Spesifikasi Bioproduk

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Transgenic Animal

Desired DNA is
added to an egg
cell.
Selective Breeding
• Breed only those plants
or animals with
desirable traits

• People have been using


selective breeding for
1000’s of years with
farm crops and
domesticated animals.
Impact of recombinant DNA technology on the
production of proteins

• Overcomes the problem of source availability : allows the


manufacture of any protein in whatever quantity it is required

• Overcomes the problem of product safety:


Transmission of blood-born pathogens such as hepatitis B, C, and
HIV
via infected blood products

• Provides an alternative to direct extraction from inappropriate or


dangerous source materials : The fertility hormones( FSH and hCG)
from the urine of pregnant women; Urokinase from urines

• Facilitates the generation of newly designed proteins:


Therapeutic proteins or enzymes with desired property
Biotechnology
 What is Biotechnology?
- Purposeful design and modification/assembly of bio-oriented
materials (e.g., proteins/enzymes, microorganisms, plant/animal
cells, tissues, stem cells etc..) and unit processes to benefit humans
or make a profit.

- Use and applications of biological system (cells, tissues etc..) or


biomolecules (enzymes/proteins, antibodies, DNA/RNA) and key
technologies to produce valuable products at commercial scale and
to treat diseases:
 Cost-effectiveness  economically feasible

 Basic Biology / Medical sciences


- To discover and understand the underlying mechanisms of behaviors
and disorders in living organisms
 Definition of Biotechnology based on the use of techniques and
methods

 Traditional Biotechnology (Before 1970)


- Broad definition of Biotech : Using a biological system to make products
- Food processing : Fermented foods, Brewery, Dairy products, etc.
The process of brewing beer : conversion of starch to sugar followed
by addition of specific yeast
- Agriculture : Modifications of living plants for improved yield of
food crops via artificial selection and hybridization
ex) Crops with reduced vulnerability to frost, draught, and the cold

 Simple process
- Direct use of or isolation from original biological sources
- Fermentation
 Modern Biotechnology (After 1970s)

• Use of recombinant DNA technology since 1973


- Stanley Cohen and Herbert Boyer perfect techniques to cut and paste DNA
(using restriction enzymes and ligases) and reproduce the new DNA in bacteria.
• Combined use of different disciplines:
-Biology-based knowledge : Cell biology, genetics, molecular biology, etc

-Knowledge linked with practical applications :Biochemical Eng,


Bioinformatics, computational design, etc.
• Use of genetically altered organisms
- Enabling the production of existing medicines or products easily and
cheaply (ex: insulin ; price between recombinant human insulin and synthetic
pig insulin ?)

• Traditional Biotechnology industries : adopts new approaches and modern


techniques to improve the quality and productivity of their products
Benefits of Recombinant Bacteria

1. Bacteria can make human insulin or human


growth hormone.

2. Bacteria can be engineered to “eat” oil


spills.
General Bioprocess

Feedstock Bioprocessing Product

GAS Cell
Cellculture
culture
GAS
Recovery PRODUCT
PRODUCT
LIQUID Biocatalyst Bioreactor
LIQUID product
LINES
LINES
SOLID Enzymatic
Enzymatic
SOLID

Feedstock Bioprocessing Product


 Gas  Immobilized Enzymes  Separation  Pharmaceuticals
− Syn. Gas − Ambient to Extreme − In situ  Fine chemicals
− CO2  Fermentation −  Specialty Chemicals
− Organic vapor − Immobilized Secondary  Feedstock
 Liquid − Free cell  Media  Bulk chemicals
− Organic − Ambient to Extreme − Gaseous
− Sugar solution  Bioreactors − Aqueous
 Solid − Continuous Systems − Organic
− Biomass − Membrane
− Consumer − Batch or Fed-batch
Waste
The DNA of plants and animals can also be altered.

PLANTS

1. disease-resistant and
insect-resistant crops

2. Hardier fruit

3. 70-75% of food in
supermarket is genetically
modified.
Genetically modified organisms are
called transgenic organisms.

