Biotechnological Aspects of Cold-Adapted Enzymes

  • Adrienne L. Huston

Beginning with the development of basic tools by our hominid predecessors, humanshave continually searched for and utilized novel materials from the natural environment to survive and thrive. Today, our knowledge of the surrounding world extends to themolecular scale as we enter the age of genomics and systems biology, enabling previously unimaginable insight into processes that promise application in the agricultural, energy, food, medical, structural material and textile industries. As environmental concerns arise, biological tools are increasingly replacing harsh chemical and physical means of processing materials and they even harbor promise for creating cost-effective sustainable energy sources. It is imperative that we continue investigating ways in which natural products can offer economical alternatives to traditional industrial processes. Due to the growing and wide-spread use of enzymes in a variety of industrial applications, this review aims to build on previous works (Brenchley 1996; Ohgiya et al. 1999; Gerday et al. 2000; Allen et al. 2001; Cavicchioli et al. 2002) by illustrating recent advances and potential opportunities for the biotechnological application of cold-adapted enzymes.


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  1. Allen D, Huston AL, Wells LE, Deming JW (2001) Biotechnological use of psychrophiles. In Bitton G (ed) Encyclopedia of environmental microbiology. Wiley, New York, pp 1–17.Google Scholar
  2. Arnold FH, Wintrode PL, Miyazaki K, Gershenson A (2001) How enzymes adapt: lessons from directed evolution. Trends Biochem Sci 26:100–106.CrossRefPubMedGoogle Scholar
  3. Asenjo JA, Andrews BA, Reyes F, Salamanca M, Burzio L (2006) Protein and nucleic acid sequence encoding a krill-derived cold adapted trypsin-like activity enzyme. Patent No. WO2006022947.Google Scholar
  4. Bansal V, Rautaray D, Ahmad A, Sastry M (2004) Biosynthesis of zirconia nanoparticles using the fungus Fusarium oxysporum. J Mater Chem 14:3303–3305.CrossRefGoogle Scholar
  5. Benesova E, Markova M, Kralova B (2005) B-glucosidase and–glucosidase from psychrotrophic strain Arthrobacter sp C2–2. Czech J Food Sci 23:116–120.Google Scholar
  6. Bentahir M, Feller G, Aittaleb M, Lamotte-Brasseur J, Himri T, Chessa J-P, Gerday C (2000) Structural, kinetic, and calorimetric characterization of the cold-active phosphoglycerate kinase from the Antarctic Pseudomonas sp. TACII18. J Biol Chem 275:11147–11153.CrossRefPubMedGoogle Scholar
  7. Bjarnason JB, Benediktsson B (2001) Protein hydrolysates produced with the use of marine proteases. Patent No. WO0128353.Google Scholar
  8. Bordusa F (2002) Proteases in organic synthesis. Chem Rev 102:4817–4867.CrossRefPubMedGoogle Scholar
  9. Brenchley JE (1996) Psychrophilic microorganisms and their cold-active enzymes. J Indust Microbiol 17:432–437.CrossRefGoogle Scholar
  10. Brenchley JE, Loveland-Curtze J, Gutshall K, Humphrey V (2001) Stain removing compositions containing particular isolated and pure proteolytic enzymes. Patent No. US6326346.Google Scholar
  11. Brutchey RL, Yoo ES, Morse DE (2006) Biocatalytic synthesis of a nanostructured and crystalline bimetallic perovskite-like barium oxofluorotitanate at low temperature. J Am Chem Soc 128:10288–10294.CrossRefPubMedGoogle Scholar
  12. Cavicchioli R (2006) Cold-adapted archaea. Nat Rev Microbiol 4:331–343.CrossRefPubMedGoogle Scholar
