thrive at higher temperature and their thermostable esterases or lipases have properties useful for biotechnological applications.

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