TRANSGENIC ANIMALS

1. Mice – used to study human


immune system

2. Chickens – more resistant to


infections

3. Cows – increase milk supply and


leaner meat

4. Goats, sheep and pigs – produce


human proteins in their milk
Transgenic Goat

.
This goat contains a human gene
that codes for a blood clotting
agent. The blood clotting agent
Human DNA in
can be harvested in the goat’s
a Goat Cell
milk.
Alur Penelitian Seed Vaksin :
Produksi Vaksin Flu Burung (Skala Industri)

Uji Klinis Fase I,II,III (RS Pendidikan)


Pilot Produksi Uji Klinis Fase I, II, & III

Interim Risk Assesment


Quality Assurance Quality Control
Pilot Formulasi & optimasi

Uji Pada Hewan


(Animal BSL-3)

Pilot Kultur Virus SEED VAKSIN Pilot Sistem Purifikasi


Whole / REVERSE
GENETIC / DNA
Specifitas Reseptor Virus H5N1 Adaptif Karakterisasi Imunologik

VIRUS H5N1 (MANUSIA); SURVEILANS; MODIFIKASI


METODA; KARAKTERISASI GENETIK & EKSPRESI FENOTIP
Ilustrasi Reassortment Vs Reverse Genetic
Reverse genetics

(Kawaoka and Webster, PNAS, 1988)


ALUR OPERASIONAL PENYEDIAAN SEED VAKSIN REVERSE GENETIC

MDCK cells

A/TDC/05 Propagasi
(WT)
TAB
Sel 293 T

A/TDC/05
(RG)

SEL Vero
Virus PR-8
Virus Seed Vaksin Flu Burung
RG
Eksternal gen : A/Indonesia/Unair/2005
HA : RG (avirulent type): NA (Riset I)
Internal gen:PR8 (ATCC No. VR-95 & VR-1469)

HA HA
NA NA
PLASMID REVERSE GENETIC

pHH-21 : - HA & NA (WT)


- PB-1; PB1; PA;
NP;
NS; M (PR-8)

pCAGGS: PB-1;PB-2;PA;NP
(PR-8)
Mutagenesis Cleavage site

RESRRKKR

RETR
Uji Patogenisitas : Telur Ayam Bertunas

A/Ind/Unair/2005 (WT) : ≤ 21 jam

A/Ind/Unair/2005 (RG) : ≥ 72 jam


Uji Patogenisitas : Ayam (IVPI)

A/Ind/Unair/2005 (WT)
Index : 2,94 (Ayam mati  hari ke 2)

A/Ind/Unair/2005 (RG)
Index : 0,00 (Ayam hidup  hari ke 10)
PROPOSAL MP3EI (2013-2015)
GARIS BESAR KERJA SAMA

Rencana Strategi
AIRC - Pusat Penelitian

Produksi Vaksin Produksi Produksi Vaksin


penyakit tanaman Anti Virus Flu Burung pada
Flu Burung Manusia

Kelapa Kakao Teh Putih Pohon


Sawit Gambung Pisang

Ganoderma VSD Multi Protein


compound Virus Flu Burung
pada Buah

Teknologi Ekstraksi Somatic


Reverse Genetic Senyawa Khusus Embryogenesis

Vaksin Vaksin Flu Burung


Tanaman dalam Pisang
Fitofarmaka
Kombinasi
Kelapa
Sawit Kakao

Kemandirian Bangsa dalam Bidang Kesehatan dan P otensi


Ekonomi Baru
Ontong pisang sebagai bahan untuk kultur dan pembuatan vaksin

Proses kultur tanaman pisang

Insersi virus flu burung

Sumber foto : PPKKI dan BPBPI


Introduction of Biotechnology
 What is Biotechnology?
- Purposeful design and modification/assembly of bio-oriented
materials (e.g., proteins/enzymes, microorganisms, plant/animal
cells, tissues, stem cells etc..) and unit processes to benefit humans
or make a profit.

