Fermentation process co-products:

Integrated protein, energy and feedstock recovery

Julio Traub23 January 2013ICBD Heriot-Watt University, Edinburgh

18th IBD Video Link Day

Contents

1. Whisky Process and Industry

2. Whisky co-products: Current and potential markets

3. Experimental Work

4. Conclusions

5. Future Work

WhiskyA Scottish success story

Source: Diageo

19461949

19521955

19581961

19641967

19701973

19761979

19821985

19881991

19941997

20002003

20062009

- 50,000,000

100,000,000 150,000,000 200,000,000 250,000,000 300,000,000 350,000,000 400,000,000

Scotch Whisky Exports - 1946 to 2011 by volume

Year

Volu

me

(M lp

a)

Whisky - Waste Fight Club...

The are only 2 Rules...

1. It’s not waste... It’s a co-product!!!

2. Don’t touch the process!

Whisky Production ProcessMalt Whisky

Extraction

Fermentation

Distillation

Evaporation

Drying

Water

Malted barley

Yeast

Draff

Pot ale

Spirit (Whisky)

Pot ale Syrup

DraffA good example of the Food vs. Fuel controversy

• Used as animal feed• Cheap, good protein source (21% in DM basis)

• Can be used as a biofuel

Pot AleTypical Properties

pH 3.5 – 4.1

Solids 3 – 5 % w/v

Yeast count ~ 10 8 cells/mL

Copper Content 2 – 6 mg/ L

BOD 12 – 35 g/ L

COD 38 – 62 g/ L • Water• PROTEINS• Yeast• Yeast Debris• Copper• Carbohydrate

s

Sources: Graham, J., et al. (2012), Russell, I. et al (2003)

Pot AleCurrent treatment technologies

Fertiliser Consented discharge

Evaporator

Co - product???

Pot Ale Syrup

• Evaporated Pot Ale (45% Solids)• High capital and running costs• Energy intensive (CO2 emissions)• Limited used (mostly Cattle) –

High copper content = toxic• Current Supply ~ 30 – 50 ktpa

– Protein ~ 5 - 8 ktpa• Commercial products available:

– Spey Syrup (AB Agri), GP feeds

20012003

20052007

20090

500

1,000

1,500

2,000

2,500

3,000

3,500

Wheat

Barley

Maize gluten feed

Wheat feed

Distillery by-products

Soya cake and meal

k to

nne

UK Animal Feed DemandTotal = 10 m tonne in 2010

8.5%

3.2%

9.2%

28.1%

Source: http://www.defra.gov.uk/statistics

1.2%

11.5%

UK- BIOETHANOL PRODUCTIONENSUS VIVERGO

Start up May 2009 Teeside

Sep 2012Hull

Investment £300 m £300 mRaw Materials 1 Mt Wheat 1 Mt WheatProduction 400 m L 420 m LAnimal Feeds 350 k tonne 545 k tonneHistory •May 2011 temp closure

•Sep 2012 reopened Dry 390 k tonne

Moist 130 k tonne

Syrup 25 k tonne

Malt WhiskyVolumes and Co-products

DistilleryCapacity

[M Lpa/ a]

Pot Ale Proteinsa

[Tonne/ a]

DraffProteinsb

[Tonne/ a]

Protein Valuec

[£/ a]

Total 287 32 K 37 K 48 m

Median 2.7 302 347 454 k

Max 10.5 1,176 1,351 1,768 k

Min 0.05 5.6 6.4 8.4 k

a 1 Lpa = 8L Pot Ale 14 g protein = 1L Pot Ale

b 1 Lpa = 2.45 kg draff DM = 25%, CP = 21%

c £700/ ton

Aquaculture FeedsScottish Salmon – the other Scottish success story

UK Salmon Production 154 k tonne (Atlantic Salmon), 3rd largest producer in the world ~200 k tonne protein , High dependency on FM/ FO

Need for Alternative Feedstuffs Economic and Sustainable Drivers

Ideal Candidate• Price and Logistics: Availability, handling, shipping, storage• Low levels of fibre, High Protein content, Amino Acid Profile,

High Digestibility

DDGS PAS SBM FMCrude Protein 29 - 35 % 34 -38% 46 - 50% 65 -72%

Lysine 0.55% 2.1% 2.69% 4.72%

Meth + Cyst 0.86% 0.35% 1.18% 3.5%

Threonine 0.81% 1.9% 1.67% 2.5%

Amino Acid Analysiswhat the fish wants...

