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BIOETHANOL FROM NON-CONVENTIONAL SOURCES

Jos A. Teixeira
IBB - Institute for Biotechnology and Bioengineering, Centre of Biological Engineering University of Minho, PORTUGAL e-mail: jateixeira@deb.uminho.pt

Raw materials and processes currently used for bioethanol production

Sugarcane

Brazil, India

Corn

United States, China, Canada China, Canada, Europe Thailand

Sugar beet

Europe (France)

Wheat

Sorghum

India

Cassava

Cheese Whey

New Zealand

Milling for sugars extraction, and fermentation of sugars

Milling, liquefaction, saccharification, and fermentation of sugars

Bioethanol production worldwide

40%

89%
60%

Sugar crops

Starch crops

Bioethanol consumption worldwide

Brazil: main exporter


Brazil United States

United States, Japan, Europe: main importers

Vehicles use ethanol in the pure form or in mixture with the gasoline, where ethanol corresponds up to 25% of the mixture Ethanol is used in two forms: mixed with gasoline in the maximum proportion of 10%, or in mixtures containing 85% ethanol and 15% gasoline, as an alternative fuel Current blend levels are 10%, but efforts are being directed to expand ethanol blends to 25% Gasoline must be blended with 10% ethanol Requires 10% ethanol blends in cities with populations over 500,000 Some regions of China use mixtures containing up to 10% ethanol in gasoline Addition of 5% ethanol to gasoline is mandatory. Efforts will be directed to increase the ethanol percentage in the mixture to 10% 5% of all motor vehicle must be ethanol or biodiesel Mixtures containing 5% ethanol in gasoline are used The replacement of 3% of gasoline by ethanol is authorized, but efforts will be done to increase this value to 10%

Bolivia Thailand Colombia China India Canada Sweden Japan

Bioethanol production from non-conventional sources

Technology under development

Rice straw
LIGNIN

Wheat straw Sugarcane bagasse


CELLULOSE

HEMICELLULOSE

Bioethanol production from non-conventional sources

- Arabinofuranose
H H OH H HO O
H

Pre-treatment: diluted acid hydrolysis


OH

Acetyl group
O CH3 O
H H H H O H OH

Xylobiose
H O H OH H H

H H OH H H O

H H OH H H

OH

H H O H H H OH

H H H OH H H O

H
O

H H O H OH H H OH

H
O

H H OH H H O

O H OH H H OH

O H OH H H OH

OH

OH

OH
O

O CH3 H3 C
O

O
H

OH
O

H3C
O

H OH H H OH

H OH H H OH

Acetyl group

Glucuronic acid

Glucuronic acid

OH OH HO O
HO

OH O OH OH HO
O O

O OH

OH HO
O

HO

O OH

HO

O OH

OH
Cellobiose

Cellulose hydrolysis: concentrated acid hydrolysis and enzymatic hydrolysis

Bioethanol production from non-conventional sources

Obtainment of a fermentable sugars solution


LIGNOCELLULOSIC BIOMASS

Fermentation of sugars

Ethanol separation and purification

Pre-treatment

(Xylose) Fermentation Ethanol

Milling

Cellulose conversion (hydrolysis)

Distillation (Glucose) Fermentation

Iogen (straw Canada); Abengoa (straw Spain, US); Etek (softwood Sweden); Elsam (straw Denmark); TMO (straw etc. UK); Tavda (wood Russia); NEDO (rice straw Japan)

Challenges to be overcome for an efficient bioethanol production from lignocellulosic biomass


1) High energy consumption for biomass pretreatment

(1)

(3)
Cellulose fibers

(2)

Challenges to be overcome for an efficient bioethanol production from lignocellulosic biomass

2) Development of a suitable and economically viable hydrolysis process step

Specific enzymes

Glucose

CELLULOSE

Challenges to be overcome for an efficient bioethanol production from lignocellulosic biomass

3) Improvement in the conversion rate and yield of hemicellulose sugars

fermentation

Pentoses (Xylose and Arabinose) and Hexoses

Ethanol Toxic compounds:

low sugars conversion yield

Challenges to be overcome for an efficient bioethanol production from lignocellulosic biomass

3) Other important considerations for the process implementation To develop microorganisms able to metabolize pentose and hexose sugars simultaneously withstanding the stress imposed by the process inhibitors To evaluate the process scalability

To perform an analysis of the costs involved for commercial production


To establish alternatives for the recovery of pretreatment chemicals and wastewater treatment

Other energy crops

Sweet potato

Sweet corn

Jerusalem artichoke

Sweet sorghum

Carbohydrate and expected ethanol yields for sweet corn, Jerusalem artichoke, sweet potato ad sweet sorghum Current ethanol yield from grain corn in the US and sugarcane in Brazil is approximately 3,500 and 6,000 L/ha, respectively.1

Sweet Sorghum as an energy crop

It
It

has a photosynthetic efficiency (~ 4 g biomass/MJ of solar radiation) two times or

more that of C3 crops (forest) is able to grow anywhere in dry climates with high yields of fermentable sugars,

grains and lignocellulosics.

In some
It

regions it is possible to obtain two plantations per year reaching full maturity

and a large production. has low water requirements 1/3 of sugar cane, 1/2 of corn, 1/4 of Short Rotation

Forestry.

