09/03/2015

Sustainable pilot-scale production of carotenoid

compounds from the green microalga
Haematococcus pluvialis

G. Feijoo & M.T. Moreira

Departamento de Ingeniera Qumica
Universidad de Santiago de Compostela
gumersindo.feijoo@usc.es
www.usc.es/biogrup
@EnxQuim_USC

09/03/2015

Contents
Introduction to microalgal processes

Goal and scope

Inventory analysis

Environmental impact assessment

Conclusions

Contents
Introduction to microalgal processes

Goal and scope

Inventory analysis

Environmental impact assessment

Conclusions

09/03/2015

USES OF MARINE RESOURCES

Marine biotechnology

Why marine resources?

USES OF MARINE RESOURCES

Marine biotechnology

Why marine resources?

With more than 70% of
the planets surface
covered by water,
oceans are probably the
most promising habitat
to explore for novel
microbial biodiversity.

09/03/2015

USES OF MARINE RESOURCES

Marine biotechnology

Why marine resources?

Marine biotechnology is an emergent sector with an estimated
global market value of 2.8 billion in 2010 and annual growth of
4.3% during the period 2007-2012.
Others 6%

Number of patents

30
25
20

Pharmacology
32%
Agriculture
2%

15

Food 6%

10
5
0

Cosmetics 1%
Chemistry
54%
Period

No. patents involving marine resources

% patents in marine biotechnological sector

Leary et al. (2009). Marine genetic resources: A review of scientific and commercial interest.
Marine Policy 33(2):183-94

SIGNIFICANCE OF MICROALGAL PROCESSES

Products from microalgae

MICROALGAE:
One resource,
many products...

Algae Oils
Carbohydrates

Proteins

Biomass

From Wilkie et al. (2011). Indigenous algae for local bioresource

production: Phycoprospecting, Energy Sustain Dev 15(4):365-71

09/03/2015

TECHNOLOGICAL OPTIONS

Cultivation systems for microalgae

How to produce microalgae?

CASE STUDY

Pilot-scale production of carotenoids by H. pluvialis

Haematococcus pluvialis
Green phase

 Green microalga
 Cultivated in two stages
 It accumulates between 1-5%
of astaxanthin

Astaxanthin
 Red carotenoid (pigment)

Red phase

 Applications as additive in food industry, cosmetics...

 Potential uses in pharmaceutical industry due to its antioxidant,
anti-inflamatory and antitumor properties

09/03/2015

Contents
Introduction to microalgal processes

Goal and scope

Inventory analysis

Environmental impact assessment

Conclusions

GOAL AND SCOPE DEFINITION

Objective of the study

 Quantification of main environmental impacts associated to the
process

 Cradle-to-gate perspective (from raw materials to extraction of the

biocompound)
 Identification of the most
problematic stages

Wild Marine Organism

Harvesting
End of
Life

Cultivation in
Reactor Systems

ASSESSMENT

HOT SPOTS

Harvesting

Product
Use

Proposal of alternative scenarios Distribution

to improve the environmental profile

Extraction and
Purification of
Biocompounds
Product
Formulation

09/03/2015

GOAL AND SCOPE DEFINITION

Functional Unit
CULTURE IN TWO PHOTOBIOREACTORS IN SERIES
(1000 L tanks, 2-4 g/L biomass)

HARVESTING
4 kg of biomass (4-5% astaxanthin)

EXTRACTION
Supercritical CO2 Extraction

800 g
astaxanthin

GOAL AND SCOPE DEFINITION

ADDITION OF
NUTRIENTS AND
INOCULUM

UV FILTER

SUBSYSTEM 3: CULTIVATION
AIR SUPPLY
GROWTH STAGE

SETTLING

SETTLING

STRESS STAGE

MACHINERY

ELECTRICITY

BACKGROUND
SYSTEM

SUBSYSTEM 4: HARVESTING

SUBSYSTEM 5: EXTRACTION

CENTRIFUGATION

SUPERCRITICAL
FLUID EXTRACTION

SPRAY DRYING

FOREGROUND SYSTEM

AIR, SOIL & WATER

EMISSIONS

REVERSE
OSMOSIS
FILTER

ASTAXANTHIN (10%
IN OLEORESIN)

