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Applied Microbiology and Biotechnology

, Volume 72, Issue 4, pp 623–634 | Cite as

Extremolytes: natural compounds from extremophiles for versatile applications

  • Georg LentzenEmail author
  • Thomas Schwarz
Mini-Review

Abstract

Extremophilic microorganisms have adopted a variety of ingenious strategies for survival under high or low temperature, extreme pressure, and drastic salt concentrations. A novel application area for extremophiles is the use of “extremolytes,” organic osmolytes from extremophilic microorganisms, to protect biological macromolecules and cells from damage by external stresses. In extremophiles, these low molecular weight compounds are accumulated in response to increased extracellular salt concentrations, but also as a response to other environmental changes, e.g., increased temperature. Extremolytes minimize the denaturation of biopolymers that usually occurs under conditions of water stress and are compatible with the intracellular machinery at high (>1 M) concentrations. The ectoines, as the first extremolytes that are produced in a large scale, have already found application as cell protectants in skin care and as protein-free stabilizers of proteins and cells in life sciences. In addition to ectoines, a range of extremolytes with heterogenous chemical structures like the polyol phosphates di-myoinositol-1,1′-phosphate, cyclic 2,3-diphosphoglycerate, and α-diglycerol phosphate and the mannose derivatives mannosylglycerate (firoin) and mannosylglyceramide (firoin-A) were characterized and were shown to have protective properties toward proteins and cells. A range of new applications, all based on the adaptation to stress conditions conferred by extremolytes, is in development.

