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Biotechnology ofBiotechnology of ExtremophilesExtremophiles
Jenny BlameyFundación Biociencia
March 2009March 2009
ExtremophilesExtremophilesPsychrophiles Thermophiles
Hyperthermophiles
T <15°C, (mín. -18°C)
yp p
T= 60°C a 80°C.T > 80°C (mín. 18 C)
(máx. 113°C).
H l hil X hil
El TatioAntarctica
Halophiles
[NaCl] > 3 5%
Xerophiles
a < 0 85Atacama Saltern Atacama Desert[NaCl] > 3,5%
A id hilAlk li hil
aw, < 0,85.
Acidophiles
pH < 5(mín pH 0)
Alkaliphiles
Alcalinidad >pH 9 (mín. pH 0)>pH 9 (máx. pH 10,5).Ascotan
Acid Lagoon, Calama
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Biotechnological importance
Biodiversity resources have a great economicalvalue.Direct application of these microorganisms to
industrial processes.Development of cellular products derived fromp pnovel microorganisms.Identification of new compounds with potentialIdentification of new compounds with potentialbiotechnological applications.
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Specialized Knowledge
Sampling of natural environments and extreme bio habitatsextreme bio-habitats.Development of specific culture media for extremophiles (i e aerobes andfor extremophiles (i.e. aerobes and strict anaerobes).Isolation and Characterization.Isolation and Characterization.Development of specific methods for DNA extraction and protein purification.p pBiochemical Characterization.
Copyright Fundación Biociencia, Chile; www.bioscience.cl
The search
Sampling Culture Enrichment Microorganisms isolation
Enzymes & ProteinsLipidsBiomolecules & BiopolymersBi ti C dBioactive Compoundsetc.Applications
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Possible applicationsPossible applicationsBiotechnologygy
Use Bioactive compoundMolecular Biology, Genomics, DNA amplification for PCR
DNA Polymerase, thermostable restriction enzymesDNA amplification for PCR restriction enzymes
DNA and protein stabilizers Compatible Solutes (halophiles)Alkaline phosphatasesMolecular Biology Alkaline phosphatases (psychrophiles)
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Possible applicationsPossible applicationsPharmaceutical and Diagnostics
Use Bioactive compound
Enantioselective resolution of Thermo/hyperthermophilicEnantioselective resolution of pharmaceutical compounds
Thermo/hyperthermophilic enzymes
Liposomes for drug administration Lipids from hyperthermophiles
Antibiotics Extremophilic bactericidesAntibiotics Extremophilic bactericides
Plastic for medical use PHB,Polyhydroxyalkenoates
Immunoenhancers Chitoologosacharides
Surfactants Halophilic membranesSurfactants Halophilic membranes
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Possible applicationsPossible applicationsAgro industry, food & feedg o dus y, ood & eedUse Bioactive compound
Bakery, glucose and fructose production
Thermophilic amylases and cellulasesproduction cellulases
Wool, feathers, chitin conversion into feed.
Thermophilic keratinases y chitinases
Dietary supplements, pigments. γ-Linoleic acid, β-carotene and other pigments from halophiles
Freezing protection Antifreeze proteins fromFreezing protection Antifreeze proteins from psychrophiles
Cheese and milk products Psychrophilic Proteases
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Possible applicationsPossible applicationsOil and FuelO a d ue
Use Bioactive compound
Surfactants for oil recuperation Hyperthermophilic oil degradersSurfactants for oil recuperation Hyperthermophilic oil degraders
Rheologic polymers, thermostable Oil recuperation
g p yand halophilic enzymes that degrade guar.