  13. Cavicchioli R, Siddiqui KS, Andrews D, Sowers KR (2002) Low-temperature extremophiles and their applications. Curr Opin Biotechnol 13:253–161.CrossRefPubMedGoogle Scholar
  14. Cha JN, Shimizu K, Zhou Y, Christiansen SC, Chmelka BF, Stucky GD, Morse DE (1999) Silicatein filaments and subunits from a marine sponge direct the polymerization of silica and silicones in vitro. Proc Natl Acad Sci USA 96:361–365.CrossRefPubMedGoogle Scholar
  15. Cherry JR, Fidanstef AL (2003) Directed evolution of industrial enzymes: an update. Curr Opin Biotechnol 14:438–443.CrossRefPubMedGoogle Scholar
  16. Coker JA, Brenchley JE (2006) Protein engineering of a cold-active ß-galactosidase from Arthrobacter sp. SB to increase lactose hydrolysis reveals new sites affecting low temperature activity. Extremophiles 10:515–524.CrossRefPubMedGoogle Scholar
  17. Collins T, Hoyoux A, Dutron A, Georis J, Genot B, Dauvrin T, Arnaut F, Gerday C, Feller G (2006) Use of glycoside family 8 xylanases in baking. J Cereal Sci 43:79–84.CrossRefGoogle Scholar
  18. D’Amico S, Gerday C, Feller G (2001) Structural determinants of cold adaptation and stability in a large protein. J Biol Chem 276:25791–25796.CrossRefPubMedGoogle Scholar
  19. D’Amico S, Marx J-C, Gerday C, Feller G (2003a) Activity-stability relationships in extremophilic enzymes. J Biol Chem 278:7891–7896.CrossRefPubMedGoogle Scholar
  20. D’Amico S, Gerday C, Feller G (2003b) Temperature adaptation of proteins: Engineering mesophilic-like activity and stability in a cold-adapted i-amylase. J Mol Biol 332:981–988.CrossRefPubMedGoogle Scholar
  21. D’Amico S, Collins T, Marx J-C, Feller G, Gerday C (2006) Psychrophilic microorganisms: challenges for life. EMBO Reports 7:385–389.CrossRefPubMedGoogle Scholar
  22. Deming JW (2002) Psychrophiles and polar regions. Curr Opin Microbiol 3:301–309.CrossRefGoogle Scholar
  23. DeVries R, Visser J (2001) Aspergillus enzymes involved in degradation of plant cell wall polysaccharides. Microbiol Mol Biol Rev 65:497–522.CrossRefGoogle Scholar
  24. Dutron A, Georis J, Dauvrin T, Collins T, Hoyoux A, Feller G (2005) Use of family 8 enzymes with xylanolytic activity in baking. Patent No. MXPA05002751.Google Scholar
  25. Felby C, Larsen D, Joergensen H, Vibe-Pederson J (2006) Enzymatic hydrolysis of biomasses having a high dry matter (DM) content. Patent No. WO2006056838.Google Scholar
  26. Feller G, Gerday C (2003) Psychrophilic enzymes: Hot topics in cold adaptation. Nat Rev Microbiol 1:200–208.CrossRefPubMedGoogle Scholar
  27. Georlette D, Jonsson ZO, Van Petegem F, Chessa J-P, Van Beeumen J, Hubscher U, Gerday C (2000) A DNA ligase from psychrophile Pseudoalteromonas haloplanktis gives insights into the adaptation of proteins to low temperatures. Eur J Biochem 267:3502–3512.CrossRefPubMedGoogle Scholar
  28. Gerday C, Aittaleb M, Arpigny JL, Baise E, Chessa J-P, Garsoux G, Perescu I, Feller G (1997) Psychrophilic enzymes: a thermodynamic challenge. Biochim Biophys Acta 1342:119–131.PubMedGoogle Scholar
  29. Gerday C, Aittaleb M, Benhatir M, Chessa J-P, Claverie P, Collins T, D’Amico S, Dumont, J, Garsoux G, Georlette D, Hoyoux A, Lonhienne T, Meuwis M-A, Feller G (2000) Cold-adapted enzymes: from fundamentals to biotechnology. TIBTECH 18:103–107.Google Scholar
  30. Gerday C, Hoyoux A, Francois J-M, Dubois P, Baise E, Jennes I, Genicot S (2005) Cold-active ß-galactosidase, the process for its preparation and the use thereof. Patent No. US2005196835.Google Scholar
