---------- Forwarded message ----------
From: Biotech-Admin <Biotech-Ad...@fao.org>
Date: 21 Oct 2008 16:48
Subject: Launch of FAO Biotech e-mail Conf. 15 (Bioenergy and
agricultural biotechnologies)

Dear Forum Members,

We wish to announce that Conference 15 of the FAO Biotechnology Forum begins
on Monday 10 November and runs for four weeks, finishing on Sunday 7 December
2008. The title of the conference is "The role of agricultural
biotechnologies for production of bioenergy in developing countries". It is
being organised in collaboration with the FAO Working Group on Bioenergy and
is a follow-up to an FAO seminar on the same subject held in Rome in October
2007 (http://www.fao.org/biotech/seminaroct2007.htm).

The conference, as usual, is open to everyone, is free and will be moderated.
The purpose of this message is to provide you with the Background Document
for the conference and to invite you to join.

The goal of this e-mail conference is to explore the role that application of
agricultural biotechnologies may play for production of bioenergy in
developing countries, with a major focus on liquid biofuels. It will focus on
the use of biotechnology processes and tools both to produce biomass for
bioenergy purposes and to convert biomass to biofuel. The conference will
cover biotechnology applications for first- and second-generation biofuels
and, to a lesser degree, for biogas production and for biodiesel production
from microalgae. It will therefore cover bioenergy production systems that
are currently a reality (first-generation biofuels and biogas) as well as
those that are still at the experimental stage (second-generation biofuels
and microalgal biodiesel).

The Background Document aims to provide information about the conference
theme that participants will find useful for the debate. First, an overview
is provided of the current status regarding bioenergy, focusing on first- and
second-generation liquid biofuels, including the major reasons for the big
interest as well as current concerns about them. Some of the potential ways
in which biotechnologies could contribute to this area are then considered,
covering production of biomass as well as conversion of the biomass to first-
or second-generation liquid biofuels, in addition to production of biodiesel
from microalgae and production of biogas. A small number of issues of
specific relevance to the debate are then briefly described while some of the
kinds of specific questions that should be addressed in the conference are
then listed. In the final section, references to articles mentioned in the
document, abbreviations and acknowledgements are provided.

As for all previous conferences hosted by the Forum, a document will be
prepared after the e-mail conference is finished to provide a summary of the
main issues that were discussed, based on the messages posted by the
participants.

Please pass this information on to other colleagues that might be interested
in joining the conference. As the Background Document sets the scene for the
conference and highlights the elements to be discussed, it should be read
carefully by members wishing to participate in the conference. The Background
Document is also available on the web - at
http://www.fao.org/biotech/C15doc.htm

NB NB NB NB NB !!!!! You, as a Forum member, are NOT automatically subscribed
to this conference. Instead, if you wish to join, you should subscribe
yourself.

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Best regards

John

John Ruane, PhD
FAO Biotechnology Forum Administrator
E-mail address: Biotech-Ad...@fao.org
FAO website http://www.fao.org
Forum website http://www.fao.org/biotech/forum.asp
FAO Biotechnology website http://www.fao.org/biotech/index.asp

**********************************************
VI. BACKGROUND DOCUMENT TO CONFERENCE 15:

The role of agricultural biotechnologies for production of bioenergy in
developing countries

1. INTRODUCTION

To put it mildly, bioenergy is currently a very hot topic. In a relatively
short time, large-scale cultivation of crops for production of liquid
biofuels has become a reality, a phenomenon that is predicted to expand,
driven by concerns about climate change, increasing petrol prices and
national energy security, among others. The potential social, economic,
environmental and human rights impacts have been much debated and have been
the subject of considerable controversy with e.g. the UN Special Rapporteur
on the Right to Food highlighting grave concerns that "biofuels will bring
hunger in their wake", arguing that "the sudden, ill-conceived, rush to
convert food - such as maize, wheat, sugar and palm oil - into fuels is a
recipe for disaster" (UN, 2007).

It is a topic that has very actively engaged governments and their policy
makers worldwide. Indeed, in June 2008, representatives from 181 countries,
including 42 Heads of State or Government, gathered at FAO Headquarters in
Rome for the High-Level Conference on World Food Security: the Challenges of
Climate Change and Bioenergy. The Summit concluded with the adoption by
acclamation of a Declaration and, regarding biofuels, the Declaration stated:
"It is essential to address the challenges and opportunities posed by
biofuels, in view of the world's food security, energy and sustainable
development needs. We are convinced that in-depth studies are necessary to
ensure that production and use of biofuels is sustainable in accordance with
the three pillars of sustainable development and takes into account the need
to achieve and maintain global food security. We are further convinced of the
desirability of exchanging experiences on biofuels technologies, norms and
regulations. We call upon relevant intergovernmental organizations, including
FAO, within their mandates and areas of expertise, with the involvement of
national governments, partnerships, the private sector, and civil society, to
foster a coherent, effective and results-oriented international dialogue on
biofuels in the context of food security and sustainable development needs"
(FAO, 2008a, p. 50)

Because of concerns about the current first-generation of liquid biofuels,
there is major interest in moving to alternative systems of biofuel
production, such as second-generation liquid biofuels based on
lignocellulosic biomass, and applications of biotechnologies will be
important if they are to become widely available in the future. Taking
therefore a topic of current global relevance and interest, the aim of this
e-mail conference hosted by the FAO Biotechnology Forum is to explore the
role that application of agricultural biotechnologies may play for production
of bioenergy in developing countries, with a major focus on liquid biofuels.

For people who are not familiar with the Forum
(http://www.fao.org/biotech/forum.asp), it was launched by FAO in 2000 with
the goal of providing access to quality balanced information and to make a
neutral platform available for all interested stakeholders to openly exchange
views and experiences on agricultural biotechnology in developing countries.
It covers applications in the crop, forestry, livestock, fisheries and
agro-industry sectors. It has hosted 14 moderated e-mail conferences so far,
and in these the messages posted have come roughly 50:50 from participants
living in developing and developed countries respectively (FAO, 2001, 2006a).

Each conference of the Forum takes one particular theme that is relevant to
agricultural biotechnology in developing countries and opens it up for debate
for a limited amount of time. The Forum covers the broad range of tools
included under the general term 'biotechnology'. Some of the technologies may
be applied to all the food and agriculture sectors, such as the use of
genomics, molecular DNA markers or genetic modification, while others are
more sector-specific, such as vegetative reproduction (crops and forest
trees) or embryo transfer and freezing (livestock). This conference is
therefore not just about genetically modified organisms (GMOs) and
discussions in this conference will not consider the issues of whether GMOs
should or should not be used per se or the attributes, positive or negative,
of GMOs themselves. Instead, the goal is to discuss the potential role that
applications of biotechnology tools (including genetic modification) can play
for production of bioenergy in developing countries. Similarly, it will not
discuss the advantages or disadvantages of bioenergy or liquid biofuels per
se.

In this context, the primary focus will be on the use of biotechnology
processes and tools both to produce biomass for bioenergy purposes and to
convert biomass to biofuel. The conference will cover biotechnology
applications for first- and second-generation biofuels and, to a lesser
degree, for biogas production and for biodiesel production from microalgae.
It will therefore cover applications of biotechnologies to crops, grasses,
forest trees and micro-organisms (bacteria, yeasts, microalgae) and to
bioenergy production systems that are currently a reality (first-generation
biofuels and biogas) as well as to those that are still at the experimental
stage (second-generation biofuels and microalgal biodiesel).

This Background Document aims to provide information about the conference
theme that participants will find useful for the debate. First, an overview
is provided of the current status regarding bioenergy, focusing on first- and
second-generation liquid biofuels (Section 2), including the major reasons
for the big interest as well as current concerns about them. Some of the
potential ways in which biotechnology could contribute to this area are then
considered, covering production of biomass as well as conversion of the
biomass to first- or second generation liquid biofuels, in addition to
production of biodiesel from microalgae and production of biogas (Section 3).
A small number of issues of specific relevance to the debate are briefly
described in Section 4 while some of the kinds of specific questions that
should be addressed in the conference are listed in Section 5. Finally,
references to articles mentioned in the document, abbreviations and
acknowledgements are provided in Section 6.

As for all previous conferences hosted by the Forum, a document will be
prepared after the e-mail conference is finished to provide a summary of the
main issues that were discussed, based on the messages posted by the
participants.