- Use and applications of biological system (cells, tissues etc..) or


biomolecules (enzymes/proteins, antibodies, DNA/RNA) and key
technologies to produce valuable products at commercial scale and
to treat diseases:
 Cost-effectiveness  economically feasible

 Basic Biology / Medical sciences


- To discover and understand the underlying mechanisms of behaviors
and disorders in living organisms
 Definition of Biotechnology based on the use of techniques and
methods

 Traditional Biotechnology (Before 1970)


- Broad definition of Biotech : Using a biological system to make products
- Food processing : Fermented foods, Brewery, Dairy products, etc.
The process of brewing beer : conversion of starch to sugar followed
by addition of specific yeast
- Agriculture : Modifications of living plants for improved yield of
food crops via artificial selection and hybridization
ex) Crops with reduced vulnerability to frost, draught, and the cold

 Simple process
- Direct use of or isolation from original biological sources
- Fermentation
 Modern Biotechnology (After 1970s)

• Use of recombinant DNA technology since 1973


- Stanley Cohen and Herbert Boyer perfect techniques to cut and paste DNA
(using restriction enzymes and ligases) and reproduce the new DNA in bacteria.
• Combined use of different disciplines:
-Biology-based knowledge : Cell biology, genetics, molecular biology, etc

-Knowledge linked with practical applications :Biochemical Eng,


Bioinformatics, computational design, etc.
• Use of genetically altered organisms
- Enabling the production of existing medicines or products easily and
cheaply (ex: insulin ; price between recombinant human insulin and synthetic
pig insulin ?)

• Traditional Biotechnology industries : adopts new approaches and modern


techniques to improve the quality and productivity of their products
Recombinant E. coli

DNA microarray

Computer-generated insulin structure Gene therapy using adenovirus


Impact of recombinant DNA technology on the
production of proteins

• Overcomes the problem of source availability : allows the


manufacture of any protein in whatever quantity it is required

• Overcomes the problem of product safety:


Transmission of blood-born pathogens such as hepatitis B, C, and
HIV
via infected blood products

• Provides an alternative to direct extraction from inappropriate or


dangerous source materials : The fertility hormones( FSH and hCG)
from the urine of pregnant women; Urokinase from urines

• Facilitates the generation of newly designed proteins:


Therapeutic proteins or enzymes with desired property
 Biotechnology focuses on :
- Development of therapeutics based on underlying mechanisms of
diseases
- Development of new methods to cure diseases : Gene and cell
(stem cells) therapies
- Production of valuable products at commercial scale
Organic acids, Antibiotics, Amino acids, Proteins(enzymes), Biofuels,
Vitamins, Hormones, Alcohols, Fermented foods, Fine chemicals, etc..

- Development of tools and methodology


Expression systems, Gene synthesis/Sequencing, Purification process,
Formulation, Bioassays, Diagnosis, Delivery
 Integration of biological sciences with Engineering principles
 cost-effectiveness
- Process development/design/optimization

 Required disciplines
- Basic biology, - Mass/Energy balance, - Thermodynamics
- Physical, organic chemistry / Pharmacology
- Biochemical engineering : Extension of chemical engineering
principles to biological system  Bioreaction engineering, plane
design, process control, optimization, and separations

Biotechnology Bio-industry
Basic Biology - Pharmaceutical
Engineering principles
- Biotech. company
 Major application areas
• Health care / Diagnostics :
- Development of therapeutics: efficacy, toxicity
- Diagnosis : early detection and prevention of diseases

• Agriculture : Crop production with high yield and quality

• Bio-based process: Pollution, CO2 emission,


global warming

• Alternative energy (Bio-energy) :


- Depletion of fossil fuels
- Use of renewable sources :Corn, sugar cane, cellulose
- Cost (?)
Key technologies and fields

• Protein engineering : Design of proteins/enzymes based on structural


and mechanistic knowledge, molecular evolution, computational design
• Metabolic pathway engineering: Design of more efficient metabolic
pathways: high yield of target product, low by-product

• Computational modeling and optimization: Systems Biology,


Genome-wide analysis
• Nano-biotechnology : Use of NPs for diagnosis and imaging

• Cell culture engineering : Microorganisms and mammalian cells


- Hybridoma technology : A technology of forming hybrid cell lines
(called hybridomas) by fusing a specific antibody-producing B cell with
a myeloma (B cell cancer) cell that is selected for its ability to grow in
tissue culture
• Separation technology : Recovery and purification of a target product

• Synthetic biology : Creation of new biosystems (Cells and


biomolecules): Systematic, hierarchical design of artificial, bio-
inspired system using robust, standardized and well-characterized
building block

