Production of cryoprotectant extracellular polysaccharide substances (EPS) by the marine psychrophilic bacterium Colwellia psychrerythraea strain 34H under extreme conditions.

Extracellular polysaccharide substances (EPS) play critical roles in microbial ecology, including the colonization of extreme environments in the ocean, from sea ice to the deep sea. After first developing a sugar-free growth medium, we examined the relative effects of temperature, pressure, and salinity on EPS production (on a per cell basis) by the obligately marine and psychrophilic gamma-proteobacterium, Colwellia psychrerythraea strain 34H. Over growth-permissive temperatures of approximately 10 to -4 degrees C, EPS production did not change, but from -8 to -14 degrees C when samples froze, EPS production rose dramatically. Similarly, at growth-permissive hydrostatic pressures of 1-200 atm (1 atm = 101.325 kPa) (at -1 and 8 degrees C), EPS production was unchanged, but at higher pressures of 400 and 600 atm EPS production rose markedly. In salinity tests at 10-100 parts per million (and -1 and 5 degrees C), EPS production increased at the freshest salinity tested. Extreme environmental conditions thus appear to stimulate EPS production by this strain. Furthermore, strain 34H recovered best from deep-freezing to -80 degrees C (not found for Earthly environments) if first supplemented with a preparation of its own EPS, rather than other cryoprotectants like glycerol, suggesting EPS production as both a survival strategy and source of compounds with potentially novel properties for biotechnological and other applications.

[1]  J. Bowman,et al.  Production of exopolysaccharides by Antarctic marine bacterial isolates , 2004, Journal of applied microbiology.

[2]  S. Giovannoni,et al.  Lentisphaera araneosa gen. nov., sp. nov, a transparent exopolymer producing marine bacterium, and the description of a novel bacterial phylum, Lentisphaerae. , 2004, Environmental microbiology.

[3]  J. Deming,et al.  High concentrations of exopolymeric substances in Arctic winter sea ice: implications for the polar ocean carbon cycle and cryoprotection of diatoms , 2002 .

[4]  E. Mercadé,et al.  Effect of incubation temperature on growth parameters of Pseudoalteromonas antarctica NF3 and its production of extracellular polymeric substances † , 2008, Journal of applied microbiology.

[5]  R. Amann,et al.  Diversity and Structure of Bacterial Communities in Arctic versus Antarctic Pack Ice , 2003, Applied and Environmental Microbiology.

[6]  G. Feller,et al.  Psychrophilic microorganisms: challenges for life , 2006, EMBO reports.

[7]  C. Fraser,et al.  The psychrophilic lifestyle as revealed by the genome sequence of Colwellia psychrerythraea 34H through genomic and proteomic analyses. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[8]  Hajo Eicken,et al.  Bacterial incorporation of leucine into protein down to -20 degrees C with evidence for potential activity in sub-eutectic saline ice formations. , 2006, Cryobiology.

[9]  J. Bowman,et al.  Bacterial Exopolysaccharides from Extreme Marine Environments with Special Consideration of the Southern Ocean, Sea Ice, and Deep-Sea Hydrothermal Vents: A Review , 2005, Marine Biotechnology.

[10]  J. Deming,et al.  Persistence of bacterial and archaeal communities in sea ice through an Arctic winter , 2010, Environmental microbiology.

[11]  P. Zanchetta,et al.  A New Bone-Healing Material: A Hyaluronic Acid-Like Bacterial Exopolysaccharide , 2002, Calcified Tissue International.

[12]  J. Deming,et al.  Characterization of a cold-active bacteriophage on two psychrophilic marine hosts , 2006 .

[13]  U. Passow Transparent exopolymer particles (TEP) in aquatic environments , 2002 .

[14]  A. Decho,et al.  Microbial exopolymer secretions in ocean environments: their role(s) in food webs and marine processes , 1990 .

[15]  J. Breezee,et al.  Subfreezing Growth of the Sea Ice Bacterium “Psychromonas ingrahamii” , 2004, Microbial Ecology.