2. BIOENERGY AND FIRST- AND SECOND-GENERATION LIQUID BIOFUELS

2.1 Bioenergy

The term bioenergy refers to energy obtained from biomass, which is the
biodegradable fraction of products, waste and residues from agriculture (of
vegetable and animal origin), forestry and related industries, as well as the
biodegradable fraction of industrial and municipal waste (FAO, 2008b). A wide
range of biomass sources can be used to produce bioenergy in a variety of
forms. For example, food, fibre and wood process residues from the industrial
sector; energy and short-rotation crops and agricultural wastes; and forest
and agroforest residues from the forestry sector can all be used to generate
electricity, heat, combined heat and power, and other forms of bioenergy
(GBEP, 2007).

Traditional biomass materials, including fuelwood, charcoal and animal dung,
continue to be important sources of bioenergy in many parts of the world and,
to date, woodfuels represent by far the most common sources of bioenergy.
Modern bioenergy relies on efficient conversion technologies for applications
at the household, small business and industrial scale. Solid or liquid
biomass inputs can be processed to be more convenient energy carriers. These
include solid biofuels (e.g. firewood, wood chips, pellets, charcoal and
briquettes), gaseous biofuels (biogas, synthesis gas, hydrogen) and liquid
biofuels (e.g. bioethanol, biodiesel) (GBEP, 2007). Among the different
segments of the bioenergy sector, the largest and most rapid growth has been
seen in liquid biofuels (FAO, 2008b). For this reason, and because of their
predicted further expansion in the future, they will be the main focus of
this conference.

The major use of liquid biofuels is for transport, where the biofuel is
either blended with traditional transport fuels (biodiesel with diesel or
bioethanol with petrol) for conventional engines or used on its own in
vehicles with specialised engines. There is also much interest in liquid
biofuels as a cooking or heating fuel, although significant barriers, such as
the need for more affordable stoves, still remain (UNCSD, 2007).

2.2 First-generation liquid biofuels

Following Larson (2008), liquid biofuels can be classified into those that
are "first-generation" and "second-generation", where the main distinction
between them is the biomass (feedstock) used. First-generation fuels are
generally made from sugars, grains or seeds, i.e. using only a specific
(often edible) portion of the above-ground biomass produced by a plant, and
relatively simple processing of the biomass is required to produce a finished
fuel (Larson, 2008).

The two main first-generation liquid biofuels are currently biodiesel and
bioethanol, representing about 15 and 85% of current global production
respectively (FAO, 2008c). A brief overview of the way they are produced is
provided here.

a) Biodiesel

For biodiesel production, the feedstocks involved include vegetable oils
(e.g. derived from oilseed crops such as soybean, sunflower, jatropha, oil
palm or rapeseed), used frying oil (e.g. from restaurants) or animal fat
(e.g. pork lard) (IEA, 2004). The major components of vegetable oils and
animal fats are triacylglycerols (TAGs, also called triglycerides), which
consist of three long-chain fatty acids linked to a glycerol backbone.
Natural oils are too viscous (i.e. thick and slow flowing) to be used in
modern diesel engines. However, in the 1980s a chemical modification of
natural oils was introduced that helped to bring the viscosity of the oils
within the range of current petroleum diesel. Thus, by reacting these TAGs
with simple alcohols such as methanol (a chemical reaction known as
"transesterification", already commonplace in the oleochemicals industry),
alkyl esters (methyl esters), generically known as biodiesel, are formed
whose properties are very close to those of petroleum diesel (Sheehan et al.,
1998, p. 6).

b) Bioethanol

Ethanol, also known as ethyl alcohol, can be produced from any biomass that
contains appreciable amounts of sugar or materials that can be converted into
sugar, such as starch or cellulose. Sugar cane, sweet sorghum and sugar beet
are examples of feedstocks that contain sugar. Maize, wheat and other cereals
contain starch (in their kernels) that can relatively easily be converted
into sugar.

In producing bioethanol from sugar crops, they are first processed to extract
the sugar (e.g. through crushing). The sugar is then fermented to yield
ethanol. (Ethanol fermentation is the biochemical process by which sugars,
such as glucose, fructose and sucrose, are converted into ethanol and carbon
dioxide using yeast or other micro-organisms. Glucose and fructose are
monosaccharides [i.e. simple sugars] with six carbon atoms, and are thus
termed 6-carbon sugars. Sucrose is a disaccharide [i.e. a sugar consisting of
two monosaccharides] made of glucose and fructose joined together). A final
step distils (purifies) the ethanol to the desired concentration and usually
removes all water to produce "anhydrous ethanol" that can be blended with
petrol. With sugar cane, the "bagasse" (i.e. the crushed stalk of the plant)
can be used as a solid fuel and burned for heat and electricity.

In producing bioethanol from starchy materials, the process is more difficult
compared to sugar crops because an additional step, hydrolysis of the
feedstock, is required. Starch is a polysaccharide (i.e. a polymer [a large
molecule created by the chemical joining of many identical or similar smaller
units] made up of linked monosaccharides) consisting of long chains of
glucose molecules. Through hydrolysis, where the starch reacts with water,
the starch is broken down to fermentable glucose molecules. Hydrolysis, also
known as saccharification, can either be enzymatic (using a mixture of
enzymes [i.e. biological catalysts used to facilitate and speed up metabolic
reactions in living organisms], known as amylases) or acid-based (Balat et
al., 2008). Once the starch is broken down to glucose syrup, the process is
similar to that for sugar crops (i.e. the sugars are fermented to ethanol
(typically using the yeast called Saccharomyces cerevisiae), followed by
distillation of the ethanol to the desired concentration and removal of
water). The process also yields several by-products, such as protein-rich
animal feed (e.g. dried distillers' grains with solubles, DDGS).

2.3 Second-generation liquid biofuels

Second-generation fuels are generally those made from non-edible
lignocellulosic (LC) biomass, either residues of forest management or food
crop production (e.g. corn stalks or rice husks) or whole plant biomass (e.g.
grasses or trees grown specifically for biofuel purposes) (Larson, 2008). LC
biomass, also called cellulosic biomass, is a complex composite material
consisting primarily of cellulose, hemicellulose and lignin bonded to each
other in the plant cell wall (USDOE, 2006).

There is major interest in moving from the current first-generation of liquid
biofuels to the second-generation biofuels. As an illustration, Larson (2008)
summarises: "By comparison to feedstocks for first-generation biofuels,
lignocellulosic biomass is generally (a) not edible and therefore does not
compete directly with food production; (b) can be bred specifically for
energy purposes, thereby enabling higher production per unit land area; and
(c) represents more of the above-ground plant material, thereby further
increasing land-use efficiency. These basic characteristics of
lignocellulosic materials translate into substantial energy and environmental
benefits for second-generation biofuels compared to most first-generation
biofuels". Similarly, at the Roundtable dedicated to 'Bioenergy and Food
Security' during the FAO Summit in June 2008, several countries "noted the
sustainability challenges related to the production of first generation
biofuels and highlighted the promise of second generation technologies to
reduce competition for natural resources" (FAO, 2008a, p. 33).

The potential importance of second-generation biofuels is clear from the
observation that most plant material is not sugar or starch but is LC
biomass. In fact, cellulose is the most abundant biological material on
earth. It is a polysaccharide that makes up about 40-50% of the weight of dry
wood. In higher plants it is organized into microfibrils, each containing up
to 36 glucan chains having thousands of glucose residues, which are largely
responsible for the plant cell wall's mechanical strength (USDOE, 2006).
Hemicellulose is also a polysaccharide, accounting for 25-35% of dry wood
(Balat et al., 2008). It is a mixture of various polymerised monosaccharides,
such as xylose and arabinose (both 5-carbon sugars, or pentoses) and glucose,
mannose and galactose (all 6-carbon sugars, or hexoses). Lignin, instead, is
not a polysaccharide and this highly branched polyphenolic macromolecule is
strongly resistant to chemical and biological degradation. It is not
fermented to produce liquid biofuels, but instead can be recovered and used
as a fuel for heat and electricity at an ethanol production facility (Larson,
2008). The relative proportions of these three materials in LC feedstocks
vary, depending on the species involved. For example, the biochemical
composition of biofuel feedstock from the pine tree is about 45% cellulose,
22% hemicellulose (mainly mannose followed by xylose sugars), 28% lignin and
6% others, while for switchgrass these proportions are 32% cellulose, 25%
hemicellulose (almost all xylose sugars), 18% lignin and 25% others (Balat et
al., 2008).