  31. Gerike U, Danson MJ, Hough DW (2001) Cold-active citrate synthase: mutagenesis of active-site residues. Protein Eng 14:655–661.CrossRefPubMedGoogle Scholar
  32. Giovannoni SJ, Britschgi TB, Moyer CL, Field KG (1990) Genetic diversity in Sargasso sea bacterioplankton. Nature 345:60–63.CrossRefPubMedGoogle Scholar
  33. Hasan AKMQ, Tamiya E (1997) Cold-active protease CP-58 and psychrotrophic bacteria. Patent No. WO9730172.Google Scholar
  34. Hasan AKMQ, Tamiya E (1998) Cold-active protease CP70. Patent No. US6200793.Google Scholar
  35. He H, Chen XL, Li JW, Zhang YZ, Gao PJ (2004) Taste improvement of refrigerated meat treated with cold-adapted protease. Food Chem 84:307–311.CrossRefGoogle Scholar
  36. Karasova-Lipovova P, Strnad H, Spiwok V, Mala S, Kralova B, Russell NJ (2003) The cloning, purification and characterization of a cold-active t-galactosidase from the psychrotolerant Antarctic bacterium Arthrobacter sp. C2–2. Enzyme Microb Technol 33:836–844.CrossRefGoogle Scholar
  37. Kisailus D, Choi JH, Weaver JC, Yang W, Morse DE (2005) Enzymatic synthesis and nanostructural control of gallium oxide at low temperature. Adv Mater 17:314–318.CrossRefGoogle Scholar
  38. Kisailus D, Truong Q, Amemiya Y, Weaver JC, Morse DE (2006) Self-assembled bifunctional surface mimics an enzymatic and templating protein for the synthesis of a metal oxide semiconductor. Proc Natl Acad Sci USA 103:5652–5657.CrossRefPubMedGoogle Scholar
  39. Krishna, SH (2002) Developments and trends in enzyme catalysis in nonconventional media. Biotech Adv 20:239–267.CrossRefGoogle Scholar
  40. Kulakova L, Galkin A, Nakayama T, Nishino T, Esaki N (2003) Improvement of thermostability of cold-active serine alkaline protease from the psychrotrophic bacterium Shewanella sp. Strain Ac10 by rational mutagenesis. J Mol Catal B Enzymatic 22:113–117.CrossRefGoogle Scholar
  41. Li JK, Lee TC (1995) Bacterial ice nucleation and its potential application in the food industry. Trends Food Sci Technol 6:259–265.CrossRefGoogle Scholar
  42. Lin Y, Tanaka S (2006) Ethanol fermentation from biomass resources; current state and prospects. Appl Microbiol Biotechnol 69:627–642.CrossRefPubMedGoogle Scholar
  43. Lonhienne T, Gerday C, Feller G (2000) Psychrophilic enzymes: revisiting the thermodynamic parameters of activation may explain local flexibility. Biochim Biophys Acta 1543:1–10.PubMedGoogle Scholar
  44. Lonhienne T, Zoidakis J, Vorgias CE, Feller G, Gerday C, Bouriotis V (2001) Modular structure, local flexibility and cold-activity of a novel chitobiase from a psychrophilic Antarctic bacterium. J Mol Biol 310:291–297.CrossRefPubMedGoogle Scholar
  45. Lorenz P, Liebeton K, Niehaus F, Eck J (2002) Screening for novel enzymes for biocatalytic processes: accessing the metagenome as a resource of novel functional sequence space. Curr Opin Biotechnol 13:572–577.CrossRefPubMedGoogle Scholar
  46. Margesin R, Feller G, Gerday C, Russell N (2002) Cold-adapted microorganisms: adaptation strategies and biotechnological potential. In Bitton G (ed) The encyclopedia of environmental microbiology. Wiley, New York, pp 871–885.Google Scholar
  47. Mavromatis K, Feller G, Kokkinidis M, Bouriotis V (2003) Cold adaptation of a psychrophilic chitinase: a mutagenesis study. Protein Eng 16:497–503.CrossRefPubMedGoogle Scholar