Sweet

sorghum (Sorghum bicolor) is frequently called as smart crop for its

ability of not only produce food but fuel as well

Agave vs sucarcane needs

Agave and sugar cane bioethanol production

Miscanthus x giganteus properties relatively high yields 8-15 t/ha (3-6 t/acre) dry weight, low moisture content (as little as 15-20% if harvested in late winter or spring), annual harvests, providing a regular yearly income for the grower, good energy balance and output/input ratio compared with some other

biomass options,
low mineral content, especially with late winter or spring harvest, which improves fuel quality. can be grown in a cool climate like that of northern Europe

Miscanthus x giganteus ethanol production yield

typical acre of corn yields around 7.6 tons of input per acre and

756 gallons of ethanol..

switchgrass,

which yields around 3-6 tons of biomass and 400-900

gallons of ethanol fuel

giant

Miscanthus is capable of producing up to 20 tons of biomass

and 3,250 gallons of ethanol fuel

Bioethanol production from non-conventional sources

Bioconversion of lactose to ethanol represents a process which can provide a value-added product from cheese manufacturing, allied with efficient bioremediation of plant effluent.

Represents 8595% of the milk volume and its world production is estimated to be over 108 tons/ year Lactose (5-6% w/v) is assumed to be responsible for 90% of the wheys BOD and COD.

Liquid remaining after the precipitation and removal of milk casein during cheese-making

CHEESE WHEY

Biological treatment by conventional aerobic process is very expensive

Bioethanol production from non-conventional sources

Obtainment of a fermentable sugars solution


CHEESE WHEY Concentrated cheese whey or cheese whey powder solution

Fermentation of sugars

Ethanol separation and purification

(Lactose) Fermentation Distillation

Ethanol

Direct fermentation of whey or whey permeate to ethanol is generally not economically feasible because the low lactose content results in low ethanol titre (23% v/v), making the distillation process too expensive.

Bioethanol production from non-conventional sources

Important process considerations:

Cheese whey concentration: by ultrafiltration and/or reverse osmosis processes. Yeasts that ferment lactose: Kluyveromyces lactis, K. marxianus, Candida pseudotropicalis, genetically modified Saccharomyces cerevisiae.

Whey to bioethanol
8 million tons of lactose (worldwide annual whey production)

~50% not transformed into added-value

sub-products

~2.3 million m3 ethanol


considering a 85% conversion yield

Worldwide production of bioethanol for fuel in 2008: ~65 million m3

Whey to Ethanol Industrial Plants


Ireland

Carbery Milk Products

since 1978, potable ethanol & ethanol for fuel (since 2005) 11 000 tons ethanol /year

New Zealand

Fonterra

Anchor Ethanol (Fonterra subsidiary)

potable ethanol & ethanol for fuel (since 2007)


17 million liters ethanol /year

United States

Golden Cheese Land OLakes

Germany

Mllermilch

near Dresden; 10 million litres ethanol /year

from dairy by-products

Whey permeate biotechnological treatment

Flocculent Saccharomyces cerevisiae strains able to metabolize lactose

Continuous high cell density systems

S. cerevisiae traditionally used in industry


Molecular biology techniques well developed Good fermentative capacity

Higher volumetric productivity


Improvement of separation processes Higher stability

A Saccharomyces cerevisiae strain that efficiently metabolizes lactose was developed, with a lactose metabolization capacity comparable to the one presented by other natural lactose users

A high ethanol productivity (10 gl-1h-1) system using cheese whey as a substrate was developed

The developed fermentation proved its long term stability

system

Pilot scale experiments validated the developed fermentation process

Fermentation of Cheese Whey powder solutions by T1-E 110150 gL-1 Lactose + Corn Steep Liquor (10 gL-1)
Repeated-batch operation with biomass recycling by flocculation

6-L Air-lift Bioreactor Aerated at 0.1 vvm pH 4.2 0.2 Temperature: 30 1 C

8% (v/v) ethanol (max.) Ethanol productivity 0.7 gL h


-1

-1

Bioethanol production from non-conventional sources

Sun light O2 O2 CO2 CO2


ETHANOL

CO2

The microalgae Chlorella vulgaris, particularly, has been considered as a promising feedstock for bioethanol production
Water

Water

nutrients

nutrients

Bioethanol production from non-conventional sources

Technology under development

Enzymatic

Fermentation

Starch
Microalgae cultivation Cell rupture

hydrolysis

sugars
ETHANOL

Some algal species are able to conduct self-fermentation

Bioethanol production from non-conventional sources

Advantages of this process:

microalgae can be harvested batch-wise nearly all-year-round they grow in aqueous media, but need less water than terrestrial crops, therefore reducing the load on freshwater sources the ability of microalgae to fix CO2

Bioethanol production from non-conventional sources

Bioethanol yield from different sources:

Source Corn stover Wheat Cassava Sweet sorghum Corn Sugar beet Sugarcane Switch grass Microalgae

Ethanol yield (gallons/acre) 112-150 277 354 326-435 370-430 536-714 662-802 1,150 5,000-15,000

Ethanol yield (L/ha) 1,050-1,400 2,590 3,310 3,050-4,070 3,460-4,020 5,010-6,680 6,190-7,500 10,760 46,760-140,290

Conclusions

World ethanol production and consumption will continue to grow strongly

Corn is the main raw material used today, but in the future.....

Dont affect the food provision Microalgae Cheese whey Lignocellulose Agave

Development of ethanol production systems all over the world