Characteristics of DNA polymerase I from an extreme thermophile, <i>Thermus scotoductus</i> strain K1

Saghatelyan, Ani; Panosyan, Hovik; Trchоunian, Armen; Birkeland, Nils Kåre

doi:10.1002/mbo3.1149

Cited by 8 publications

(6 citation statements)

References 24 publications

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“…Similar A-family thermophilic DNAPs have been identified from other Thermus strains, such as Tfi DNAP from Thermus filiformis [ 62 ], Tth DNAP from Thermus thermophilus [ 28 ], Tfl DNAP from Thermus flavus [ 63 ], Tca DNAP from Thermus caldophilus [ 30 ], and TsK1 DNAP from Thermus scotoductus [ 31 ]. Like Taq DNAP, these DNAPs possess 5′-3′ exonuclease activity but no 3′-5′ exonuclease activity.…”

Section: Thermophilic and Hyperthermophilic Nucleic Acid Polymerasesmentioning

confidence: 84%

Thermophilic Nucleic Acid Polymerases and Their Application in Xenobiology

Wang

et al. 2022

IJMS

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Thermophilic nucleic acid polymerases, isolated from organisms that thrive in extremely hot environments, possess great DNA/RNA synthesis activities under high temperatures. These enzymes play indispensable roles in central life activities involved in DNA replication and repair, as well as RNA transcription, and have already been widely used in bioengineering, biotechnology, and biomedicine. Xeno nucleic acids (XNAs), which are analogs of DNA/RNA with unnatural moieties, have been developed as new carriers of genetic information in the past decades, which contributed to the fast development of a field called xenobiology. The broad application of these XNA molecules in the production of novel drugs, materials, and catalysts greatly relies on the capability of enzymatic synthesis, reverse transcription, and amplification of them, which have been partially achieved with natural or artificially tailored thermophilic nucleic acid polymerases. In this review, we first systematically summarize representative thermophilic and hyperthermophilic polymerases that have been extensively studied and utilized, followed by the introduction of methods and approaches in the engineering of these polymerases for the efficient synthesis, reverse transcription, and amplification of XNAs. The application of XNAs facilitated by these polymerases and their mutants is then discussed. In the end, a perspective for the future direction of further development and application of unnatural nucleic acid polymerases is provided.

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Section: Thermophilic and Hyperthermophilic Nucleic Acid Polymerasesmentioning

confidence: 84%

Thermophilic Nucleic Acid Polymerases and Their Application in Xenobiology

Wang

et al. 2022

IJMS

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“…In total, more than 130 strains of 40 distinct species belonging to 22 different genera, namely Bacillus, Geobacillus, Parageobacillus, Anoxybacillus, Paenibacillus, Brevibacillus Aeribacillus, Ureibacillus, Thermoactinomyces, Sporosarcina, Thermus, Rhodobacter, Thiospirillum, Thiocapsa, Rhodopseudomonas, Methylocaldum, Desulfomicrobium, Desulfovibrio, Treponema, Arcobacter, Nitrospira , and Methanoculleus , have been isolated from the hot springs in Armenia and Nagorno-Karabakh. Among these isolates, many prospective strains have been selected for future studies to explore their potential in white (industrial), green (agricultural), and red (pharmaceutical) biotechnology [ 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 ].…”

Section: Microbiological Analysismentioning

confidence: 99%

“…T. scotoductus strain SA-01, recovered from a deep gold mine in South Africa, was found to be the closest relative species of strain K1, with 80% genome sequence similarity. The DNA polymerase of strain K1 ( Ts K1) has been cloned and subsequently expressed in Escherichia coli host cells [ 43 ]. The purified enzyme could efficiently amplify 2.5 kb DNA products under the optimal reaction conditions (74 °C and pH 9), and required 3–5 mM Mg 2+ for optimal activity.…”

Section: Microbiological Analysismentioning

confidence: 99%

“…The purified enzyme could efficiently amplify 2.5 kb DNA products under the optimal reaction conditions (74 °C and pH 9), and required 3–5 mM Mg 2+ for optimal activity. Ts K1 polymerase was stable for at least 1 h at 80 °C, with half-lives of 30 and 15 min at 88 °C and 95 °C, respectively [ 43 ].…”

Section: Microbiological Analysismentioning

confidence: 99%

“…The Ts K1 DNA polymerase obtained from the thermophile T. scotoductus strain K1 also has the potential to be commercialized. The better base insertion fidelity of Ts K1 is a feature that demonstrates an advantage over Taq DNA polymerase [ 43 ] and can be used in various high-temperature polymerization reactions.…”

Section: Microbiological Analysismentioning

confidence: 99%

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

et al. 2021

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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.

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Mining thermophiles for biotechnologically relevant enzymes: evaluating the potential of European and Caucasian hot springs

Burkhardt,

Baruth,

Neele Meyer-Heydecke

et al. 2023

Extremophiles

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The development of sustainable and environmentally friendly industrial processes is becoming very crucial and demanding for the rapid implementation of innovative bio-based technologies. Natural extreme environments harbor the potential for discovering and utilizing highly specific and efficient biocatalysts that are adapted to harsh conditions. This review focuses on extremophilic microorganisms and their enzymes (extremozymes) from various hot springs, shallow marine vents, and other geothermal habitats in Europe and the Caucasus region. These hot environments have been partially investigated and analyzed for microbial diversity and enzymology. Hotspots like Iceland, Italy, and the Azores harbor unique microorganisms, including bacteria and archaea. The latest results demonstrate a great potential for the discovery of new microbial species and unique enzymes that can be explored for the development of Circular Bioeconomy.Different screening approaches have been used to discover enzymes that are active at extremes of temperature (up 120 °C), pH (0.1 to 11), high salt concentration (up to 30%) as well as activity in the presence of solvents (up to 99%). The majority of published enzymes were revealed from bacterial or archaeal isolates by traditional activity-based screening techniques. However, the latest developments in molecular biology, bioinformatics, and genomics have revolutionized life science technologies. Post-genomic era has contributed to the discovery of millions of sequences coding for a huge number of biocatalysts. Both strategies, activity- and sequence-based screening approaches, are complementary and contribute to the discovery of unique enzymes that have not been extensively utilized so far.

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Characteristics of DNA polymerase I from an extreme thermophile, Thermus scotoductus strain K1

Cited by 8 publications

References 24 publications

Thermophilic Nucleic Acid Polymerases and Their Application in Xenobiology

Thermophilic Nucleic Acid Polymerases and Their Application in Xenobiology

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

Mining thermophiles for biotechnologically relevant enzymes: evaluating the potential of European and Caucasian hot springs

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