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NEW  MICROALGAE  BIOREFINERY  CONCEPT  

   Laura  Azócar,  Hernán  Díaz,  Robinson  Muñoz,  Leonel  González,  David  Jeison,  Rodrigo  

Navia  

Financiado  por  

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IntroducEon  

Lam  and  Lee,  2012  

Brennan  and  Owende,  2010  

Downstream  

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Mata  et  al.  2012  

Ahmad  et  al.  2012  

Brennan  and  Owende,  2010  

Downstream  

Waste  biomass  (70%)  Solvents  

Harvest  

Biomass  procesing    

Lipid  extracEon  

TransesterificaEon  

Protein  extracEon  

Anaerobic  digesEon  

Biodiesel  upgrading    

Harvest  

Carbohidrate  extracEon  to  bioethanol  producEon  

Direct  transesterificaEon  

Protein  extracEon  

Anaerobic  digesEon  

Biodiesel  upgrading    

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SequenEal  opEmizaEon    

Variables  pH=  9-­‐10-­‐11-­‐  12-­‐13  

Temperature  (°C)=  30-­‐50-­‐70  Eme  (min)=  10-­‐  20-­‐  30    

SEr  (rpm)=  100-­‐  200-­‐  300    

Chemical  proteins  solubilizaEon  &  EnzymaEc  hydrolysis    

PROTEIN  EXTRACTION  

CHEMICAL  PROTEIN    SOLUBILIZATION  

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pH

9 10 11 12 13

% P

rote

in s

olub

ility

0

10

20

30

40

50

temperature [°C]

20 30 40 50 60 70

% P

rote

in s

olub

ility

0

10

20

30

40

50

60

Results  ü 50°C  ü 20min    ü 200  rpm  

ü pH  12  ü 20min    ü 200  rpm  

Figure   1.   %   Protein   solubility   from  microalgae  biomass  to  different  pH  condiEons      

Figure   2.   %   Protein   solubility   from   microalgae  biomass  to  different  reacEon  temperature      

time [min]

10 20 30 40 50 60

% P

rote

in s

olub

ility

0

10

20

30

40

50

60

70

Stir [rpm]

50 100 150 200 250 300 350

% P

rote

in s

olub

ility

0

20

40

60

80

ü pH  12  ü 50°C    ü 200  rpm  

ü pH  12  ü 50°C    ü 40  min  

OpEmal  condiEons:  pH  12  50°C    40  min  200rpm    Yield:  64%  

Figure   3.   %   Protein   solubility   from   microalgae  biomass  to  different  reacEon  Eme        

Figure  4.  %  Protein  solubility  from  microalgae  biomass  to  different  sEr  condiEons  

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 1)  EnzymaEc   hydrolysis   of   microalgae   biomass   through   successive   steps  

addiEon  of:  Viscozyme,  Alcalasa  and  Flavourzyme    2)    EnzymaEc  hydrolysis  of  solubilized  protein  by  chemical  treatment  by  using  

Alcalasa.    3)  EnzymaEc  hydrolysis  of  solubilized  protein  by  chemical  treatment  by  using  

Flavourzyme.    Raw  material    1)  Pretreated  Microalgae  150g/L  (similar  to  biomass  post  harvest)      

ENZYMATIC  HYDROLYSIS  

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Reactor   On  line  traking  Thermoregulated  bath  

Set  up  

EnzymaEc  hydrolysis  of  microalgae  biomass  

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Time [h]

0 50 100 150 200 250 300

% H

ydro

lysi

s de

gree

30

40

50

60

70

80

90Disadvantage  Carbohydrates  loss  (65%)  

Hydrolysis  degree:  84%    

Viscozyme  Alcalasa  

Flavourzyme  

Figure   5.   Hydrolysis   degree   aner   enzymaEc   hydrolysis   of  microalgae  biomass  

Flavourzyme  

Time [h]

0 20 40 60 80 100

Hyd

roly

sis

degr

ee [%

]

20

30

40

50

60

70

80

90

Time [h]

0 50 100 150 200 250 300

Hyd

roly

sis

degr

ee (%

)

40

50

60

70

80

90

Flavourzyme  

Alcalasa  EnzymaEc  hydrolysis  of  previously  solubilized  protein  by  chemical  process  

Low  carbohydrates  loss      Carbohydrates  extracEon  to  bioethanol  producEon???  