  48. Michel V, Lehoux I, Depret G, Anglade P, Labadie J, Hebraud M (1997) The cold shock response of the psychrotrophic bacterium Pseudomonas fragi involves four low-molecular-mass nucleic acid binding proteins. J Bacteriol 23:7331–7342.Google Scholar
  49. Miyazaki K, Wintrode PL, Grayling RA, Rubingh DN, Arnold FH (2000) Directed evolution study of temperature adaptation in a psychrophilic enzyme. J Mol Biol 297:1015–1026.CrossRefPubMedGoogle Scholar
  50. Muryoi N, Sato M, Kaneko S, Kawahara H, Obata h, Yaish MW, Griffith M, Glick BR (2004) Cloning and expression of afpA, a gene encoding an antifreeze protein from the arctic plant growth-promoting rhizobacterium Pseudomonas putida GF12–2. J Bacteriol 186:5661–5671.CrossRefPubMedGoogle Scholar
  51. Nakagawa T, Nagaoka T, Taniguchi S, Miyaji T, Tomizuka N (2004) Isolation and characterization of psychrophilic yeasts producing cold-adapted pectinolytic enzymes. Lett Appl Microbiol 38:383–387.CrossRefPubMedGoogle Scholar
  52. Narinx E, Baise E, Gerday C (1997) Subtilisin from psychrophilic Antarctic bacteria: characterization and site-directed mutagenesis of residues possibly involved in the adaptation to cold. Protein Eng 10:1271–1279.CrossRefPubMedGoogle Scholar
  53. Nichols CM, Lardiere SG, Bowman JP, Nichols PD, Gibson JAE, Guezennec J (2005) Chemical characterization of exopolysaccharides from Antarctic marine bacteria. Microb Ecol 49:578–589.CrossRefPubMedGoogle Scholar
  54. Novozymes (2005) Presentation from the ninth chemical industry finance and investments conference. URL:–4B62–A1EB-5BBE6A55468E/0/Chemicals_Conference_ML_2005.pdf.
  55. Ohgiya S, Hoshino T, Okuyama H, Tanaka S, Ishizaki K (1999) Biotechnology of enzymes from cold-adapted microorganisms. In: Margesin R, Schinner F (eds) Biotechnological applications of cold-adapted organisms. Springer, Berlin, pp 17–34.Google Scholar
  56. Ohtani N, Haruki M, Morikawa M, Kanaya S (2001) Heat labile ribonuclease HI from a psychrotrophic bacterium: gene cloning, characterization and site-directed mutagenesis. Protein Eng 14:975–982.CrossRefPubMedGoogle Scholar
  57. Owusu-Apenten RK (1999) Low temperature organic phase biocatalysis using cold-adapted enzymes. In: Margesin R, Schinner F (eds) Biotechnological applications of cold-adapted organisms. Springer, Berlin, pp 35–48.Google Scholar
  58. Panasik N (2002) Structural basis for thermostability and thermal dependence of activity in P// barrel glycosyl hydrolases. PhD Thesis, Pennsylvania State University, USA.Google Scholar
  59. Peck LS (2002) Ecophysiology of Antarctic marine ectotherms: limits to life. Polar Biol 25:31–40.CrossRefGoogle Scholar
  60. Pomeroy LR, Wiebe WJ (2001) Temperature and substrates as interactive limiting factors for marine heterotrophic bactera. Aquat Microb Ecol 23:187–204.CrossRefGoogle Scholar
  61. Rasmussen BF, Stock AM, Rings D, Petsko GA (1992) Crystallin ribonuclease A loses function below the dynamical transition at 220 K. Nature 357:423–424.CrossRefPubMedGoogle Scholar
  62. Rina M, Pozidis C, Mavromatis K, Tzanodaskalaki M, Kokkinidis M, Bouriotis V (2000) Alkaline phosphatase from the Antarctic strain TAB5. Eur J Biochem 267:1230–1238.CrossRefPubMedGoogle Scholar
  63. Robertson GH, Wong DWS, Lee CC, Wagschal K, Smith MR, Orts WJ (2006) Native or raw starch digestion: a key step in energy efficient biorefining of grain. J Agric Food Chem 54: 353–365.CrossRefPubMedGoogle Scholar
