1. BIOPRODUCTS & BIOMATERIALS
Prof. Giovanni Sannia

2. Evolution

3. Industrial/White Biotechnology uses enzymes and/or micro-organisms in tailored processes/bioreactors to produce:
Conventional chemicals and materials through more efficient and sustainable ways
Compounds not obtainable through chemicals routes (I & II metabolites, chiral compounds, etc..)
New or conventional fine- and specialty-chemicals, biofuels and biomaterials from biomass, agroindustry by-products, wastes, etc
Industrial/White Biotechnology can ensure reduction of both energy and H2O consumption, waste and CO2 generation as well as the dependency of current industry from expensive and pollutting petrochemicals feedstock.
Thus, Industrial/White Biotechnology can markedly contribute to increase the sustainability & competitiveness of the current chemical, pharmaceutical, textile and energy industry.

4. Environmentally Sound
What are the benefits of industrial Biotechnology?
Drivers
Cost reduction
Novel products
Hurdles
Regulation
Further technology development
Feedstock prices
Investments
Profit
Economically Viable
Drivers
Less energy
Less waste
Hurdles
Further technology development
Waste management
Drivers
Knowledge-based quality jobs
Responsibility
Hurdles
Unawareness
Acceptance
SUSTAINABILITY
People
Socially Responsible
Planet
Environmentally Sound
The Triple-P Bottom Line

5. RESOURCES PROCESS PRODUCTS BIOPRODUCTS BIOENERGIES Bioplastics
Cash bags in maize starch, disposable
forks, spoons and knives made
of beetroot, food recipients…
Biolubricants
Chain oil for chainsaw, engine
lubricants, hydraulic fluids…
Vegetal ink
Biomaterials
Insulation materials, gardening
pots and miscanthus…
Chemical products
Vegetal solvents, paints, additives…
Biodetergent
Washing-up liquid, shampoo,
washing powder, floors and surfaces
cleaning detergent…
BIOPRODUCTS
BIOENERGIES
Green heat
Wood pellet\s stoves and central heating
systems, wood chips central heating
systems, heat produced by biomethanation
of breeding effluents, cereals stoves and
central heating systems…
Biofuel
Biodiesel from rapeseed, pure plant oil,
bioethanol from wheat or beetroot
Green electricity
Electricity produced from wood,
electricity produced from biomethanation
of breeding effluents and other matters…
Starch, colza, cereals, maize, forestry wastes, co-products of wood industry, beetroots, potatoes, breeding effluents, flax, hemp, miscanthus, other renewable materials…
White biotechnologies (micro-organism use in the production process),
Vegetal chemistry (esterification…),
Mechanic treatments (crushing, grinding up …),
Combustion (in an engine or a heating device),
Biomethanation (methane production from breeding effluents and other matters),
Cogeneration (combined heat and electricity production),
Gasification, pyrolysis…

6. Bioproducts Terminology
Bioprocess
Technology
Renewable
Bioresource
Feedstock
Plants
crops
trees
algae
Animals, fish
Microorganisms
Organic residues
municipal
industrial
agricultural
forestry
aquaculture
Industrial
Bioproducts
Bioenergy and Biofuels
Manufactured products:
biochemicals
biosolvents
bioplastics
‘smart’ biomaterials
biolubricants
biosurfactants
bioadhesives
biocatalysts
biosensors
Biocatalysis (Enzymes)
Fermentation (Microorganisms)
Physico – Chemical Process Technology
Extraction
Pyrolysis
Gasification

7. Biomass applications
Green: plant biotechnology
bring together industrial (bioproducts and biomolecules) or energy (bioenergy) applications
use renewable resources from vegetal or animal origin
reduce greenhouse gas emissions
Products issued from biomass are generally less toxic and more biodegradable than equivalent products issued from petrol
have technology qualities expected by the industry
use material and human local resources, contributing to the industrial development and to the region rural activity.
limit the waste quantity to handle by the straight utilisation and by the products good biodegradability.

