Thermus aquaticus
| Thermus aquaticus | |
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| Species: | T. aquaticus
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| Thermus aquaticus Brock & Freeze, 1969
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Thermus aquaticus is a species of bacterium that can tolerate high temperatures, one of several thermophilic bacteria that belong to the Deinococcus-Thermus group. It is the source of the heat-resistant enzyme Taq DNA polymerase, one of the most important enzymes in molecular biology because of its use in the polymerase chain reaction (PCR) DNA amplification technique.
History
When studies of biological organisms in hot springs began in the 1960s, scientists thought that the life of thermophilic bacteria could not be sustained in temperatures above about 55° Celsius (131° Fahrenheit).[2] Soon, however, it was discovered that many bacteria in different springs not only survived, but also thrived in higher temperatures. In 1969, Thomas D. Brock and Hudson Freeze of Indiana University reported a new species of thermophilic bacterium which they named Thermus aquaticus.[3] The bacterium was first discovered in the Lower Geyser Basin of Yellowstone National Park, near the major geysers Great Fountain Geyser and White Dome Geyser,[4] and has since been found in similar thermal habitats around the world.
Biology
It thrives at 70°C (160°F), but can survive at temperatures of 50°C to 80°C (120°F to 175°F). This bacterium is a chemotroph — it performs chemosynthesis to obtain food. However, since its range of temperature overlaps somewhat with that of the photosynthetic cyanobacteria that share its ideal environment, it is sometimes found living jointly with its neighbors, obtaining energy for growth from their photosynthesis.
Enzymes from T. aquaticus
T. aquaticus has become famous as a source of thermostable enzymes, particularly the "Taq" DNA polymerase, as described below.
- Aldolase
- Studies of this extreme thermophilic bacterium that could be grown in cell culture was initially centered on attempts to understand how protein enzymes (which normally inactivate at high temperature) can function at high temperature in thermophiles. In 1970, Freeze and Brock published an article describing a thermostable aldolase enzyme from T. aquaticus.[5]
- RNA polymerase
- The first polymerase enzyme isolated from T. aquaticus in 1974 was a DNA-dependent RNA polymerase,[6] used in the process of transcription.
- Taq I restriction enzyme
- Most molecular biologists probably became aware of T. aquaticus in the late 1970s or early 1980s because of the isolation of useful restriction endonucleases from this organism.[7] Use of the term "Taq" to refer to Thermus aquaticus arose at this time from the convention of giving restriction enzymes short names, such as Sal and Hin, derived from the genus and species of the source organisms.
- DNA polymerase ("Taq pol")
- DNA polymerase was first isolated from T. aquaticus in 1976.[8] The first advantage found for this thermostable (temperature optimum 80°C) DNA polymerase was that it could be isolated in a purer form (free of other enzyme contaminants) than could the DNA polymerase from other sources. Later, Kary Mullis and other investigators at Cetus Corporation discovered this enzyme could be used in the polymerase chain reaction (PCR) process for amplifying short segments of DNA,[9] eliminating the need to add enzyme after every cycle of thermal denaturation of the DNA. The enzyme was also cloned, sequenced, modified (to produce the shorter 'Stoffel fragment'), and produced in large quantities for commercial sale.[10] In 1989 Science magazine named Taq polymerase as its first "Molecule of the Year".[11] In 1993, Dr. Mullis was awarded the Nobel Prize for his work with PCR.
- Other enzymes
- The high optimum temperature for T. aquaticus allows researchers to study reactions under conditions for which other enzymes lose activity. Other enzymes isolated from this organism include DNA ligase, alkaline phosphatase, NADH oxidase, isocitrate dehydrogenase, amylomaltase, and fructose 1,6-disphosphate-dependent L-lactate dehydrogenase.
Controversy
The commercial use of enzymes from T. aquaticus has not been without controversy. After Dr. Brock's studies, samples of the organism were deposited in the American Type Culture Collection, a public repository. Other scientists, including those at Cetus, obtained it from there. As the commercial potential of Taq polymerase became apparent in the 1990s,[12] the National Park Service labeled its use as the "Great Taq Rip-off".[13] Researchers working in National Parks are now required to sign "benefits sharing" agreements that would send a portion of later profits back to the Park Service.
See also
References
- ^ "Kingdom Archaea". Plant Physiology Information Website. Ross Koning. 1994. Retrieved September 1 2009.
{{cite web}}: Check date values in:|accessdate=(help) - ^ Thomas Brock's essay "Life at High Temperatures", available at
- ^ Brock TD and Freeze H (1969). "Thermus aquaticus, a Nonsporulating Extreme Thermophile". J. Bact. 98 (1): 289–97. PMC 249935. PMID 5781580.
- ^ Bryan, T. Scott (2008). Geysers of Yellowstone, The (4th ed.). University Press of Colorado. ISBN 978-0-87081-924-7.
- ^ Freeze H and Brock TD (1970). "Thermostable Aldolase from Thermus aquaticus". J. Bact. 101 (2): 541–50. PMC 284939. PMID 4984076.
- ^ Air GM and Harris JI (1974). "DNA-Dependent RNA Polymerase From the Thermophilic Bacterium Thermus aquaticus". FEBS Letters. 38 (3): 277–281. doi:10.1016/0014-5793(74)80072-4. PMID 4604362.
- ^ Sato, S (1978). "A single cleavage of Simian virus 40 (SV40) DNA by a site specific endonuclease from Thermus aquaticus, Taq I". J. Biochem (Tokyo). 83 (2): 633–5. PMID 204628.
{{cite journal}}: Unknown parameter|month=ignored (help) - ^ Chien, A (September 1, 1976). "Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus". J. Bact. 127 (3): 1550–7. PMC 232952. PMID 8432.
{{cite journal}}: Unknown parameter|coauthors=ignored (|author=suggested) (help) - ^ Saiki, RK (1988). "Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase". Science. 239 (4839): 487–91. doi:10.1126/science.2448875. PMID 2448875.
{{cite journal}}: Unknown parameter|coauthors=ignored (|author=suggested) (help) - ^ Lawyer FC; et al. (1993). "High-level expression, purification, and enzymatic characterization of full-length Thermus aquaticus DNA polymerase". PCR Methods Appl. 2 (4): 275–87. PMID 8324500.
{{cite journal}}: Unknown parameter|author-separator=ignored (help) - ^ Guyer RL, Koshland DE (1989). "The Molecule of the Year". Science. 246 (4937): 1543–6. PMID 2688087.
{{cite journal}}: Unknown parameter|month=ignored (help) - ^ Fore J, Wiechers IR, Cook-Deegan R (2006). "The effects of business practices, licensing, and intellectual property on development and dissemination of the polymerase chain reaction: case study". J Biomed Discov Collab. 1: 7. doi:10.1186/1747-5333-1-7. PMC 1523369. PMID 16817955.
{{cite journal}}: CS1 maint: multiple names: authors list (link) CS1 maint: unflagged free DOI (link) — Detailed history of Cetus and the commercial aspects of PCR. - ^ Robbins J (28 November 2006). "The Search for Private Profit in the Nation's Public Parks". The New York Times.