Thermococcus litoralis

Mukund S, MWW Adams (1993) Characterization of a novel tungsten-containing formaldehyde oxidoreductase from the extremely thermophilic archaeon. Thermococcus litoralis. J Biol Chem 268 13592-13600. 

M. Takacs, G. Rakhely, K. L. Kovacs (2001) Molecular characterization and heterologous expression of hypCD, the first two [NiFe] hydrogenase accessory genes of Thermococcus litoralis. Arch. Microbiol., 176 231-235... 

Diederichs, K, Diez, J., Greller, G., Muller, C., Breed, J., Schnell, C., Vonrhein, C., Boos, W. and Welte, W. (2000). Crystal structure of MalK, the ATPase subunit of the trehalose/maltose ABC transporter of the archaeon Thermococcus litoralis, EMBO J., 19, 5951-5961. 

C Thermococcus litoralis hydrogenase D Bidirectional hydrogenase from Cyanobacteria... 

In the hyperthermophilic Archaea, NAD(P)-reactive enzymes are involved in recycling the reduced cofactors to produce H2 as a waste product as in the case of the NADPH oxidising hydrogenases from the hyperthermophilic Archaea, e.g. Pyrococcus species (Bryant and Adams 1989 Pedroni et al. 1995) and Thermococcus litoralis (Rakhely et al. 1999). These enzymes are also heterotetramers (Fig. 2.2C) with an apparently similar organisation of subunits and prosthetic groups to the Eubacterial examples of Group 5. 

Rakhely, G., Zhou, Z. FI., Adams, M. W. and Kovacs, K. L. (1999) Biochemical and molecular characterization of the [NiEe] hydrogenase from the hyperthermophilic archaeon. Thermococcus litoralis. Eur. J. Biochem., 266, 1158-65. 

An especially intriguing pair of products obtained from marine organisms in recent years are Vent and Deep Vent DNA polymerase. These products are used in DNA research studies. Their special feature is that they are at least 10 times as efficient as other similar products in polymerase chain reactions because they can tolerate temperatures just below the boiling point of water, a characteristic that comparable research tools lack. Vent and Deep Vent DNA polymerases are obtained from the bacterium Thermococcus litoralis, which is found around deep-sea hydrothermal vents at the bottom of the ocean. 

S. Ohshima, T. Biochemical characterization, cloning, and sequencing of ADP-dependent (AMP-forming) glucokinase from two hyperthermophilic archaea, Pyrococcus furiosus and Thermococcus litoralis. J. Biochem., 128, 1079-1085 (2000)... 

Kong, H., R.B. Kucera and W.E. Jack 1993. Characterization of a DNA polymerase from the hyperthermophile archaea Thermococcus litoralis Vent DNA polymerase, steady state kinetics, thermal stability, processivity, strand displacement, and exonuclease activities. J. Biol. Chem. 268 1965-1975. 

Section 4.3), which had to be replenished at every new cycle, the application of heat-stable polymerases from organisms such as Thermus aquations (Taq polymerase), Pyrococcus woesii (Pwo polymerase), Pyrococcus juriosus (Pfu polymerase), or Thermococcus litoralis (Vent polymerase) facilitated automation of the thermal cycling process. 

Xavier, K. B., L. O. Martins, R. Peist, M. Kossmann, W. Boos, and H. Santos. 1996. High-affinity maltose/trehalose transport system in the hyperthermophilic archaeon Thermococcus litoralis. Journal of Bacteriology 178 4773—4777. 

S. Mukund, Biochemical and Biophysical Characterization of Novel Tungsten-Containing Enzymes from Hyperthermophilic Archaea (Pyrococcusfuriosus, Thermococcus litoralis). Ph.D. Thesis, University of Georgia, Athens (1996). 

Imamura, H., Fushinobu, S., Yamamoto, M., Kumasaka, T., Wakagi, T., and Matsu-zawa, H. 2001. Reaction mechanism and crystal structure of 4-a-Glucanotransferase from a hyperthermophilic archaeon, Thermococcus litoralis. J. Appl. Glycosci., 48, 171-175. 

Xavier, K. B., Peist, R., Kossmann, M., Boos, W., and Santos, H. 1999. Maltose metabolism in the hyperthermophilic archaeon Thermococcus litoralis Purification and characterization of key enzymes. J. Bacteriol, 181, 3358-3367. 

Thermococcus litoralis gains energy fiom the fermentation of peptides. The tungstoenzyme formaldehyde ferredoxin oxidoreductase (FOR) catalyzes the formaldehyde oxidation to formate ... 

More recently, the discovery and commercialization of L-aminoacylase from Thermococcus litorali was a product of the LINK project between Chirotech Technology and the University of Exeter. The L-aminoacylase of T. litoralis had broad substrate specificity for the hydrolysis of N-acylated a-amino acids, with respect to both the side chain and the N-acyl group. It is especially useful for the enantiospecific hydrolysis of acyl groups, particularly N-benzoyl groups of a-amino acids. This can be used to advantage in synthetic processes that require the enantiospecific deprotection of racemates [29]. 

The most widely used DNA polymerase in PCR is from Thermus acquaticus, the Taq polymerase. This enzyme has an optimal temperature for polymerization in the range of 70 to 75°C. It extends DNA chains at a rate of about 2 kb per minute. It is fairly resistant to the continual cycles of heating and cooling required for PCR. The half-life for thermal denaturation of Taq polymerase is 1.6h at 95°C. When very high denaturation temperatures are needed, as in the PCR amplification of very G+C-rich DNA, more thermal-stable polymerase such as the enzyme from Thermococcus litoralis with a half-life of 1.8h at 100°C or the enzyme from Pyrococcus furiosis with a half-life of 8 h at 100°C, can be employed. However, these enzymes are not as processive as Taq polymerase, which makes it more difficult to amplify long templates. 

Although, especially for the 1 -isomer, it is possible to use commercial aminoacylases for the purpose, we felt that for the technology to be industrially ready, we had to be assured of the catalyst supply. To achieve this, we wanted to produce the catalyst ourselves and embarked on a program of biocatalyst discovery and development. Through this we identified and characferized our own cloned proprielary 1 - and d-aminoacylase enzymes, which we can now use to manufaclure products in ton quantities if needed. Collaboration with Littlechild at the University of Exeter characterized a particularly useful theromo-stable and therefore robust 1 -aminoacylase from Thermococcus litoralis that can optionally be immobilized to generate a reusable industrial biocatalyst. ... 

Fig. 1. Amino acid sequence alignments of some 4Fe-type feiredoxins to show consensus sequence of cysteinyl residues. The duster-binding ligands are indicated in bold and with an asterisk. The vertical lines indicate the two additional Cys residues found in many of these proteins. The abbreviations are Pf, Pyrococcus furiosus Tl, Thermococcus litoralis Tm, Thermotoga maritima Da, Desulfovibrio africanus Dg, Desulfovibrio gigas Mb, Methanosarcina barkeri Ct, Clostridium thermoaceticum. The sequences were taken from P. S. Brereton, R. E. Duderstadt, C. R. Staples, M. K. Johnson, and M. W. W. Adams, Biochemistry 38, 10594 (1999). Whether these proteins contain a [4Fe-4S] or a [3Fe-4S] cluster when they are purified is indicated by (4Fe) and (3Fe), respectively.

GDH from P. furiosus is not the only enzyme stabilized by high pressure. Other enzymes found to be stabilized by high pressure in the authors laboratory include GDHs from the hyperthermophiles Thermococcus litoralis and... 

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