Branches of Biotechnology
• Blue biotechnology : Marine and aquatic
applications of biotechnology
• Green biotechnology : Agricultural applications
• Red biotechnology : Medical applications
• White biotechnology : Industrial applications
Typical examples of Bio-Products

Company Procucts
 Vitamin B-2
 Methoxy isopropyl amine (chiral intermediate)
BASF  Styrene oxide
Amino acids

Eastman Chemical / Genencor  Ascorbic acid


 Acrylamide
 Fatty acid – derived esters
Degussa  Polyglycerine ester
 Organo modified silicones and oleochemicals
 Acetic acid
 Polyunsaturated fatty acids
Celanese / Diversa  Non-digestible starch
 Polylactic acid (PLA)

Cargill  Polylactic acid (PLA) (140,000 MT/yr)


 1,3-Propanediol
DuPont / Genencor  Terephthalic acid
 Adipic acid

Chevron / Maxygen  Methanol


General Bioprocess

Feedstock Bioprocessing Product

GAS Cell
Cellculture
culture
GAS
Recovery PRODUCT
PRODUCT
LIQUID Biocatalyst Bioreactor
LIQUID product
LINES
LINES
SOLID Enzymatic
Enzymatic
SOLID

Feedstock Bioprocessing Product


 Gas  Immobilized Enzymes  Separation  Pharmaceuticals
− Syn. Gas − Ambient to Extreme − In situ  Fine chemicals
− CO2  Fermentation −  Specialty Chemicals
− Organic vapor − Immobilized Secondary  Feedstock
 Liquid − Free cell  Media  Bulk chemicals
− Organic − Ambient to Extreme − Gaseous
− Sugar solution  Bioreactors − Aqueous
 Solid − Continuous Systems − Organic
− Biomass − Membrane
− Consumer − Batch or Fed-batch
Waste
New paradigms in Biotechnology
 Advent of Bio-based Economy
 Genome and proteom wide analyses: Global analysis
 Integration of high throughput analysis system

• Massive and high speed analysis system


- Genome and proteom-wide approach : Systemic approach
- High amounts of relevant knowledge

• Genomics (Gene chips) : 1 million genes / chip


- Gene (mRNA) expression profiling in high throughput way
-Single nucleotide polymorphism (SNP)

• Proteomics (2-D gel, LC/MS, protein microarray)


- Functional genomics
- Bio-molecular interactions (Interactom)

• Development and commercialization of target products


- Bioinformatics
- Drug target identification via high throughput screening
Bio-based economy: Impact on global economy

 Shift from petroleum-based economy


- Exhaustion and soaring price of petroleum (> $ 100 /gallon)
- Environmental issue
Global warming (greenhouse gas, CO2 , emission)
Pollution

• Development of renewable source-based Bioprocess


• Replacement of chemical processes with Bio-based ones

White Biotechnology
Value chains from renewable sources
Alternative energy sources

 Production of biofuels from natural resources


• Increase in the yield and alcohol tolerance
- Redesign of pathway for the ethanol production in yeast to use
raw materials : corn starch, cellulose, soybean, sugar cane
- Elucidation of enzyme mechanisms
- Redesign of pathway to increase the yield and to reduce
by-products
- Design of critical enzymes in the pathway
• Process development : Fermentation process
• Separation and concentration

 Role of Agricultural Biotech in the production of biofuels ?


 Adverse effects due to the production of biofuels from corn ?
Key role of enzymes in Bio-based economy
Energy and Environmental issues
- Depletion of fossil fuels
- Limitation to CO2 emission (Kyoto protocol)

Petrochemical-based economy Renewable source-based economy


Chemical process Bio-based process

Enzymes

Use of enzymes in Biofuel production from renewable biomass


such as starch and cellulose  amylase, cellulase etc.
Enzymes
• Most proficient catalysts with high specificity
• Competitive and cost-effective processes

• Cleaning (Detergents) Use for biosciences


• Textiles  DNA polymerase: Thermostability, fidelity
• Starch Processing  Restriction enzymes: Specificity
• Brewing  Alkaline phosphatase
• Leather  Peroxidase
• Baking
• Pulp and Paper
• Food and Specialties Industrial use for specialty chemicals
• Animal feeds  Chiral drugs
• Cosmetics
 Chiral intermediates
 Semi-synthetic antibiotics
 Organic acids

