Non-food uses:Opportunities for the Australian oilseed industry

Phil Salisbury & Allan Green

The petrochemical factory

Unlockingancient carbon

(non-renewable)

AOF Innovations Committee Report:Industrial Use of Vegetable Oils

Report evaluates:

Current and future uses of edible vegetable oils for industrial purposes, either directly or after fractionation

Current use of existing industrial (non-edible) vegetable oils

The further domestication of existing wild species that produce industrial oils

The development of novel industrial oils, using gene technology to breed new fatty acid compositions in existing, high yielding oilseed species

Industrial Use of Vegetable Oils

Vegetable oils can be directly used for industrial purposes or can be split into different derivatives then utilized

Oleochemicals refer to chemicals derived from natural oils and fats of both plant and animal origins

Essentially, they refer to the fatty acids and glycerol derived from splitting of the triglyceride structure of oils and fats.

They also include those derivatives derived from the subsequent modification of the carboxylic acid group of the fatty acid by chemical or biological means and further derivatives

Industrial Use of Vegetable Oils

Oleochemicals are often categorized into basic groups including:

fatty acids

fatty acid methyl esters

fatty alcohols

fatty amines

glycerol

other compounds derived from further chemical modification of these groups

Direct industrial use of edible vegetable oils

Direct industrial uses of edible vegetable oils include use as surfactants and adjuvants,

drying agents and surface coatings,

lubricants,

dust suppressants,

cosmetics,

industrial raw materials

specialty chemicals

adhesives

biocomposites

Direct industrial use of edible vegetable oils

Direct use of vegetable oils in many industrial products is limited by:

the restricted carbon chain length variation in the major cultivated oilseed species (mostly C16–C18) and

the lack of very high levels (>80%) of individual fatty acids in the major vegetable oils

An exception to this general trend is the high oleic acid oils (75-85% oleic acid) developed in sunflower, canola and other oilseed species.

HO oils have significant industrial application in lubricants, plastics and cosmetics.

Further processing into oleochemicals is generally required to enable current vegetable oil products to be more efficiently utilized in industrial products

Use as oleochemicals

Total oleochemical production

010002000300040005000600070008000

1995 2000 2010

Oleo

chem

ical k

T)Europe AmericaAsia Others

Use as oleochemicals

World oleochemical production

0100020003000400050006000700080009000

1995 2000 2010

Oleo

chem

ical (k

T)Fatty acids Fatty Methyl Esters

Fatty alcohols – natural Fatty amines

Glycerol – natural Basic oleochemicals

Use of existing industrial (non-edible) vegetable oils

High Erucic Acid RapeseedHEAR typically contain 50-60% erucic acid.

In 2000, around 90,000t of HEAR seed was harvested across France, Germany, UK and Italy.

Australia has also grown small commercial areas (1-3,000 ha) of HEAR in recent years using locally developed and adapted cultivars, but markets for the seed have not been readily available.

HEAR oil has special properties including high smoke and flash points, oiliness and stability at high temperatures, ability to remain fluid at low temperatures and durability

The principal end use is erucamide, a slip agent used in injection moulded plastics and polythene manufacture as an industrial lubricant, reducing surface tension and preventing adhesion between film surfaces

HEAR oil is also used in printing inks, lubricants and a range of other applications

Use of existing industrial (non-edible) vegetable oils

Crambe Crambe abyssinica has reached commercial status in North America where it grown for its erucic acid for industrial purposes

In the UK it has also reached commercial status on a small scale, with several thousand hectares of production

In Australia, Crambe has yielded well (>1t/ha) in Mallee type environments

Commercial adoption limited by high glucosinolate meal

Use of existing industrial (non-edible) vegetable oils

Linseed Linseed (Linum usitatissimum) is grown worldwide as a source of oil for industrial use in the manufacture of paints, varnishes, resins, inks and linoleum, because of its drying and hardening properties when exposed to air and sunlight.

World production of linseed is currently around 1 million tonnes.

A few thousand hectares of linseed are grown in Australia annually, but much of this is used in bakery applications.

Domestication of wild industrial oil species

One limitation to the use of current vegetable oils as oleochemicals is that most oils predominantly consist of C16 and C18 fatty acids. This restricts their use as oleochemicals.

Many industrial products require shorter or longer chain fatty acids and often more complex fatty acids (e.g. with functional groups).

This has led to investigations of less domesticated plant species as potential sources of these required shorter chain, longer chain & functionalised fatty acids.

Among the fatty acids that have been identified are:

unusual chain lengths

hydroxy-, epoxy-, acetylenic-,

conjugated fatty acids with double bonds at unusual positions.

