MBT Fall2015
MicroalgalBiotechnology:
CultivationandProcessing
HectorDelaHozSiegler,Ph.D.,P.Eng.
DepartmentofChemicalandPetroleumEngineering
UniversityofCalgary
h.siegler@ucalgary.ca
October29th,2015
HectorDelaHozSiegler
Outlineoftodaystalk
I. Introduction to microalgae
What and why
Applications
PartI
INTRODUCTIONTOMICROALGAL
BIOTECHNOLOGY
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Microalgae:whatarethey?
Microalgaeareplantlikeunicellularorganisms:apolyphyletic group
ofphotosyntheticeukaryotes.
Sunlight-driven cell factories able to convert carbon dioxide to
potential biofuels, food, and high-value products
ProductsfromMicroalgae
Image source: Rosenberg, J.N., Oyler, G.A., Wilkinson, L., Betenbaugh, M.J. A green light for engineered algae: redirecting metabolism to fuel a biotechnology
revolution ,Current Opinion in Biotechnology, 19 (5), pp. 430-436 (2008)
Applications
Humanand
animalfood
Pharma and
nutraceuticals
CO2 Capture
Microalgae
Biofuels
Wastewater
treatment/
Remediation
Finechemicals:
e.g.antioxidants
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Microalgaeascellfactories
Benefits:
Highlyefficient
Simplenutritionalrequirements
Easilyadaptabletoenvironmentalstresses
Produceandstorehighamountsofoil
Othervaluablebyproducts
Eukaryotic!
Challenges
Lowculturedensity
Slowgrowth:lowproductivity(comparedtobacteriaandyeast)
Highproductioncosts
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SomeCommercialApplications
Species/group
Product
Application areas
Production
facilities
References
Haematococcus
pluvialis /
Chlorophyta
Carotenoids,
astaxanthin
Open ponds,
PBR
Odontella aurita
/ Bacillariophyta
Fatty acids
Pharmaceuticals,
cosmetics, baby food
Open ponds
Isochrysis
galbana /
Chlorophyta
Fatty acids
Animal nutrition
Open ponds,
PBR
Phaedactylum
tricornutum /
Bacillariophyta
Lipids, fatty
acids
Open ponds,
basins, PBR
Muriellopsis sp.
/ Chlorophyta
Carotenoids,
Lutein
Open ponds,
PBR
Crypthecodinium
cohnii
DHA
Food additive
Fermenters
(heterotrophic)
Carvalho et al.
(2006)
Currently,applicationsofmicroalgalbiotechnologyarelimitedtoniche(small)markets.Though
highvalue!Weexpecttomoveintolargescalemarkets.
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PartII
ALGAEASASOURCEOF
RENEWABLEENERGY
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Energy,economy,andGlobalwarming
100
PrimaryEnergyConsumption(kWyear/person)
Poland
Ghana
Uzbekistan
Canada
US(2002)
10
Russia
South
Korea
France
UK
China
China(2002)
Norway
US
Japan
Brazil
India(2002)
India
0.1
Vanuatu
Chad
0.01
100
1,000
10,000
100,000
GDP($/person/year)
Energyreserves/Energyconsumption
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Climatechangedebate
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Thenaturalcarboncycle
Carbon CaptureandUtilisation(CCU)
T. Shirvani, X. Yan, O. R. Inderwildi, P. P. Edwards and D. A. King, Energy Environ. Sci., 2011, 4, 37733778
Landrequiredforsatisfydemand
Timetorealworldapplication
Biofuels
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Themicroalgalbiofuelsportfolio
Feedstocks
Photosynthesis
CO2
Algal Biomass:
Water
Intermediates
Fuels
Biodiesel
- Oil/Lipids
Green Diesel
Hydrodeoxygenation
Sunlight
Trace elements
- Sugars/Starch
Hydrolysis
- Lignocellulose
Sugars
Bio-oil
Pyrolysis
Excreted products:
Gasoline
Hydrotreating
SynGas
- Hydrogen
Hydrogen
- Alcohols
Alcohols
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BiodieselfromMicroalgae
Biodieselderivedfromoilcropsisapotentialrenewableand
carbonneutralalternativetopetroleumfuels.
Biodieselfromoilcrops,wastecookingoilandanimalfatcannot
realisticallysatisfythedemandfortransportfuels.
Crop
Oil yield
(L/Ha)
% of existing US
cropping area
Corn
172
1540
846
Soybean
446
594
326
Canola
1190
223
122
Oil Palm
5950
45
24
136900
1.1
58700
4.5
2.5
Microalgae
(70% oil w/w)
Microalgae
Croplandrequirement
bydifferentoilcropsto
replace50%ofall
transportfuelneedsof
theUS.Chisti(2007).
Toooptimistictobetrue!
