Post on 13-Jan-2016
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The Sugarcane Biofactory The Sugarcane Biofactory
Why biomaterials?Why biomaterials?Why sugarcane?Why sugarcane?
Which targets?Which targets?SugarsSugars
Bio-Bio-polymerspolymers
HurdlesHurdles ActionsActions
Sucrose
Sorona
Isomaltulose
Ethanol
for a sustainable for a sustainable future future
Most people in the world are here
Individual Energy Use
Challenges to SustainabilityChallenges to Sustainability
We are all here
Atmospheric CO2
We are all here Most want to be here
Exponential increase in (non-renewable) resource consumptionExponential increase in (non-renewable) resource consumption
Huge imbalances in resource consumptionHuge imbalances in resource consumption Limited agricultural area - global loss of biodiversityLimited agricultural area - global loss of biodiversity
Our World TodayOur World TodayGlobal ImpactsGlobal ImpactsGlobal ImpactsGlobal Impacts
Allowing 12 percent for biodiversity, only 1.7 hectares of biologically productive area per capita is available for human use.
Ecological Footprint
Our World Our World TodayToday
Resource depletionResource depletion uneven consumptionuneven consumption
Environmental impactsEnvironmental impacts loss of biodiversityloss of biodiversity climate changeclimate change
Global Global communicationscommunications rich or poorrich or poor
Global conflicts Global conflicts over resourcesover resources
AbundantAbundant ‘‘guns, germs & steel’guns, germs & steel’
Global ImpactsGlobal ImpactsGlobal ImpactsGlobal Impacts
Industry Response (e.g. DuPont):* 10% of research budget into
renewables from CHO ($130M)
* 25% of sales in renewables
by 2010 ($8B revenues by 2015)
* 100% of vehicle fleet using
renewable fuels by 2015
* Major international biofuels
producer (partner BP)
* Major international bio-based
materials producer (partner MIT)
Global Global ChallengeChallengeAchieve Achieve sustainability sustainability
The greatest scientific challenge The greatest scientific challenge in human historyin human history
Renewable bioenergy Renewable bioenergy & biomaterials& biomaterials
Plant biotechnology Plant biotechnology is a keyis a key Sugarcane is one of the Sugarcane is one of the
most promising cropsmost promising crops
Harvesting SunlightHarvesting Sunlight
At an energy transformation efficiency of 2%, solar energy collectors covering 1% of land surface with would provide the equivalent of world current oil usage.
?
OR BIOMATERIALS!
Photosynthesis? Photosynthesis?
Photosynthesis: Photosynthesis: Harvesting SunlightHarvesting Sunlight
Why Sugarcane?Why Sugarcane?
High biomass productionHigh biomass production» 40-80 tons DW / ha / yr40-80 tons DW / ha / yr
Simple extractionSimple extraction» Soluble sugars Soluble sugars (20 tons sucrose ≈ 10 kL ethanol/ha/yr)(20 tons sucrose ≈ 10 kL ethanol/ha/yr)
» Fibre provides energy for processing Fibre provides energy for processing (excess)(excess)
Established gene transfer systemEstablished gene transfer system» Into elite cultivarsInto elite cultivars
Inbuilt containmentInbuilt containment» No survival outside cultivationNo survival outside cultivation
» No pollination of native plants or other cropsNo pollination of native plants or other crops
» Extraction removes all genes and proteinsExtraction removes all genes and proteins
Advantages for sustainable biomaterialsAdvantages for sustainable biomaterials
The OpportunityThe OpportunityEconomic and environmental sustainabilityEconomic and environmental sustainability