LC biomass can be converted to biofuels by thermo-chemical or biochemical
(biological) processing and many efforts are being made worldwide to
commercialise second-generation biofuels through both routes (e.g. Larson,
2008). The thermo-chemical processes generally use much higher temperatures
and pressures, begin with gasification (where the biomass is converted into
synthesis gas, also called syngas, that is a mixture of hydrogen and carbon
monoxide) or pyrolyis (heating of organic material in the absence of oxygen),
and can produce a wider variety of fuels than biochemical conversion
processes (see e.g. Larson, 2008; Royal Society, 2008). Many of the
second-generation thermo-chemical fuels, such as demethyl ether, refined
Fischer-Tropsch liquid (FTL) and methanol, are fuels that are already made
commercially from fossil fuels. For example, FTL is a mixture of hydrocarbon
compounds, resembling a semi-refined crude oil, that can be refined to
produce different hydrocarbon fuels, the primary one being a diesel-like fuel
for compression ignition engines. In addition to LC biomass, coal and natural
gas can also be used as feedstocks for FTL production (Larson, 2008).
Thermo-chemical processing of LC biomass is not described in any detail in
this document as, with few exceptions (see Section 3.2.d), they do not depend
on applications of biotechnology.

In biochemical processing of LC biomass to produce bioethanol, the process is
more complicated than converting starch to bioethanol. There are two key
parts. First, the cellulose and hemicellulose portions of the biomass must be
broken down into sugars. This is a major challenge, and a variety of thermal,
chemical and biochemical methods are being developed to carry out this
saccharification step in an efficient and low-cost manner (IEA, 2004).
Second, these sugars must be fermented to make bioethanol. The yielded
sugars, however, are a complex mixture of 5-carbon and 6-carbon sugars and
this provides a greater challenge for complete fermentation into bioethanol.

2.4 Global production of liquid biofuels

Although some pilot plants currently exist, second-generation biofuels still
remain a product for the future. Larson (2008) predicts that substantial
commercial production using biochemical processing will only begin in 10-20
years (versus 5-10 years for thermo-chemical processing). Such estimates
vary, depending on factors such as expected private sector investments and
oil prices, but it seems that it will take a minimum of five years (Rotman,
2008). First-generation biofuels, on the other hand, are already being
produced in significant commercial quantities in a number of countries. World
production has increased steadily in recent years, with production currently
dominated by two countries, the United States and Brazil, and one type of
fuel, bioethanol.

Estimates for global production of fuel ethanol indicate that it has tripled
from 2000 to 2007 to reach 52 billion litres. Although the United States and,
to a lesser degree, Brazil accounted for most of this growth, a large number
of other countries also began or increased production in this period
(OECD-FAO, 2008). Globally, most bioethanol production is from two crops,
maize and sugar cane, although other significant crops include cassava, rice,
sugar beet and wheat. In Brazil, most of the bioethanol is produced from
sugar cane while in the United States it is from maize. Of the estimated 52
billion litres of bioethanol produced in 2007, 26.5 billion (i.e. 51%) were
from the United States, 19 (37%) from Brazil, 2.3 (4%) from the European
Union (EU), mainly France and Germany, 1.8 (4%) from China, 1.0 (2%) from
Canada, 0.4 (1%) from India and 1.0 (2%) from other countries (FAO, 2008c,
Table 1).

For biodiesel, there has also been a major rise in global production over the
same period, increasing from less than 1 billion litres in 2000 to over 10
billion litres in 2007. Until 2004, the EU accounted for over 90% of global
biodiesel production, a proportion which has dropped to about 60% in 2007
because of increased production in other countries, especially the United
States (OECD-FAO, 2008). The most popular feedstocks used for biodiesel
production are rapeseed in the EU, soybean in Brazil and the United States
and palm, coconut and castor oils in tropical and subtropical countries, with
growing interest in jatropha. Of the estimated 10.2 billion litres of
biodiesel produced in 2007, 6.1 billion (i.e. 60%) were from the EU (led by
Germany), 1.7 (17%) from the United States, 0.4 (4%) from Indonesia, 0.3 (3%)
from Malaysia, 0.2 (2%) from Brazil, 0.1 (1%) from China and 1.2 (12%) from
other countries (FAO, 2008c, Table 1).

What about the future? The latest OECD-FAO Agricultural Outlook report
provides an assessment of future prospects in the major agricultural
commodity markets over the period 2008 to 2017 and, for the first time,
includes an analysis of and projections for global biofuel markets for
bioethanol and biodiesel (OECD-FAO, 2008). While noting that a number of
uncertainties (such as oil prices and government policies) affect their
projections, they predict that global ethanol production will continue to
increase so that the quantity produced in 2017 will double that of 2007. It
predicts that the United States and Brazil will continue to be the largest
ethanol producers through to 2017 but also that production in several other
countries, including China, India and Thailand, will grow rapidly. Regarding
global biodiesel production, the report suggests that it will grow at
slightly higher rates than for bioethanol to reach 24 billion litres by 2017
and that production in 2017 will continue to be dominated by the EU (over
50%), followed by Indonesia, Brazil, the United States and Malaysia
respectively.

Policy interventions, especially in the form of subsidies and mandated
blending of biofuels with fossil fuels, are driving the rush to liquid
biofuels (FAO, 2008c). For example, the EU decided in March 2007 to set
mandatory targets of a 20% share of renewable energies in overall EU energy
consumption by the year 2020, and a mandatory 10% minimum target for the
share of biofuels in overall EU transport petrol and diesel consumption by
2020. Also, the United States Congress in December 2007 passed the Energy
Independence and Security Act which, inter alia, sets required minimum annual
levels of renewable fuel (biofuel) in United States transportation fuel,
beginning at about 34 billion litres in 2008 and rising to about 137 billion
litres in 2022 (Zarrilli, 2008). Indeed, a recent study of bioenergy
development in the G8+5 countries (i.e. the G8 countries - Canada, France,
Germany, Italy, Japan, Russia, United Kingdom and United States - plus
Brazil, China, India, Mexico and South Africa), shows that all except one
(Russia) have set either mandatory or voluntary biofuel transport targets
(GBEP, 2007, Table 2.2).

Biofuel development in Organisation for Economic Co-operation and Development
(OECD) countries has therefore been promoted and supported by government
policies and a growing number of developing countries are also beginning to
introduce policies to promote biofuels (FAO, 2008c). Analysis indicates that,
with the exception of bioethanol from sugar cane in Brazil, biofuels are
generally not economically competitive with fossil fuels without subsidies
(FAO, 2008c). In most countries, biofuel production is therefore dependent on
public support and the ongoing discussion about the potential and actual
benefits of supporting biofuel production/use will have a major influence on
biofuel production in the future. As summarised by OECD-FAO (2008), "changes
in biofuel policies, either to raise or to lower domestic targets or to
review current policy incentives downwards, could be of major importance for
agricultural markets given that biofuel production is one of the important
factors lending strength to these markets over the medium term".

2.5 Reasons for major current focus on liquid biofuels

As mentioned above, many governments are actively encouraging liquid biofuel
production in their countries. A number of policy mechanisms are being used
by governments for this purpose, including feed-in tariffs (where a
regulatory minimum guaranteed price is paid for renewable power fed into the
electricity grid), taxes, guaranteed markets, compulsory grid connections for
renewable energy producers and other direct support for bioenergy production,
such as grants, loans and subsidies (GBEP, 2007). Government policies are
therefore playing a key role in influencing investment in bioenergy.
Following GBEP (2007), there are four main factors driving the current
interest in liquid biofuels:

a) High energy prices

In recent years, the average price per barrel of oil has increased steadily
e.g. from 27 US dollars (USD) in 2002 to 62 USD in 2006 and it continues to
be high (rising to new records, peaking at almost 150 USD in July 2008 -
http://omrpublic.iea.org/). Over the next decade, OECD-FAO (2008) predicts
that world oil prices are also likely to remain high relative to historical
levels and their report assumes prices will increase from 90 USD in 2008 to
104 USD in 2017, while noting that there is major uncertainty about future
oil prices.

b) Energy security

For countries dependent on oil or natural gas imports, biofuels offer a way
to diversify energy supplies and reduce their reliance on a few exporting
countries.

c) Climate change

Anthropogenic (human-induced) climate change has recently become a well
established fact and the resulting impact on the environment is already being
observed (e.g. FAO, 2006b). Because of increased atmospheric concentrations
of so-called greenhouse gases (GHGs, such as carbon dioxide (CO2), methane
(CH4), nitrous oxide (NO2) and chlorofluorocarbons), the average temperature
of the earth's surface has increased. To mitigate climate change, countries
have committed themselves to varying degrees to reducing GHG release into the
atmosphere. For example, the Kyoto Protocol, ratified by 182 countries and
the EU, which entered into force in 2005, sets legally binding targets and
timetables for cutting GHG emissions for the world's leading economies which
have accepted it (http://unfccc.int/kyoto_protocol/items/2830.php).

