Abstract
The study of enzymes isolated from organisms inhabiting unconventional ecosystems has led to the realization that biocatalysis need not be constrained to mild conditions and can be considered at pH's, temperatures, pressures, ionic and solvent environments long thought to be destructive to biomolecules. Parallel to this, it has been demonstrated that even conventional enzymes will catalyze reactions in solvents other than water. However, the intrinsic basis for biological function under extreme conditions is only starting to be addressed, as are associated applications. This was the focus of a recent NSF/NIST-sponsored workshop on extremozymes. Given the information acquired from the study of extremozymes, modification of enzymes to improve their ranges of stability and activity remains a possibility. Ultimately, by expanding the range of conditions suitable for enzyme function, new opportunities to use biocatalysis will be created.
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References
Flam, F. 1994. The chemistry of life at the margins. Science 265: 471–472.
Hodgson, J. 1994. The changing bulk catalyst market. Bio/Technology 12: 789–790.
Matin, A. 1990. Keeping a neutral cytoplasm; the bioenergetics of obligate aci-dophiles. FEMS Microbiol. Rev. 75: 307–318.
Peeples, T.L. and Kelly, R.M. 1995. Bioenergetic response of the extreme thermoaddophile Metallosphaera sedula to thermal and nutritional stresses. Appl. Environ. Microbiol. 61: 2314–2321.
Fiala, G. and Stetter, K.O. 1986. Pyrococcus furiosus sp. nov. represents a novel genus of marine heterotrophic archaebacteria growing optimally at 100°C. Arch. Microbiol. 145: 56–61.
Davail, S., Feller, G., Narinx, E. and Gerday, C. 1994. Cold adaptation of proteins. Purification, characterization, and sequence of the heat-labile sub-tilisin from the Antarctic psychrophile Bacillus TA41. J. Biol. Chem. 269: 17448–17453.
Jones, W.J., Leigh, J.A., Leigh, J.A., Mayer, E., Woese, C.R., and Wolfe, R.S. 1983. Methanococcus jannaschii sp. nov., an extremely thermophilic methanogen from a submarine hydrothermal vent. Arch. Microbiol. 136: 254–61.
Li, Y., Mandelco, L. and Wiegel, J. 1993. Isolation and characterization of a moderately thermophilic anaerobic alkaliphile, Clostridium paradoxum, sp. nov. Int. J. Sys. Bacteriol. 43: 450–460.
Huber, R., Spinnler, C., Gambacorta, A. and Stetter, K.O. 1989. Metallosphaera sedula gen. and sp. nov. represents a new genus of aerobic, metal-mobilizing, thermoacidophilic archaebacteria. Syst. Appl Microbiol. 12: 38–47.
Kushner, D.J. 1978. In: Microbial Life in Extreme Environments. Kushner, D. J. (Ed.). Academic Press, London, p. 317–368.
Bartlett, D., Wright, D. and Yayanos, A.A. 1989. Isolation of a gene regulated by hydrostatic pressure in a deep-sea bacterium. Nature 342: 572–574.
Miller, J.E., Shah, N.N., Nelson, C.M., Ludlow, J.M. and Clark, D.S. Pressure and temperature effects on growth and gas production of the extreme thermophile Methanococcus janaschii. Appl. Environ. Microbiol. 54: 3039–3042.
Trent, J.D., Osipiuk, J. and Pinkau, T. 1990. Acquired thermotolerance and heat shock in the extremely thermophilic archaebacterium Sulfolobus sp. Strain B12. J. Bact. 172: 1478–1484.
Trent, J.D., Nimmesgern, E., Wall, J.S., Hartl, F.U. and Horwich, A. 1991 A molecular chaperone from a thermophilic archaebacterium is related to the eukaryotic protein T-complex polypeptide-1. Nature 354: 490–493.
Holden, J.F. and Baross, J.A. 1993. Enhanced thermotolerance and temperature-induced changes in protein composition in the hyperthermophilic archaeon ES-4. J. Bacteriol. 175: 2839–2843.
Adams, M.W.W. and Kelly, R.M. (Eds.) 1992. Biocatalysis at Extreme Temperatures: Enzyme Systems Near and Above 100°C, Washington, D.C, American Chemical Society, 215 pp., Series No. 498.