A profitable future based on:A profitable future based on: Value-added biomaterials & Value-added biomaterials &
bioenergybioenergy from renewable resources, from renewable resources, in sustainable & efficient in sustainable & efficient
production systems production systems
Competitive edge from IP Competitive edge from IP ownershipownership built on collaborative R&Dbuilt on collaborative R&D
Delivering benefits valued by Delivering benefits valued by customers & consumerscustomers & consumers health & quality of lifehealth & quality of life environmentenvironment
SucroseSucrose
Power & FuelPower & Fuel
Value-added sugars
Value-added sugars
Bio-polymers
Bio-polymers
IPIP
The Sugarcane Renewable Biomaterials Industry
RequirementsRequirementsfor economic viabilityfor economic viability
PlatformsPlatforms gene expression patternsgene expression patterns
ProductsProducts with enhanced valuewith enhanced value
MarketsMarkets & market development& market development
SucroseSucrose
Power & FuelPower & Fuel
Value-added sugars
Value-added sugars
Bio-polymers
Bio-polymers
IPIP
The Sugarcane Renewable Biomaterials Industry
Which Biomaterials?Which Biomaterials?For sustainable profitabilityFor sustainable profitability
Sugarcane Sugarcane metabolic metabolic
engineeringengineering
BiopolymersBiopolymers
Industrial Industrial enzymesenzymes
Enhanced sucrose
yield
Waxes, pigments,
antioxidants
Improved fibre quality
High-value sugars
Biofuel feedstock
Aromatics Aromatics
Suit non-food Suit non-food cultivars cultivars
Suit food cultivarsBy-products with
sugar
Which Biomaterials?Which Biomaterials?For sustainable profitabilityFor sustainable profitability
Target Compound Sucrose Candidate
Required Production Scale Projected world demand (tonnes / year) 2020 Yield (tonnes / ha) / Area (ha)
>130,000,00020 / 6,500,000 ?
Value Price (AU$ / ton wholesale); margin; stability Indirect benefits for environment, industry or consumers
<500; lownil ?
Production Method Co-production : food / non-food cv? Effect on sucrose yield? Alternative to sucrose
+ / +nil ?
Technical Feasibility New biological / industrial process needs / likely constraints Anticipated capital costs for new industrial facilities Research costs and timeline
nilnilnil
?
Potential to Capture Value Competitors? / Partners? FTO / Protected competitive advantage? Timing to market
many+ / -now
?
A high-value sucrose isomerA high-value sucrose isomerIsolated genes for sucrose isomerase Isolated genes for sucrose isomerase (SI)(SI)
Sucrose ( 1-2 GF)Isomaltulose ( 1-6 GF)
(= Palatinose)
SI
Engineering Sucrose ConversionEngineering Sucrose ConversionA pilot study: IsomaltuloseA pilot study: Isomaltulose
Which plant is more efficient?
Which is more sustainable?
Benefits from BiofactoriesBenefits from Biofactories
Consumer benefitsConsumer benefits» Naturally occurring, widely approved Naturally occurring, widely approved
» Non-cariogenic, ‘slow-release’ sugarNon-cariogenic, ‘slow-release’ sugar
» Not fermented by most microbesNot fermented by most microbes
» Non-hygroscopic, acid stableNon-hygroscopic, acid stable
Industry compatibilityIndustry compatibility» Existing infrastructureExisting infrastructure
Downstream potentialDownstream potential» Growing market Growing market (potential for value-added blends)(potential for value-added blends)
» Precursor for ‘isomalt’Precursor for ‘isomalt’ low calorie sweetenerlow calorie sweetener
» Potential precursor for petrochemical replacements Potential precursor for petrochemical replacements
Why Isomaltulose?Why Isomaltulose?