In this context, replacement of fossil fuels, such as petrol, by biofuels for
transport purposes has been advocated as an option for a country to reduce
its GHG emissions. This is because most life-cycle studies indicate that the
use of biofuels instead of fossil fuels reduces GHG emissions, as biofuels
sequester carbon through growth of the feedstock (e.g. Searchinger et al.,
2008). These life-cycle studies typically estimate that when biofuel and
fossil fuel GHG emissions are compared during the steps of making the
feedstocks (e.g. maize or crude oil), refining them into fuel and burning the
fuel in the transport vehicle, the combined GHG emissions from bioethanol (of
maize or LC origin) exceed or match those from fossil fuels. However, when
the calculations also include the fact that the growing biofuel feedstocks
remove (sequester) carbon dioxide from the atmosphere through photosynthesis,
the overall GHG emissions from biofuels are typically estimated to be lower
than those from fossil fuels (Searchinger et al., 2008).

d) Rural development

Cultivation of biofuels can contribute to maintaining employment and creating
new jobs in rural areas, avoiding land abandonment and reducing migration to
cities. The importance for developing countries is underlined by Zarrilli
(2008): "For both developed and developing countries alike, the bio-based
economy may boost employment and revenues in rural areas and revitalize them.
It may offer new end-markets for agricultural products and therefore add
value to them. All these goals are crucially important for all countries, but
particularly for developing countries". Similarly, FAO (2008c) points out
that the increasing demand for biofuels may offer opportunities for farmers
and rural communities in developing countries and thus contribute to rural
development. However, the report cautions that their capacity to take
advantage of these opportunities depends on the existence of an enabling
environment and that, at the global level, current trade policies do not
favour developing country participation or an efficient international pattern
of biofuel production while at the domestic level, farmers depend critically
on the existence of an appropriate policy framework and the necessary
physical and institutional infrastructure (FAO, 2008c).

2.6 Current concerns about production of liquid biofuels

As noted in the Introduction, production of liquid biofuels for transport
purposes is currently quite controversial and has led to concerns about a
number of issues, such as:

a) Increasing food prices

Agricultural commodity prices rose sharply towards the end of 2006 and in
2007 and continued to rise even more sharply in early 2008 before stabilising
and then declining to below January 2008 levels. The FAO Food Price Index
therefore rose on average 8% in 2006 compared with the previous year, in 2007
it increased by 24% compared to 2006 and in September 2008 (the latest data
available) it was up 11% from its value in September 2007 and 51% from
September 2006 (http://www.fao.org/worldfoodsituation/FoodPricesIndex/en/).
The surge in prices has been seen in almost all major food and feed
commodities. The driving forces behind the soaring food prices are many and
complex, where both supply-side and demand-side factors play a part. One of
the demand-side factors underlying the current state of the markets is the
demand from the biofuel industry for agricultural commodities such as sugar,
maize, cassava, oilseeds and palm oil (FAO, 2008d). Increased demand for
these commodities has been one of the leading reasons for the increase in
their prices in world markets, which in turn has led to higher food prices.
It is estimated that about 100 million tonnes of cereals (nearly 5% of global
cereal production) are being used for biofuels production in 2007-08 (FAO,
2008d).

For the future, OECD-FAO (2008) projects that food commodity prices will
continue to be higher than in the past. Compared to the period 1998-2007, it
predicts that average agricultural commodity prices will be substantially
higher for the period 2008-2017 (e.g. 40-60% higher for wheat, maize and skim
milk powder, over 60% higher for butter and oilseeds and over 80% higher for
vegetable oils). The demand for biofuels is one of the main factors
underlying their projections as "biofuel demand is the largest source of new
demand in decades and a strong factor underpinning the upward shift in
agricultural commodity prices" (OECD-FAO, 2008).

b) Land use changes

The Earth's land surface covers about 13.4 billion hectares. Of these,
roughly 1.5 are used for crop production, 3.5 as grassland, 3.9 for forests,
0.2 for urban settlements and the remaining 4.2 billion hectares consist of
desert, mountains and otherwise land that is unsuitable for productive use
(Doornbosch and Steenblik, 2007; FAO, 2003). The large demand for liquid
biofuels has led to increasing proportions of certain crops being used for
biofuel instead of for food/feed. For example, in the United States the
estimated proportion of maize cultivated that is used for biofuels has
steadily increased from less than 5% in 1997 to about 30% in 2008 (Larson,
2008; FAO, 2008e). It has also resulted in farmers switching from non-biofuel
crops (such as rice) to biofuel crops. It has also led to forests, peatlands,
savannas and grasslands being converted to agricultural lands for biofuel
production (or for non-biofuel crop production, to replace agricultural land
that has already been diverted to biofuel production) (Fargione et al.,
2008).

The conversion of natural lands, such as wetlands and natural forests, for
biofuel production represents an important threat to biodiversity through the
loss of habitats, their biodiversity components and the loss of essential
ecosystem services. In addition, the large-scale ploughing of
non-agricultural land and pasture land as well as peatland degradation could
result in substantive release of carbon emissions into the atmosphere
(SBSTTA, 2007). For example, Fargione et al. (2008) looked at six different
cases of habitat conversion situations currently taking place as a result of
biofuel production: conversion of 1) Brazilian Amazon rainforest to soybean
2) Brazilian Cerrado (i.e. tropical savannah) to soybean 3) Brazilian Cerrado
to sugar cane 4) Indonesian or Malaysian lowland tropical rainforest to oil
palm 5) Indonesian or Malaysian peatland tropical rainforest to oil palm and
6) United States central grassland to maize. Their results suggested that if
produced on converted land, then biofuels could, for long periods of time, be
much greater net emitters of GHGs than the fossil fuels they typically
displace.

c) Increased pressure on scarce water resources

As described in the Background Document to the previous e-mail conference of
this Forum, scarcity of water is one of the major global problems facing
humankind at the moment and it is likely to be an ever increasing problem in
the future. Furthermore, there will be more intense competition from the
industrial and municipal sectors for the water resources available for
agriculture in the future, despite the fact that there will also be an
ever-increasing demand for water in agriculture to meet the needs of the
growing world population (FAO, 2007). In water-short countries where
agriculture relies essentially on irrigation, increasing production of
biofuels will simply add to the strain on stressed water resources because of
the large quantities of water required for production of the feedstock and
its conversion to biofuel. Sugar cane and oil palm have high water
requirements (1500-2500 mm per year), while cassava, castor bean, cotton,
maize and soybean, all crops considered suitable for biofuels, require medium
levels of water (500-1000 mm per year) (FAO, 2008b). However, it is the share
of irrigation water used to meet these requirements which will influence
pressure on water resources. De Fraiture (2007) notes that the biomass needed
to produce one litre of liquid biofuel evaporates between 1000 and 4000
litres of water, depending on the type of feedstock and conversion techniques
used, and argues that "pursuing biofuel production in water-short countries
will put pressure on an already stretched resource and will turn green energy
into a major threat to resources".

3. BIOTECHNOLOGIES AND BIOENERGY PRODUCTION

As described in the Introduction, a wide range of biotechnologies are
available and many of them can be applied for bioenergy production in
developing countries. They include, among others, fermentation, genomics and
genetic modification and cover applications to micro-organisms, crops and
forest trees. In the context of bioenergy production, they can be used to
increase the efficiency of both parts of the production cycle i.e. the
production of biomass for bioenergy purposes and the conversion of the
biomass to biofuels.

Here, we will briefly consider some of the kinds of areas where
biotechnologies are or can be applied for production of first-generation
biofuels (Section 3.1), second-generation biofuels (3.2) as well as
microalgal biodiesel and biogas (3.3). Greatest attention is paid to
second-generation biofuels because of the large expectations they have
created and because of the significant role that biotechnology applications
are likely to play in their development.

3.1 Application of biotechnologies for first-generation biofuels

Apart from a range of factors including the amount of rainfall etc., yields
of liquid biofuel also depend on the crop that is cultivated and the part of
the world where it is grown. Estimated bioethanol yields per hectare have
been calculated to be about 5500 and 4500 litres (L) from sugar cane in
Brazil and India respectively, 3800 and 2000 L from maize in the United
States and China respectively and about 1900 and 1500 L from cassava in
Brazil and Nigeria respectively, while for biodiesel, estimated yields per
hectare are 4700 and 4100 L from oil palm in Malaysia and Indonesia
respectively and 550 and 500 L from soybean in the United States and Brazil
respectively (FAO, 2008c, Table 2).

a) Production of biomass

One way in which biotechnologies (or, indeed, conventional plant breeding)
could contribute is by improving biomass production. The plant varieties
currently being used for first-generation biofuels worldwide have been
genetically selected for agronomic characteristics relevant to food and/or
feed production and they have not been developed considering their
characteristics as potential feedstocks for biofuel production. Varieties
could be selected with increased biomass per hectare, increased yields of
oils (biodiesel crops) or fermentable sugars (bioethanol crops) or with
improvements in characteristics relevant for their conversion to biofuels. As
little genetic selection has been carried out in the past for biofuel
characteristics in most of these species, considerable genetic improvement
should be possible.