Reeve, J.N. 1992. Molecular biology of methanogens. Annu. Rev. Microbiol. 46: 165–191.
Reeve, J.N. 1994. Thermophiles in New Zealand. ASM News. 60: 541–545.
Stetter, K.O., Fiala, G., Huber, R. and Segerer, A. 1990. Hyperthermophilic microorganisms. FEMS Microbiol. Rev. 75: 117–124.
Kelly, R.M. and Adams, M.W.W. 1994. Metabolism in hyperthennophilic microorganisms. Antonie van Leeuwenhoek 66: 247–270
Kelly, R.M., Brown, S.H., Blumentals, I.I. and Adams, M.W.W. 1992. Characterization of Enzymes from High Temperature Bacteria, in Biocatalysis at Extreme Temperatures, Adams, M.W.W. and Kelly, R. M. (Eds.). ACS Symposium Series No. 498, p. 23–42.
Hochachka, P.W. and Somero, G.N. 1984. Biochemical Adaptations. Princeton University Press, Princeton, NJ.
Craik, C.S. 1994. Natural adaptation analysis for engineering proteases. Presented at NSF Workshop on Extremozymes, Washington, DC, May, 1994.
Muriana, F.J.G., Alvarez-Ossorio, M.C. and Relimpio, A.M. 1991. Purification and characterization of aspartate aminotransferase from the halophile archaebacterium Haloferax mediterranei. Biochem. J. 278: 149–154.
Ryu, K., Kim, J. and Dordick, J.S. 1994. Catalytic properties and potential of an extracellular protease from an extreme halophile. Enzyme Microbiol. Technol. 16: 266–275.
Adams, M.W.W. and Kelly, R.M. 1994. Thermostability and thennoactivity of enzymes from hyperthermophilic microorganisms. Bioorg. Med. Chem. 2: 659–667.
Kelly, R.M. and Brown, S.H. 1993. Enzymes from high temperature microorganisms. Curr. Opin. Biotechnol. 4: 188–192.
Adams, M.W.W. 1993. Enzymes and proteins from organisms that grow near and above 100°C. Ann. Rev. Microbiol. 47: 627–658.
Kelly, R.M., Adams, M.W.W. and Baross, J.A. 1994. Biotechnology and life in boiling water. Chemistry in Britain 30: 555–558.
Costantino, H.R., Brown, S.H. and Kelly, R.M. 1990. Purification and characterization of an a-glucosidase from a hyperthermophilic archaebacterium, Pyrococcus furiosus, exhibiting a temperature optimum of 105 to 115°C. J. Bacteriol. 172: 3654–60.
Kegen, S.W.M., Luesink, E.J., Stams, A.J.M. and Zehnder, A.J.B. 1993. Purification and characterization of an extremely thermostable β-glucosidase from the hyperthermophilic archaeon Pyrococcus furiosus. Eur. J. Biochem. 213: 305–312.
Laderman, K.A., Davis, B.R., Krutzsch, H., Krutzsch, H.C., Lewis, M.S., Griko, Y.V., Privalov, P.L. and Anfinsen, C. B. 1993. The purification and characterization of an extremely thermostable α-amylase from the hyperthermophilic archaebacterium Pyrococcus furiosus. J. Biol. Chem. 268: 24394–24401.
Blumentals, I.I., Robinson, A.S. and Kelly, R.M. 1990. Characterization of SDS-resistant proteolytic activity in the hyperthermophilic archaebacterium, Pyrococcus furiosus. Appl. Environ. Microbiol. 56: 1992–1998.
Brown, S.H. and Kelly, R.M. 1993. Characterization of amylolytic enzymes, having both α-1,4 and α-1,6-hydrolytic activity, from the thermophilic archaea Pyrococcus furiosus and Thermococcus litoralis. Appl. Environ. Microbiol. 59: 2614–2621.
Mukund, S. and Adams, M.W.W. 1991. The novel tungsten-iron-sulfur protein of the hyperthermophilic archaebacterium, Pyrococcus furiosus, is an aldehyde ferredoxin oxidoreductase: evidence for its participation in a unique glycolytic pathway. J. Biol. Chem. 266: 14208–14216.
