Bioprocessing opportunities for

Australia- from mega to nano

BPN Conference

Wednesday 30th September, 2009

Peter Gray

The financial support of the Atlantic Philanthropies and the Queensland Government is gratefully

acknowledged.

Two main reasons for optimism:

1.! Power of new disruptive technologies:

–! Synthetic Biology (Genetic Engineering)

–! Stem and iPS cells

2. Sustainability issues:

Synthetic Biology:

Rapidly developing field which designs and constructs new microorganisms, using bioengineering and molecular biology.

Synthetic Biology:

Made possible because we can use DNA synthesis machines to write cheaply and rapidly in DNA code, instead of getting the genes from biological sources.

Year

Ca

pa

city

Genomics Synthetic Biology

Moore’s law (transistors per chip)

DNA sequencing

DNA synthesis

Electronics

Biology

(bp per person per day)

R. Kelly – QB3

Systems & synthetic biotechnology

Manipulate

Mine

Model

Measure

‘Omics’ to characterise host cell Genomics, proteomics, metabolomics

‘In-silico’ redesign of MO

Genetic construction Of improved MO

Testing productivity, yield New products and

microbial processes

Foundational Technologies

Applications of Synthetic Biology

Bioenergy

Therapeutics

Advanced Textiles

Drug Production

R. Kelly – QB3

Drug Production

Keasling Lab, UC-Berkeley

•! 1-3 million people die of malaria every year

•! 300-500 million people infected

•! Artemisinin from Artimesia annua

Current cost of API $1.00 / g

Cost with new process $0.10 / g

30 grams per liter in engineered yeast!

PMK MPD MK idi ispA

HMGS atoB tHMGR

AMO ADS CPR

Genes from plants, fungi, and bacteria

•! Classical biotechnology

–! Random mutations

–! Process optimisation

–! Fixed product range

Synthetic Biology:

•! Genetic engineering

–! New products

•! Enzyme engineering

–! Improved kinetics

–! New products

•! Metabolic engineering

–! Pathway redesign

–! Control redesign

PDO

! !

!

!

!

!

L. Nielsen- AIBN

Scale of Bioprocessing Opportunities:

Mega Liquid fuels

Biocommodities- chemical building blocks

Biologics

Stem, iPS cells, growth and controlled differentiation-

patterning of surfaces.

Nano

2008 World Fuel Ethanol Production

(Millions of Gallons)

Queensland Bio Jet –A1 initiative

Why:

•! Jet fuels are 5-8% of the world’s transportation fuels- 15% in Australia.

•! Aviation depends on liquid fuels with high energy- ground transportation has electric or hydrogen option.

•! Replacement of petroleum derived aviation fuels with bio-derived fuels would mitigate greenhouse gas emissions and provide security of supply.

•! Industry policy- IATA’s goal is to have 10% bio-derived jet fuels by 2017.

Queensland Bio Jet –A1 initiative (II)

How:

Three Potential Biological Feedstocks for

conversion to Jet Fuel:

•! Alkane fermentations from sucrose.

•! Algal oil production.

•! Oilseed plants.

Queensland Bio Jet –A1 initiative (III)

How:

Two key questions for each bio-derived feedstock:

1.! Composition and suitability of feedstock

2.! Cost of production of feedstock.

Initially economic process modelling and life cycle analysis will be used to determine production cost for each feedstock based on current technology – this information will then be used to direct research into economic bottlenecks.

Queensland Bio Jet –A1 initiative (II)

Who:

•! University of Queensland: –! AIBN – Synthetic Biology; economic process modelling;

bioprocess development.

–! IMB – Algal systems.

•! James Cook University- Townsville:

–! Algal technology – pilot plant facilities

•! Queensland Department of Primary Industries & Fisheries.

–! Agronomy and processing of oil-seed plants.

Hutt Lagoon; WA

Whyalla; SA

A little calculation:

•! At a productivity of 30 g.m-2.d-1 and with a lipid content of 40% = lipid productivity of 12 g lipid.m-2.d-1

•! Therefore for 1 L lipid/oil per day need 83.3 m2 of pond

•! Now to produce 1 barrel (158.987 L) of oil per day need 1.325 hectares

( M. Borowitzka- Murdoch University, WA)

A little more:

•! Oil consumption

~!Australia 796,500 barrels.d-1

~!North America 19.6 x 106 barrels.d-1

•! Therefore, to produce 1% of Australia’s daily demand would need

~!10,554 ha ( = 105.54 km2)

( M. Borowitzka- Murdoch University, WA)

Not quite so mega:

‘ Industrial or white biotechnology.’’