Algaeasasourceofoil
Species
Oil content
(% dw)
Reference
Ankistrodesmus TR-87
28 40
Botryococcus braunii
25 75
Sheehan et al. (1998); Banerjee et al. (2002); Metzger and Largeau (2005)
Chlorella sp.
28 32
Chlorella protothecoides
15 55
Xu et al. (2006)
Cyclotella DI-35
Dunaliella tertiolecta
Hantzschia DI-160
42
36 42
66
Isochrysis sp.
7 33
Nannochloris
20 - 35 (6 - 63)
Nannochloropsis
46 (31 - 68)
Nitzschia TR-114
28 50
Phaeodactylum tricornutum
20 31
Scenedesmus TR-84
Stichococcus
45
33 (9 - 59)
Tetraselmis suecica
15 32
Thalassiosira pseudonana
(21 - 31)
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OilandBiodiesel
Triglycerides:
Glycerol
FattyAcids
BiodieselProduction:
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PolyunsaturatedFattyAcids(PUFA)
Fattyacidswithmultipledoublebonds
EPA: C20:5
DHA: C22:6
C18:3andlongerareessential:mammalscannotsynthesize
C18:3.Needtotakethemfromtheirdiet
Multiplebiologicalfunctionsassignallingmoleculesorbuilding
blocks
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MicroalgaeasaSourceof3PUFA
FishoilhasbeenusedforthecommercialproductionofEPA
andDHA.
Factorsthatlimitfishoilasasourceof3fattyacidsinclude:
taste,odourandstabilityproblems.Highpurificationcost.
Fishobtain3fattyacidsfromtheirdiet.
Severalspeciesofmicroalgaeareprimaryproducersoflong
chainPUFA.
US$1.5billion/yeargeneratedfromtheproductionofDHA
(Pulz andGross,2004).
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PUFAproportionsinMicroalgae(%TFA)
Organisms
ARA (20:4)
EPA (20:5)
DHA (22:6)
Gymnodinium splendens
30
Cricosphaera elongata
28
Isochrysis galbana
15
7.5
Monodus subterraneus
4.7
33
Nannochloropsis sp.
35
Schizochytrium sp.
1.0
2.3
40.9
Chlorella minutissima
5.7
45
Hetermastrix rotundra
28
Chromonas sp.
12.0
6.6
Cryptomonas sp.
16
10
Rhodomonas sp.
8.7
4.6
Asterionella japonica
11
20
Biddulphia sinensis
24
Crypthecodinium cohnii
30
Nitzschia laevis
6.2
19.1
Phaeodactylum Tricornutum
34.5
Skeletonema costatum
29.2
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GeneralProcessDiagram
Solvent
recovery
Culture
Extraction
Crude
Product
Harvesting
debris
Cell disrupter
Dryer
S/L Separator
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PartIII
MICROALGALCULTURING
TECHNIQUES
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Nutritionalrequirements
Dependsonapplication
Foodorhealthoils:foodgradechemicals
Otherwiseindustrialchemicalsorseawater/wastewater
Carbonsource:CO2,sugars,acetate,ethanol
Macronutrients:Nitrogenandphosphorus
Micronutrients:Fe,Mg,Si,S,K
Traces:Ca,Mn,Zn,Co,Se,Cu,Mo
Vitamins:B1,B12,B6,B2
Seawater:Na,K,Mg,Ca,Cl,SO4,HCO3,BO3
Br,F,IO3,Li,Rb,Sr,Ba,Mo,V,Cr,As,Se
NO3,PO4,Fe,Zn,Mn,Cu,Co,Si,Ni
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Multiplewaysofgrowing
Phototrophic
Heterotrophic
Mixotrophic
EnergySource
Applications
FixCO2
Slowgrowth
Noactivityduring
night
CO2 capture
Fast growth
Productiveallday
long
Dontfixgreenhouse
gases
ProduceCO2
Wastewatertreatment
Highvalueproducts
Proteins
FixCO2
Fast growth
Productiveallday
long
Complexand
unknowncell
regulation
CO2 capture
Wastewatertreatment
Highvalueproducts
Proteins
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SolarradiationinCanada
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SolarradiationinAlberta
FortMcMurray:
Edmonton:
MedicineHat:
4181MJ/m2y
4510MJ/m2y
5221MJ/m2y
Munich(GE):
Naples(IT):
KualaLumpur:
Orlando(FL):
Acapulco(MX):
Phoenix(AZ):
4044MJ/m2y
5293MJ/m2y
5622MJ/m2y
5922MJ/m2y
7261MJ/m2y
7621MJ/m2y
Solarradiationdatatakenfrom:U.S.DepartmentofEnergy EnergyPlusWeatherData.
http://apps1.eere.energy.gov/buildings/energyplus/cfm/weather_data.cfm
Culturingtechniques:OpenPonds
Byfar,themostcommon
productionsystem.