8. Glossary
Bioproducts is a catchall term for products manufactured using energy, chemicals or processes derived from biological materials, or biomass.
Bioenergy technologies use renewable biomass resources to produce an array of energy related products including electricity, liquid, solid, and gaseous fuels, heat, chemicals, and other.
Biomass is the total mass of matter generated by the growth of living organisms, including plants, animals, and microorganisms. 100 to 200 billion tons of new biomass are produced each year. Most biomass now simply returns to the ecosystem through natural processes of decay, without being exploited for practical use.
Biomaterial is "any substance (other than drugs) or combination of substances synthetic or natural in origin, which can be used for any period of time, as a whole or as a part of a system which treats, augments, or replaces any tissue, organ, or function of the body".
Biocompatibility is the ability of a material to perform with an appropriate host response in a specific application.
Bioeconomy: a Revolution in getting industrial products of Commercial Value (Bioproducts) from biorenewable resources.

9. Primary drivers for the bioproducts sector
Value-added Markets
Energy issues (cost and security)
Climate change
Social development and job creation
Issues surrounding the disposal of municipal, agricultural and industrial waste
The accelerating development of enabling technology
National Security
Greneer environment
Reduced dependency on petroleum
PURE ECONOMICS!

10. Some of the disciplines required…..
Evolution & screening
Advanced analysis
Industrial fermentation
Biochemistry/enzymology
Bioprocess development
Down stream processing
Chemistry
Bioinformatics
Proteomics
Metabolic engineering
Microbiology
Molecular biology

11. BioMaterial Science and Engineering
Novel biomaterials, novel bioproduct design,
Structure-functional properties, modifications, bioconversions, reactions, catalysts, etc.
Property measurements, performance evaluations,
Processing design, control, scale-up, etc.
Plant/Microorganism/enzyme Science
Genomics/breading, DNA/gene analysis, genetic engineering, protein cloning, bioinformatics,etc.
Chemistry/Biochemistry
Biopolymer structures, Biopolymer reaction pathways, analytical analysis, etc
Agriculture Economics
Economic analysis, life cycle analysis
Integrated Research Team

12. Renewable Bioproducts Vision
Achieve 10% of basic chemical building blocks from plant-derived sources, with concepts in place to achieve 50% by 2050
Vision Goals:
Vision goals strive toward a viable industry platform for renewable bioproducts by 2020.
Industry views 10% of basic chemical building blocks as a very ambitious goal -- this represents a 5-fold increase over the current situation.
Plant-derived resources are expected to supplement, not replace petroleum-based resources. It is hoped they will capture a percentage of the increasing market demand for consumer products.

13. The thousands of different industrial bioproducts produced today can be categorized into four major areas:
Classes of Bioproducts
Sugar and starch bioproducts derived through fermentation and thermochemical processes include alcohols, acids, starch, xanthum gum, and other products derived from biomass sugars. Primary feedstocks include sugarcane, sugarbeets, corn, wheat, rice, potatoes, barley, sorghum grain, and wood.
Oil- and lipid- based bioproducts include fatty acids, oils, alkyd resins, glycerine, and a variety of vegetable oils derived from soybeans, rapeseed, or other oilseeds.
Gum and wood chemicals include tall oil, alkyd resins, rosins, pitch, fatty acids, turpentine, and other chemicals derived from trees.
Cellulose derivatives, fibers and plastics include products derived from cellulose, including cellulose acetate (cellophane) and triacetate, cellulose nitrate, alkali cellulose, and regenerated cellulose. The primary sources of cellulose are bleached wood pulp and cotton linters.
Industrial enzymes are used as biocatalysts for a variety of biochemical reactions in the production of starch and sugar, alcohols, and oils. They are also used in laundry detergents, tanning of leathers and textile sizing.

14. Life cicle of bioproducts CO2 balance The cycle of CO2 is closed!
Source: A.J. Ragauskas et al., Science 311, 484 (2006);
Source: A. K: Mohanty et al., Natural Fibers, Biopolymer and Biocomposites, CRC Press (2006)
CO2 balance The cycle of CO2 is closed!