Therapeutics
• Treatment of Gaucher’s disease
Chemical company devoting to Biotechnology : BASF

Emphasis on Bio-products mainly using Enzymes

Ecoflex®
Therapeutic proteins
 High specificity and less toxicity  high safety and efficacy
 Therapeutic proteins
- Antibodies, proteins, enzymes, peptides etc.
ex) EPO, Interferon, Insulin, Avastin, Enbrel, Remicade, Herceptin,
EPO (Erythropoietin) : Stimulating the proliferation of red blood cells
Herceptin : Mab against EGFR2(Epidermal growth factor receptor 2)
Avastin : Mab against VEGF (Vascular endothelial growth factor)
Remicade: Mab against TNF-α (Tumor necrosis factor- α)

 World market
- EPO alone : ~ $ 11 Billion per year
- $ 50 Billion (2007) $ 190 Billion (2015)
- Antibodies > 50 %
- Intensive investment in monoclonal antibodies: Biosimilar

Therapeutic proteins will form the back-born of future medicinal therapy


Structural and functional features of antibodies

CDRS 1 2
3 1
3

FR

VL VH
Blockbuster Therapeutic Antibodies
Approved Market Antibody
Product Target Indication Company
Year size(07)* Type
Non-Hodgkin's
1997 Rituxan CD20 Genentech 4,603 Chimeric
lymphoma

1998 Herceptin Her2/neu Breast cancer Genentech 4,047 Humanized

1998 Synagis RSV RSV prophylaxis MedImmune 1,100 Humanized

RA, Chron’s
1998 Remicade TNF-  J&J 5,234 Chiemric
disease

2002 Humira TNF- RA Abbott 3,064 Human

Genentech
2003 Raptiva CD11a Psoriasis 211 Humanized
Xoma
Imclone
2004 Erbitux EGFR Colorectal cancer Bristol- 1,336 Chiemric
Myers

2004 Avastin VEGF Colorectal cancer Genentech 3,335 Humanized

2006 Vectibix EGFR Colorectal cancer Amgen 170 Human

* Million $ (Data Monitor ‘Monoclonal 2008)


Drawbacks of immunoglobulin antibodies

• Complicated process for selecting cell lines and


the production using mammalian cells  very expensive
• Intellectual property barriers
• Tend to aggregate due to large size (~ 150 KDa)
• Difficult to penetrate inside the cells
• Limited binding affinity due to confined binding surface

Ideal scaffold for alternative therapeutics


High-level soluble expression in bacteria
High stability (thermodynamic, pH)
Easy design of binders with high affinity for a target

Low immunogenicity and cytotoxicity


Therapeutics based on non-antibody scaffold

New paradigm in therapeutic proteins


 Development of new therapeutics with high efficacy and
low side effect from non-antibody protein scaffold
Designer therapeutic proteins
IP issue and cost-effectiveness

GlaxoSmithKline,
Amgen Strategic alliance or merger
Bristol-Myers-Squibb,
Boehringer Ingelheim between big pharma and
Eli Lilly, biotech companies
Roche,
Avidia, Ammunex. Affibody,
Ablynex, Adnexus Therapeutics
…….
Non-antibody scaffolds

Human lipocalin
Ankyrin
Human fibronectin Z domain of Staphylococcal protein A
Therapeutic Enzymes

Sales
Disease Product Developer (US$Millions) Features
2004 2007
 Glucocerebrosidase
Ceredase® Genzyme 443 N/A
 Purified from human placenta
Gaucher’s  Produced in CHO cells
932
Cerezyme ® Genzyme 1,048  3 Exoglycosidases process
(2005)
for Terminal Mannose
Fabrazyme® Genzyme 209 397  alpha-galactosidase
Fabry’s  Mannose-6-phosphate for
Replagal TKT 57 168 Glycotargeting

MPS-1 Aldurazyme® Genzyme 12 204  alpha –L-iduronidase

Approved
Pompe Myozyme® Genzyme  alfa-glucosidase
(2006)

Treatment of Gaucher’s disease by Cerezyme costs up to $550,000 annually

Most of therapeutic enzymes : glycoproteins


Gaucher’s Disease : Lysosomal Storage Disease
Caused by a recessive mutation in a gene located on
chromosome 1, affecting both males and females.