CHEMICALS
(Nutrients & Solvents)

REACTOR
STERILIZATION
(ozonization or
chemicals)

SUBSYSTEM 2: PREPARATION OF THE

CULTURE MEDIUM

BY- PRODUCT
(FERTILIZER)

WATER SUPPLY

SUBSYSTEM 1:
CLEANING OF THE
REACTOR

WASTE TO
TREATMENT

RAW MATERIALS, WATER & FOSSIL FUELS

System boundaries

09/03/2015

GOAL AND SCOPE DEFINITION

System boundaries
EXTRACTION WITH ORGANIC SOLVENTS

DMSO Extraction
(Heating + vortex mixing
+ centrifugation)

Centrifugation
and drying
Microalgal cells
following cultivation

Microalgal
powder

DMSO extract
containing carotenoids

Consideredsolvents
Example:
Microalgae

Macroalgae

- Dimethyl sulfoxide (DMSO) - Dichloromethane

for Haematococcus
(DCM) for essential
astaxanthin
terpene oils from O.
- Chloroform:methanol for secundiramea
PUFAs from P. tricornutum
- Hexane as alternative
cleaner solvent for PUFAs

DMSO
Sponges

extraction
for the Marine
separation
Epiphytic bacteria
fungi & of
protists
- Methanol:water,
- Chloroform:methanol
astaxanthin
from H. pluvialis biomass
n-hexane and
as extraction solvent - Chloroform:methanol
carbon
(acetonitrile, formic
as extraction solvent
tetrachloride
acid and isopropanol - Hexane for washing
as
LC-MS-MS
elution
- Methanol:water,
solvents)
n-hexane and
chloroform

GOAL AND SCOPE DEFINITION

System boundaries

09/03/2015

Contents
Introduction to microalgal processes

Goal and scope

Inventory analysis

Environmental impact assessment

Conclusions

INVENTORY ANALYSIS

Data sources
GENERAL INFORMATION
Partner

Field data (Surveys to

producers)





Databases
Bibliographic sources

Objective

Production

 Extraction/Purification

Type of marine organism (please tick the appropriate box

)
 Microalgae

 Marine sponges

 Macroalgae

 Epiphytic bacteria

 Marine fungi and marine protists

Scientific name
Bioactive compound
Alternative producer/producers of the compound
Cultivation method (please tick the appropriate box
)
 Open raceway ponds

 Shore base farming

 Photobioreactor

 Traditional biofermenter

 Tubular reactor

 Tissue culture

 Other:...........................................................................