15. 2010 consumption “4 million ton”
Emergence of “Green Plastics”
2010 consumption “4 million ton”
Source: Biopolymers from crops: Proceedings of the Australian Agronomy Conference
At present Europe share 40% of global bioplastics consumption
Norocon Innovation Consulting Kaeb, Denmark. The Bioplastics Industry in Europe

16. 1,31,3--propanediol (PDO) Dupont Sorona Polyhydroxy alkonates/PHAs
Bio-plastic
Bioplastics Innovation and Commercialization Drive
Biotechnologically produced high volume commercial or potentially commercial polymers:
Polylactide (PLA)
1,31,3--propanediol (PDO) Dupont Sorona
Polyhydroxy alkonates/PHAs
Thermoplastics starch
Soy Polyol-PU/Soy resin

17. Polylactide (PLA)
PLA
In Bioproducts, the accomplishments of BioInitative participants include:
The OIT AG IOF helped fund the development of a cost effective process for converting corn into plastic. Cargill Dow is expected to break ground on a plant capable of producing 300 million pounds of PLA in November 2001.
Other Products Accomplishments
Development of a new separation technology to remove organic products from fermentation.
Production of Chemicals from Hog Manure
Production of Purified cellulose for the manufacture of Rayon, acetate fibers, and other thermoplastics.

18. Polylactide (PLA)
PLA in packaging
Corn-based plastic polylactic acid (PLA) is used in several kinds of packaging, including these beverage containers from NatureWorks.

19. Polylactide (PLA)
Applications of PLA include (a) clear thermoformed articles for disposable food service items and other containers, and (b) fibers for bedding products, clothing, diapers, wipes, carpets, sheets and towels, and wall coverings.
Starch-PLA eating utensils (not yet commercialized).
The utensils on the left are made of 55% cornstarch and 45% poly(lactic acid). They are biodegradable and compostable.
Non-degradable polystyrene utensils are shown on the right.
PLA diapers and baby-wipe

20. A biodegradable thermoplastic
Polylactide (PLA)
PLA in Automotive
Poly Lactic Acid in Automotive Existing Technology: Limited Scope
Corn
A biodegradable thermoplastic
Functional properties match petroleum plastics
High Price
Poor durability
Toyota Tire Cover
APPLICATIONS: Tire Cover, Carpet, Antenna
Cargill-USA, - Apack Germany/Fortum Oyj.Finland, Mitsui Chemicals-Japan, Brimingham Polymers USA/Phusi France

21. Polyhydroxy alkonates/PHAs
PHAs in Automotive
Corn & Diverse Grass
Outlook - More Promising than PLA
Exceeds PP Properties
Price Claimed < $1.00/lb
Projected Production 2008
No Commercial Availability
Small Player
(R&D + Investment Risk)
Metabolix to ADM, (USA); Procter & Gamble Tech to Kaneka Corporation (Japan), Biomatera Inc., Quebec

22. Polyhydroxy alkonates/PHAs
PHAs in packaging
Under specific conditions, bacteria produces PHA polymer inside cell (fermentation process)
PHA is then extracted from the bacteria and cleaned (purification-extraction process)
The result is a PHA resin which can be transformed into various products currently manufactured using petroleum-based plastic
Petroleum-based plastic takes centuries to degrade and sometimes never degrades
PHA biodegrades within 6 to 12 months when buried in soil, such as in landfills, and in less than 12 weeks in composting conditions
Biomatera Inc., Quebec

23. diverse material properties depending on polymer composition
Polyhydroxy alkonates/PHAs
General structure of polyhydroxyalkanoates; R1/R2= alkyl groups C1-C13; x=1-4; n=
The wide variety of monomers yields PHAs with
diverse material properties depending on polymer composition
and expands the range of PHA applications to various fields
nCmon≤5, short chain length (scl-PHA); nCmon≥ 66, medium chain length (mcl-PHA)
PHAs Vs Polypropylene
Properties
scl-PHAs
mcl-PHAs
PP (crystal)
Crystallinity (%)
40 / 80
20 / 40 70
Melting point (°C)
53 / 80
30 / 80
176
Density (g/cm-3)
1.25
1.05
0.91
Tensile strenght (Mpa)
43 / 104 20 34
Glass transition temperature (°C)
-148 / 4
-40 / 150
-10
Extension to break (%)
6 / 1000
300 / 450
400
UV light resistant
Good
Poor
Solvent resistant
Biodegradability
None
UV light resistant
Good
Poor
Solvent resistant
Biodegradability
None

24. Polyhydroxy alkonates/PHAs
PHAs composition is influenced by the type of supplied Carbon source:
Sugars (unrelated)
Fatty acids (related)
according to the biosynthetic pathways shown in figure
Poly 3-hydroxybutyrate
Poly 3-hydroxyalkanoates

