Biotechnology ofBiotechnology of...

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.


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.


The search

Sampling Culture Enrichment Microorganisms isolation

Enzymes & ProteinsLipidsBiomolecules & BiopolymersBi ti C dBioactive Compoundsetc.Applications


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)


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


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


Possible applicationsPossible applicationsOil and FuelO a d ue


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


Possible applicationsPossible applications Chemical IndustryC e ca dus y


D

Lipases, pullulanases, amylases y proteases:

thermostableDetergents thermostablepsychroactiveacidophilicpalkaline

Synthesis of ester, organic Esterasesbiosynthesis


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


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.


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.


Chile: a privileged country21º S

Atacama Desert57º S



Moon Valley 57º SCopyright Fundación Biociencia, Chile; www.bioscience.cl


El T tiEl Tatio57º S



Acid lagoon, Chuquicamata 57º SCopyright Fundación Biociencia, Chile; www.bioscience.cl

Chile: a privileged country

AntarcticaCopyright 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.


Isla Decepción

Expedition 2009ECA-45

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.


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


Results: Antarctic bioprospection

2 Antioxidant activity on psychrophiles


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.


Scale up cultures Purification of SOD


3. Hyperthermophilic microorganisms and thermostable enzymes


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


•Source of highly thermostable enzymes

Chile and the Antarctic Territory

Project open for international collaboration on j pbiotechnological development.


Thank youJenny Blamey

Fundación BiocienciaS i t l Bi t hSwissaustral Biotech

[email protected]

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