DDGS: Wheat Distillers Dried Grain with SolublesPAS: Pot Ale SyrupSBM: Soy bean MealFM: Fish Meal

The Horizon Protein Project

Identify• Inefficiencies• Proteins• Markets

Optimize

• Process• Product

Scale Up

• Lower Energy Consumption

• Reduce CO2 emissions• Value added By-

products

Research WorkAnalytical Process

Design &Optimization

EconomicalModelling

•Solid content

•pH

•Yeast count

•Particle size

•Protein content

•Metal Analysis (Cu,

Fe, Zn, Mn)

•Cell disruption

•Protein Purification

and Concentration

•Solid/ liquid

separation

•Cost and Capital

estimation

•Economical

Analysis (NPV, IRR,

etc.)

•Protein Markets

Process SelectionPotential processes identified:

1. Pot Ale – Barley Protein Fraction

2. Pot Ale – Yeast Protein Fraction

3. Pot Ale – Barley + Yeast Fractions

4. Pot Ale + Draff – Barley Proteins

5. Pot Ale + Draff – Barley + Yeast Fractions

Process OverviewBarley + Yeast Fractions

PROTEINS

EnzymaticTreatment

High Pressure Homogeniser

Centrifugation

Filtration

Ion Exchange Chromatography

Pot Ale

• Water• PROTEINS• Yeast• Yeast Debris• Copper• Carbohydrates

ExtractionPurification

Experimental WorkProtein Content and Distribution

Lowland PA (Feb)

Lowland PA (Aug)

Spent Wash (Jun)

Speyside PA (Nov)

02468

10121416

PelletSoluble

Prot

ein

Cont

ent (

g/ L)

Experimental WorkCopper analysis

Lowland PA (Feb)

Lowland PA (Aug)

Spent Wash (Jun)

Speyside PA (Nov)

0.00

0.50

1.00

1.50

2.00

2.50

3.00

PelletSoluble

Copp

er C

onte

nt (m

g/ L)

To disrupt or to not disruptThat is the question...

Yes No“Wasted” Protein •~30% of total protein content•Favourable AA profile •Yeast high in Lysine (close to FM)•Better end-product price

Cost + CapitalHomogeniser, enzymes, bioreactor

“Wasted” Minerals Downstream processes•Micronisation, nucleic acids, etc.•Buffers•More cost and capital!

Other value added chemicals•Glucans, chitins, phenols

Toxicity •Copper: ~70% bound to cells

Experimental Work Protein Purification and Concentration

• Buffers: PBS, lactic acid, citric acid

• Cation and anion exchange

• pH elution: 4-7

• Elution mode: Gradient and Step

• Flowrate: 1 – 5 ml/ min

Conclusionsso far...

Pot Ale is a co-product Valuable proteins are currently underused at least £20 million pa across Scotland

Current whisky co-product treatment technologies are unsustainable Expensive and energy intensive Increasing supply with static demand New markets: Salmon (?!)

Conclusions cont’dso far...

More understanding of what is needed for Salmon Feeds AA, minerals, fibres, nutritional properties

Yeast cell fraction has not yet proved to be a viable option High copper, Low protein but it must NOT be “wasted”

Draff proteins may increase feasibility of the project, but with some controversies and problems

Sustainable protein sources

Carbon Emissions

Energy Efficiencies

Added Value Co-products

Cost Savings

Industrial Symbiosis

Academic Research

Future work

Proteins• IEC: Columns (media, size), pH, flow rate• Protein Identification (SDS-Page, AA analysis)

Cell disruption • Combined method• Scale up Enzymatic Treatment

Metal Analysis• Protein and metal binding affinity

AcknowledgmentsThe Horizon Proteins Team

Nik Willoughby Lydia Campbell Paul Hughes Alan Harper Dawn Maskell Jane White Sara Bagés

www.horizonproteins.com

Acknowledgments

Heriot-Watt University (SLS + EPS) Eileen McEvoy James Bryce Margaret Stobie Sean McMenamy Steve Euston Vicky Goodfellow

Glenkinchie and North British Distilleries

Industrial Partners, Collaborators and Sponsors

QUESTIONS?

