Polar Biology

, Volume 28, Issue 4, pp 319–325

Properties of extracellular proteases from three psychrotolerant Stenotrophomonas maltophilia isolated from Antarctic soil

  • Susana Vazquez
  • Lucas Ruberto
  • Walter Mac Cormack
Original Paper


Three Antarctic psychrotolerant Stenotrophomonas maltophilia were isolated and the characteristics of their extracellular serine proteases were described. The isolates were able to grow at 14 and 34°C, but grew better between 20 and 28°C. The highest protease secretion was reached at 20–24°C. The purified enzyme preparations had maximal activity at 55–60°C and alkaline pH. They showed high pH stability, retaining more than 60% of residual activity after 3 h of incubation at a pH range of 4–12. The thermal stability was slightly lower compared with a commercial mesophilic protease, with 74–79% residual activity after 90 min at 40°C and 50% inactivation at 50°C between 43 and 69 min. These properties suggest that the Antarctic isolates could be adapted to cold by means of synthesising more enzymes with high activity but that the proteases they produce are not truly cold-active, being more similar to mesophilic enzymes.


  1. Cole JR, Chai B, Marsh TL, Farris RJ, Wang Q, Kulam SA, Chandra S, McGarrell DM, Schmidt TM, Garrity GM, Tiedje JM (2003) The Ribosomal Database Project (RDP-II): previewing a new autoaligner that allows regular updates and the new prokaryotic taxonomy. Nucleic Acids Res 31:442–443CrossRefPubMedGoogle Scholar
  2. Deming JW (2002) Psychrophiles and polar regions. Curr Opin Microbiol 5:301–309CrossRefPubMedGoogle Scholar
  3. Feller G (2003) Molecular adaptations to cold in psychrophilic enzymes. CMLS Cell Mol Life Sci 60:648–662CrossRefGoogle Scholar
  4. Feller G, Gerday C (1997) Psychrophilic enzymes: molecular basis of cold adaptation. CMLS Cell Mol Life Sci 53:830–841CrossRefGoogle Scholar
  5. Helmke E, Weyland H (1991) Effect of temperature on extracellular enzymes occurring in permanently cold marine environments. Kiel Meeresforsch Sonderh 8:198–204Google Scholar
  6. Heussen C, Dowdle EB (1980) Electrophoretic analysis of plasminogen activators in polyacrilamide gels containing sodium dodecyl sulfate and copolymerised substrates. Anal Biochem 102:196–202PubMedGoogle Scholar
  7. Hoshino T, Ishizaki K, Sakamoto T, Kumeta H, Yumoto I, Matsuyama H, Ohgiya S (1997) Isolation of a Pseudomonas species from fish intestine that produces a protease active at low temperature. Lett Appl Microbiol 25:70–72CrossRefPubMedGoogle Scholar
  8. Huston AL, Krieger-Brockett BB, Deming JW (2000) Remarkably low temperature optima for extracellular enzyme activity from Arctic bacteria and sea ice. Environ Microbiol 2:383–388CrossRefPubMedGoogle Scholar
  9. Irwin JA, Alfredsson GA, Lanzetti AJ, Gudmundsson HM, Engel PC (2001) Purification and characterisation of a serine peptidase from the marine psychrophile strain PA-43. FEMS Microbiol Lett 201:285–290CrossRefPubMedGoogle Scholar
  10. Kärst U, Woehl M, Czempinski K, Schmid RD (1994) Characterization of extracellular hydrolases from marine psychrophilic bacteria. ECB6 Proceedings of the 6th European congress in biotechonologyGoogle Scholar
  11. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227:680–685Google Scholar
  12. Margesin R, Schinner F (1991) Characterization of a metalloprotease from psychrophilic Xanthomonas maltophilia. FEMS Microbiol Lett 79:257–262CrossRefGoogle Scholar
  13. Margesin R, Schinner F (1992) A comparison of extracellular proteases from three psychrotrophic strains of Pseudomonas fluorescens. J Gen Appl Microbiol 38:209–225Google Scholar
  14. Morita RY (1975) Psychrophilic bacteria. Bacteriol Rev 39:144–167PubMedGoogle Scholar
  15. Nichols D, Bowman J, Sanderson K, Mancuso Nichols C, Lewis T, McMeekin T, Nichols PD (1999) Developments with Antarctic microorganisms: culture collections, bioactivity screening, taxonomy, PUFA production and cold-adapted enzymes. Curr Opin Biotechnol 10:240–246CrossRefPubMedGoogle Scholar
  16. Oikawa T, Kazuoka T, Soda K (2003) Paradoxical thermostable enzymes from psychrophile: molecular characterization and potentiality for biotechnological application. J Mol Cat B Enzymatic 23:65–70CrossRefGoogle Scholar
  17. Palleroni NJ, Bradbury JF (1993) Stenotrophomonas, a new bacterial genus for Xanthomonas maltophilia (Hugh 1980) Swings et al. 1983. Int J Syst Bacteriol 43:606–609PubMedGoogle Scholar
  18. Pirt SJ (1985) Effects of temperature. In: Pirt SJ (ed) Principles of microbial and cell cultivation. Blackwell, London, pp 137–142Google Scholar
  19. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring HarborGoogle Scholar
  20. Secades P, Alvarez B, Guijarro JA (2003) Purification and properties of a new psychrophilic metalloprotease (Fpp2) in the fish pathogen Flavobacterium psychrophilum. FEMS Microbiol Lett 226:273–279CrossRefPubMedGoogle Scholar
  21. Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchonic acid. Anal Biochem 150:76–85PubMedGoogle Scholar
  22. Vazquez SC, MacCormack WP (2002) Effect of isolation temperature on the characteristics of extracellular proteases produced by Antarctic bacteria. Polar Res 21:63–71Google Scholar
  23. Vazquez SC, Rios Merino LN, Mac Cormack WP, Fraile ER (1995) Protease-producing psycrotrophic bacteria isolated from Antarctica. Polar Biol 15:131–135CrossRefGoogle Scholar
  24. Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Susana Vazquez
    • 1
  • Lucas Ruberto
    • 2
  • Walter Mac Cormack
    • 3
  1. 1.CONICETBuenos AiresArgentina
  2. 2.Cátedra de Biotecnología, FFyBUniversidad de Buenos AiresBuenos AiresArgentina
  3. 3.Instituto Antártico ArgentinoBuenos AiresArgentina

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