09/03/2015

INVENTORY ANALYSIS

Inputs and outputs

INPUTS from TECHNOSPHERE
Transport
Distance (km)
1,982.37
kWhTECHNOSPHERE
Cleaning of the reactor
INPUTS from
OUTPUTS
to TECHNOSPHERE
1,982.49
kWh
(OPT.
2)
OPTION
1
Lorry,
3.5-7.5
t,
Euro
4
3.207
tkm
600
Materials
Products
Chemicals
Ship
5.613 tkm
1400
Cleaning of the reactor
Cleaning
of the reactor
OPTION
2
Lorry, 3.5-7.5 t, Euro 4 0.120 tkm
800
OLEORESIN
(10%
astaxanthin)
8.00
kg OPTION
OPTION
1 for reactor sterilization
2 for reactor sterilization
OPTION 2 for reactor sterilization
Equipments
Ship
0.281 tkm
1400
Tap water
(cleaning)
4,009
steel
0.200
kg
Pure astaxanthin
kg Stainless
Reactor
sterilization
0.120.80
kWh L
Preparation
of the
culture medium
NaClO
4.009
kg
Lorry, 3.5-7.5 t, Euro 4 6.504 tkm
600
Co-solvent Preparation of the culture medium
7.16
kgChemicals
OUTPUTS
to ENVIRONMENT
Preparation
of the
culture
medium
Preparation of Ship
the culture medium
Reverse
osmosis
filtration
7.71
kWh
11.382 tkm
1400
Water emissions
ImpuritiesNaNO3
0.04
kg Stainless steel
4.4651
UV filtration
0.35
kWhkg
Equipments
Lorry, 3.5-7.50.3446
t, Euro 4 0.300 kgtkm
800
Cleaning of the reactor (OPTION
Harvesting
Extraction
K2HPO
kg
PVC
0.9121 1)
0.0213
4
Avoided product
(fertilizer)
Cultivation
Ship
0.700 kgtkm
1400
Wastewater
4,0091,539.43
Wastewater
L
Wastewater
0.93
L
KH2PO
kg Cultivation
UV1,001.30
lamps
kg
0.4041 L
0.0175
Reactor
4
lighting
kWh
Cell paste
20.516
kg
NaClO
4.009
kg
NaNO
1.7522
g
NaNO
mg
CaCl2Air blowing
Polyamide Lorry, 3.5-7.50.1169
kgtkm
0.2914
3 4
96.21
kWhkg 3 Equipments
t, Euro
5.0911.6329
800
N fertilizer
0.431
kg
ammonium
sulfate,
as
N
Cultivation
K
HPO
0.0080
g
K
HPO
0.0075
mg
MgSOAgitation
kg
Cultivation
1.3473
96.21
kWh2
4
Ship
2
4
11.879 tkm
1400
4
L
KHkg
0.0035
KH8.36
0.0033
mg
NaCl
steel gphosphate,
Kg
0.1194
Harvesting Wastewater 1,783.98
2PO4 Harvesting
2PO
4 O
diammonium
as P
P fertilizer
0.282
kg Stainless
2 5
NaNO
87.3400
CaCl
0.6108
g 3.5-7.5CaCl
mg
C 6H8O
kg 2 Equipments
Reactor
lamps
Kgtkm
0.0287
Centrifugation
(harvesting)
1.50 g
kWh
Lorry,
t,0.13
Euro
4 2.3050.5692
800
3
2
7
Waste treatment
K2HPO4
19.4050
MgSO
2.8237 Shipg
MgSO4
mg
C6H5+4y
FexNdrying
kg 4
Harvesting
Spray
82.70 g
kWh
5.3792.6313
tkm
1400
0.0287
yO7
Steel, to inert
landfill
kg Stainless
KH2PO4
8.59700.4778
g
NaCl
0.2503steel g
NaCl
0.2333
mg
Na2Extraction
CO3
kg Extraction
3.84
kg
13.29
Supercritical
CO
158.25 g
kWh
4 32.0980.0560
tkm
600
3.48480.0263
C6kg
H8O7 Chemicals
0.0601 Lorry,
g 3.5-7.5Ct,6HEuro
mg
C10H16
Nto
2 extraction
2
8O7
2O8sanitaryCaCl
Polyvinyl chloride,
landfill
kg Extraction
0.02
56.171kgtkm
1400
C6kg
H5+4yFexNyODrying agent Ship
C35.46
H BO3
0.0137
6H5+4yFexNyO
Fluorescent 3lamps,
to specific
treatment for
Equipments
Lorry,
3.5-7.5
t,
Euro
4
0.330
tkm
800
MgSO
16.1100
g
0.0601
g
0.0560
mg
ZnSO4
kg
Co-solvent
4.67
kg
0.0011
kg
4
7
7
electronics wastes
0.7710.9331
1400
NaCl
1.42820.00040.15
g
Nakg
1.0013steelShipg
Na0.55
mg
CuSO4
Stainless
kgtkm
2CO3
2CO3
Textiles, to Co(NO
municipal
incineration
kg disposal
C6H8O7
0.34280.00020.12
g
C10kgH16NFinal
0.0551
g
C10H16N2O8
0.0513
mg
2O8
3 )2
Solid
waste
Lorry,
3.5-7.5
t,
Euro
4
2.442
tkm
50
C
H
Fe
N
FeCl
kg
0.0028
6 5+4y x y
Diatomaceous earth,
to inert landfill
kg
3
35.46
2.4520.0267
tkm (OPT. 2)mg
O7
0.34280.0001 g
H3kg
BO3
0.0286
g
H3BO3
ZnCl2
INPUTS
from
ENVIRONMENT
Na2CO3
5.71270.0001 g
ZnSO
0.0022
g
ZnSO4
0.0021
mg
CoCl2
kg 4
of
culture
medium
C10the
H16N
0.31420.0098 g
CuSO
g
CuSO4
0.0007
mg
MnClPreparation
kg 4 Cultivation0.0008
2O8
2
Biomass inoculum
0.02 g
kgCo(NO ) CO2 0.0005
kg
H3BO3
0.16340.0012
g 26.44Co(NO
)
0.0005
mg
Na2MoO4
kg 3 2
3 2
River/rain water
LFeCl
ZnSO4
0.0126 2,786 g
0.0058
g
FeCl3
0.0054
mg
3
CuSO4
0.0046
g
ZnCl2
0.0003
g
ZnCl2
0.0003
mg
Co(NO3)2
0.0029
g
CoCl2
0.0001
g
CoCl2
0.0001
mg
FeCl3
0.0332
g
MnCl2
0.0206
g
MnCl2
0.0192
mg
ZnCl2
0.0017
g
Na2MoO4
0.0024
g
Na2MoO4
0.0023
mg
CoCl
0.0007
g
Energy
Total electricity from Irish grid

