Cassava breeding potential for bioethanol

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Cassava breeding potential Cassava breeding potential

for bioethanol for bioethanol for bioethanol for bioethanol

Becerra López

Becerra López‐‐Lavalle L A Dufour DLavalle L A Dufour D Becerra López

Becerra López Lavalle, L.A. , Dufour, D., Lavalle, L.A. , Dufour, D., Sánchez, T. and H. Ceballos

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Outline Outline

I t d ti

• Introduction

• High stable and reliable productivityHigh, stable and reliable productivity • Novel traits

• Processing methods X root quality interactions • _Perspectives

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I t d ti

• High stable and reliable productivityHigh, stable and reliable productivity

• Novel traits

• Processing methods X root quality interactions

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Cassava origin Cassava origin

Flooded land Flooded land

Low

Low Soil Soil FFertilityertility

Degraded Soils Degraded Soilsgg

Slopped Land Slopped Land

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Cassava modern production Cassava modern production

Sub

Sub--humid environmenthumid environment Acid Acid ––Soil environmentSoil environment

19’000,000 hectares 19’000,000 hectares

Near Hanoi, Vietnam

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Cassava modern production Cassava modern production

Sub

Sub--humid environmenthumid environment Acid Acid ––Soil environmentSoil environment

19’000,000 hectares 19’000,000 hectares 233 000 233 000 TT 233,000 233,000 TonnesTonnes

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Main uses of Cassava Main uses of Cassava

Fresh

Fresh -- boiledboiled Farinha Farinha -- GariGari

Human consumption Human consumption

Cassava leaves

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Chicken factory

Chicken factory Dry chips for animal feedDry chips for animal feed

Animal feedstock Animal feedstock

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Bio

Bio--EthanolEthanol StarchStarch

Industrial use of Cassava Industrial use of Cassava

Fried

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Tropical/Sub

Tropical/Sub--tropical croptropical crop

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I t d ti

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Crop Potential Crop Potential

South China 5 Breeding successfully increased

fresh-root (FR) productivity & dry-matter (DM) content. We now need

South-China 5

matter (DM) content. We now need

STABLE -DM contents

SM 1433-4

84 t/ha FR in a 9.5 ha commercial field (~25 t/ha DM)

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The case of “watery” roots for ethanol

Fresh root yield (t/ha) Dry matter content (%) Dry matter yield (t/ha) (%) SM 2775‐2 53.8 32.1 17.3 At two location: Codazzi (Cesar) and Barrancas (Guajira) SM 2775‐4 35.3 35.9 12.7 SM 2775 2 37 3 30 7 11 5 At five location: Patalito, Sto Thomas & Molinero (Atlantico), La Union (Sucre) and Chinu (Cordoba) SM 2775‐2 37.3 30.7 11.5 SM 2775‐4 27.1 36.9 10.0

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The case of “watery” roots for ethanol

Fresh root yield (t/ha) Dry matter content (%) Dry matter yield (t/ha) (%) SM 2775‐2 53.8 32.1 17.3 At two location: Codazzi (Cesar) and Barrancas (Guajira) SM 2775‐4 35.3 35.9 12.7 SM 2775 2 37 3 30 7 11 5 At five location: Patalito, Sto Thomas & Molinero (Atlantico), La Union (Sucre) and Chinu (Cordoba) High Dry Matter content does not seems critical to ethanol production SM 2775‐2 37.3 30.7 11.5 SM 2775‐4 27.1 36.9 10.0

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I t d ti

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Cassava “Novel” traits Cassava “Novel” traits

Amylose-free (“waxy”)

Amylose free ( waxy )

starch mutation

•Amylose is difficult to

degrade

•Amylose-free starch

should cost less to convert into ethanol

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Less amylose = Less amylose = more ethanol

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F bili

Fermentability:

assess their potential in bio-ethanol, bio-plastics, sweeteners

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Cassava starch fermentation: with and without starch 400 250 300 350 Total et Total ethanol 100 150 200 Total et (ml eth kg sta Total ethanol (mL/Kg of starch) CM 523 7 With enzyme 0 50 100 CM 523-7 Rayong 60 NEP WAXY Without enzyme Clone 4 1/3 days

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Small granule/high amylose Small granule/high amylose

h m al Starc h Nor m e Starch a ll granul e Sm a

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h m al Starc h Nor m

• A small granule and a rough surface

e

Starch

A small granule and a rough surface

facilitate the action of enzymes (less consumption of enzymes, lower costs of conversion).

a

ll granul

e )

• But higher amylose content would

increase costs….