Oil Biodesulfuration Thermophilic microorganismsOil Biodesulfuration Thermophilic microorganisms
Cleaning of oil spilling Psychrophiles which degrade oil
Hydrogen production Hydrogenases
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Possible applicationsPossible applications Chemical IndustryC e ca dus y
Use Bioactive compound
D
Lipases, pullulanases, amylases y proteases:
thermostableDetergents thermostablepsychroactiveacidophilicpalkaline
Synthesis of ester, organic Esterasesbiosynthesis
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Possible applicationsPossible applicationsElectronic, Optic, Nanotechnology, p , gy
Use Bioactive compoundUse Bioactive compound
Optical “Switch” 500.000 Ghz (500x10-15 s)
Bacteriorhodopsin from halophiles and xerophiles(500x10 s) and xerophiles.
Re writeable films of high resolution (5μm)
Bacteriorhodopsin from halophiles and xerophiles.
Autoensambled substrate for l t i t t f 1 t 10
Protein obtained from an id hilelectronic structures of 1 to 10 nm. acidophile
Copyright Fundación Biociencia, Chile; www.bioscience.cl
OpportunitiesLess than 1% of the existent microorganisms have been cultivated and gcharacterizedThe percent of extremophiles discoveredThe percent of extremophiles discovered is even less due to:
Inaccessibility to the extreme habitats.Novel techniques and methods are required.q q
Chile has all extreme habitats and most of them completely unexploredof them completely unexplored.
Copyright Fundación Biociencia, Chile; www.bioscience.cl
New goalsOptimization of growth conditions to use extremophiles as complete organisms in some industrial processes.Development of new methods for cell disruption that will allow to obtain nucleic acids, proteins and other metabolites in its optimal conditions.Design of appropriate new vectors that allow cloning and expression of extremophilic g p penzymes and biocompounds.
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Chile: a privileged country21º S
Atacama Desert57º S
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Chile: a privileged country21º S
Moon Valley 57º SCopyright Fundación Biociencia, Chile; www.bioscience.cl
Chile: a privileged country21º S
El T tiEl Tatio57º S
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Chile: a privileged country21º S
Acid lagoon, Chuquicamata 57º SCopyright Fundación Biociencia, Chile; www.bioscience.cl
Antarctic Bioresources Platform.
Creating a platform to facilitate access to Antarcticbioresources, microorganisms and plants, allowingits valorization for scientific and biotechnologicalits valorization for scientific and biotechnologicaldevelopment.
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Areas of studyAreas of study
Visited sites: 15 differentVisited sites: 15 different geographic places, including South Shetland
d th A t ti ti tand the Antarctic continent.
More than 170 samples.More than 170 samples.
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Results: Antarctic bioprospectionResults: Antarctic bioprospection
1. Hydrocarbon degradation at low temperature by psychrophiles.
Enzymes involved in hydrocarbon metabolism
Surfaction and crude oil
Search of oxygenase activitySampling
Surfaction and crude oil degrading at 3ªC
Isolation and characterization of hydrocarbon degrading microorganisms
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Results: Antarctic bioprospection
2 Antioxidant activity on psychrophiles
Results: Antarctic bioprospection
2. Antioxidant activity on psychrophiles.
Search of superoxide dismutase (SOD)
Isolation and characterization dismutase (SOD)
O2 + H2O2SOD2O - + 2H+
characterization
O2 + H2O22O2- + 2H+
Antarctic Territory: exposed to high UV radiation.
Copyright Fundación Biociencia, Chile; www.bioscience.cl
Scale up cultures Purification of SOD
Results: Antarctic bioprospection
3. Hyperthermophilic microorganisms and thermostable enzymes
Results: Antarctic bioprospection
Enrichments at 80ºC.
Enriquecimiento y cultivo de microorganismosde microorganismos
a 80°C.
Biotech Potential:
•Molecular adaptation mechanismsDeception Island: unique diversity of habitats.
•Molecular adaptation mechanisms
•Source of novel microorganisms
•Source of highly thermostable enzymes
Copyright Fundación Biociencia, Chile; www.bioscience.cl
•Source of highly thermostable enzymes
Chile and the Antarctic Territory
Project open for international collaboration on j pbiotechnological development.
Copyright Fundación Biociencia, Chile; www.bioscience.cl