Contents
Introduction to microalgal processes

Goal and scope

Inventory analysis

Environmental impact assessment

Conclusions

10

09/03/2015

ENVIRONMENTAL IMPACT ASSESSMENT

Selected methodology and impact categories

 Use of LCI results to quantify environmental potential impacts
 Assignment of impact categories according to CML 2001 method
Abiotic depletion

POFP

Photochemical oxidants
formation

AP

Acidification

HTP

Human toxicity

EP

Eutrophication

MEP

Freshwater aquatic
ecotoxicity

GWP

Global warming

MEP

Marine ecotoxicity

ODP

Ozone layer depletion

TEP

Terrestrial ecotoxicity

ADP

ENVIRONMENTAL IMPACT ASSESSMENT

Characterization results
Functional unit =
800 g astaxanthin
Impact category
ADP

Unit

Chemical
disinfection

Ozone
sterilization

kg Sb eq

13.6

13.6

AP

kg SO2 eq

12.1

12.1

EP

kg PO4-3 eq

1.90

1.88

GWP

kg CO2 eq

1.86

1.86

ODP

kg CFC-11 eq

0.130

0.129

HTP

kg 1,4-DBeq

321

319

FEP

kg 1,4-DBeq

261

259

MEP

kg 1,4-DBeq

190

188

TEP

kg 1,4-DBeq

0.088

0.087

POFP

kg C2H4 eq

0.490

0.490

11

09/03/2015

ENVIRONMENTAL IMPACT ASSESSMENT

Relative contribution per stage

Relative contributions

100%
80%
60%

Cultivation stage
80% of impacts

40%
20%
0%
ADP

AP

EP

GWP

ODP

HTP

FEP

MEP

TEP

POFP

Cleaning of the reactor

Preparation of the culture medium

Cultivation

Harvesting

Extraction

ENVIRONMENTAL IMPACT ASSESSMENT

Relative contribution per involved activity

Relative contributions

100%
80%
60%

Electricity
>75% of impacts

40%
20%
0%
ADP

AP

EP

GWP

ODP

HTP

FEP

MEP

TEP

POFP

Water

Chemicals

Materials

Transport

Electricity

Waste treatment

12

09/03/2015

ENVIRONMENTAL IMPACT ASSESSMENT

Relative contribution of electricity requirements

Agitation in stress
stage 2.4%
Spray drying 4.2%
Aeration in stress
stage 2.4%
Supercritical fluid
extraction 8.0%

Reactor lighting in
growth stage 19.5%
Reactor lighting in
stress stage 58.5%

Aeration in growth
stage 2.4%
Agitation in growth
stage 2.4%

ENVIRONMENTAL IMPACT ASSESSMENT

Improvement scenarios

How to reduce electricity

consumption

13

09/03/2015

ENVIRONMENTAL IMPACT ASSESSMENT

Improvement scenarios

Improvement
alternatives

Annular PBR
with sunlight

Flat-panel PBR
with artificial
lighting

Flat-panel
PBR with
sunlight

ENVIRONMENTAL IMPACT ASSESSMENT

Improvement scenarios
 Sc 1 (base case): annular PBR with artificial
light and 800 g astaxanthin produced.
 Sc 2: annular PBR with sunlight and 400 g
astaxanthin produced.
 Sc 3: flat-panel PBR with artificial light
and 800 g astaxanthin production.
 Sc 4: flat-panel PBR with sunlight and
400 g astaxanthin produced.