The field of genomics is likely to play an important role here. Genomics is
the study of an organism's genome i.e. the entire complement of its genetic
material (genes plus non-coding sequences). The goal of modern plant genomics
is to understand how plants do what they do i.e. to discover the function of
each gene; the cells in which each gene functions (and when); the
relationship each gene has with all other genes; and the consequences of
altered gene function (Tuskan, 2007). Draft genomes of several
first-generation feedstocks, such as maize, sorghum and soybean, are in the
pipeline or have already been published. For example, the project to sequence
the genetic code of soybean (Glycine max) began in 2006 and is expected to be
completed in 2008. Using the information this will provide on the genetic
make-up of soybean, research can aim to produce better varieties for biofuel
production by changing the type, quantity and/or location of the oil produced
by the plant (JGI, 2008). Apart from genomics, a range of other
biotechnologies can also be used, such as marker-assisted selection and
genetic modification. For example, Murphy (2007) describes how the task of
oil palm breeders can be facilitated by biotechnologies such as
marker-assisted selection (where DNA markers can be used to identify
genetically-superior individuals when they are just weeks old rather than
when the trees are 5-7 years old, after they produce the fruits that are the
source of the oil) or tissue culture (applied to multiply up genetically
superior trees).

b) Conversion of biomass to liquid biofuels

Another area where biotechnology can be applied is in improving the
conversion of biomass to liquid biofuels. For example, as the yeast
Saccharomyces cerevisiae cannot directly ferment starchy materials (e.g. corn
starch), the feedstock must first be hydrolysed using acids or enzymes, in
particular a family of enzymes called amylases, normally alpha-amylase and
glucoamylase. In the past, enzymes were isolated primarily from plant and
animal sources, and thus a relatively limited number of enzymes were
available. Today, bacteria and fungi are exploited and used for the
commercial production of a diversity of enzymes. Several strains of
micro-organisms have been selected or genetically modified to increase the
efficiency with which they produce enzymes. In most cases, the modified genes
are of microbial origin, although they may also come from different kingdoms
(FAO, 2006a, section 6.1.2). Many of the current commercially available
enzymes, including amylases, are produced using genetically modified (GM)
micro-organisms where the enzymes are produced in closed fermentation tank
installations (e.g. Novozymes, 2008). The final enzyme product does not
contain GM micro-organisms. Royal Society (2008) suggests that as the current
usage of GM micro-organisms within fermentation systems involves keeping them
in contained environments such as fermentation vats, then genetic
modification is a far less contentious issue here than with GM crops.

To reduce costs and increase the efficiency of bioconversion, research is
also ongoing to develop GM yeast strains which produce the amylases
themselves so that the saccharification and fermentation steps can be
combined, as well as to develop GM maize plants which can produce the
amylases (Royal Society, 2008). After fermentation, the ethanol produced
needs to be separated from the dilute solution using distillation. The step
requires a lot of energy and could be made more efficient by genetically
improving the micro-organisms used in the fermentation process so that the
ethanol concentration is increased prior to distillation (Royal Society,
2008).

3.2 Application of biotechnologies for second-generation biofuels

Because of the kinds of concerns mentioned in Section 2.6, there is great
interest in moving from first-generation biofuels towards use of LC biomass
for second-generation biofuels. This has brought with it major investments in
research and development (R+D) in this area, where e.g. in 2007 venture
entities invested an estimated 2.9 US billion dollars in the biofuel industry
sector in the United States alone and where investments worldwide are
expected to increase significantly in coming years (Kamis and Joshi, 2008).
If second-generation biofuels are to become a reality in the future some
technological breakthroughs are needed, and applications of biotechnology in
this context are discussed here.

However, it should be noted that these biotechnology breakthroughs alone will
not be enough. Second-generation biofuels will also have to be economically
viable and environmentally sustainable, which will depend on a series of
factors, including the logistical challenge of collecting and transporting
large amounts (in quantity and volume) of LC biomass to the biofuel
production facilities (Doornbosch and Steenblik, 2007). This may require that
the LC biomass is produced close to the processing site, which can be a
disadvantage for developing countries who at the moment have the option of
producing feedstock that can be shipped, processed or semi-processed, for
further conversion in the country of use. Also, competition for land and
other inputs will remain a challenge and it is not certain that all the
concerns related to use of first-generation biofuels will be alleviated by
second-generation biofuels. For example, Fargione et al. (2008) suggest that,
like first-generation biofuels, second-generation biofuels may also result in
land clearing and land use changes. FAO (2008c) also notes that excessive
withdrawal of agricultural residues for bioenergy purposes could negatively
impact soil fertility and quality by removing decomposing biomass.

The LC biomass needed for second-generation biofuels can come from two main
sources. The first source is from by-products, such as agricultural residues
like sugar cane bagasse, corn stover, straws from barley, oats, rice, wheat
and sorghum; residues from the pulp and paper industry; and municipal
cellulosic solid wastes. For example, Bon and Ferrara (2007) predict that in
Brazil there will eventually be significant production of bioethanol from
sugar cane bagasse and straw, materials that are available on a large-scale.
The second source is from dedicated biomass feedstocks, grown specifically
for the purpose of biofuel production, such as perennial grasses and
short-rotation forest trees (Tuskan, 2007). As with first-generation
biofuels, applications of biotechnologies can be considered separately for
production of biomass and for conversion of the biomass to biofuels.

a) Production of LC biomass from by-products

Concerning the by-products of crop production, relatively little R+D has yet
been carried out with biofuels in mind. For example, cereal production has
been optimised for grain yield but the crops have not been bred for straw
quality in relation to its use as biomass for biofuel purposes (Royal
Society, 2008). Substantial room for genetic improvement therefore exists.
Thus, information from genomic projects of first-generation biofuel crops,
such as those mentioned in Section 3.1.a, can also be used in genetic
improvement programmes to breed varieties with LC biomass characteristics
that are more suitable for biofuel purposes (JGI, 2008). Some examples of
ongoing research projects in this area include attempts to: identify and
isolate genes in sweet sorghum that control the high stalk sugar trait and a
decreased stalk lignin trait, in order to combine both traits within the same
plant; identify genes that regulate cell wall synthesis in rice, in order to
genetically manipulate them to change the cell wall composition for cost
efficient ethanol fermentation; and optimise the use of DNA markers to
simultaneously breed for high grain yield (for energy or non-energy purposes)
and high stover quality (for ethanol production) in maize (USDOE, 2007).

b) Production of LC biomass from dedicated feedstocks

Concerning dedicated biomass feedstocks, a range of potential candidates are
of interest. They include perennial grasses (i.e. which flower for several
years) such as switchgrass, miscanthus, reed canary grass and giant reed
(Royal Society, 2008). They also include tree species such as the poplar and
eucalyptus. As for some of the first-generation biofuel species, the genomes
of a number of second-generation species are also being sequenced. For
example, the recent announcement that the eucalyptus tree genome is to be
sequenced is important because eucalyptus species are the most widely planted
hardwood trees in the world (occupying more than 18 million hectares),
supplying woody biomass for several industrial applications. The challenges
and potential of applying new molecular techniques and approaches to
eucalyptus breeding for traits such as those relevant to biofuel purposes
have recently been reviewed by Grattapaglia (2007).

The eucalyptus genome will be the second tree genome to be sequenced
following that of the poplar already published in 2006. Tuskan (2007)
describes how the genomics information of the poplar can be used in
combination with the extensive knowledge already available about important
identified genes of other species, such as rice or the model species
Arabidopsis, to identify equivalent (homologous) genes in the poplar so that
trees with desirable properties for biomass production can be developed.
Among other things, the trees would ideally: accumulate greater carbon
allocation in the stem, through the development of a less extensive root
system and through reduced height and minimal perennial branch formation and
growth; be relatively short but with a large stem diameter, generating lower
amounts of low quality wood, more harvested biomass and improved
harvesting/handling efficiencies; display higher productivity per unit area
and drought/stress tolerance; not produce flowers; and be modified to produce
optimal feedstocks for energy conversion (e.g. by increasing the
polysaccharide component in the wood at the expense of lignin, for
biochemical conversion of the biomass to liquid biofuels) (Tuskan, 2007).
Apart from genomics, other biotechnologies can also be applied. For example,
Sticklen (2006) reviews some of the ways in which genetic modification can be
applied to improve the biomass characteristics of plants for biofuels,
including development of crop varieties that produce less lignin; that
self-produce cellulase enzymes for cellulose degradation and ligninase
enzymes for lignin degradation; or that have increased cellulose or overall
biomass yields.

c) Conversion of LC biomass to liquid biofuels

As mentioned earlier, LC biomass can be converted to biofuels in two main
ways, by thermo-chemical or biochemical processing (Larson, 2008) and here we
will discuss biochemical processing, because of the extensive applications of
biotechnology involved. Depending on factors such as the kind of feedstock
available, biochemical conversion of LC biomass to liquid biofuels can follow
a number of different pathways, in which four major steps can generally be
identified (Balat et al., 2008).

