Cassava breeding potential Cassava breeding potential for bioethanolfor bioethanolfor bioethanolfor bioethanol

Becerra LópezBecerra López‐‐Lavalle L A Dufour DLavalle L A Dufour DBecerra LópezBecerra López Lavalle, L.A. , Dufour, D., Lavalle, L.A. , Dufour, D., Sánchez, T. and H. CeballosSánchez, T. and H. Ceballos

OutlineOutline

I t d ti• Introduction

• High stable and reliable productivityHigh, stable and reliable productivity

• Novel traits

• Processing methods X root quality interactions

• Perspectives

OutlineOutline

I t d ti• Introduction

• High stable and reliable productivityHigh, stable and reliable productivity

• Novel traits

• Processing methods X root quality interactions

• Perspectives

Cassava originCassava originFlooded landFlooded land

Low Low Soil Soil FFertilityertility

Degraded SoilsDegraded Soilsgg

Slopped LandSlopped Land

Cassava modern productionCassava modern productionSubSub--humid environmenthumid environment Acid Acid ––Soil environmentSoil environment

19’000,000 hectares19’000,000 hectares

Near Hanoi, VietnamNear Hanoi, Vietnam GuanGuan--Xi Province, ChinaXi Province, China

Cassava modern productionCassava modern productionSubSub--humid environmenthumid environment Acid Acid ––Soil environmentSoil environment

19’000,000 hectares19’000,000 hectares

233 000233 000 TT233,000 233,000 TonnesTonnes

Near Hanoi, VietnamNear Hanoi, Vietnam GuanGuan--Xi Province, ChinaXi Province, China

Main uses of CassavaMain uses of CassavaFresh Fresh -- boiledboiled Farinha Farinha -- GariGari

Human consumptionHuman consumption

Cassava leavesCassava leaves FufuFufu

Main uses of CassavaMain uses of CassavaChicken factoryChicken factory Dry chips for animal feedDry chips for animal feed

Animal feedstockAnimal feedstock

Near Hanoi, VietnamNear Hanoi, Vietnam Pressed cake Pressed cake

Main uses of CassavaMain uses of CassavaBioBio--EthanolEthanol StarchStarch

Industrial use of CassavaIndustrial use of Cassava

FriedFried--ChipsChips GorenGoren--KrupukKrupuk

Tropical/SubTropical/Sub--tropical croptropical crop

Main cassava production regions in the world

OutlineOutline

I t d ti• Introduction

• High stable and reliable productivityHigh, stable and reliable productivity

• Novel traits

• Processing methods X root quality interactions

• Perspectives

Crop PotentialCrop PotentialSouth 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 1433-484 t/ha FR in a 9.5 ha commercial field

(~25 t/ha DM)

Crop PotentialCrop Potential

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

Crop PotentialCrop Potential

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

OutlineOutline

I t d ti• Introduction

• High stable and reliable productivityHigh, stable and reliable productivity

• Novel traits

• Processing methods X root quality interactions

• Perspectives

Cassava “Novel” traitsCassava “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

Cassava “Novel” traitsCassava “Novel” traits

Less amylose = more ethanol

Less amylose =Less amylose =more ethanol

Cassava “Novel” traitsCassava “Novel” traits

F biliFermentability: assess their potential in bio-ethanol, bio-plastics, sweeteners

Cassava starch fermentation: with and without starch

400

250

300

350

Total etTotal ethanol

100

150

200

Total et(ml eth

kg sta

Total ethanol (mL/Kg of starch)

CM 523 7

With enzyme0

50

100

CM 523-7Rayong 60

NEPWAXY

Without enzyme

Clone 4 1/3 days

Small granule/high amyloseSmall granule/high amyloseh

mal

Sta

rch

Nor

me

Star

chal

l gra

nule

Sma

Small granule/high amyloseSmall granule/high amyloseh

mal

Sta

rch

Nor

m

• A small granule and a rough surface

e St

arch

A small granule and a rough surfacefacilitate the action of enzymes (less consumption of enzymes, lower costs of conversion).

all g

ranu

le

)

• But higher amylose content would increase costs….