14

09/03/2015

ENVIRONMENTAL IMPACT ASSESSMENT

Improvement scenarios
Reductions of impact in all
proposed scenarios,
ranging between 15-75%

Relative contribution (%)

100
80
60
40
20
0
ADP

AP

EP

GWP

ODP

HTP

FEP

MEP

TEP

Sc. 1: 2 annular, artificial

Sc. 2: 2 annular, sunlight

Sc. 3: 2 flat-panel, artificial

Sc. 4: 2 flat-panel sunlight

POFP

ENVIRONMENTAL IMPACT ASSESSMENT

Conventional vs BAMMBO process

3 alternative routes for
astaxanthin production
According to Baker
and Saling (2003)

Chemical
synthesis
 Petrochemical

raw materials
 Solvent loss
neglected

Yeast
fermentation
 0.4-1%
astaxanthin content
in Phaffia
rhodozyma

Algal astaxanthin
 2% astaxanthin
content
 Cultivated in
open pond reactors

15

09/03/2015

ENVIRONMENTAL IMAPCT ASSESSMENT

Conventional vs BAMMBO process

ENVIRONMENTAL IMPACT ASSESSMENT

Conventional vs BAMMBO process

BAMMBO process:
 4-5% astaxanthin
 Annular PBR (closed) + conventional lamps
Additional scenario:
 Flat-panel PBR + LEDs

16

09/03/2015

ENVIRONMENTAL IMPACT ASSESSMENT

Normalized impacts of
astaxanthin production

Conventional vs BAMMBO process

1,0
0,9
0,8
0,7
0,6
0,5
0,4
0,3
0,2
0,1
0,0
Energy use

Emissions

Raw
material
use

Fermentation
Lucantin Pink
Algae (flat-panel+LED BAMMBO)

Toxicity
potential

Risk
potential

Surface
use

Algae (Open pond, 2003)

Algae (annular BAMMBO)

Contents
Introduction to microalgal processes

Goal and scope

Inventory analysis

Environmental impact assessment

Conclusions

17

09/03/2015

Conclusions
Environmental performance of a pilot-scale
microalgal process for the production of
high value-added molecules was evaluated
with LCA methodology.

Electricity consumption was identified as the

main responsible for the total impacts in all
categories, especially due to lighting
requirements in the cultivation stage.

Conclusions
Other steps of the process chain, such as
production of chemicals for culture medium,
materials for equipment or air supply present
secondary contributions in all the assessed
impact categories.

Results suggest that significant improvements

may be achieved by the use of alternative reactor
configurations, thanks to the lower electricity
requirements of these options.

18

09/03/2015

General conclusions of comparative analysis

Annular reactor with conventional lights:
 Improvement in 3 categories.
 High energy use (lights, aeration, agitation).

Flat-panel reactor with LEDs (improved option):

 Improvement in all categories with respect to
conventional scenarios.
 Similar impacts as synthetic, but synthetic is not
approved for human consumption and solvent loss
is not taken into account.

La mer est tout! Elle couvre

les sept diximes du globe
terrestre. Son souffle est pur
et sain. Cest limmense
dsert o lhomme nest
jamais seul, car il sent frmir
la vie ses cts
(Vingt mille lieues sous les mers, 1870)

19

09/03/2015

Acknowledgements
Sustainable Production of Biologically Active
Molecules of Marine Based Origin

FP 7 - KBBE-2010-4 Collaborative Project

www.bammbo.eu

Sustainable pilot-scale production of carotenoid

compounds from the green microalga
Haematococcus pluvialis

G. Feijoo & M.T. Moreira

Departamento de Ingeniera Qumica
Universidad de Santiago de Compostela
gumersindo.feijoo@usc.es
www.usc.es/biogrup
@EnxQuim_USC

20