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LAB-SCALE TECHNOLOGIES FOR THE CULTIVATION AND BIOREFINERY OF MICROALGAE SPEAKER: DAVIDE DE FRANCISCI TECHNICAL UNIVERSITY OF DENMARK This project has received funding from the European Union’s Seventh Programme for research, technological development and demonstration under grant agreement No 280756.

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Page 1: Lab-scale technologies for the Cultivation and biorefinery ...e4water.eu/_media/1625_DTU E4Water copy.pdf · EXPONENTIAL GROWTH RATE . ... Cyclic Autotrophic/heterotrophic Constant

LAB-SCALE TECHNOLOGIES FOR THE CULTIVATION AND BIOREFINERY OF MICROALGAE

SPEAKER: DAVIDE DE FRANCISCI TECHNICAL UNIVERSITY OF DENMARK

This project has received funding from the European Union’s Seventh Programme for research, technological development and demonstration under grant agreement No 280756.

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E4Water Final Conference, Brussels, April 19-20, 2016 2

MICROALGAE CHARACTERISTICS STRENGHTS AND BENEFITS

Diameter: 0,5 µm – 2 mm

Salt, brakish and fresh water Amount to 0.2 % of the total global standing biomass Produce app. 55 % of the total global annual biomass

1 ton of algae can fixate 1.8 tons CO2

High lipid content (up to 77% of DW)* *Chistie, 2007

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E4Water Final Conference, Brussels, April 19-20, 2016 3

ADDED VALUE FROM MICROALGAE

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E4Water Final Conference, Brussels, April 19-20, 2016 4

Algal biomass = high added value products and bioenergy

(Bruton et al, 2009)

ADDED VALUE FROM MICROALGAE

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E4Water Final Conference, Brussels, April 19-20, 2016 5

Trend alert: Biofuel companies have shifted focus to high-value products: Algenol Sapphire Synthetic Genomics Aurora Cellana Solix

ADDED VALUE FROM MICROALGAE

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E4Water Final Conference, Brussels, April 19-20, 2016 6

Production is costly (cultivation , harvesting) - Open pond - Photobioreactor

> Economically unfeasible

CHALLENGE

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E4Water Final Conference, Brussels, April 19-20, 2016 7

Algal biomass

Harvesting

Bioprocessing Biofuels

• Bioethanol • BioH2 • BioCH4 • Biodiesel

Extraction processing

• Food (dietary) • Biochemicals (vitamins,

minerals, pigments, enzymes, etc.)

Biofertilizer

Rice, wheat, cultivation

• Feed (fish, poultry)

N, P, K nutrients recycling Recycle exhaust gas for a carbon source

Algal biomass = high added value products and bioenergy

BIOREFINERY

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E4Water Final Conference, Brussels, April 19-20, 2016 8

POSSIBLE SOLUTION

Integration of algae production and wastewater treatment

16.04.2016

Remove pollutants Obtain bio-products

Utilities - CO2 - Power

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E4Water Final Conference, Brussels, April 19-20, 2016 9

STRATEGY

Pond-like Lab scale

Photobioreactor

Flat panel Lab scale

Photobioreactor

Comparison of potential yield

vs. cost

Cultivation system choice

Raceway pond

Photobioreactor

Biomass separation

Biomass characterization

High added value products

Biogas potential

Productivity comparison

Best algal species for

each wastewater

1. Different species are screened for their potential to grow in different wastewaters

2. Upscaling strategy

Candidates

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E4Water Final Conference, Brussels, April 19-20, 2016 10

• Variety of microalgal species can be screened for their potential to grow in waste-waters

Best algal species for

each wastewater

Microplate Reader

MICROPLATE SCREENING

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E4Water Final Conference, Brussels, April 19-20, 2016 11

SYMBIOSIS

11

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E4Water Final Conference, Brussels, April 19-20, 2016 12

Fluorescence measurements have a lower limit of quantification and can therefore extend the observable period of exponential growth

Fluorescence has less variation among triplicates and a signal more quickly separated from the blank. Chlorella vulgaris growth curve.

FLOURESCENCE TO DETERMINE EXPONENTIAL GROWTH RATE

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E4Water Final Conference, Brussels, April 19-20, 2016 13

VALIDATION OF THE SCREENING METHOD

Microplate-based measurements enable predictions comparable to reactor-based ones

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E4Water Final Conference, Brussels, April 19-20, 2016 14

Microplate screening is a fast and cost effective

method to test hundreds of different waste-waters/strains combinations

The microplate screening method fits perfectly into the Symbiosis scenario

SCREENING – STRENGTHS AND BENEFITS

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E4Water Final Conference, Brussels, April 19-20, 2016 15

APPLICATION

Effluent TN: 180 mg L-1

TP: 10 mg L-1

VFA: ~300 mg L-1

Novozymes Kalundborg WWTP

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E4Water Final Conference, Brussels, April 19-20, 2016 16

FRESHWATER

Bothryococcus braunii

Chlorella sp

Euglena gracilis

Galdieria sulphuraria

Haematococcus pluvialis

Nitzschia palea

Scenedesmus acutus

MARINE

Brachiomona submarina

Dunaliella tertiolecta

Isochrysis sp.

Nannochloropsis oculata

Phaeodactylum tricornutum

Porphyridium cruentum

Rhodomonas salina

MICROALGAE SPECIES CANDIDATES

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E4Water Final Conference, Brussels, April 19-20, 2016 17

NOVOZYMES WASTEWATER

C. protothecoides

S. acutus

H. pluvialis

C. vulgaris

Unidentified diatom

The best candidate resulted to be Chlorella spp. S. obliquus and H. pluvialis also shows good growth on 100% WW.