Total  yield:  88%    

Figure   6.   Hydrolysis   degree   aner   enzymaEc  hydrolysis  of  solubilized  protein  by  chemical  process  

Figure  7.  Hydrolysis  degree  aner  enzymaEc  hydrolysis  of  solubilized  protein  by  chemical  process  

12  Biomass  to  biodiesel  producEon  (high  lipid  content)  

Carbohydrates  to  bioethanol  producEon  

CARBOHYDRATES  EXTRACTION  TO  BIOETHANOL  PRODUCTION  

BIODIESEL  PRODUCTION  

Lipids  extracIon  and  transesterificaIon  

Figure   8.   Chromatogram   faqy   acid   profile   (a)  microalgae  1  and  (b)  microalgae  2    

(a)  

(b)  

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BIODIESEL  PRODUCTION  

Direct  transesterificaIon  

CondiEons  invesEgated  RSM    ü Methanol  to  oil  molar  raEo  ü Catalyst  ü Time  ü Temperature  

Figure  9.  Biodiesel  produced  by  direct  transesterificaEon  

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BIOGAS  PRODUCTION  

Figure  10.  Biomass  during  microalgae  biorefinery  process  (a)Protein    extracEon,  (b)  Drying,  (c)   Dry   biomass,   (d)   Biomass   milled   to   the     lipids   extracEon,   (e)   Biomass   aner   lipids  extracEon,  (f)  Biomass  milled  to  biogas  producEon    

Biochemical  methane  potenIal  by  using  microalgae  biomass    aQer  lipids  extracIon  

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BIOGAS  PRODUCTION  Biochemical  methane  potenIal  by  using  microalgae  

biomass  aQer  transesterificaIon  reacIon  

Figure  10.  Biomass  aner  transesterificaEon  reacEon  

117  serum  boqles  50  mL  reacEon  Nutrients  Bicarbonate  35°C    

Assays  

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BIOGAS  PRODUCTION  

Parámetro Microalgae 1 Spent microalgae 1 Spent microalgae 2 Spent microalgae 2

Humidity content [%] 79,58 7,66 3,99 3,13Lipids [%]* 19,20 5,23 9,61 7,4Protein [%]* 33,00 22,80 39,42 41Fiber [%]* 3,33 7,94 4,34 3,54Ash [%]* 31,00 40,13 10,34 13,75Carbohidrates [%] 13,47 16,24 32,3 31,18C/N [gC/gN] 5.85 6.97 - -

*dry  basis  

Table  1.  Microalgae  characterizaEon  

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BIOGAS  PRODUCTION  

Time [d]

0 20 40 60 80

Bio

chem

ical

met

hane

pot

entia

l BM

P [m

L C

H4/g

VS

]

0

50

100

150

200

250

300

350

Spent microalgae post transesterification 1Spent microalgae post transesterification 2

Figure   12.   BMP   using   spent   microalgae   aner   direct  transesterificaEon  

Time [d]

0 20 40 60 80

Bio

chem

ical

met

hane

pot

entia

l BM

P [m

L C

H4/g

VS

]

0

100

200

300

400

500

Spent microalgae + residual culture mediumSpent microalgae + water + nutrients

Figure  11.  BMP  using  spent  microalgae  aner  protein  and  lipids  extracEon    

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BIOGAS  PRODUCTION  

Figure  13.  TheoreEcal  and  real  BMP  usiing  spent  microalgae  

Microalga agotadaMicroalgae 1.1

Microalgae 1.2Microalgae 2.1

Microalgae 2.2

BM

P [m

LCH

4/gV

S]

0

200

400

600

800

Theoretical BMPReal BMP

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NEW  MICROALGAE    BIORREFINERY  CONCEPT    

*BIOCOMPOSITS  *PYROLYSIS  PRODUCTS  

Wastewater

Wastewater

Spent biomass

Spent biomass

BIODIESEL

Microalgae

(

Direct transesterification

Spent biomass

1. Harvest

Solvent recovery

BIOGAS

5. Upgrading

3. Bioethanol production

2. Protein solubilization

4. Biodiesel production

AMINO ACIDS

BIOETHANOL

Residual culture medium

6. Anaerobic digestion

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ACKNOWLEDGMENT  

ü   Consorcio  Desert  Bioenergy  S.A.,  Innova-­‐CORFO  ü   CONICYT  Project  78110106  ü   Chilean  Fondecyt  project  3120171  ü   PIA  project  DI12-­‐7001  from  University  of  La  Frontera    

Grupo  de  Biotecnología  de  Microalgas  Marinas  Universidad  de  Almería  (G.  Acién,  P.  García)  

 Center  of  Waste  Management  and  Bioenergy      

ScienEfic  and  Technological  Bioresource  Nucleus  

NEW  MICROALGAE  BIOREFINERY  CONCEPT  

   Laura  Azócar,  Hernán  Díaz,  Robinson  Muñoz,  Leonel  González,  David  Jeison,  Rodrigo  

Navia  

www.desertbioenergy.cl  www.  bioren.cl  

lazocar@desertbioenergy.cl