Express an introduced SI gene:Express an introduced SI gene:
promoter determines which cells express promoter determines which cells express NTPP directs the protein to the vacuoleNTPP directs the protein to the vacuole SI enzyme converts some sucrose to isomaltulose (IM)SI enzyme converts some sucrose to isomaltulose (IM)
Progress with IsomaltuloseProgress with Isomaltulose
SI geneNTPPPromoter
Sucrose ( 1-2) Isomaltulose ( 1-6)
SI
→
Storage parenchyma
cytosol
vacuole
apoplasm
Engineered sugarcane
Some Plants Accumulate IMSome Plants Accumulate IMWithout corresponding decrease in sucrose
Q117 controlSI-expressing Q117 transformants
N3.2 N3.2H
Results consistent over generations in containment glasshouseResults consistent over generations in containment glasshouse
High Total Sugar ContentHigh Total Sugar ContentWithout corresponding decrease in fibre
Q117
N3.2
N3.2H
Water
Sugar
Fibre
Some lines accumulate isomaltuloseSome lines accumulate isomaltulose Some lines show enhanced sucrose accumulationSome lines show enhanced sucrose accumulation
implications for biomaterials & bioenergyimplications for biomaterials & bioenergy stability and field performance are key considerationsstability and field performance are key considerations
Transgenic sugarcane expressing Transgenic sugarcane expressing SISIS
ug
ar C
on
ten
t (m
M s
uc
rose
eq
uiv
alen
ts i
n j
uic
e)
Controls SI transformed lines
High Total Sugar ContentHigh Total Sugar Content
Results in containment glasshouse tests
High Total Sugar ContentHigh Total Sugar ContentHow does it work?How does it work?
cytosol
storage vacuole
apoplasm
Vascular bundles
Storage parenchyma
Sun light
Photosynthesis in source tissues:primarily leaf parenchyma
Sucrose storage in sink tissues:
primarily mature stem parenchyma
Sucrose transport:phloem, symplasticand apoplasticpaths
CO2
Futile cycle and mobilization
IM
Sun lightSun light
Photosynthesis in source tissues:primarily leaf parenchyma
Sucrose storage in sink tissues:
primarily mature stem parenchyma
Sucrose transport:phloem, symplasticand apoplasticpaths
CO2
Futile cycle and mobilization
IM
Enhanced PhotosynthesisEnhanced Photosynthesisand sucrose transport
Electron transport CO2 assimilationSucrose transport into leaf plasma membrane vesicles
Increased Sink Strength?Increased Sink Strength?Cell wall invertase in storage parenchyma
Q117 N3.2 N3.2H
Lines
Central parenchyma-rich zone
Peripheral vascular-rich zone
Dissected tissues from central zone
Reference: Wu L, Birch RG (2007) Doubled sugar content in sugarcane plants modified to produce a Reference: Wu L, Birch RG (2007) Doubled sugar content in sugarcane plants modified to produce a sucrose isomer. sucrose isomer. Plant Biotechnology JournalPlant Biotechnology Journal 5, 109-117. 5, 109-117.
3 5 7 10 15 20 25 30 35 40 45 50 55 580
200
400
600
800
1000
Q117 Parent
Sucrose
Glucose + Fructose
Isomaltulose (IM)
Internode # from TVD
To
tal s
ug
ar c
on
cen
trat
ion
(m
M)
3 5 7 10 15 20 25 30 35 40 45 50 55 580
200
400
600
800
1000
IM-producing transformant
Internode # from TVD
Working to establishWorking to establish optimal implementationoptimal implementation
stability & efficacy in the fieldstability & efficacy in the field
genotype specificitygenotype specificity
applicability across speciesapplicability across species
PotentialsPotentials enhanced sugar accumulationenhanced sugar accumulation
enhanced food productionenhanced food production
enhanced biofuel productionenhanced biofuel production
enhanced understanding of enhanced understanding of source-sink relationshipssource-sink relationships
SugarBooster TechnologySugarBooster TechnologyContinuing effort with government & industry Continuing effort with government & industry
TM
Progress with other TargetsProgress with other TargetsSugar derivatives – e.g. Sorbitol
World marketPrice
~ 900,000 tons / yr~ $1,500 / ton
Technology Conversion of cytosolic G-6-P into sorbitol by apple sorbitol-6-P dehydrogenase
Yields Leaf: 12% DWStem: 1% DW
Challenges Toxicity – leaf necrosis and stuntingSubstrate-limited yield?