First is pre-treatment of the biomass, which promotes the physical disruption
of the LC matrix. This is necessary because the LC materials are structured
for strength and resistance to biological, physical and chemical attack (Bon
and Ferrara, 2007). Pre-treatment can be carried out in a number of ways e.g.
using dilute acids (such as sulphuric or hydrochloric acid), alkalines (such
as calcium hydroxide), liquid ammonia (the ammonia fibre explosion
pre-treatment) or steam explosion (Balat et al., 2008).

Second is hydrolysis i.e. breakdown of the polysaccharides to their simple
sugars, which is carried out using either acid (dilute or concentrated) or
enzymes. According to Royal Society (2008), the current trend is towards
enzymatic hydrolysis to avoid costly recovery and wastewater treatment
requirements resulting from the use of acid. Balat et al. (2008) also
indicate that enzymatic hydrolysis is attractive because it produces better
yields and that enzyme manufacturers have recently reduced costs
substantially using biotechnology.

The importance and interest in enzymatic hydrolysis has renewed and increased
the focus on several aspects of cellulases (i.e. enzymes, such as
endoglucanases, cellobiohydrolases and beta-glucosidases, which break down
cellulose) and hemicellulases (i.e. enzymes, such as xylanases, mannanases,
xylosidases, glucosidases or arabinosidases, that break down hemicelluloses).
These include the search for high cellulase-producing organisms; the
production of hypercellulolytic mutants (i.e. which are highly efficient at
degrading cellulose) of organisms suitable for cellulase production; genetic
modification to develop high cellulase-producing organisms with high specific
activity; and theoretical studies on the mechanism of action of a
multi-enzyme system on a complex polymer (Bon and Ferrara, 2007). Engineering
of enzymes using advanced biotechnologies is ongoing to develop enzymes with
improved characteristics such as higher efficiencies, increased stability at
elevated temperatures and at certain pH levels and higher tolerance to
end-product inhibition (Bon and Ferrara, 2007).

Regarding the search for efficient biomass-degrading organisms, a wide range
of micro-organisms can produce cellulases and hemicellulases in nature and
are at the centre of major R+D initiatives. Among others, these include
strains of fungi (of Trichoderma, Penicillium or Chrysosporium species) and
bacteria (of Bacillus, Clostridium or Cellulomonas species). For example,
Tuskan (2007) describes some genome sequencing projects that are aiming
ultimately to find genes to produce new enzymes for plant cell wall
breakdown. These include projects focusing on specific micro-organisms known
to have desirable biomass-degrading characteristics, such as the bacterium
Clostridium thermocellum (which degrades cellulosic materials using a large
extracellular cellulase system called the cellulosome) or the white rot
fungus Phanerochaete chrysosporium  (which produces enzymes that degrade
lignin) or the bacterial community resident in the hindgut of a wood-feeding
termite.

The third step is fermentation of the sugars, resulting from the breakdown of
cellulose and hemicellulose, to bioethanol. Unlike production of bioethanol
from first-generation sugar crops or starchy materials, fermentation is more
complicated here as it is a mixed-sugar fermentation (involving pentose and
hexose sugars) and it takes place in the presence of inhibiting compounds
released and formed during the first two steps of the process, i.e.
pre-treatment and hydrolysis. Because of their larger sizes, thicker cell
walls, better growth at low pH, less stringent nutritional requirements and
greater resistance to contamination, yeasts are preferred to bacteria for
commercial fermentations (Jeffries, 2006). However, LC biomass, in particular
hardwood and agricultural raw materials, can contain 5-20% (or more) of the
pentose sugars xylose and arabinose which are not fermented to ethanol by the
yeast Saccharomyces cerevisiae, the most commonly used industrial
fermentation micro-organism (Hahn-Hagerdal et al., 2006).

To overcome these problems, several different approaches are being explored.
One is to develop efficient xylose-fermenting strains of Saccharomyces
cerevisiae using a range of biotechnologies, including genetic modification
(where genes to enable xylose fermentation are transferred from the yeast
Pichia stipitis, the bacteria Thermus thermophilus or the fungus Piromyces
species) and global gene expression analysis combined with targeted deletion
or altered expression of key genes (Jeffries, 2006). Another approach is to
focus on yeast species that naturally ferment xylose. For example, Pichia
stipitis is a well-studied natural xylose-fermenting yeast. The recent
reporting of its genome sequence, predicting over 5800 genes, is important in
this context as the genetic information can be employed to improve usefulness
of this yeast for commercial fermentation operations (Jeffries et al., 2007).
The optimism regarding these approaches is summarised by Jeffries (2006):
"Genomic and expression analysis of Pichia stipitis along with new strains
from nature should continue to drive this field forward. The eventual goal is
a yeast that is capable of efficiently fermenting glucose, xylose and other
minor sugars to ethanol, and progress is being made on multiple fronts".

Another approach is to focus on bacteria instead of yeast. Three bacterial
species that have received much attention are Escherichia coli, Klebsiella
oxytoca and Zymomonas mobilis and GM strains have been produced for each of
them for bioethanol purposes. For example, Zymomonas mobilis has been shown
to have higher ethanol yields and productivity than traditional yeast
fermentations. However, like Saccharomyces cerevisiae it cannot naturally
ferment pentose sugars. To overcome this, GM strains using genes from
Escherichia coli have been developed which can also ferment xyloses and/or
arabinoses (Balat et al., 2008).

The fourth step is removal of the bioethanol. The step involves distillation
which separates the bioethanol from water in the liquid mixture. See Balat et
al. (2008) for more details.

For simplicity, the process above has been described in four sequential
steps. In practice, enzymatic hydrolysis (2nd step) and fermentation (3rd
step) can also be carried out together, called simultaneous saccharification
and fermentation (SSF). This has a number of advantages, as they take place
in the same reactor, thus reducing costs, and increase the hydrolysis rate,
since the sugars resulting from hydrolysis are fermented and thus do not
inhibit cellulase activity. On the negative side, the ideal pH or temperature
conditions for the saccharification step may differ from those for the
fermentation step (Balat et al., 2008). For the future, it would also be
desirable to combine cellulase production, enzymatic hydrolysis and
fermentation - called consolidated bioprocessing (CBP) (Lynd et al., 2008).

Through the steps described above, two of the three main components of LC
biomass, i.e. cellulose and hemicellulose, are converted to bioethanol. The
third component, lignin, as well as its by-products, need to be removed
before fermentation takes place as they are often toxic to micro-organisms
and the enzymes used for hydrolysis, which can reduce the conversion
efficiency. According to Royal Society (2008), this could be partly addressed
by using low lignin feedstocks or developing new strains of lignin tolerant
and lignin degrading micro-organisms. Lignin can be burnt to provide a source
of heat and power for the conversion process. Alternatively new developments
may make it valuable as a chemical feedstock.

The importance of processes for converting LC biomass to liquid biofuels was
recently underlined by Lynd et al. (2008) in an analysis of the economics of
second-generation bioethanol production in the United States, concluding that
"the immediate factor impeding the emergence of an industry converting
cellulosic biomass into liquid fuels on a large scale is the high cost of
processing rather than the cost or availability of feedstock". In their
analysis, they also looked at the different steps involved in converting the
biomass to bioethanol and estimated that the cost savings of improving the
conversion of LC biomass to sugars (e.g. by eliminating pre-treatment,
reducing the amount of cellulase needed, using CBP) were in general much
larger than from improving the conversion of sugars to bioethanol (e.g. by
increasing fermentation yield).

d) Biotechnology in thermo-chemical conversion of LC biomass

As mentioned in Section 2.3, in addition to biochemical conversion, LC
feedstocks can also be converted to biofuels using a number of
thermo-chemical conversion processes. In one of these processes, i.e. the
production of alcohol fuels (ethanol or butanol) from syngas, biotechnology
may also play an important role as one option is to use micro-organisms to
ferment the syngas (Larson, 2008, section 3.2). Production of ethanol from
fermentation of syngas has already been demonstrated but considerable
improvements could be made (e.g. isolation of micro-organisms that act well
in hot temperatures, as gasification results in syngas with a high
temperature) and approaches such as metabolic engineering will be important
here (Henstra et al., 2007). (Metabolic engineering involves the targeted and
purposeful manipulation of the metabolic pathways of an organism, typically
involving alteration of cellular activities by the manipulation of the
enzymatic, transport and regulatory functions of the cell using recombinant
DNA and other genetic techniques). If this process could be made commercially
viable, it would be particularly advantageous as, unlike biochemical
conversion, the lignin in LC biomass, as well as cellulose and hemicellulose,
would be converted to a liquid biofuel (Larson, 2008).

