Life at the Lower Limit:

1
Life at the Lower Limit

• Microbial Survival in Cryoenvironments

Elizabeth Connors
2
Questions

• What are some of the challenges facing
  microorganisms living in cryoenvironments?
• What are some of the methods used by
  psychrophiles to overcome these challenges?
• How can we benefit from understanding survival in
  cryoenvironments?

3
Overview Definitions

• Psycrophiles microorganisms that can survive,
  grow, and reproduce at temperatures of 5C or
  lower
• Cryoenvironments environments having average
  temperatures of 5C or lower

4
Cryoenvironments

• 90 marine environments
• below -1000m, oceans have a constant temperature
  of 5C
• 14 land surface is polar
• polar regions are located at latitudes greater
  than 66.33

National Geophysical Data Center Global Relief
Model (http//www.ngdc.noaa.gov/mgg/global/global.
html)
NASA Earth Observations (http//neo.sci.gsfc.nasa.
gov)
5
Challenges

• Decreased membrane fluidity
• 2. Decreased enzyme function
• 3. Ice crystal formation

Jean-Paul Baubet (http//fr.wikipedia.org/wiki/Fic
hierFleurgivre-1.jpg)
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Challenge 1 Decreased Membrane Fluidity

• The Problem rigid membranes, lower permeability,
  slower trans-membrane transport
• Solution alter membrane composition for maximum
  fluidity

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Challenge 1 Decreased Membrane Fluidity

• Case Study Pseudomonas syringae Lz4w (Seshu
  Kumar et al. 2002)
• -grown at 4C and 22C
• -composition of LPS compared
• -amount of hydroxy fatty acids in LPS increased
  at lower temperature

Acyl chain composition of LPS from low (4C)- and
high (22C)-temperature-grown cells of P.
syringaea
Amt (, mol/mol) grown at
a The values (average of three measurements) have
been rounded off to the nearest whole number.
8
Challenge 2 Decreased Enzyme Function

• The Problem cold denaturation and rigidity
• Solution lessen the stability of enzymes

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Challenge 2 Decreased Enzyme Function

• Case study proteome analysis was done on
  psychrophilic and mesophilic species (Metpally
  and Reddy 2009)
• -comparison of amino acid composition
• Observed significant differences
• 1. increased frequency of amino acids that
  avoid helices
• 2. increase in small/tiny and neutral group
  residues
• 3. decrease in aliphatic residues

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Challenge 3 Ice crystal formation

• The Problem ice crystal formation inhibits cell
  function and can rupture the cell
• Solution anti-freeze proteins, thermal hysteresis

11
Challenge 3 Ice crystal formation

• Case study Gilbert et al. (2005) studied the
  thermal hysteresis protein in the Antarctic
  bacterium Marinomonas primoryensis
• -hyperactive protein freeze tolerance vs.
  freeze avoidance
• -location intercellular, periplasmic space?

Ice crystal morphologies obtained with MpAFP. (a)
and (b) two different ice crystals obtained in
the presence of M. primoryensis crude lysate
supernatant at 0.3 C of under-cooling. (c) a
dendritic crystal growing extremely rapidly below
the non-equilibrium freezing point. Size marker
50 µm and relates to (a) and (b) only.
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Potential Applications?
13
References