Key groups BSES / CRCSIIB, Australia
Reference Chong BF, Bonnett GD, Glassop D, O'Shea MG, Brumbley SM (2007) Growth and metabolism in sugarcane are altered by the creation of a new hexose-phosphate sink. Plant Biotechnology Journal 5, 240-253.
Progress with other TargetsProgress with other TargetsPolymers – e.g. PolyhydroxyAlkanoates
World marketPrice
~ 100,000 tons / yr~ $1,000 / ton
Technology Conversion of plastid acetyl-coA via 3 bacterial genes into pHB
Yields Leaf: 2% DWStem: <0.01% DW
Challenges Toxicity in other plants from higher yields Substrate-limited yield in other plantsExtraction costs (needs >15% DW)
Key groups BSES / CRCSIIB, Australia
Reference Petrasovits LA, Purnell MP, Nielsen LK, Brumbley SM (2007) Production of polyhydroxybutyrate in sugarcane. Plant Biotechnology Journal 5, 162-172.
Progress with other TargetsProgress with other TargetsAromatics – e.g. paraHydroxyBenzoate
World marketPrice
~ 10,000 tons / yr~ $2,400 / ton
Technology Conversion of cytosolic phenylpropanoid into pHBA by bacterial HCHL enzyme
Yields Leaf: ~2% DWStem: ~1% DW
Challenges Toxicity in other plants from higher yields Substrate-limited yield in other plants
Key groups BSES / CRCSIIB, Australia
Reference McQualter RB, Chong BF, Meyer K, Van Dyk DE, O'Shea MG, Walton NJ, Viitanen PV, Brumbley SM (2005) Initial evaluation of sugarcane as a production platform for p-hydroxybenzoic acid. Plant Biotechnology Journal 3, 29-41.
Progress with other TargetsProgress with other TargetsProteins – e.g. Cytokine GM-CSF
Collagen?World marketPrice
~ 200 g @ $1 million / g to 100,000 tons / yr @ $2,000 / ton?
Technology ER-targeted accumulation of constitutively expressed protein
Yields 0.02 – 1% soluble protein (~ 1 g / ton cane?)
Challenges Low yieldEconomic extraction
Key groups USDA – TAMUHSPA- HARC
Reference Wang ML, Goldstein C, Su W, Moore PH, Albert HH (2005) Production of biologically active GM-CSF in sugarcane: a secure biofactory. Transgenic Research 14, 167-178.
Sugarcane BiofactorySugarcane Biofactory
PlatformsPlatforms reliable transgene expression patterns reliable transgene expression patterns
Priority targetsPriority targets technical feasibilitytechnical feasibility protected competitive advantageprotected competitive advantage market appealmarket appeal
Partnerships -Partnerships - major industry major industry market developmentmarket development competitive investment levelcompetitive investment level
» for delivery & sustainable advantagefor delivery & sustainable advantage
Policy - Policy - government leadershipgovernment leadership» corrects historical anomaliescorrects historical anomalies
» provides initial marketsprovides initial markets
» permits industry investmentpermits industry investment
Needs to capture value in AustraliaNeeds to capture value in Australia
SucroseSucrose
Power & FuelPower & Fuel
Value-added sugars
Value-added sugars
Bio-polymers
Bio-polymers
IPIP
The Sugarcane Renewable Biomaterials Industry
SRDCSRDC CSRCSR ARC – UQARC – UQ
Visionary support and continuing collaboration Visionary support and continuing collaboration
Collaborations inCollaborations inplatform scienceplatform science
Luguang WuLuguang Wu Steve MudgeSteve Mudge Mick GrahamMick Graham Dennis HamerliDennis Hamerli Lianhui ZhangLianhui Zhang
ThanksThanks
BSESBSES CSIROCSIRO
Leading the teams that do all the hard workLeading the teams that do all the hard work Terry MorganTerry Morgan Doug ChamberlainDoug Chamberlain Jirri StillerJirri Stiller Annathurai Annathurai
GnanasambandamGnanasambandam
AusIndustry REDIAusIndustry REDI