- Found by Phillipe
Glucosyl Ceramide
Gaucher in 1882
- Biochemical basis O=C-CH2-CH2-CH2-(CH2)n-CH3
for the disease in N OH
1965 by Brady et al..
CH2-CH-CH-CH=CH-(CH2)12-CH3

Glucocerebrosidase
Glucocerebrosidase(b-glucosidase)
(b-glucosidase)

O=C-CH2-CH2-CH2-(CH2)n-CH3
N OH
OH-CH2-CH-CH-CH=CH-(CH2)12-
CH3
Autosomal recessive inheritance Glucose Ceramide
Lysosomal storage diseases (LSDs): Lysosomal
Enzymes  Lysosomes: Cellular organelles containing
acid hydrolase enzymes to break down waste
materials and cellular debris
 Cells’ garbage disposal system
• Digestive organelle in the cell
• Contains ~40 hydrolytic enzyme
(1) The ER and Golgi apparatus make a lysosome
• Acidic pH (about pH4.8) within the lysosome
(2) The lysosome fuses with a digestive vacuole is required for lysosomal enzymes to be
(3) Activated acid hydrolases digest the contents active

(LSD)
Lysosome Lysosome with
substrate
accumulation

Nucleus

Mitochondria

(Normal cell) (LSD cell)


Gaucher’s disease : Occurrence and symptoms

 1/ 40,000~60,000 (Jew 1/~500)


 Swollen vacuoles  Gaucher cells
 Accumulation in spleen, liver, kidney, brain
 Enlarged spleen and liver, liver malfunction,
neurological complications etc..

Glucocerebrosides Exocytosi
Normal cells s
Glucocerebrosidase

+
Digestive glucose ceramide Residual
vacuole vacuole
Distended abdomen

Gaucher cells Glucocerebrosides


Incomplete
digestion
Residual vacuole
accumulated

Digestive
vacuole No exocytosis
Diagnostics
• Diagnosis of disease as early as possible :
Best solution compared to treatments

• Prediction and treatment of diseases based on


individual genomes
- personalized medicine
- treatment with appropriate therapeutic agents

• Analysis / Detection of disease biomarkers:


- Invasive or non-invasive analysis
Perspectives
 Biotechnology will have the greatest impact on
humans in the future in terms of health, life-style,
and economy.
- Therapeutic proteins
- Bio-based economy : Bioprocess and Bio-Energy
- Diagnostics

 Modern Biotechnology constitutes a variety of


diverse areas and technologies, requiring
interdisciplinary collaborations.
Bio Platform

Industrial
Residual

Anaerobic
Digestor

Bio
Methane CHP Heat & Power €.14/KWh
Fertilise
Digestate
r Fertiliser €250/MT
Leachate
Process
Bio Platform
Lactic Acid
€400/MT
Silage Press
Agri Press Juice Separation Amino Acid
Residue Feed €350/MT
Industrial Press Cake Supplement
Residual
C5/C6 Sugars
Enzyme
Separation Food or €400/MT
Hydrolysis Fermentation

Separati C5 Sugar
on Fraction

Anaerobic
Digestor

Bio
Methane CHP Heat & Power €.14/KWh
Fertilise
Digestate
r Fertiliser €250/MT
Leachate
Process
Bio Platform
Lactic Acid
€400/MT
Silage Press
Agri Press Juice Separation Amino Acid
Residue Feed €350/MT
Industrial Press Cake Supplement
Residual
C5/C6 Sugars
Enzyme
Separation Food or €400/MT
Hydrolysis Fermentation

Sugars

Bio Plastics €750/MT


Anaerobic
Fermentation
Digestor Bio Ethanol €515/MT

Bio
Methane CHP Heat & Power €.14/KWh
Fertilise
Digestate
r Fertiliser €250/MT
Leachate
Process
Bio Platform
Lactic Acid
€400/MT
Silage Press
Agri Press Juice Separation Amino Acid
Residue Feed €350/MT
Industrial Press Cake Supplement
Residual
C5/C6 Sugars
Enzyme
Separation Food or €400/MT
Hydrolysis Fermentation