Sma

Small granule/high amyloseSmall granule/high amylose

h ( )

RVA Amylogram

Starch Viscosity (5%) 

1000

1200

80

100

Waxy

600

800

osity (cP)

60

200

400

Visco

20

40

Small granules00 5 10 15 20

Temperature (minutes)

0

StarchStarch--lessless mutationmutation

Source:L. CarvalhoEMBRAPABrazil

OutlineOutline

I t d ti• Introduction

• High stable and reliable productivityHigh, stable and reliable productivity

• Novel traits

• Processing methods X root quality interactions

• Perspectives

BioBio--ethanol productionethanol productionEthanol factory in Thai Nguan near Khon Kaen (Thailand)

BioBio--ethanol productionethanol productionEthanol factory in Thai Nguan near Khon Kaen (Thailand)

5.27 kg of fresh root produce one liter of ethanol1.4 – 1.5 bath / kg fresh root1.4 1.5 bath / kg fresh root

25 bath / lt of ethanol produced

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 lEthanolMaize or Cassava

Distillation &dehydration

Starch degradationLiquefaction & saccharification

Fermentation

y

SugarcaneSugarcane juices

Sorce ofsatrch

Thermo-stableAlpha-amylase Yeasts

GrindingJet cooker>100 °C

Alpha amylase(Liquefacction)

YeastsStorage

tank

Grinding >100 C (5-8’)

onatio

nor

as)

tank Secondary

Liquefaction erm

enta

tio

acch

arifi

ca°C

(8-1

0 ho

latio

n &

ydra

tion

Slur

ry t Liquefaction

(95 °C – 90’)

FeSa 60 °

Dis

tilde

h y

Glucoamylase(Saccharification)

Solids

Sorce ofsatrch

Thermo-stableAlpha-amylase Yeasts

GrindingJet cooker>100 °C

Alpha amylase(Liquefacction)

YeastsStorage

tank

Grinding >100 C (5-8’)

onatio

nor

as)

tank Secondary

Liquefaction erm

enta

tio

acch

arifi

ca°C

(8-1

0 ho

latio

n &

ydra

tion

Slur

ry t Liquefaction

(95 °C – 90’)

FeSa 60 °

Dis

tilde

h y

New enzymesGlucoamylase(Saccharification)

Solids

yLiquefaction + saccharification

Sorce ofsatrch Yeasts

Grinding

Storagetank

Yeasts

Grinding

onatio

nor

as)

tank

erm

enta

tio

latio

n &

ydra

tion

acch

arifi

ca°C

(8-1

0 ho

Slur

ry t Fe

Dis

tilde

h ySa 60 °

New enzymesNew enzymes

+ yeasts

Solids

yLiquefaction + saccharificationLiquefaction + saccharification

+ fermentation

Storagetank

latio

n &

drat

ion

Dis

till

dehy

d

Solids

Medium throughput Medium throughput fermentersfermenters

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

rolis

is In

d

0

20Hid

~30%

0 10 20 30 40 50 60

Time (minutes)

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

ex (%

)

~60%

40

Hidro

lisis in

de

20 ~30%

0

0 10 20 30 40 50 60

Time (minutes)

Root processing vs. qualityRoot 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.

• What is the potential of “sugary” cassava?

OutlineOutline

I t d ti• Introduction

• High stable and reliable productivityHigh, stable and reliable productivity

• Novel traits

• Processing methods X root quality interactions

• Perspectives

Cassava BioCassava 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 enzyme and yeast. y y y

• Advances in microbiology and enzymology can significantly reduceenzymology can significantly reduce ethanol production cost from starches

Cassava BioCassava Bio--ethanol ethanol prespectiveprespective

• There are clones with low dry mattercontent 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”?)

Energy crops: farms of 1-100 ha Cassava Banana Coffee residues Ethanol

(99,5%)

Sweet potato Sweet sorghumSugar cane

(99,5%)

Small rural communities Central Plant (dehydration)

Micro-plants1.000 – 2.000 lt/day

5 – 10 t crop/day< 1 ha crop/day< – 1 ha crop/day

Ethanol (50%)

Transport

( )

Cassava BioCassava Bio--ethanol perspectiveethanol perspective

• For ethanol production a key issue is the continuous supply of feedstock all year round.

• Processing of fresh roots (low dry g ymatter?) at harvest time and dried chips during off-season is one potential l ialternative.

Cassava BioCassava Bio--ethanol perspectiveethanol perspective

• 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 byproducts and their potential use for animal feeding.g

Thank youThank you