Sm

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h ( ) RVA Amylogram Starch Viscosity (5%) 1000 1200 80 100 Waxy 600 800 o si ty (c P ) 60 200 400 Vi sc o 20 40 Small granules 0 0 5 10 15 20 Temperature (minutes) 0

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Starch

Starch--lessless mutationmutation

Source:

L. Carvalho EMBRAPA Brazil

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I t d ti

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Bio

Bio--ethanol productionethanol production

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Bio

Bio--ethanol productionethanol production

Ethanol factory in Thai Nguan near Khon Kaen (Thailand)

5.27 kg of fresh root produce one liter of ethanol 1.4 – 1.5 bath / kg fresh root

1.4 1.5 bath / kg fresh root 25 bath / lt of ethanol produced

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Ethanol from corn or cassava is

more expensive because starch

Boiler

more expensive because starch

need to be degraded to the equivalent of sugar cane juices

Eth l Ethanol Maize or Cassava Distillation & dehydration Starch degradation

Liquefaction & saccharification

Fermentation y

Sugarcane Sugarcane juices

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Sorce of satrch Thermo-stable Alpha-amylase _Yeasts Grinding Jet cooker >100 °C Alpha amylase

(Liquefacction) Yeasts Storage

tank Grinding >100 C (5-8’) o n a tion _oras) tank Secondary Liquefaction ermentati o a ccharific a °C (8-10 h o

lation & _ydration

Slurry t _Liquefaction (95 °C – 90’) F e S a 60 ° Distil deh y Glucoamylase (Saccharification) Solids

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Sorce of satrch Thermo-stable Alpha-amylase _Yeasts Grinding Jet cooker >100 °C Alpha amylase

(Liquefacction) Yeasts Storage

tank Grinding >100 C (5-8’) o n a tion _oras) tank Secondary Liquefaction ermentati o a ccharific a °C (8-10 h o

Slurry t _Liquefaction (95 °C – 90’) F e S a 60 ° Distil deh y New enzymes Glucoamylase (Saccharification) Solids y Liquefaction + saccharification

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Sorce of satrch _Yeasts Grinding Storage tank Yeasts Grinding o n a tion _oras) tank ermentati o

a ccharific a °C (8-10 h o Slurry t F e Distil deh y S a 60 ° New enzymes New enzymes + yeasts Solids y Liquefaction + saccharification Liquefaction + saccharification + fermentation

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Storage tank

lation & dration

Distil

l

dehy

d

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Medium throughput

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Digestion rate of different cassava starches (1 0 ml of pacreatic α-amilase) 100 (1.0 ml of pacreatic α amilase) pH 6.9 at 37°C 80 ex (% ) ~80% 40 60 ro lis is In d 0 20 Hi d ~30% 0 10 20 30 40 50 60 Time (minutes)

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Digestion rate of different cassava starches (0 5 ml of StargenTM ₂₎ 80 (0.5 ml of Stargen 2) pH 4.0 at 37°C 60 e x (% ) ~60% 40 H id ro lis is in d e 20 ~30% 0 0 10 20 30 40 50 60 Time (minutes)

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Root processing vs. quality Root processing vs. quality • Starch degrading enzymes and yeast are being improved.

h h i h l l h

• The process to convert starch into ethanol constantly changes.

• As in maize, there are genetic differences in cassava for ethanol production (small starch granule).

• We are in a unique position to analyze the best germplasm – processing method to maximize economic benefit and reduce negative impact on the environment.

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I t d ti

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Cassava Bio

Cassava Bio--ethanol perspectiveethanol perspective

• Cassava is a competitive raw material

for bio-ethanol production in Asia

(Thailand, China, Vietnam, Indonesia?, Australia?)

• A large % of the ethanol production cost

is constitute by the y enzyme and yeasty y .

• Advances in microbiology and

enzymology can significantly reduce enzymology can significantly reduce

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Cassava Bio

Cassava Bio--ethanol ethanol prespectiveprespective

• There are clones with low dry matter

content but maximum productivity per hectare that can now be used in ethanol production

• Different mutants could reduce costs of

conversion from root to ethanol (including “sugary”?)

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Energy crops: farms of 1-100 ha

Cassava Banana _{Coffee residues} _Ethanol

(99,5%)

Sweet potato Sugar cane Sweet sorghum

(99,5%)

Small rural communities _{Central Plant (dehydration)}

Micro-plants 1.000 – 2.000 lt/day 5 – 10 t crop/day < 1 ha crop/day < – 1 ha crop/day Ethanol (50%) Transport ( )

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Cassava Bio

• For ethanol production a key issue is the

continuous supply of feedstock all year round.

• Processing of g fresh roots (low dry y

matter?) at harvest time and dried chips

during off-season is one potential

l i

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Cassava Bio

• Combining feedstock from different

crops. For instance, cassava/sweet-sorghum has proved advantageous.

• We need to further analyze the by-• We need to further analyze the by

products and their potential use for

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Cassava breeding potential for bioethanol

Thank you

Thank you