  64. Russell NJ (1997) Psychrophilic bacteria-molecular adaptations of membrane lipids. Comp Biochem Physiol Physiol 118:489–493.CrossRefGoogle Scholar
  65. Schoemaker HE, Mink E, Wubbolts MG (2003) Dispelling the myths—biocatalysis in industrial synthesis. Science 299:1694–1697.CrossRefPubMedGoogle Scholar
  66. Sellek GA, Chaudhuri JB (1999) Biocatalysis in organic media using enzymes from extremophiles. Enzyme Microb Technol 25:471–482.CrossRefGoogle Scholar
  67. Senapati S, Ahmad A, Khan MI, Sastry M, Kumar R (2005) Extracellular biosynthesis of bimetallic Au-Ag alloy nanoparticles. Small 5:517–520.CrossRefGoogle Scholar
  68. Shahidi F, Kamil YVAJ (2001) Enzymes from fish and aquatic invertebrates and their application in the food industry. Trends Food Sci Technol 12:435–464.CrossRefGoogle Scholar
  69. Shetty JK, Lantero OJ, Dunn-Colemen N (2005) Technological advances in ethanol production. Int Sugar J 107:605.Google Scholar
  70. Shimizu K, Cha J, Stucky GD, Morse DE (1998) Silicatein h: Cathespin L-like protein in sponge biosilica. Proc Natl Acad Sci USA 95:6234–6238.CrossRefPubMedGoogle Scholar
  71. Stinson SC (1998) Counting on chiral drugs. Chem Eng News 76:83–96.Google Scholar
  72. Suen W-C, Zhang N, Xiao L, Madison V, Zaks A (2004) Improved activity and thermostability of Candida antarctica lipase B by DNA family shuffling. PEDS 17:133–140.PubMedGoogle Scholar
  73. Sumerel JL, Yang W, Kisailus D, Weaver JC, Choi JH, Morse DE (2003) Biocatalytically templated synthesis of titanium oxide. Chem Mater 15:4804–4809.CrossRefGoogle Scholar
  74. Tahir MN, Eberhardt M, Therese HA, Kolb U, Theato P, Muller WEG, S H-C, Tremel W (2006) From single molecules to nanoscopically structured functional materials: Au nanocrystal growth on TiO2 nanowires controlled by surface-bound silicatein. Angew Chem Int Ed 45:4803–4809.CrossRefGoogle Scholar
  75. Takaiwa M, Saeki K, Okuda M, Kobayashi T, Ito S, Kubota H, Ota Y, Fujimori N (1997) Cold alkaline protease, microorganism producing the same, process for producing the same, and detergent compositions and food processing enzyme preparations containing the same. Patent No. WO9743406.Google Scholar
  76. Trytek M, Fiedurek J (2005) A novel psychrotrophic fungus, Mortierella minutissima, for D-limonene biotransformation. Biotechnol Lett 27:149–153.CrossRefPubMedGoogle Scholar
  77. van den Burg B (2003) Extremophiles as a source for novel enzymes. Current Opin Microbiol 6:213–218.CrossRefGoogle Scholar
  78. Wells LE, Deming JW (2006) Characterization of a cold-active bacteriophage on two psychrophilic marine hosts. Aquat Microb Ecol 45:15–29.CrossRefGoogle Scholar
  79. Wyman CE (2003) Potential synergies and challenges in refining cellulosic biomass to fuels, chemicals, and power. Biotechnol Prog 19:254–262.CrossRefPubMedGoogle Scholar
  80. Zecchinon L, Oriol A, Netzel U, Svennberg J, Gerardin-Otthiers N, Feller G (2005) Stability domains, substrate-induced conformational changes, and hinge-bending motions in a psychrophilic phosphyglycerate kinase—A microcalorimetric study. J Biol Chem 280:41307–41314.CrossRefPubMedGoogle Scholar

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© Springer-Verlag Berlin Heidelberg 2008

Authors and Affiliations

  • Adrienne L. Huston
    • 1
  1. 1.2006–2007 AAAS Science and Technology Policy FellowAmerican Association for the Advancement of Science (AAAS)WashingtonUSA

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