25. PHA synthases subunit/ Transcriptional regulator?
Polyhydroxy alkonates/PHAs
Recipient E.coli
Isolation of phaRBC biosynthetic operon
PhaR
PhaB
PhaC
ketoacyl-CoA reductase
PHA synthases subunit
PHA synthases subunit/
Transcriptional regulator?
Construction of recombinant E.coli biopolymer producing systems
Case study

26. Additional carbon source
Polyhydroxy alkonates/PHAs
Strain
Growth medium
Additional carbon source
CDW (mg/L)
PHA (%)
Composition (%)
3HB
3HHx LB
0.2% Octanoate
380.0
0.3
0.2
99.8 MM
156.2
9.9
0.05
99.95
0.1% Octanoate
128.0
7.0
0.1
99.9
0.1% Decanoate
185.7
5.7
0.05% Decanoate
334.9
4.9
0.05% Dodecanoate
344.7
6.65
6% Corn oil
553.6
0.9
0.6
99.4
6% Coconut oil
365.5
0.4
0.5
99.5
Results provide a “proof of concept” of the potential exploitation of a recombinant system for valorisation of waste oils into valuable products, such as a near P(3HHx) homopolymer
Case study

27. Additional Carbon Source Additional Carbon Source
Polyhydroxy alkonates/PHAs
Sustainable mcl-PHAs production from Waste Frying Oils (WFOs)
Medium
Additional Carbon Source
D.C.W. [mg/L]
PHA amount [mg/L]
PHA % MM
Frying oil A
709,75
0,0
0,0%
Frying oil B
466,25
Medium
Additional Carbon Source
Extraction water
D.C.W. [mg/L]
PHA amount [mg/L]
PHA % MM
Frying oil A -
709,75
0,0
0,0%
Tap water
359,00
0,6
0,2%
Demineralised W.
377,63
Elix
362,75
1,4
0,4%
Frying oil B
466,25
409,87
3,5
0,8%
437,50
4,5
1,0%
495,63
3,7
Prefered C-sources
Aqueous extraction
“Waste water valorization”
Case study

28. Isolation of PhaA coding gene
PHAs production from glucidic carbon sources
Polyhydroxy alkonates/PHAs
To this aim, a B. cereus PhaA already characterized for the production of P(3HB) when expressed in E. coli (Davis et al. 2008) has been chosen. Reported two promoters vectors has been constructed.
phaRBC
Isolation of PhaA coding gene
β-ketothiolase
Implementation of biosynthetic gene(s) to complete pathways in order to obtain more flexible E. coli PHAs producer
phaARBC
Case study

29. Thermoplastic Starch in Automotive
Rapidly Replacing PS Foam in Auto-part Packaging
Fastest Growing
Least expensive biopolymer
Stable price vs Oil (low cost)
Proven technology and Process versatility
Poor performance
Potato, Corn
CornBIOTECH®-Germany., COHPOL™-Finland, ECOPLAST®Holland, VEGEMAT®France; US, Mich. State; Canada, UofTInnovation

30. Soy-Polyol Foam in Automotive
Promise with Competitive Technologies
Price Neutral
Improved properties over synthetic polyol
Adaptable with minimum adjustment of existing process
100% Polyol replacement possible
Brookestone
Cargill, Soy Polyurethane Systems, Dow and 6 others; BASF?

31. Soy Based Plastics in Automotive
Soy-Polyol- Promise with Competitive Technologies. Seven Technologies Commercialized.
Most Common:
Epoxidation of Soy-oil & hydrolysis to Polyol (Less capital, high operating)
Hydroformylation Process (high capital, low operating)
Ozonolysis (Complex, high investment, low capital)
Cargill, Dow, Soy Polyurethane Systems, and 6 others

32. New DaimlerChrysler Mercedes S-Class
Use of renewables dramatically increased
27 Components
43 kg bioparts:
door& pillar inners, head liner, rear cargo shelf & trunk components, thermal insulation & isolation mats

33. Toyota’s Green Car Drive
Toyota–A Global Model (Sweet Potato)

34. A historical note
Henry Ford experimented with soy meal in the manufacture of automobiles. In 1941 he produced an entire prototype "soybean plastic automobile," including a plastic body. World War II interrupted Ford’s development of soybean plastics.
Ford's “soybean plastic” automobile.