Some unusual industrial fatty acids

lauric acid(detergents)

conjugated fatty acids(superior drying oils)

petroselenic acid(polymers, detergents)

ricinoleic acid(lubricants, cosmetics

pharmaceuticals)

vernolic acid(resins, coatings,

plasticisers)

erucic acid(polymers, cosmetics,inks, pharmaceuticals)

Domestication of wild industrial oil species

Exploitation of these new sources requires either:

the domestication of these wild species

OR

genetic engineering to transfer these traits into already adapted crop species

Domestication of wild industrial oil species

Advantages

Reduced reliance on traditional crops/products

Extended range of crop options for farmers

Reduced genetic vulnerability through diversification

Domestication of wild industrial oil species

DisadvantagesUndomesticated or wild species can have a number of agronomic problems which need to be addressed, including :

low yield

bienniality/perenniality

very long flowering period

pod shattering

harvesting difficulty associated with poor plant type/architecture

toxins

Indicative oil yields (t/ha) of traditional and potential new oilseed crops

0.2

0.2

0.3

0.3

0.4

0.7

0.9

1.3

1.7

1.0

Sunflower

Linseed

Lesquerella

Dimorphotheca

Euphorbia

Calendula

Rapeseed

Crambe

Lunaria

Limnanthes

Premium prices obtainable forspecialty oils are insufficient to offset current low oil yieldsof most candidate new crops

Domestication of wild industrial oil species

DisadvantagesOften a lack of understanding of breeding systems, phenology andareas of adaptation

e.g. jojoba

Long term breeding programs (20+ years) can be required to domesticate new crops. Need long term funding

Success is dependent on being able to identify (or create) the required genetic variation

New industrial oils in existing oil crops

This strategy involves extending the range of useful oils in existing high yield oil crops using gene transfer technologies

Four major oil crops account for almost three-quarters of globally traded vegetable oils:

soybean

oil palm

rapeseed

sunflower

It is an attractive option to use the recently developed GMO technology to manipulate these dominant oil crops so that they produce an enhanced range of valuable products

Where to in Australia?

Need to identify (higher value) product areas where Australia is competitive or could develop a competitive advantage:

Current and future uses of edible vegetable oils for industrial purposes, either directly or after fractionation - NO

Current use of existing industrial (non-edible) vegetable oils - NO

The further domestication of existing wild species that produce industrial oils – SOME NEW CROPS WORK WORTHWHILE, PROVIDED LONG TERM FUNDING AVAILABLE

The development of novel industrial oils, using gene technology to breed new industrial oil compositions in existing, high yielding oilseed species – AUSTRALIA WELL PLACED WITH SEVERAL POTENTIAL PATENT POSITIONS

Crop Biofactories Initiative

Strategic alliance between CSIRO and GRDC Plant Industry, Entomology, Molecular & Health Technologies

Collaboration between materials scientists and biotechnologists to focus on future materials that could be made from future plants

Focus areasIndustrial Oils

Complex Monomers

Protein Biopolymers

Three stagesStage 1 from 04/05 to 07/08 ($13M co-investment)

Stage 2 & Stage 3 conditional on outcomes of Stage 1

Expected time of up to 10 years until commercialisation

Crop Biofactories Initiative

CSIRO~GRDC~RIRDC national survey of expertise & interest in building an Australian biotransformation industry (2002)

International Workshop on Biotransformation (2002) held to explore Australia’s competitive niche

advanced agriculture and downstream processing capability

diversity of crops and growing seasons

strong R&D capacity in biotechnology and materials science

sound regulatory infrastructure

Crop Biofactories Initiative

CSIRO~GRDC~RIRDC national survey of expertise & interest in building an Australian biotransformation industry (2002)

International Workshop on Biotransformation (2002) held to explore Australia’s competitive niche

advanced agriculture and downstream processing capability

diversity of crops and growing seasons

strong R&D capacity in biotechnology and materials science

sound regulatory infrastructure

Detailed scoping of proposal and development of project plans (2003)

CBI agreed, funded and research commenced (2004/05)

Focus on combined value and volume

Volume (MT)

Price$/T

1000

2000

3000

Specialty chemicals

Pharma

COMMODITY

SPECIALTY

1 20 3

Biodiesel

The plant biofactory

Protein

Oil

CHO

CHO

Locking upcurrent carbon

Low-value commodity products

Protein

Oil

CHO

CHO

FAMEs

“Biodiesel”

Petrol additiveEtOH

Locking upcurrent carbon

High-value specialty products

Protein

Oil

CHO

CHO

Industrial oils & lubricants

Oleochemicals

Polymers(PHB, PLA)

Monomers( HB, LA)

Locking upcurrent carbon

“BioNylon”

Monomers(C6, C9)

CBI research stages

Stage 1Develop proprietary platform technologies for enzymatic synthesis in three focus areas

Achieve proof of concept production in model plants

Stage 2Evaluate range of candidate products from Stage 1

Select first generation products

Stage 3Select most valuable product-crop combinations & achieve economic production in selected crop plants