Robb, F.T., Park, J.B. and Adams, M.W.W. 1992. Characterization of an extremely thermostable glutamate dehydrogenase: A key enzyme in the primary metabolism of the hyperthermophilic archaebacterium Pyrococcus furiosus. Biochem. Biophys. Acta 1120: 267–272.
Ma, K. and Adams, M.W.W. 1994. Sulfide dehydrogenase from the hyperthermophilic archaeon, Pyrococcus furiosus: a new multifunctional enzyme involved in the reduction of elemental sulfur. J. Bacteriol. 176: 6509–6517.
Bryant, F.O. and Adams, M.W.W. 1989. Characterization of hydrogenase from the hyperthermophilic archaebacterium, Pyrococcus furiosus. J. Biol. Chem. 264: 5070–9.
Badr, H.R., Sims, K.A. and Adams, M.W.W. Purification and characterization of a sucrose a-glucohydrolase from Pyrococcus furiosus exhibiting a temperature optimum above 100°C. Syst. Appl. Microbiol. 17: 1–6.
Lundberg, K.S., Shoemaker, D.D., Short, J.M., Sorge, J.A., Adams, M.W.W. and Mathur, E. 1991. High fidelity amplification with a thermostable DNA polymerase isolated from Pyrococcus furiosus. Gene 108: 1–6.
Chi, E. and Bartlett, D.H. 1993. Use of a reporter gene to follow high-pressure signal transduction in the deep-sea bacterium Photobacterium sp. Strain S9. 175: 7533–
Heremans, K. 1980. Biophysical chemistry at high pressure. Rev. Phys. Chem. Jpn. 50: 256–273.
Hei, D.J. and Clark, D.S. 1994. Pressure stabilization of proteins from extreme thermophiles. 60: 932–939.
Reiter, W., Hudepohl, U. and Zillig, W. 1990. Mutational analysis of an archae-bacterial promoter: Essential role of a TATA box for transcription efficiency and start-site selection in vitro. Proc. Natl. Acad. Sci. USA 87: 9509–13.
Thomm, M., Wich, G., Brown, J.W., Frey, G., Sherf, B.A. and Beckler, G.S. 1989. An archaebacterial promoter sequence assigned by RNA polymerase binding experiments. Can. J. Microbiol. 35: 30–35.
Hudepohl, U., Reiter, W.D. and Zillig, W. 1990. In vitro transcription of two rRNA genes of the archaebacterium Sulfolobus sp. B12 indicates a factor requirement for specific initiation. Proc. Natl. Acad. Sci. USA 87: 5851–5855.
Grayling, R.A., Sandman, K. and Reeve, J.N. 1994. Archaeal DNA binding proteins and chromosome structure. System. Appl. Microbiol. 16: 582–590.
Kjems, J. and Garrett, R.A. 1988. Novel splicing mechanism for the riboso-mal RNA intron in the archaebacterium Desulfumcoccus mobilis. Cell 54: 693–703.
Daniels, C.J., Gupta, R. and Doolittle, W.F. 1985. Transcription and excision of a large intron in the tRNA Trp gene of an archaebacterium, Halobacterium volcanii. J. Biol. Chem. 260: 3132–34.
Marguet, E. and Forterre, P. 1994. DNA stability at temperatures typical for hyperthermophiles. Nucleic Acids. Res. 22: 1681–86.
Scholz, S., Sonnenbichler, J., Schäfer, W. and Hensel, R. 1992. Di-myo-inositol-1,1′-phosphate: a new inositol phosphate isolated from Pyrococcus woesei. FEBS Lett. 306: 239–42.
Xu, M., Southworth, M.W., Mersha, F.B., Hornstra, L.J. and Perler, F.B. 1993. In vitro protein splicing of purified precursor and the identification of a branched intermediate. Cell 75: 1371–1377.
Barnes, W.M. 1994. PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates. Proc. Natl. Acad. Sci. USA 91: 2216–2220.
Cheng, S., Focker, C., Barnes, W.M. and Higuchi, R. 1994. Effective amplification of long targets from cloned inserts and human genomic DNA. Proc. Natl. Acad. Sci. USA 91: 5695–9.