3.3 Applications of biotechnologies for some other biofuels

Apart from using sugars, grains, or seeds (first-generation biofuels) or LC
biomass (second-generation biofuels), a number of other biomass sources can
be used to produce biofuels. Two of them will be briefly mentioned here, as
well as the role that biotechnologies may play for them. The first one,
involving microalgae, is an option for the future while the second one,
involving biogas production, is currently available in both developed and
developing countries.

a) Biodiesel from microalgae

The potential importance that microalgae might have for production of
biodiesel has long been recognised (see e.g. Sheehan et al. (1998) for some
historical background). Microalgae are unicellular photosynthetic
micro-organisms, living in saline or freshwater environments, that convert
sunlight, water and carbon dioxide to algal biomass. They are categorised
into four main classes: diatoms, green algae, blue-green algae and golden
algae. Like higher plants, they produce storage lipids in the form of TAGs.
Many species exhibit rapid growth and high productivity, and many microalgal
species can be induced to accumulate substantial quantities of lipids, often
greater than 60% of their dry biomass (Sheehan et al., 1998).

The advantages of using microalgae instead of crops for biodiesel production
include that they represent much higher potential biodiesel yields per
hectare (so that they could theoretically, unlike biodiesel crops, meet the
global demand for transport fuels); can be harvested throughout most of the
year, thus giving a regular supply of biomass; and use less freshwater
(Chisti, 2008; Schenk et al., 2008). As microalgal production can also take
place in ponds or bio-reactors on non-arable land or in a marine environment,
it need not compete with food production for land or water. Apart from high
efficiency production of TAGs for biodiesel, Schenk et al. (2008) argue that
microalgae are also well suited for the production of feedstocks for other
biofuels, including biohydrogen, bioethanol and biogas. Algae can also be
efficiently grown when coupled with CO2-emitting flue gases from power plants
and can contribute to atmospheric CO2 reductions when the biomass remaining
after extracting the oil for biodiesel is fed into carbon sequestration
processes (Schenk et al., 2008).

There are however, some serious hurdles to be overcome before the process
becomes a realistic alternative. For example, Chisti (2008) estimates that
the price of biomass production needs to fall about 9-fold for it to become
feasible, underlining the importance of improving the production technology
through e.g. developing efficient methods for recovering algal biomass from
the dilute broths produced in the bioreactors. He also argues that genetic
modification and metabolic engineering are likely to have the greatest impact
on improving the economics of production of microalgal biodiesel and that
some specific applications of biotechnologies that might be considered
include increasing the biomass yield; increasing the biomass growth rate as
well as the oil content in the biomass; and improving temperature tolerance
of the microalgae so that there is a reduced need for cooling, which is
expensive. In a similar vein, Schenk et al. (2008) argue that the biggest
challenge over the next few years will be to reduce costs for cultivation and
to further improve the biology of oil production from the microalgae. They
also emphasise the role that advanced biotechnologies will play in this
context, where "future algal strain improvement will utilize methodologies
such as lipidomics, genomics, proteomics, and metabolomics to screen for and
develop new strains that exhibit high growth and lipid biosynthesis rates,
broad environmental tolerances, and that produce high value-add by-products".
Currently, there are a number of companies setting up pilot plants with
anticipation to scale up to commercial biodiesel production from algae within
the next few years.

b) Biogas

In the absence of oxygen, certain bacteria will ferment biomass into methane
and carbon dioxide, a mixture called biogas. In this anaerobic digestion
process (i.e. without oxygen), the feedstocks used to obtain biogas may
include sewage sludge, agricultural by-products and wastes (especially animal
manure), industrial wastes (e.g. organic solid wastes) or municipal solid
wastes. They may also include dedicated feedstocks grown for the purpose of
biogas production. The resulting fuel can be used for heat, electricity and
as a vehicle fuel (after the gas has been compressed and using the same
engine and vehicle configuration as natural gas). As methane is a GHG, its
capture and use as a biofuel prevents the release of methane into the
atmosphere.

Biogas can be produced at landfill sites, centralised co-digestion units
(co-operative units, using different biomass sources) or in farm-scale units.
These farm-scale digestion plants, mainly using animal wastes, are widespread
throughout the developing and developed world (IEA, 2005). For example, in
China 17 million biogas users are reported in 2005, in India there are an
estimated 3.8 million household-scale biogas plants while in Nepal over 170
000 plants, using cattle and buffalo manure, are in operation (Nepal, 2008).
These plants (also called digesters) are generally used to provide gas for
cooking and lighting for a single household.

At the biochemical level, anaerobic digestion is complex, consisting of a
series of reactions catalysed by a mixture of different bacterial species.
Four stages of anaerobic digestion are generally distinguished: hydrolysis,
acidogenesis, acetogenesis and methanogenesis. In the first stage, bacteria
secrete enzymes which hydrolyse polymers, such as lipids, proteins and
carbohydrates, to smaller molecules such as fatty acids, amino acids and
glucose. For example, proteins are generally hydrolysed to amino acids by
protease enzymes secreted by Bacteroides, Butyrivibrio, Clostridium,
Fusobacterium, Selenomonas and Streptococcus species. Second, in
acidogenesis, these products are metabolised by groups of bacteria and
fermented to produce organic acids, such as butyric acid, propionic acid and
acetic acid, as well as hydrogen and carbon dioxide. Third, acetogenic
bacteria (i.e. that make acetic acid as their sole or primary metabolic
end-product) convert organic acids to acetic acid plus hydrogen and carbon
dioxide. Fourth, methanogenic bacteria produce methane from acetic acid or
from hydrogen and carbon dioxide (FAO, 1997, Chapter 4).

Anaerobic digestion happens slowly in nature and could be accelerated in
several ways, such as using more efficient micro-organisms in these
processes, although knowledge of these microbial communities is generally
still quite basic. However, to improve the understanding and efficiency of
biogas production, some studies on the roles of the different populations of
micro-organisms have been carried out, on specific types of micro-organisms
such as cellulolytic bacteria (that break down cellulose) and methanogenic
bacteria in specific environments like landfill sites or solid waste or
sewage sludge digesters (Cirne et al., 2007).

As an example of one such study, Cirne et al. (2007) looked at the microbial
community involved in the first stage (hydrolysis) of anaerobic digestion of
two different kinds of organic substrates (sugar beets and grass/clover)
using a technique called fluorescence in situ hybridisation (FISH), where
fluorescently labelled DNA sequences are added to bacterial cells, making it
possible to identify, quantify and localise different bacterial species in
complex microbial communities without having to actually cultivate the
microbes. From the study they were able to identify the general bacterial
groups involved, concluding that their results "could be considered as a
first step towards the development of strategies to stimulate hydrolysis
further and ultimately increasing the methane production rates and yields
from reactor-based digestion of these substrates". A range of other
biotechnologies are also being applied in this context, such as the use of
metagenomics (i.e. isolating, sequencing and characterising DNA extracted
directly from environmental samples) to study the micro-organisms involved in
a biogas producing unit in order to improve its operation (e.g.
http://www.jgi.doe.gov/sequencing/why/99203.html).

4. SOME ISSUES RELEVANT TO THE DEBATE

As with each conference hosted by this FAO Biotechnology Forum, the focus is
on application of biotechnology in developing countries. For this debate, a
small number of issues of specific relevance to application of
biotechnologies for bioenergy purposes in developing countries are briefly
described below.

4.1 Technology relevance

Considering second-generation biofuel technologies, Larson (2008) argues that
since they are primarily being developed in industrialised countries, there
are important issues that must be considered about their relevance for
developing countries. Thus, he writes: "Technologies developed for
industrialized country applications will typically be capital-intensive,
labour-minimizing, and designed for large-scale installations to achieve best
economics. In addition, the biomass feedstocks for which technologies are
designed may be quite different from feedstocks that are suitable for
production in developing countries. To capitalize on their comparative
advantages of better growing climates and lower labour costs, developing
countries will need to be able to adapt such technologies. Tailoring
feedstocks to local biogeophysical conditions will be important for
maximizing biomass productivity per hectare and minimizing costs. In
addition, adapting conversion technologies to reduce capital intensities and
increase labour intensities will be important for providing greater
employment opportunities and reducing the sensitivity of product cost to
scale".