Lowinstallationcost
Lagoonsorartificialponds
Highriskofcontamination:
Unwantedalgae
Grazers
Applicationlimitedtofewspecies
(extremophiles).
Unmixedponds:arearangefrom1
200Ha,depth2030cm
Racewaypondsareupto1Ha.
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Culturing:ClosePondsandTanks
Simplerdesignssimilartoopen
ponds,withacover
(greenhouses).
Aimtoreducecontamination
risks.
ControlCO2 looses.
Tanksareusuallymixedby
aeration.
Deeptanksareinefficient.Bad
lighttransmission.
Easytooperate,lowcost.
30
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Culturing:Photobioreactors
FlatPanelphotobioreactor
ArizonaCenterforAlgalTechnologyandInnovation
TubularPhotobioreactor AlgaeandBiofuelsFacility,South
AustralianResearchandDevelopmentInstitute
FlexibleplasticfilmPhotobioreactor Algenol,Florida
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Culturing:Photobioreactors
Betterculturecontrol
Higherproductivity,andculture
density
Minimalcontaminationrisk
Wellmixed
Excellenttemperaturecontrol
Oxygencontrolisanissue
Highcapitalinvestment
Frequentcleaningrequired
Coolingrequired
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HeterotrophicProductionofAlgae
Somealgaespeciescangrow
usinganorganiccarbonsource.
Conventionalbioreactorscanbe
used.
33
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Phototrophicvs.Heterotrophic?
Oil content
(%)
Cell conc.
(g/L)
Oil Prod.
(mg/L d)
Ettlia oleoabundans
36 42
2.9
164
Griffiths et al (2009);
Li et al. (2008)
Nannochloropsis sp.
31 68
2.1
204
Amphora
40 51
593
Chlorella sp.
28 32
1.1
139
Chlorella vulgaris
25 42
1.7
54
Chlorella zofingiensis
25.8
1.9
35
Chlorella zofingiensis
51.1
9.6
354
Nitzschia laevis
16.5
22.1
914
S. Limacinum (DHA)
17.3
37.9
656
A. protothecoides
38.3 53.0
8.4
820
A. protothecoides
50.3 57.8
51.1
3320
Phototrophic
References
Heterotrophic
Specie
34
PartIV
MODELBASEDOPTIMIZATIONOF
HETEROTROPHICALGALCULTURES
35
BioprocessOptimization
Continuous/
Realtime
Processconditions
Mediaformulation
Strainselection
Geneticmodification
36
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TheObjectiveforOptimization
Stress
Nitrogen
37
Understandingalgalgrowth
Nitrogen uptake
Cellular growth
Lipid production
38
Analgalgrowthmodel
Cellular growth
Nitrogen uptake
Oil production
39
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Macroscopicbalances
40
Optimization:Problemformulation
Subject to:
41
Simulationresults
Biomassproductivityin
continuouscultures
Lipidproductivityin
continuouscultures
42
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Experimentalresults
43
Biomassproductivityandgrowthrate
44
Lipidproductivityandproductionrate
45
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Oil Productivity
(g/L h)
Bacteria
Specie
E. coli (gen. modified)
25.4
0.246
Elbahoul et al (2010)
R. opacus PD630
38.4
0.171
Kurosawa et al (2010)
M. ramanniana
67.7
0.17
Hiruta et al (1997)
C. echinulata
26.9
0.07
Kosa et al (2011)
R. toruloides
67.5
0.54
Li et al. (2007)
L. starkeyi
56.0
0.04
C. curvatus
82.7
0.47
Schizochytrium sp.
30
0.096
Ganuza et al (2007)
C. vulgaris
9.7
0.12
A. protothecoides
50.3
0.14
A. protothecoides
49.4
0.43 0.84
Microalgae
Yeasts
Lipid content
(%, w/w)
Molds
Comparativestudy:growthonglucose
References
De la Hoz et al (2012)
46
Optimization:closingremarks
Modelbased optimization of heterotrophic microalgal cultures
allowed to reach very high densities, with biomass productivity
greater than 30 g/L d, and as high as 70 g/L d.
High oil content (4060% w/w) can be sustained with a lipid
productivity around 20 g/L d.
High quality monitoring and control is essential to achieve high
productivities.
Better control / sensors = higher productivity.
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Summary
Algaearepromisingorganisms:highlyefficient
Goodsourceofoil:PUFA,biodieselprecursor
Algaecangrowthonsimpleinexpensivemedia
Severalreactortypesandgeometry.Applicationwilllimit
reactorchoice
Severalsuccessfulcommercialapplicationscurrentlyworking.
Alotofresearchisstillneeded!
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