Keywords

Ectoine Compatible solutes Osmolytes Extremophiles Extremolytes Cytoprotection 

References

  1. Andersson MM, Breccia JD, Hatti-Kaul R (2000) Stabilizing effect of chemical additives against oxidation of lactate dehydrogenase. Biotechnol Appl Biochem 32(Pt 3):145–153CrossRefGoogle Scholar
  2. Arakawa T, Timasheff SN (1985) The stabilization of proteins by osmolytes. Biophys J 47:411–414Google Scholar
  3. Arora A, Ha C, Park CB (2004) Inhibition of insulin amyloid formation by small stress molecules. FEBS Lett 564:121–125CrossRefGoogle Scholar
  4. Baliarda A, Robert H, Jebbar M, Blanco C, Deschamps A, Marrec C (2002) Potential osmoprotectants for the lactic acid bacteria Pediococcus pentosaceus and Tetragenococcus halophila. Int J Food Microbiol 2607:1–8Google Scholar
  5. Baluna R, Vitetta ES (1997) Vascular leak syndrome: a side effect of immunotherapy. Immunopharmacology 37:117–132CrossRefGoogle Scholar
  6. Barth S (2000) Pharmaceutical preparation. European Patent EP000001183047Google Scholar
  7. Barth S, Huhn MF, Matthey B, Klimka A, Galinski EA, Engert A (2000) Compatible-solute-supported periplasmic expression of functional recombinant proteins under stress conditions. Appl Environ Microbiol 66:1572–1579CrossRefGoogle Scholar
  8. Bersch S, Vangala M, Schwarz T, Kaufmann M (2000) Protection of antibodies against proteolytic degradation by compatible solutes. In: 2nd international conference on protein stabilisation/biomolecule stabilisation, LissabonGoogle Scholar
  9. Beyer N, Driller H, Bünger J (2000) Ectoin—a innovative, multi-functional active substance for the cosmetic industry. SÖFW-J 126:27–29Google Scholar
  10. Borges N, Ramos A, Raven ND, Sharp RJ, Santos H (2002) Comparative study of the thermostabilizing properties of mannosylglycerate and other compatible solutes on model enzymes. Extremophiles 6:209–216CrossRefGoogle Scholar
  11. Brown AD (1976) Microbial water stress. Bacteriol Rev 40:803–846Google Scholar
  12. Bünger J (1999) Ectoine added protection and care for the skin. Eurocosmetics 7:22–24Google Scholar
  13. Bünger J, Driller H (2004) Ectoin: an effective natural substance to prevent UVA-induced premature photoaging. Skin Pharmacol Physiol 17:232–237CrossRefGoogle Scholar
  14. Bünger J, Degwert J, Driller H (2001) The protective function of compatible solute ectoine on the skin cells and its biomolecules with respect to UV-radiation, immunosuppression and membrane damage. IFSCC Mag 4:1–6Google Scholar
  15. Buommino E, Schiraldi C, Baroni A, Paoletti I, Lamberti M, De Rosa M, Tufano MA (2005) Ectoine from halophilic microorganisms induces the expression of hsp70 and hsp70B′ in human keratinocytes modulating the proinflammatory response. Cell Stress Chaperones 10:197–203CrossRefGoogle Scholar
  16. Chen L, Roberts MF (1998) Cloning and expression of the inositol monophosphatase gene from Methanococcus jannaschii and characterization of the enzyme. Appl Environ Microbiol 64:2609–2615Google Scholar
  17. Chen L, Spiliotis ET, Roberts MF (1998) Biosynthesis of di-myo-inositol-1,1′-phosphate, a novel osmolyte in hyperthermophilic archaea. J Bacteriol 180:3785–3792Google Scholar
  18. Ciulla RA, Diaz MR, Taylor BF, Roberts MF (1997) Organic Osmolytes in Aerobic Bacteria from Mono Lake, an Alkaline, Moderately Hypersaline Environment. Appl Environ Microbiol 63:220–226Google Scholar
  19. Cruz PE, Silva AC, Roldao A, Carmo M, Carrondo MJ, Alves PM (2006) Screening of novel excipients for improving the stability of retroviral and adenoviral vectors. Biotechnol Prog 22:568–576CrossRefGoogle Scholar
  20. Dobson CM (2003) Protein folding and disease: a view from the first Horizon Symposium. Nat Rev Drug Discov 2:154–160CrossRefGoogle Scholar
  21. Empadinhas N, Albuquerque L, Costa J, Zinder SH, Santos MA, Santos H, da Costa MS (2004) A gene from the mesophilic bacterium Dehalococcoides ethenogenes encodes a novel mannosylglycerate synthase. J Bacteriol 186:4075–4084CrossRefGoogle Scholar