Putting these considerations into the specific context of the subject of this
conference, biotechnologies (for production of biofuel feedstocks and/or for
conversion of feedstocks to biofuels) developed in industrialised countries
may need to be adapted for appropriate use in developing countries. This may
involve the need to adapt biotechnologies or biotechnology products developed
elsewhere to different crop/tree species and agro-ecological zones or to
different kinds of biomass production and/or bioconversion systems in
developing countries. The ability of developing countries to adapt complex
and sophisticated technologies to local needs and capacities may, however, be
problematic (Zarrilli, 2008).

4.2 Intellectual property rights

Related to the previous issue, major investments are being made today in R+D
in biofuels, primarily by the private sector and in developed countries. This
is clearly reflected in the major increase in biofuel-related patents. A
recent analysis by Kamis and Joshi (2008) indicates that the number of
biofuel-related patents (defined as U.S. patents applied for or granted, plus
Patent Cooperation Treaty (PCT) international applications) has risen from
about 150 in 2002, to about 400 in 2005 and to over 1,000 in 2007, when the
number of patents was higher than the combined total of patents related to
solar and wind power. For 2006-2007, the major focus of patents was on
biodiesel rather than bioethanol and most patents were owned by the private
sector, with 57% owned by corporate entities, 11% by universities or other
academic institutions and 32% undesignated. The authors predict that for the
future, the number of biofuel patents will continue to increase steadily; the
number of agricultural biotechnology biofuel patents will significantly
increase as transgenic plant technology is directed to biofuel applications;
and that LC biofuel patents will increase in number.

Zarrilli (2008) notes that forthcoming biofuel technology will be proprietary
and points out that strong intellectual property rights (IPR) regimes may
mean that access to technology is problematic, especially for developing
countries. The issue of IPR has frequently been raised in previous
conferences of this Forum, with participants indicating that they are an
issue for both GMOs and non-GMO biotechnologies. Their consequences were
generally seen as negative, with concerns expressed that they might for
example, act as a constraint to biotechnology research in developing
countries (FAO, 2006a).

4.3 Non-transport biofuels

While the major focus today is on production of liquid biofuels for transport
purposes, it is also important to keep in mind that the production of
biofuels for non-transport needs (lighting, heating, cooking) could have
tremendous advantages for developing countries. For example, for regions such
as sub-Saharan Africa, breakthroughs in the area of liquid biofuels for
cooking would be very important as energy for cooking is a priority since 95%
of all staple foods must be cooked and traditional cookstoves, powered by
fuelwood and dung, have negative health and social impacts. According to
UNCSD (2007), "transition to improved cookstoves using biobased feedstocks
could free women and children from the collection and transport of wood and
dung which can account for up to one-third of their productive time, and
reduce the effects of indoor air pollution which is responsible for more
deaths of women and children than malaria and tuberculosis combined".

5. POTENTIAL TOPICS TO BE DISCUSSED IN THE CONFERENCE

Some of the kinds of specific questions that participants might wish to
address in the e-mail conference are given below:

5.1 For first-generation liquid biofuels (i.e. from grains, seeds, oils):

- As described in Section 2.6, there are currently a number of concerns about
first-generation biofuels. Can applications of biotechnology substantially
alleviate any of these concerns? If so, how?

- For R+D programmes in developing countries, should applications of
biotechnology focus on production of biomass for biofuel purposes or on
bioconversion of biomass to liquid biofuels?

5.2 For second-generation liquid biofuels (i.e. from LC biomass):

- Second-generation biofuels, although not yet commercially available, are
likely to be a reality in the future. How important is it for developing
countries to be involved in the biotechnology-based R+D that will play a key
role in their eventual availability? Alternatively, should developing
countries prioritise other activities now and use the biotechnology
tools/products for second-generation biofuels developed elsewhere (probably
in developed countries) when they are eventually available on the market?

- Most of the world's industrial enzymes (60%) are produced in Europe, while
the remaining 40% come from the United States and Japan, although countries
like China, India and South Korea are likely to play a greater role in the
future (Bon and Ferrara, 2007). For conversion of LC biomass to liquid
biofuels, use of cellulases plays a key role in the economics of the
operation. How realistic is it for developing countries to produce their own
cellulases? Can regional co-operation be important here?

- As mentioned in Section 2.3, LC biomass can be converted to biofuels
through two major routes, by thermo-chemical or biochemical processing, where
only the latter involves extensive applications of biotechnology. For
developing countries wishing to produce second-generation liquid biofuels,
are there strong arguments in favour of one of the processing routes over the
other?

5.3 For other kinds of biofuels

- Production of biodiesel from microalgae is not currently feasible but may
be so in the future. As for other future biofuels, such as second-generation
biofuels based on LC biomass, should developing countries invest their
(generally scarce) biotechnology R+D resources in this area or should they
wait until commercial products are available in the future? If so, which
aspects should be prioritised?

- Small-scale biogas units are already operating in developing countries. Is
there a role for biotechnologies in improving the operation/efficiency of
these units? If so, how?

5.4 General questions

- The issue of the relevance for developing countries of bioenergy-related
biotechnologies produced in developed countries was discussed briefly in
Section 4.1. How important is this issue and what can developing countries do
about it?

- Regarding IPR mentioned in Section 4.2, how big of an issue is this in
relation to biotechnologies for bioenergy production in developing countries
and how should developing countries act to ensure they have access to
appropriate biotechnologies for bioenergy production?

- Regarding biofuel production for non-transport purposes (Section 4.3), can
applications of biotechnology contribute in a significant way to the
non-transport energy needs of people living in developing countries? If so,
how?

- First generation biofuels and biogas are currently available, while
second-generation biofuels and microalgal biodiesel are still in the
pipeline. Should developing countries prioritise their biotechnology
resources (people, money etc.) on the range of biofuels currently available
or on those showing great promise but which will only be available in the
future?

- In the biofuel sector today, some developing countries, in particular
Brazil, are key players. In the context of applying biotechnology for
bioenergy production, how important can South-South co-operation be so that
technicians and experts in developing countries can help each other?

- Are certain applications of biotechnology for bioenergy purposes of major
specific relevance/benefit to rural smallholders in one or more regions of
the developing world? If so, which ones?

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*** Paper prepared for the FAO seminar on "The role of agricultural
biotechnologies for production of bioenergy in developing countries", held in
Rome, Italy, 12 October 2007. http://www.fao.org/biotech/seminaroct2007.htm

NB: When submitting messages (which should not exceed 600 words),
participants are requested to ensure that their messages address the kinds of
issues mentioned in Section 5 of this document. Before sending a message,
members of the Forum are requested to have a look at the Rules of the Forum
and the Guidelines for Participation in the E-mail Conferences. These were
provided when joining the Forum, and they can also be found at
http://www.fao.org/biotech/forum.asp. One important rule is that participants
are assumed to be speaking in their personal capacity, unless they explicitly
state that their contribution represents the views of their organisation.

ABBREVIATIONS: CBP = Consolidated bioprocessing; EU = European Union;
FTL = Fischer-Tropsch liquid; GHGs = Greenhouse gases; GM =
Genetically modified;
GMOs = Genetically modified organisms; IPR = Intellectual property rights; LC
= lignocellulosic; OECD = Organisation for Economic Co-operation and
Development; R+D = Research and development; TAGs = Triacylglycerols.

ACKNOWLEDGEMENTS: The FAO Working Group on Biotechnology expresses its
grateful appreciation to the following people for their comments on
parts or
all of the document: To the external referees Elba P.S. Bon, Federal
University of Rio de Janeiro, Brazil (http://www.iq.ufrj.br), Maria Antonieta
Ferrara, Oswaldo Cruz Foundation, Brazil (http://www.far.fiocruz.br), Denis
J. Murphy, University of Glamorgan, United Kingdom
(http://people.glam.ac.uk/view/184) and Peer Schenk, University of
Queensland, Australia (http://profiles.bacs.uq.edu.au/Peer.Schenk.html) as
well as to our FAO colleagues, in particular Astrid Agostini, as well as
Jean-Marc Faurès, Hannes Johnson, Preetmoninder Lidder, Tony Piccolo and Wim
Polman.

FAO, 21 October 2008.

Recommended reference for this publication:
FAO, 2008. The role of agricultural biotechnologies for production of
bioenergy in developing countries. Background Document to Conference 15 of
the FAO Biotechnology Forum (10 November to 7 December 2008):
http://www.fao.org/biotech/C15doc.htm