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E4Water Final Conference, Brussels, April 19-20, 2016 18

REACTOR CONFIGURATION

High light Low light

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E4Water Final Conference, Brussels, April 19-20, 2016 19

Flat-panel photobioreactors (0.4 L) 200 and 2100 µmol m-2s-1 PAR Optimal temp and pH maintained by build in feedback systems C. sorokiniana 100 % waste-water Optical density, dry weight and nutrients

REACTOR CONFIGURATION

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E4Water Final Conference, Brussels, April 19-20, 2016 20

Photosynthetic yield (n, g mol-1 ), and percentage of light (PAR) absorbed (p, % ).

REACTOR CONFIGURATION

High light Low light

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E4Water Final Conference, Brussels, April 19-20, 2016 21

NUTRIENT REMOVAL

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E4Water Final Conference, Brussels, April 19-20, 2016 22

Legend. Pigment content in Chlorella sorokiniana biomass. Dilution rate in decreasing order.

BIOREFINERY

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E4Water Final Conference, Brussels, April 19-20, 2016 23

Dilution rate is in decreasing order Linoleic acid (18:2) is an omega-6 fatty acid α-linolenic acid (18:3) (ALA) is an omega 3 fatty acid

BIOREFINERY

Fatty acids

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E4Water Final Conference, Brussels, April 19-20, 2016 24

UTILIZATION OF ORGANIC CARBON

waste streams offer a low cost, sustainable source of N, P and other minerals

they may also contain organic carbon, which can me metabolized by microalgae

acetate is characteristic of AD effluents

Aim: develop a strategy for adding AD effluent to bioreactor in order to maximize productivity

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E4Water Final Conference, Brussels, April 19-20, 2016 25

UTILIZATION OF ORGANIC CARBON

• Strategy 1. Autotrophic: M8a Dilution during light cycle 2. Mixotrophic: + 1 g/L Acetate during light cycle 3. Cyclic Autotrophic/heterotrophic Constant rate: +2 g/L during dark cycle, + acetate free medium during the light. 4. Cyclic Autotrophic/heterotrophic only diluting at night: +2 g/L during dark cycle, no dilution during the day. “enhanced heterotrophy”

Conditions: Mimic Kalundborg, Ecoduna + Anaerobic Digestor effluent 16:8 light:dark cycle, 190-200 μmol m-2 sec-1

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E4Water Final Consortium Meeting, Antwerp, April 18, 2016 26

UTILIZATION OF ORGANIC CARBON

Day

0 2 4 6 8 10 0

1

2

3

4

5

6

2 mixotrophic reactor 3 cylcic reacfor

tota

l har

vest

(g)

Acetate addition increased productivity Highest substrate efficiency came when adding acetate

at night

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E4Water Final Consortium Meeting, Antwerp, April 18, 2016 27

CONCLUSIONS

identified a waste-water stream that can replace culture medium

identified the preferred vertical orientation demonstrated a new method to characterize

growth, composition and nutrient removal in microalgae cultivation in waste-waters

identified a possible route to optimize biomass productivity mixotrophically

validated a microplane-based screening method

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E4Water Final Conference, Brussels, April 19-20, 2016 28

• Due to high protein and lipid content (over 70% in total), microalgae can be dedicated for biofuels production, such as biogas (methane).

• Biogas derived from the digestibility of microalgae has a high methane content, i.e. over 60%

• Methane content is 7–13% higher compared to biogas from energy crops, like maize silage.

• Biogas derived from the digestibility of microalgae does normally not contain sulphur.

• Co-digestion of manure with several organic wastes is a common practice applied in the Danish centralised biogas plants.

• Biogas plants currently utilise around 8 % of available manure and a large part of the organic wastes from the Danish industries.

BIOENERGY

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E4Water Final Conference, Brussels, April 19-20, 2016 29

SUBSTRATE CHARACTERISATION

Parameters Unit Microalgae Swine manure

TS g/L 24.60 33.50 VS g/L 22.69 23.00 pH - 5.87 8.50 TKN g/L 1.78 4.31 Ammonium nitrogen g/L 0.18 3.09 Organic N g/L 1.60 1.22 Proteins g/L 10.00 7.63 Lipids g/L 2.63 1.30 Carbohydrates g/L 8.19 6.53 Total VFA g/L 0.27 6.33 Acetate g/L 0.13 4.91 Propionate g/L 0 1.07

Iso-butyrate g/L 0 0 Butyrate g/L 0 0 Iso-valerate g/L 0 0.35 Valerate g/L 0 0 n-hexanoate g/L 0.14 0

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E4Water Final Conference, Brussels, April 19-20, 2016 30

BATCH ASSAY

0

175

350

525

700

0 5 11 16 21

Met

hane

yie

ld

mLC

H4/

gVS

Days

Experimental BMP of swine manureExperimental BMP of microalgae

Theoretical BMP of swine manureTheoretical BMP of microalgae

0

175

350

525

700

0 5 11 16 21

Met

hane

yie

ld

mLC

H4/

gVS

Days

80/20 60/40

40/60

Manure /algae ratio

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E4Water Final Conference, Brussels, April 19-20, 2016 31

Batch experiments

• The addition of 40% wastewater microalgae in swine manure exhibited the highest methane yield (38% higher compared to swine manure). Further addition affected negatively the biomethanation process.

• Possible synergistic effect (further investigation needed)

Continuous experiments

• Co-digestion of SM/M at a ratio of 60/40 presented 21.6% higher methane yield compared with the corresponding one from the degradation of SM as mono-substrate.

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E4Water Final Conference, Brussels, April 19-20, 2016 32

Thank you for your attention

This project has received funding from the European Union’s Seventh Programme for research, technological development and demonstration under grant agreement No 280756.