  22. Faria TQ, Lima JC, Bastos M, Macanita AL, Santos H (2004) Protein stabilization by osmolytes from hyperthermophiles: effect of mannosylglycerate on the thermal unfolding of recombinant nuclease a from Staphylococcus aureus studied by picosecond time-resolved fluorescence and calorimetry. J Biol Chem 279:48680–48691CrossRefGoogle Scholar
  23. Flock S, Labarbe R, Houssier C (1996) 23Na NMR study of the effect of organic osmolytes on DNA counterion atmosphere. Biophys J 71:1519–1529Google Scholar
  24. Foord RL, Leatherbarrow RJ (1998) Effect of osmolytes on the exchange rates of backbone amide protons in proteins. Biochemistry 37:2969–2978CrossRefGoogle Scholar
  25. Frings E, Sauer T, Galinski EA (1995) Production of hydroxyectoine: high cell-density cultivation and osmotic downshock of Marinococcus strain M52. J Biotechnol 43:53–61CrossRefGoogle Scholar
  26. Furusho K, Yoshizawa T, Shoji S (2005) Ectoine alters subcellular localization of inclusions and reduces apoptotic cell death induced by the truncated Machado–Joseph disease gene product with an expanded polyglutamine stretch. Neurobiol Dis 20:170–178CrossRefGoogle Scholar
  27. Galinski EA, Pfeiffer HP, Truper HG (1985) 1,4,5,6-Tetrahydro-2-methyl-4-pyrimidinecarboxylic acid. A novel cyclic amino acid from halophilic phototrophic bacteria of the genus Ectothiorhodospira. Eur J Biochem 149:135–139CrossRefGoogle Scholar
  28. Galinski EA, Stein M, Amendt B, Kinder M (1997) The kosmotropic (structure-forming) effect of compensatory solutes. Comp Biochem Physiol 117A:357–365CrossRefGoogle Scholar
  29. Göller K, Galinski EA (1999) Protection of a model enzyme (lactate dehydrogenase) against heat, urea and freeze-thaw treatment by compatible solute additives. J Mol Catal B Enzym 7:37–45CrossRefGoogle Scholar
  30. Grammann K, Volke A, Kunte HJ (2002) New type of osmoregulated solute transporter identified in halophilic members of the bacteria domain: TRAP transporter TeaABC mediates uptake of ectoine and hydroxyectoine in Halomonas elongata DSM 2581(T). J Bacteriol 184:3078–3085CrossRefGoogle Scholar
  31. Grether-Beck S, Timmer A, Felsner I, Brenden H, Brammertz D, Krutmann J (2005) Ultraviolet A-induced signaling involves a ceramide-mediated autocrine loop leading to ceramide de novo synthesis. J Invest Dermatol 125:545–553CrossRefGoogle Scholar
  32. Harjes S, Scheidig A, Bayer P (2004) Expression, purification and crystallization of human 3′-phosphoadenosine-5′-phosphosulfate synthetase 1. Acta Crystallogr D Biol Crystallogr 60:350–352CrossRefGoogle Scholar
  33. Hensel R, Jakob I (1994) Stability of Glyceraldehyde-3-Phosphate Dehydrogenases from Hyperthermophilic Archaea at High Temperature. Syst Appl Microbiol 16:742–745Google Scholar
  34. Hinrichsen LL, Montel MC, Talon R (1994) Proteolytic and lipolytic activities of Micrococcus roseus (65), Halomonas elongata (16) and Vibrio sp. (168) isolated from Danish bacon curing brines. Int J Food Microbiol 22:115–126CrossRefGoogle Scholar
  35. Inbar L, Lapidot A (1988) The structure and biosynthesis of new tetrahydropyrimidine derivatives in actinomycin D producer Streptomyces parvulus. Use of 13C- and 15N-labeled L-glutamate and 13C and 15N NMR spectroscopy. J Biol Chem 263:16014–16022Google Scholar
  36. Jebbar M, Talibart R, Gloux K, Bernard T, Blanco C (1992) Osmoprotection of Escherichia coli by ectoine: uptake and accumulation characteristics. J Bacteriol 174:5027–5035Google Scholar
  37. Jebbar M, von Blohn C, Bremer E (1997) Ectoine functions as an osmoprotectant in Bacillus subtilis and is accumulated via the ABC-transport system OpuC. FEMS Microbiol Lett 154:325–330CrossRefGoogle Scholar
