Table 9.

Examples of thermophilic and hyperthermophilic enzymes with applications as molecular biology reagents

EnzymeOriginApplicationsPropertiesSource or reference(s)
TaqpolymeraseT. aquaticusPCR technologiesOptimal activity at 75°C, pH 9.026, 199
Vent DNA polymeraseT. litoralisOptimal activity at 75°C, proofreading activity2, 271
Deep Vent DNA polymeraseP. furiosusOptimal activity at 75°C, proofreading activity; t1/2, 23 h (95°C)2, 343
C. therm DNA polymeraseCarboxydothermus  hydrogenoformansReverse transcription-PCRReverse transcriptase activity, 3′→5′ proofreading activity; optimal activity as 60–70°CRoche Molecular Biochemicals
DNA polymeraseThermus  thermophilusReverse transcriptase activityRoche Molecular Biochemicals
Pfu DNA ligaseP. furiosusLigase chain reaction and DNA ligationsActive at 45–80°C;t1/2 >60 min (95°C)Stratagene
Tcs DNA ligaseThermus  scodoductusLigase chain reactionOptimal activity at 45°CRoche Molecular Biochemicals
DNA binding protein Ssod7S. solfataricusTime-reducing and specificity-enhancing in DNA-DNA hybridizations; locking of antisense oligonucleotide to target sequenceSequence-aspecific DNA binding; ATP-independent, homology-dependent DNA annealing at 60°C123
Serine protease (PreTaq)Thermus strain  Rt41ADNA and RNA purifications; cellular structures degradation prior to PCROptimal activity at 90°C, pH 8.0;t1/2, 20 min (90°C) (+CaCl2)Life Technologies
Protease SP. furiosusProtein fragmentation for sequencingOptimal activity at 85–95°C, pH 6.0–8.0; 80% active after 3 h (95°C)TaKaRa Biomedicals
Methionine aminopeptidaseP. furiosusCleavage of the N-terminal Met in proteinsOptimal activity at 85–95°C, pH 7.0–8.0; stable for 1 h (75°C)TaKaRa Biomedicals
Pyroglutamate aminopeptidaseP. furiosusCleavage of the N-terminall-pyroglutamate in proteinsOptimal activity at 95–100°C, pH 6.0–9.0; 95% active after 2.5 h (75°C)TaKaRa Biomedicals
CarboxypeptidaseS. solfataricusC-terminal sequencingBroad specificity (can release basic, acidic, and aromatic residues); stable in solvents at 40°C66
Alkaline phosphataseT. neapolitanaEnzyme-labeling applications where high stability is requiredOptimal activity at 85°C, pH 9.9;t1/2, 4 h (90°C) (+ Co2+)87