Holmes, M.L. and Dyall-Smith, M.L. 1990. A plasmid vector with a selectable marker for halophilic archaebacteria. J. Bacteriol. 172: 756–61.
Schleper, C., Kubo, K. and Zillig, W. 1992. The particle SSV1 from the extremely thermophilic archaeon Sulfolobus is a virus: Demonstration of infectivity and of transfection with viral DNA. Proc. Natl. Acad. Sci. USA 89: 7645–7649.
Mather, M.W. and Fee, J.A. 1992. Development of plasmid cloning vectors for Thermus thermophilus HB8: expression of a heterologous, plasmid-borne kanamycin nucleotidyltransferase gene. Appl Environ Microbiol. 58: 421–5.
Perler, F.B., Comb, D.G., Jack, W.E., Moran, L.S., Qiang, B., Kucera, R.B., Benner, J., Slatko, B.E., Nwankwo, D.O., Hempstead, S.K. and et al. 1992. Intervening sequences in an Archaea DNA polymerase gene. Proc. Natl. Acad. Sci. USA 89: 5577–81.
Heltzel, A., Smith, E.T., Zhou, Z.H., Blarney, J.M. and Adams, M.W.W. 1994. Cloning, characterization and expression of the gene for the [4Fe-4S] ferredoxin from the hyperthermophilic archaeon Pyrococcus furiosus. J. Bacteriol. 176: 4790–4793.
Mroczkowski, B.S., Huvar, A., Lernhardt, W., Misono, K., Nielson, K. and Scott, B. 1994. Secretion of thermostable DNA polymerase using a novel Baculovirus vector. J. Biol. Chem. 269: 13522–13528.
DiRuggiero, J. and Robb, F.T. 1995. Expression and in vitro assembly of recombinant glutamate dehydrogenase from the hyperthermophilic archaeon Pyrococcus furiosus. Appl. Environ. Microbiol. 61: 159–164.
Sandman, K., Perler, F.B. and Reeve, J.N. 1994. Histone-encoding genes from Pyrococcus: Evidence for members of the HMf family of archaeal histones in a non-methanogenic Archaeon. Gene 150: 207–208.
Pedroni, P., Dellavolpe, A., Galli, G., Mura, G.M., Pratesi, C. and Grandi, G. 1995. Characterization of the locus encoding the [Ni-Fe] sulfhydrogenase from the archaeon Pyrococcus furiosus: Evidence for a relationship to bacterial sulfite reductases. Microbiology 141: 449–458.
Robinson, K.A., Bartley, D.A., Robb, F.T. and Schreier, H.J. 1995. A gene from the hyperthermophile Pyrococcus furiosus whose deduced product is homologous to members of the prolyl oligopeptidase family of proteases. Gene 152: 103–106.
Robinson, K.A. and Schreier, H.J. 1995 Isolation, sequence and characterization of the maltose-regulated mlrA gene from the hyperthermophilic archaeum Pyrococcus furiosus. Gene 151: 173–176.
Zwickl, P., Fabry, S., Bogedain, C., Haas, A. and Hensel, R. 1990. Glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic archaebacterium Pyrococcus woesei: characterization of the enzyme, cloning and sequencing the gene, and expression in Escherichia coli. J. Bacteriol. 172: 4329–38.
Tiboni, O., Cammarano, P. and Sanangelantoni, A.M. 1993. Cloning and sequencing of the gene encoding glutamine synthetase I from the archaeum Pyrococcus woeseii: anomalous phytogenies inferred from analysis of archaeal and bacterial glutamine synthetase I sequences. J. Bacteriol. 175: 2961–2969.
DiRuggiero, J., Robb, F.T., Jagus, R., Klump, H.K., Borges, K.M., Mai, X., Kessel, M. and Adams, M.W.W. 1993. Characterization, cloning, and in vitro expression of an extremely thermostable glutamate dehydrogenase from the hyperthermophilic archaeon ES4. J. Biol. Chem. 268: 17767–17774.