Using synthetic biology to construct specialised micro-organisms which will to produce the chemical building blocks and specialty chemicals of the future.

We are studying microbes as "programmable"

manufacturing factories to make chemicals,

monomers and polymers from different nutrient feedstocks. Current feedstocks for these materials

Are petrochemicals from oil. We are programming

microbes to make very sophisticated polymer

building blocks and molecules out of simple,

renewable feedstocks, like glucose and methane. Chad Holliday, Chairman & CEO – DuPont, Boston Chief Executive Club, Sept 99

Synthetic Biology:

Industrial or White Biotechnology

Australian Integrated Biorefinery Concept

NSW: 3 Mt

Northern: 7 Mt

QUEENSLAND: 32+ Mt cane

Herbert/Burdekin: 12 Mt

Central: 9 Mt

Southern: 4 Mt

AUSTRALIA: 35+ Mt cane 4.5 Mt raw sugar 3500 Cane Growers 24 Sugar Mills 7 Bulk-storage export ports ______________________ TOTAL VALUE OF PRODUCTION A$1.2 billion Second largest export crop

The Sugar Industry

Australian Integrated Biorefinery Complex: The Dimensions

Scale of Bioprocessing Opportunities:

Mega Liquid fuels

Biocommodities- chemical building blocks

Biologics

Stem, iPS cells, growth and controlled differentiation-

patterning of surfaces.

Nano

Biotechnology in Healthcare- Red Biotechnology:

Global Pharmaceuticals and Biologics Sales

•! Biologics – US $63 billion 2006

•! Pharmaceuticals - $US 650 billion 2006

•! Biologics – growth 20%

•! Pharmaceuticals – growth 7%

Ernst and Young, Beyond Boarders, Global Biotechnology Report 2006

Monoclonal Antibodies

•! Ability to treat a range of diseases including cancer, inflammation and infectious disease

•! Account for approximately

•! 30-50% of new biologics in development.

•! Source: Ernst and Young

•! Approvals include only new molecular entities and include label approvals, new formulations and combinations.

•! Certain drugs partnered between biotech and big pharma companies are counted in both groups.

•! Big pharma is defined as the 15 largest global pharmaceutical companies by market cap.

High-Throughput Cell Line Development

!"#$%&'()'*+

,--.+

/0+,.#%12*%3+

Select cells

eGFP (mAb HC)

eY

FP

(m

Ab

LC

)

BD FACS AriaII

~ 70,000 cells/sec

ClonepixFL

~140,000 cells/hour

Expansion and Production

96-well plate culture. Screen for growth and productivity

Cloneselect Imager

Biopharmaceuticals Australia Pty Ltd (BPA)

Australia‘s GMP scale-up facility

•! Footprint of 65 metre by 30 metres (1,950 m2)

•! Two working floors

–! Total area approx 4000m2

•! Two upstream fermentation suits –! One 2000L microbial

bioreactor

–! One 2000L mammalian cell bioreactor

•! Two downstream purification suites

•! Designed to comply with the most stringent cGMP systems –! Australian TGA

–! EMEA

–! US FDA

–! ICH

Design phase drawing – Level 1, BPA facility

Science precincts – future planning

•! BPA and TRI (Translational Research Institute)

•! Part of strategic vision for Qld bioscience – “109 central” concept

BPA LOCATED ADJACENT TRI

Scale of Bioprocessing Opportunities:

Mega Liquid fuels

Biocommodities- chemical building blocks

Biologics

Stem, iPS cells, growth and controlled differentiation-

patterning of surfaces.

Nano

Pluripotency

HES cells

Pluripotent maintenance and propagation of hES cells

(trophectoderm)

Theme Objectives

•! Defined, scalable expansion of pluripotent hES cell

Novel Surfaces Cell Immobilisation

Recombinant Growth Factors

Pluripotency Reporter Lines

Human ES cells

Controlled Bioreactors

Spin EB

3D Beads and encapsulation

2D ! ! ! ! ! ! !3D

hESC Tailored

Microbeads

hESC Tailored

Culture-ware

Scalable hESC and EB

bioprocesses

Commercial Outcomes:

Lineage specific hESC

reporters

Nishikawa et al. Nature Reviews Molecular Cell Biology 9, 725-729 (2008)

Oct3/4, Sox2, Klf4 and c-Myc (Yamanaka)

Oct3/4, Sox2, Nanog, and LIN28 (Thomson)

Generating iPS Cells

The promise of iPS cells

E. Wolvetang-AIBN

Bris1-6 HiPS, 6 lines from human control fibroblast lines generated at AIBN

Brightfield DAPI Oct4

dish Colony

E. Wolvetang-AIBN