  38. Jebbar M, Sohn-Bosser L, Bremer E, Bernard T, Blanco C (2005) Ectoine-induced proteins in Sinorhizobium meliloti include an Ectoine ABC-Type transporter involved in osmoprotection and ectoine catabolism. J Bacteriol 187:1293–1304CrossRefGoogle Scholar
  39. Kanapathipillai M, Lentzen G, Sierks M, Park CB (2005) Ectoine and hydroxyectoine inhibit aggregation and neurotoxicity of Alzheimer’s beta-amyloid. FEBS Lett 579:4775–4780CrossRefGoogle Scholar
  40. Kanodia S, Roberts MF (1983) Methanophosphagen: Unique cyclic pyrophosphate isolated from Methanobacterium thermoautotrophicum. Proc Natl Acad Sci USA 80:5217–5221Google Scholar
  41. Karsten U, West JA, Ganesan EK (1993) Comparative physiological ecology of Bostrychia moritziana (Ceramiales, Rhodophyta) from freshwater and marine habitats. Phycologia 32:401–409Google Scholar
  42. Kolp S, Pietsch M, Galinski EA, Gütschow M (2003) Protection against proteolytic cleavage by compatible solutes. In: Polish–Austrian–German–Hungarian–Italian joint meeting on medicinal chemistry, KrakowGoogle Scholar
  43. Kraegeloh A, Kunte HJ (2002) Novel insights into the role of potassium for osmoregulation in Halomonas elongata. Extremophiles 6:453–462CrossRefGoogle Scholar
  44. Lamosa P, Burke A, Peist R, Huber R, Liu MY, Silva G, Rodrigues-Pousada C, LeGall J, Maycock C, Santos H (2000) Thermostabilization of proteins by diglycerol phosphate, a new compatible solute from the hyperthermophile Archaeoglobus fulgidus. Appl Environ Microbiol 66:1974–1979CrossRefGoogle Scholar
  45. Lamosa P, Turner DL, Ventura R, Maycock C, Santos H (2003) Protein stabilization by compatible solutes. Effect of diglycerol phosphate on the dynamics of Desulfovibrio gigas rubredoxin studied by NMR. Eur J Biochem 270:4606–4614CrossRefGoogle Scholar
  46. Lapidot A, Iakobashvili R, Malin G (1999) Methods for DNA amplification and sequencing. Patent WO1999041410Google Scholar
  47. Lentzen G, Schwarz T (2005) Kompatible Solute: Mikrobielle Herstellung und Anwendung. In: Anthranikian G (ed) Angewandte Mikrobiologie. Springer, Berlin Heidelberg New York, pp 355–372Google Scholar
  48. Lippert K, Galinski EA (1992) Enzyme stabilization by ectoine-type compatible solutes: protection against heating, freezing and drying. Appl Microbiol Biotechnol 37:61–65CrossRefGoogle Scholar
  49. Louis P, Trüper HG, Galinski EA (1994) Survival of Escherichia coli during drying and storage in the presence of compatible solutes. Appl Microbiol Biotechnol 41:684–688Google Scholar
  50. Malin G, Lapidot A (1996) Induction of synthesis of tetrahydropyrimidine derivatives in Streptomyces strains and their effect on Escherichia coli in response to osmotic and heat stress. J Bacteriol 178:385–395CrossRefGoogle Scholar
  51. Manzanera M, Garcia de Castro A, Tondervik A, Rayner-Brandes M, Strom AR, Tunnacliffe A (2002) Hydroxyectoine is superior to trehalose for anhydrobiotic engineering of Pseudomonas putida KT2440. Appl Environ Microbiol 68:4328–4333CrossRefGoogle Scholar
  52. Manzanera M, Vilchez S, Tunnacliffe A (2004) High survival and stability rates of Escherichia coli dried in hydroxyectoine. FEMS Microbiol Lett 233:347–352CrossRefGoogle Scholar
  53. Martins LO, Santos H (1995) Accumulation of mannosylglycerate and di-myo-inositol-phosphate by Pyrococcus furiosus in response to salinity and temperature. Appl Environ Microbiol 61:3299–3303Google Scholar
  54. Martins LO, Carreto LS, Da Costa MS, Santos H (1996) New compatible solutes related to di-myo-inositol-phosphate in members of the order Thermotogales. J Bacteriol 178:5644–5651Google Scholar
  55. Matussek K, Moritz P, Brunner N, Eckerskorn C, Hensel R (1998) Cloning, sequencing, and expression of the gene encoding cyclic 2, 3-diphosphoglycerate synthetase, the key enzyme of cyclic 2, 3-diphosphoglycerate metabolism in Methanothermus fervidus. J Bacteriol 180:5997–6004Google Scholar