Some (Scottish) thoughts....

“Some hae meat and canna eat,

And some wad eat that want it,

But we hae meat and we can eat,

And sae the Lord be thankit.”

Robert Burns (1759 –1796)

Experimental WorkCell Disruption

Source: Sara Bagés (2012)

Malt WhiskyDistillery size distribution

Less

than

500

500

- 100

0

1,00

1 - 2

,500

2,50

1 - 5

000

5,00

1 -7

,500

7,50

1 - 1

0,00

0

Mor

e th

an 1

0,00

0

0

10

20

30

40

0%20%40%60%80%100%

Frequency

% Cumulative Distillery Capacity (k Lpa/ a)

Num

ber o

f Disti

llerie

s

Source: SWA

Protein MarketsFish Meal and Soybean Meal

Source: http://www.indexmundi.com

Last 12 monthsFM: up 55%SBM: up 48%

Feb-07Dec-07Oct-08Aug-09Jun-10 Apr-11Feb-120

200

400

600

800

1000

1200

1400

Fishmeal PriceSoybean Meal

Poun

d St

erlin

g pe

r M

etric

Ton

Aug-07

Dec-07

Apr-08

Aug-08

Dec-08

Apr-09

Aug-09

Dec-09

Apr-10

Aug-10

Dec-10

Apr-11

Aug-11

Dec-11

Apr-12

Aug-12

Dec-12

0.0

1.0

2.0

3.0

4.0

5.0

6.0

Poun

d st

erlin

g pe

r Kg

Protein Markets Salmon Price

Last 12 months: up 17%

Down 30% since Apr-11

Source: http://www.indexmundi.com

Experimental WorkCell Disruption

Particle Diameter (µm.)

Volume (%)

0

10

0

10

20

30

40

50

60

70

80

90

100

0.1 1.0 10.0 100.0 1000.0 10000.0

Particle Diameter (µm.)

Volume (%)

0

10

0

10

20

30

40

50

60

70

80

90

100

0.1 1.0 10.0 100.0 1000.0 10000.0

Distribution of copper in soluble fraction of pot ale

Supernatant separated into low and high MW fractions using Amicon Ultra-15 Centrifugal filter tubes

Retentate (>3 or 10 KDa)

Filtrate (<3 or 10 KDa)

Pot Ale (Aug) Pot Ale (Feb)

3 K 0.92 1.13 0.95 0.78

10 K 0.92 1.19 0.96 0.90

initialfiltrateV

initialfiltrateV

initialfiltrateCu

initialfiltrateCu

15 ml

Dialysis with 15 ml water

SDS-PAGE analysis

Copper analysis

SDS-PAGE analysis of proteins

1 2 3 4 5 6 7 8 9 10

38-42 kDa

12-14 kDa

Spent wash and pot ale (Feb) – no pretreatment

175

8058463025

177

5.3 3.5 9.9 6.6 Spent wash

Pot ale

• Gel: 4-20% Bio-Rad Mini-Protean TGX precast gel (Cat No. 456-1094)• Sample buffer: Laemmli 2x sample buffer (Sigma, S3401) • Running buffer: Tris-Glycine• Protein marker: Prestained protein marker, New England Biolabs (P7709V)• Stain: Colloidal Coomassie Brilliant Blue (Kang et al., 2002)

‘Lysed’ Pot ale

‘Lysed’ cells

1 2 3 4 5 6 7 8 9 10

175

8058463025

177

Pot ale (Feb) – Retentate

µg protein: µg protein: 9.9Pot ale

3K Retentate

10K Retentate

Pot ale

9.9131.1 65.6 26.2 66.3132.6 26.5 26.5

72-80 kDa

MWKDa

MWKDa

Horizon ProteinsPAS Pot Ale Pot Ale + Draff

Barley Yeast MixedBarley +

DraffMixed +

DraffProtein Yield % 71% 29% 100% 87% 100% 100%Protein output ton/ a 216 86 302 563 650 302 Price* £/ ton 545 1,391 787 545 658 545Total Cost £/ ton 364 987 568 314 509 629

Process Selection – Price vs. Cost

*Bulk Protein Price. SBM: £230/ ton, FM : £830/ ton

Process Selection – Investment + NPV

Source: FAO

Source: FAO