[16]  J. Deming,et al.  The Role of Exopolymers in Microbial Adaptation to Sea Ice , 2008 .

[17]  I. Sutherland Novel and established applications of microbial polysaccharides. , 1998, Trends in biotechnology.

[18]  M. Gosselin,et al.  Seasonal study of sea-ice exopolymeric substances on the Mackenzie shelf: implications for transport of sea-ice bacteria and algae , 2006 .

[19]  A. Engel,et al.  Abundance and variability of microorganisms and transparent exopolymer particles across the ice–water interface of melting first-year sea ice in the Laptev Sea (Arctic) , 2001 .

[20]  E. Oner,et al.  Exopolysaccharides from extremophiles: from fundamentals to biotechnology , 2010, Environmental technology.

[21]  B. Methé,et al.  Purification, Characterization, and Sequencing of an Extracellular Cold-Active Aminopeptidase Produced by Marine Psychrophile Colwellia psychrerythraea Strain 34H , 2004, Applied and Environmental Microbiology.

[22]  J. Deming,et al.  Elevated bacterial abundance and exopolymers in saline frost flowers and implications for atmospheric chemistry and microbial dispersal , 2010 .

[23]  A. Poli,et al.  Bacterial Exopolysaccharides from Extreme Marine Habitats: Production, Characterization and Biological Activities , 2010, Marine drugs.

[24]  F. Smith,et al.  COLORIMETRIC METHOD FOR DETER-MINATION OF SUGAR AND RELATED SUBSTANCE , 1956 .

[25]  P. Gervais,et al.  Cell Size and Water Permeability as Determining Factors for Cell Viability after Freezing at Different Cooling Rates , 2004, Applied and Environmental Microbiology.

[26]  J. Deming,et al.  Psychrophiles and polar regions. , 2002, Current opinion in microbiology.

[27]  Mark V Brown,et al.  Diversity and association of psychrophilic bacteria in Antarctic sea ice , 1997, Applied and environmental microbiology.

[28]  E. Delong,et al.  Phylogenetic diversity of aggregate‐attached vs. free‐living marine bacterial assemblages , 1993 .

[29]  J. Deming,et al.  A microscopic approach to investigate bacteria under in situ conditions in sea-ice samples , 2001, Annals of Glaciology.

[30]  P. Zanchetta,et al.  Systemic Effects on Bone Healing of a New Hyaluronic Acid-Like Bacterial Exopolysaccharide , 2003, Calcified Tissue International.

[31]  J. Deming Extremophiles: Cold Environments , 2009 .

[32]  J. Lundberg,et al.  An extraordinary example of photokarren in a sandstone cave, Cueva Charles Brewer, Chimantá Plateau, Venezuela: Biogeomorphology on a small scale , 2010 .

[33]  N. Bhosle,et al.  Microbial extracellular polymeric substances in marine biogeochemical processes , 2005 .

[34]  John A. Baross,et al.  Planets and life : the emerging science of astrobiology , 2007 .

[35]  J. Deming,et al.  Modelled and measured dynamics of viruses in Arctic winter sea-ice brines. , 2006, Environmental microbiology.

[36]  J. Costerton,et al.  Biofilms, bacterial signaling, and their ties to marine biology , 2003, Journal of Industrial Microbiology and Biotechnology.

[37]  J. Deming Extreme High-Pressure Marine Environments , 2007 .

[38]  J. Deming,et al.  Spatial heterogeneity and temporal dynamics of particles, bacteria, and pEPS in Arctic winter sea ice , 2008 .

[39]  J. Guézennec Deep-sea hydrothermal vents: A new source of innovative bacterial exopolysaccharides of biotechnological interest? , 2002, Journal of Industrial Microbiology and Biotechnology.

[40]  R. Amann,et al.  Predominance of β‐proteobacteria in summer melt pools on Arctic pack ice , 2004 .

[41]  J. Deming,et al.  Remarkably low temperature optima for extracellular enzyme activity from Arctic bacteria and sea ice. , 2000, Environmental microbiology.