biotecnología aplicada a cultivos de microalgas

BIOTECNOLOGIA APLICADA AL CULTIVO DE MICROALGAS:

BUSQUEDA, SELECCIÓN Y OPTIMIZACION DE MICROALGAS PARA LA PRODUCCION DE ACEITE

COMO BIOCOMBUSTIBLE.BIOTECHNOLOGY APPLIED TO MICROALGAE

CULTURES: SELECTION AND OPTIMIZATION OF MICROALGAE CULTURES FOR BIODIESEL

PRODUCTION.

Instituto Biomar is a 14 year old biotechnological company located in Parque

Tecnológico de León

The company

1996: Spin off from PharmaMar

1997: Move to facilities in biotech business incubator

2000: Start of work in Industrial Applications: Biospesticides

2001: Start of the Fermentation plant

2005: Start of the Microalgae project

2007: Start of the “Microalgae for biodiesel” project.

2009: Opening of new facilties in Leon Technological Park

The company

TECHNOLOGIES APPLICATIONS

The company

Instituto Biomar: fields of interest

MICROBIOLOGY

CHEMISTRY

FERMENTATION

HUMAN HEALTH

FOOD

BIOPESTICIDES

AQUICULTURE

BIODIESEL

The company

The company

The company

BIODIESEL

Metanol + KOH

Trans-esterification Crude

biodiesel Glicerol

Metanol

• Proteins• Sugars• High value

compounds

Photosynthetic

microorganisms

(Biomass)

Acylglicerides

Microorganisms

ActinobacteriaFungi

Biodiesel Project

Biodiesel Project

STRAIN SELECTION

GROWTH RATE OIL CONTENT

OPTIMIZATING PRODUCTION

CLASSIC METHODS GENETIC MANIPULATION

INDUSTRIAL SCALEOPEN SYSTEMS

(RACEWAYS)CLOSED- SYSTEMS(PHOTO-REACTORS)

CO2 Lipids

Chemical transformation

Water

Salts

MICROALGAEBIOMASS

BIODIESEL

EXTRACTION

Sugars, proteins

High value compounds

Biodiesel Project

Isolation from marine invertebrate organisms, water, sediments or soils and plant samples of different sites of the world (Europe, America, Africa…)

Australia

Mediterranean Sea Japan

Ecuatorial Guinea

Canarias

Escocia

HawaiiRed Sea

Spain

Senegal

Argentina

Puerto RicoPanamáGuatemala

MartinicaMéxico

Alaska

NewZeland

NorwayIsland

Chile

Perú

Groenland

Biodiesel Project

Laboratory of Microalgae

ISOLATIONMarine macroorganisms, sediments, rocks, soils…are crushed and cultured on different media (BG-11, ASNIII,…).Isolation process: Picking colonies by hand under stereomicroscopes.

(3000 isolated monocultures per year)

STRAIN SELECTIONDereplication process by sample and sampling point, using stereomicroscope and microscope. This process leads to different strains (aproximately 1200-1400 different monocultures per year).

Laboratory of Microalgae

Current collection of microalgae and cyanobacteria is approximately 2.500 monocultures

STRAIN CULTUREPRIMARY CULTURES on 50 ml flasks are cultured with controlled light and temperature (1200-1400 monocultures per year).

Laboratory of Microalgae

STRAIN CULTURE (2)SECONDARY CULTURES on 200 mL tubes, aireated and with temperature and light controlled (1200-1400 monocultures per year).

Laboratory of Microalgae

STRAIN CONTROLAll strains are photographed, growth is measured, dry weight is estimated, HPLC (UV-vis) profile is determined and DNA is obtained.

With these secondary cultures we obtain biomass for our screening programs:- Drug discovery screening program.- Biodiesel screening program.

Laboratory of Microalgae

SCREENINGAcylglicerides measurementsSamples are extracted with hexane and measured for acylgliceride content (not lipid content) by colorimetry (595 nm) on 96 multiwell plates.

Laboratory of Microalgae

SCREENING Positive samples are re-tested using TLC in order to

diferentially detect acylglicerides (tri-, di-, monoacylglicerides).

TAG

DAGMAG

Laboratory of Microalgae

We select 10-20 strains per year to initiate studies

in order to improve growth and/or productivity.

0,00

5,00

10,00

15,00

20,00

25,00

% mg AG/mg DW

mg DW · L-1/100

SELECTING STRAINS

Laboratory of Microalgae

IMPROVEMENT- Growth: Testing different conditions on 24 well plates.

- Acylglicerides content: searching for conditions leading to accumulation but with no effects on growth.

Laboratory of Microalgae

SCALE-UPFrom these studies we aim to select 1-2 strains with appropiate characteristics to initiate open air cultures (200-400 L) during 2010, as a step previous to industrial scale up.

Laboratory of Microalgae

1

2 Isolating

3

4

Culturing5

Caracterizing

8

7

6Selecting

Improving

Producing strain

DYNAMIC PROCESS

0,00

5,00

10,00

15,00

20,00

25,00

% mg TG/mg DW

mg dw/L cult entre 100

Bioprospecting

Profiling

Biodiesel project

Biodiesel project

STRAIN SELECTION

GROWTH RATE OIL CONTENT

OPTIMIZATING PRODUCTION

CLASSIC METHODS GENETIC MANIPULATION

INDUSTRIAL SCALEOPEN SYSTEMS

(RACEWAYS)CLOSED- SYSTEMS(PHOTO-REACTORS)