Develop supply chains

Complete regulatory and registration R&D

Conduct extension and marketing

CBI Stage 1 project portfolio

Industrial oils

Complex monomers

Protein biopolymers

Novel oils with unique functional groups and high-

value direct uses

Chemically complex monomers enable sophisticated polymers with

innovative properties

Novel structural proteins as advanced biomaterials

and adhesives

Market analysis and business development

CBI Stage 1 project portfolio

Industrial oils

Complex monomers

Protein biopolymers

VernolicOil

EpoxyOils

HydroxyOils

Poly-acetylenic

Oils

Fatty AcidMonomers Dicarboxylic

NovelSilks

RepeatMotif Adhesives

Market analysis and business development

Oilseed projects in CBI Stage 1 (PI)

VernolicOil

EpoxyOils

HydroxyOils

Poly-acetylenic

Oils

Market analysis and business development

CBI Project 2: Vernolic oil (pathfinder)

OH

O

CC

CC

CC

CC

O

CC CC

CC

CCC

C

Vernolic

Aim is to develop a “pathfinder” oilseed plant having high levels of vernolic acid (∆12-epoxy linoleic acid)

oils rich in vernolic acid are highly reactive and make excellent drying agents for use in oil-based or alkyd-resin paints

paints containing vernolic-based drying solvents have much lower levels of VOCs (market estimated to grow to US $3.5 billion globally by 2007)

vernolic acid also can be used in manufacture of novel interpenetrating plastics, metal coatings, and other high value compounds

bombykol (a sex pheromone which can be used as insecticide)

traumatic acid (an intermediate in prostaglandin synthesis)

Seed oils from Euphorbia lagascae and Vernonia galamensis are rich in vernolic acid (50-70%) but yields are low

CBI Project 2: Vernolic oil (pathfinder)

∆12-epoxygenase gene (Cpal2) cloned

from Crepis palaestina

70% vernolic acid

OH

O

CC

CC

CC

CC

O

CC CC

CC

CCC

C

Vernolic

CBI Project 2: Vernolic oil (pathfinder)

Cpal2 gene introduced into Arabidopsis and linseed

Cpal2

∆12-epoxygenase gene (Cpal2) cloned

from Crepis palaestina

70% vernolic acid 5% vernolic acid

20% vernolic acid

OH

O

CC

CC

CC

CC

O

CC CC

CC

CCC

C

Vernolic

General problem for ∆12-modified FAs

Low accumulation is a general phenomenon for plants expressing unusual ∆12-modified fatty acids

0

20

40

60

80

100

% in

see

d oi

ls

Wild plantTransgenic plant

Vernolic Ricinoleic Crepenynic Eleostearic

CBI Project 3: Poly-epoxy oils

Can poly-epoxy fatty acids be synthesised in plants?main product of interest with high value in glues, resins, surface coatings

currently made from petroleum, and by chemical epoxidation of poly-unsaturated vegetable oils (soy, linseed)

requires genes for epoxygenases that act on C=C bonds at other positions in acyl chain (e.g. ∆9 & ∆15)

these enzymes will need to be able to work sequentially on previously epoxygenated acyl substrates

OH

O O

CC

CC

O

CC CC

CC

CCC

C

O

CCC C

OH

O

CC

CC

CC CC

CC

CCC

C CCC C

Linolenic (18:3)

∆12 epoxy∆9 epoxy ∆15 epoxy

Correct biological functioning

To be commercially viable as crop biofactories, plants engineered with novel oils will need to have:

normal vegetative and reproductive performance

high grain yield

normal oil content in the grain

high concentrations of desired fatty acid in the oil

… and be able to break downunusual oils during germination

genes for specialised TAG lipases may need to be obtained from wild species containing these oils

Delivery and adoption challenges

Product segregationindustrial products produced from traditional food crops (e.g. oilseeds) will require strict segregation from food-grade products because they:-

will not be approved for food use

may actually be toxic

genetic isolation & Identity Preservation will be important crop and product management tools

dedicated industrial (non-food) crop plants may be preferred

Freedom to operatelong term large investment requires some certainty about ability to eventually commercialise products

access to enabling plant biotechnology could be an obstacle

international collaborations developing to overcome this

IP sharing + product competition

Proprietary product traits

Hydroxy AcetylEpoxy

Open-access platform and enabling technology

Transformation methods

Elite germplasm

Agronomic traits

Seed promoters

Planned EC-US Cooperation in Bio-based Product Research

Oilseed Crop Flagship Project … planned to focus on production of novel plants oils as industrial feedstocks.

EC-US Task Force on Biotechnology Research

Planned EC-US Cooperation in Bio-based Product Research

“The time is now right for the public sector to invest in team-based approach to discover the nature of the barriers to successful production of novel fatty acids in plants”

Working Group report (March, 2005)

EC-US Task Force on Biotechnology Research

A word of advice for plant biotechnology GRADUATES