Sugars Organic Acids €400/MT

Phased Anaerobic Bio Plastics €750/MT


Anaerobic
Digestor
Digestor
(Short
Organic
Acid Fermentation
Chem Refined
Syn Bio Diesel €600/MT
Residence)
Bio
Methane CHP Heat & Power €.14/KWh
Fertilise
Digestate
r Fertiliser €250/MT
Leachate
Process
Thermo Chem Platform
Agri
Residue
Wastes
Energy
Crops

Reagent H2 Hydrogen Gas

Levullinic Acid €450/MT


Feedstock Acid HM Chemical
Optimisation Hydrolysis Furfural Refining

€400

Gasificatio
Bio Char Heat & Power €.14/KWh
n
Thermo Chem Platform
Agri
Residue
Wastes
Energy
Crops

Reagent H2 Hydrogen Gas

Levullinic Acid €450/MT


Feedstock Acid HM Chemical
Optimisation Hydrolysis Furfural Refining

Bio Char Levulinic Acid

Ethyl Levulinate
Esterification €600/MT
Hydro Bio Diesel

thermal
Pyrolysis
PH
Crude Stabilisation
Bio Oil Deoxygenation Bio Oil €350/MT

Gasificatio
Bio Char Heat & Power €.14/KWh
n

Developing Expertise to Optimise Value of Alternatives


Biorefining Platform
Lactic Acid
€400/MT
Silage Press
Agri Press Juice Separation Amino Acid
Residue Feed €350/MT
Industrial Press Cake Supplement
Residual
C5/C6 Sugars
Enzyme
Separation Food or €400/MT
Hydrolysis Fermentation

Sugars
Separati C5 Sugar Acids €400/MT
Organic
on Fraction
Anaerobic Bio Plastics €750/MT
Digestion
(Short Fermentation
Bio Ethanol €515/MT
Residence)

Bio
Methane CHP Heat & Power €.14/KWh
Fertilise
Digestate
r Fertiliser €250/MT
Leachate
Process

Low cost production of multiple high value products from low cost biomass =
Economic Viability
BIOLOGICAL PROCESS CATEGORIES

Microbial Animal Biological


Production Production Waste
Fermented Poultry, Closed
bio- swine, fish, system
Agri-Bio pesticides, shellfish aquaculture
APPLICATION SEGMENTS

Products fungicides,
growth
lifecycle
growth
or irrigation
pond
enhancers enhancement remediation

Remediation
Bio-Energy, Cellulosic Slaughter or byproducts
ethanol fecal waste (microbial
Renewable fermentation digestion to lipids, omega
Chemicals , algal lipids bio-gas 3,6 fatty
acids)
Corn

Source: Nicolle Rager Fuller, National Science Foundation


Supply Chain
Sun
Water

Seed
Grow Harvest
Fertilizer

Store Transport
to Bioprocessing
Bioprocessing
Enzymes Yeast
Glucose

Pretreatment Hydrolysis xylose Fermentation

Fuel Ethanol Distillation

Delivery to markets
Infrastructure
Corn Stover: 1 to 2 tons /acre

Leaves

Cobs

Stalks

Roots
Bioethanol Production

Feedstock
Preparation

Pretreatment

Hydrolysis Ethanol
of Solids Fermentation
Cellulose Pretreatment gives
Lignin enzyme accessible
substrate

Amorphous Pretreatment
Region

Crystalline
Region

Hemicellulose
Components of plant cell walls
Cellulose
Fermentable sugars obtained
from cellulose in 1819

Lignin

Hemicellulose Extractives
(need special yeast to Ash
convert to ethanol)

Chapple, 2006; Ladisch, 1979


Yeast Metabolism: pentose fermentation

Xylose Glucose
NAD(P)H
NAD(P)+
Xylitol Glucose-6-P
NAD+

NADH Xylulose Fructose-6-P

Glyceraldehyde-3-P
Xylulose-5-P NAD+
NADH
3-Phosphoglycerate
PPP Ethanol
NADH NAD+
Phosphoenolpyruvate
Ho et al

TCA Cycle Pyruvate Acetaldehyde

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