• Amato, P. and Brent C. Christner. 2009. Energy
  metabolism response to low-temperature conditions
  in Psychrobacter cryohalolentis. Applied and
  Environmental Microbiology 75 711-718.
• Bakermans, C., Ayala-del-Rio, H.L., Ponder, M.A.,
  Vishnivetskaya, T., Gilichinsky, D., Thomashow,
  M.F., and J.M. Tiedje. 2006. Psychrobacter
  cryohalolentis sp. Nov. and Psychrobacter
  arcticus sp. Nov., isolated from Siberian
  permafrost. International Journal of Systematic
  and Evolutionary Microbiology 561285-1291.
• Bakermans, C., Tollaksen, S.L., Giometti, C.S.,
  Wilkerson, C., Tiedje, J.M., and M.F. Thomashow.
  2007. Proteomic analysis of Psychrobacter
  cryohalolentis K5 during growth at subzero
  temperatures. Extremophiles 11343-354.
• DAmico, S., Collins, T., Marx, J-C., Feller, G.,
  and C. Gerday. 2006. Psychrophilic
  microorganisms Challenges for life. EMBO Reports
  7385-389.
• Feller, G., and C. Gerday. 2003. Psychrophilic
  enzymes Hot topics in cold adaptation. Nat Rev
  Microbiol 1 200-208.
• Georlette, D., Blaise, V., Collins, T., DAmico,
  S., Gratia, E., Hoyoux, A., Marx, J-C., Sonan,
  G., Feller, G., and C. Gerday. 2004. Some like it
  cold Biocatalysis at low temperatures. FEMS
  Microbiology Reviews 2825-42.
• Gilbert, J.A., Davies, P.L., and J.
  Laybourn-Parry. 2005. A hyperactive
  Ca2-dependent antifreeze protein in an Antarctic
  bacterium. FEMS Microbiology Letters 24567-72.
• Hoyoux, A., Blaise, V., Collins, T., DAmico, S.,
  Gratia, E., Huston, A.L., Marx, J-C., Sonan, G.,
  Zeng, Y., Feller, G., and C. Gerday. 2004.
  Extreme catalysts from low-temperature
  environments. Journal of Bioscience and
  Bioengineering 98317-330.
• Janech, M.G., Krell, A., Mock, T., Kang, J-S.,
  and J.A. Raymond. 2006. Ice-binding proteins from
  sea ice diatoms (Bacillariophyceae). Journal of
  Pycology 42410-416.
• Kawamoto, J., Kurihara, T., Yamamoto, K.,
  Nagayasu, M., Tani, Y., Mihara, H., Hosokawa, M.,
  Baba, T., Sato, S.B., and N. Esaki. 2009.
  Eicosapentaenoic acid plays a beneficial role in
  membrane organization and cell division of a
  cold-adapted bacterium, Shewanella
  livingstonensis Ac10. Journal of Bacteriology
  191632-640.
• Metpally, R.P.R., and B.V.B. Reddy. 2009.
  Comparative proteome analysis of psychrophilic
  versus mesophilic bacterial species Insights
  into the molecular basis of cold adaption
  proteins. BMC Genomics 1011.
• Priscu, J.C. and B.C. Christner. 2004. Earths
  icy biosphere, p. 130-145. In T.A. Bull (ed.),
  Microbial diversity and bioprospecting. ASM
  Press, Washington, DC.
• Seshu Kumar, G., Jagannadham, M.V., and M.K. Ray.
  2002. Low-temperature-induced changes in
  composition and fluidity of lipopolysaccharides
  in the Antacrtic psychrotrophic bacterium
  Pseudomonas syringae. Journal of Bacteriology
  1846746-6749.
• Shcherbakova, V.A., Chuvilskaya, N.A., Rivkina,
  E.m., Pecheritsyna, S.A., Laurinavichius, K.S.,
  Suzina, N.E., Osipov, G.A., Lysenko, A.mgt,
  Gilichinsky, D.A., and V.K. Akimenko. 2005. Novel
  psychrophilic anaerobic spore-forming bacterium
  from the overcooled water brine in permafrost
  description Clostridium algoriphilum sp. Nov.
  Extremeophiles 9239-246.
• Smith, J.J., Howington, J.P., and G.A. McFeters.
  1994. Survival, physiological response, and
  recovery of enteric bacteria exposed to a polar
  marine environment. Applied and Environmental
  Microbiology 60 2977-2984.
• Steven, B., Leveille, R., and W.H. Pollard. 2006.
  Microbial ecology and biodiversity in permafrost.
  Extremeophiles 10259-267.
• Yang, C., and K.A. Sharp. 2005. Hydrophobic
  tendency of polar group hydration as a major
  force in type I antifreeze protein recognition.
  Proteins Structure, Function, and Bioinformatics
  59266-274.
• Zhou, X-X., Wang, Y-B., and W-E. Li. 2008.
  Differences in amino acids composition and
  coupling patterns between mesophilic and
  thermophilic proteins. Amino Acids 3425-33.

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Image Credits

• Title Page
• http//resurgere.deviantart.com/art/Package-Ice-7
  -48144161
• Cryoenvironments
• Nasa Earth Observatory (NEO) http//neo.sci.gsfc.
  nasa.gov
• NOAA National Geophysical Data Center Global
  Releif Model http//www.ngdc.gov/mgg/global/global
  .html
• Challenges
• Cristaux de glace sur une branche darbre by
  Jean-Paul Baubet http//fr.wikipedia.org/wiki/Fich
  ierFleurgivre-1.jpg
• Challenges 3 Ice Crystal Formation
• Gilbert, J.A., Davies, P.L., and J.
  Laybourn-Parry. 2005. A hyperactive
  Ca2-dependent antifreeze protein in an Antarctic
  bacterium. FEMS Microbiology Letters 24567-72.
• Potential Applications
• Mt Herschel, Antarctica, Jan 2006 by Andrew
  Mandemaker http//commons.wikimedia.org/wiki/File
  Mt_Herschel,_Antarctica,_Jan_2006.jpg
• Challenge 1 Decreased Membrane Fluidity
• Table Acyl chain composition of LPS from low
  (4C)- and high (22C)-temperature-grown cells of
  P. syringaea from Seshu Kumar, G., Jagannadham,
  M.V., and M.K. Ray. 2002. Low-temperature-induced
  changes in composition and fluidity of
  lipopolysaccharides in the Antacrtic
  psychrotrophic bacterium Pseudomonas syringae.
  Journal of Bacteriology 1846746-6749.