Busse, S.A., La Mar, G.N., Yu, L.P., Howard, J.B., Smith, E.T., Zhou, Z.H. and Adams, M.W.W. 1992. Proton NMR investigation of the oxidized three-iron clusters in the ferredoxins from the hyperthermophilic archaea, Pyrococcus furiosus and Thermococcus litoralis. Biochemistry 31: 11952–11962.
Jaenicke, R. 1991. Protein stability and molecular adaption to extreme conditions. Eur. J. Biochem. 202: 715–28.
Böhm, G. and Jaenicke, R. 1994. Relevance of sequence statistics for the properties of extremophilic proteins. Int. J. Pept. Prot. Res. 43: 97–106.
Blake, P.R., Park, J.B., Bryant, F.O., Aono, S., Magnuson, J.K., Eccleston, E., Howard, J.B., Summers, M.F. and Adams, M.W.W. 1991. Determinants of protein hyperthermostability. 1. Purification, amino acid sequence, and secondary structure from NMR of the rubredoxin from the hyperthermophili-carchaebacterium, Pyrococcus furiosus. Biochemistry 30: 10885–10891.
Day, M.W., Hsu, B.T., Joshua-Tor, L., Park, J.-B., Zhou, Z.H., Adams, M.W.W. and Rees, D. C. 1992. X-ray crystal structure of the oxidized and reduced forms of the rubredoxin from the marine hyperthermophilic archaebacterium, Pyrococcus furiosus. Protein Science 1: 1494–1507.
Blake, P.R., Park, J.-B., Zhou, Z.H., Hare, D.R., Adams, M.W.W. and Summers, M.F. 1992. Solution state structure by NMR of zinc-substituted rubredoxin from the marine hyperthermophilic archaebacterium, Pyrococcus furiosus. Protein Science 1: 1508–1521.
Chan, M.K., Mukund, S., Kletzin, A., Adams, M.W.W. and Rees, D.C. 1995. Structure of the hyperthermophilic tungstoprotein enzyme aldehyde ferredoxin oxidoreductase. Science 267: 1463–1469.
Korndörfer, I., Steipe, B., Huber, R., Tomschy, A. and Jaenicke, R. 1995. The crystal structure of holo-glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic bacterium Thermotoga maritima at 2.5 angstrom resolution. J. Mol. Biol. 246: 511–521.
Ragone, R. and Colonna, G. 1995. Do globular proteins require some structural peculiarity to best function at high temperatures? J. Am. Chem. Soc. 117: 16–20.
Wick, C.B. 1994. Enzymology advances offer economical and environmentally safe ways to make paper. GEN 14: 1
McCutchen, C.M., Duffaud, G.D., Leduc, P., Petersen, A., Tayal, A., Khan, S.A. and Kelly, R.M. 1995. Purification, biochemical characterization and use of β-1,4-mannanase and α-1,6-galactosidase from the hyperthermophilic eubacterium Thermotoga neapolitona 5068 for hydrolysis of galactomannans in hydraulic fracturing fluids. Biotechnol. Bioeng. Submitted.
Klibanov, A.M. 1986. Enzymes that work in organic solvents. Chemtech 16: 354–359.
Russell, A.J., Beckman, E.J. and Chaudhary, A. Studying enzyme activity in supercritical fluids. Chemtech 24: 33–38.
Margolin, A.L. 1991. Enzymes: Use them. Chemtech 21: 160–167.
Dordick, J.S. 1989. Enzymic catalysis in monophasic organic solvents. Enzyme Microb. Technol. 11: 194–211.
Arnold, F.H. 1993. Protein engineering for unusual environments. Tibtech 4: 450–455.
Mozhaev, V.V., Heremans, K., Frank, J., Masson, P. and Balny, C. 1994. Exploiting the effects of high hydrostatic pressure in biotechnological applications. Tibtech 12: 493–501.
Kunugi, S. 1993. Modification of biopolymer function by high pressure. Prog. Polym. Sci. 18: 805–838.
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Adams, M., Perler, F. & Kelly, R. Extremozymes: Expanding the Limits of Biocatalysis. Nat Biotechnol 13, 662–668 (1995). https://doi.org/10.1038/nbt0795-662
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DOI: https://doi.org/10.1038/nbt0795-662
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