  56. Melo EP, Faria TQ, Martins LO, Gonçalves AM, Cabral JMS (2001) Cutinase unfolding and stabilization by trehalose and mannosylglycerate. Proteins 42:542–552CrossRefGoogle Scholar
  57. Moritz P (2003) Die Bedeutung der cyclischen 2,3-Diphosphoglycerat Synthetase für die temperaturabhänige Regulation der intrazellulären Konzentration von cyclischem 2,3-Diphosphoglycerat im hyperthermophilen Archaeum Methanothermus fervidus. Ph.D. thesis, Universität Duisburg-EssenGoogle Scholar
  58. Nakayama H, Yoshida K, Ono H, Murooka Y, Shinmyo A (2000) Ectoine, the compatible solute of Halomonas elongata, confers hyperosmotic tolerance in cultured tobacco cells. Plant Physiol 122:1239–1247CrossRefGoogle Scholar
  59. Pais TM, Lamosa P, dos Santos W, Legall J, Turner DL, Santos H (2005) Structural determinants of protein stabilization by solutes. The important of the hairpin loop in rubredoxins. Febs J 272:999–1011Google Scholar
  60. Ramos A, Raven ND, Sharp RJ, Bartolucci S, Rossi M, Cannio R, Lebbink J, Oost Jvd, Vos WMd, Santos H (1997) Stabilization of enzymes against thermal stress and freeze-drying by mannosylglycerate. Appl Environ Microbiol 63:4020–4025Google Scholar
  61. Rattray FP, Fox PF (1999) Aspects of enzymology and biochemical properties of brevibacterium linens relevant to cheese ripening: a review. J Dairy Sci 82:891–909CrossRefGoogle Scholar
  62. Rothschild LJ, Mancinelli RL (2001) Life in extreme environments. Nature 409:1092–1101CrossRefGoogle Scholar
  63. Santos H, da Costa MS (2002) Compatible solutes of organisms that live in hot saline environments. Environ Microbiol 4:501–509CrossRefGoogle Scholar
  64. Santos H, Lamosa P, Burke A, Maycock C (1998) Thermostabilization, osmoprotection and protection against desiccation of enzymes, cell components and cells by di-glycerol-phosphate. European Patent EP0965268Google Scholar
  65. Sauer T, Galinski EA (1998) Bacterial milking: a novel bioprocess for production of compatible solutes. Biotechnol Bioeng 57:306–313CrossRefGoogle Scholar
  66. Scholz S, Sonnenbichler J, Schafer W, Hensel R (1992) Di-myo-inositol-1,1′-phosphate: a new inositol phosphate isolated from Pyrococcus woesei. FEBS Lett 306:239–242CrossRefGoogle Scholar
  67. Silva Z, Borges N, Martins LO, Wait R, da Costa MS, Santos H (1999) Combined effect of the growth temperature and salinity of the medium on the accumulation of compatible solutes by Rhodothermus marinus and Rhodothermus obamensis. Extremophiles 3:163–172CrossRefGoogle Scholar
  68. Steger R, Weinand M, Kramer R, Morbach S (2004) LcoP, an osmoregulated betaine/ectoine uptake system from Corynebacterium glutamicum. FEBS Lett 573:155–160CrossRefGoogle Scholar
  69. Valentao P, Fernandes E, Carvalho F, Andrade PB, Seabra RM, de Lourdes Bastos M (2002) Antioxidant activity of Hypericum androsaemum infusion: scavenging activity against superoxide radical, hydroxyl radical and hypochlorous acid. Biol Pharm Bull 25:1320–1323CrossRefGoogle Scholar
  70. Vermeulen V, Kunte HJ (2004) Marinococcus halophilus DSM 20408T encodes two transporters for compatible solutes belonging to the betaine-carnitine-choline transporter family: identification and characterization of ectoine transporter EctM and glycine betaine transporter BetM. Extremophiles 8:175–184CrossRefGoogle Scholar
  71. Vreeland RH, Litchfield CD, Martin EL, Elliot E (1980) Halomonas elongata, a new genus and species of extremely salt-tolerant bacteria. Int J Syst Bacteriol 30:485–495Google Scholar
  72. Yasuhiko Y, Yoshio K, Eiichiro K, Yasuhiro T, Seichi S (1997) Production of l-amino acid by fermentation method using ectoine. Japanese Patent JP9271382, 21 Oct 1997Google Scholar
  73. Yu I, Nagaoka M (2004) Slowdown of water diffusion around protein in aqueous solution with ectoine. Chem Phys Lett 388:316–321CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  1. 1.bitop AGWittenGermany

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