35. Bio-dyes
The colour industry’s difficult situation creates the need to find new ways of synthesising that are more environmentally friendly and economic for the production factories. The micro-organisms and enzymes studied by the SOPHIED project are capable of synthesising coloured compounds in softer production conditions.

36. Bio-Materials BIOMATERIALS
The field of biomaterials is highly multidisciplinary and involves principles from medicine, materials science and engineering, chemistry and biology. It involves the engineering and testing of the materials into useful devices for therapies. Its multidisciplinary nature often means that materials engineers work closely with surgeons, microbiologists, ethicists, and lawyers to name a few.
BASIC SCIENCES
ENGINEERING SCIENCES
BIOMATERIALS
MEDICAL SCIENCES
FORENSIS SCIENCES

37. Commercially, biomaterials are of enormous importance
Commercially, biomaterials are of enormous importance. It is thought that biomaterials based devices cost some $AUS400 billion dollars per year which constitutes roughly 8% of money spent on health-related issues.
Under FP5 and FP6 the EU has funded biomaterials research by a total of € million.

38. A Little History on Biomaterials
Romans, Chinese, and Aztecs used gold in dentistry over 2000 years ago, Cu not good.
Ivory & wood teeth
Aseptic surgery 1860 (Lister)
Bone plates 1900, joints 1930
Turn of the century, synthetic plastics came into use
WWII, shards of PMMA unintentionally got lodged into eyes of aviators
Parachute cloth used for vascular prosthesis
1960- Polyethylene and stainless steel being used for hip implants
Kolff Artificial Kidney, 1943

39. Applications include:
implants eg. titanium hip joints, artificial lenses
tissue engineering for the regeneration of damaged or diseased tissues eg. nerve regeneration for spinal cord injuries
‘smart’ surfaces for the culture of embryonic stem cells
artificial muscles using electroactive polymers
using peptides and DNA molecules as building blocks to build new nanostructures
‘bionanotech’ engineering approaches to manipulate cell function
site specific drug delivery using nanostructured materials
improved dressings for chronic wounds

40. Evolution of Biomaterials
Structural
Soft Tissue Replacements
Functional Tissue Engineering Constructs

41. Advances in Biomaterials Technology
Cell matrices for 3-D growth and tissue reconstruction
Biosensors, Biomimetic , and smart devices
Controlled Drug Delivery/ Targeted delivery
Biohybrid organs and Cell immunoisolation
New biomaterials - bioactive, biodegradable, inorganic
New processing techniques

42. Classes of Synthetic Biomaterials
Metals
stainless steel, cobalt alloys, titanium alloys
Ceramics
aluminum oxide, zirconia, calcium phosphates
Polymers
silicones, poly(ethylene), poly(vinyl chloride), polyurethanes, polylactides
Natural polymers
collagen, gelatin, elastin, silk, polysaccharides
Semiconductor Materials

43. Application of Synthetic Biomaterials
Metals
Dental Implants, Orthopedic screws/fixation
Ceramics
Bone replacements, Heart valves, Dental Implants
Polymers
Drug Delivery Devices, Skin/cartilage, Ocular implants
Semiconductor Materials
Implantable Microelectrodes, Biosensors

44. Artificial Hip Joints
Basic Material:
Stainless Steel, titanium and its alloys, and UHMWPE.
Challenges:
Prevention of wear & loosening over extended periods (10-15 yrs.).

45. Vascular Grafts
Basic Material:
Polyurethane, Teflon & Dacron
Challenges:
Maintenance of mechanical integrity
Long term blood compatibility (avoidance of blood clotting).

46. Substitute Heart Valves

47. Bio-sensors
Biosensors are devices used to monitor living systems. Biosensors have been applied to a wide variety of analytical problems including in medicine, drug discovery, the environment, food, security and defense. The potential growth in the world biosensor industry is remarkable, the emerging Biosensor market is expected to grow at over 9% in the coming years thus becoming one of the fastest growing sectors in the World. Biosensor industry is thus a key spin-off of the biological, materials & electronics discipline fusion.

48. Block Diagram of a Biosensor
Olfactory Membrane
Olfactory Nerve Cell
Sample (Analyte or Substrate)
Signal Processing Device
BiorecognitionElement
Transducer

49. Glucose Biosensor
The user carries a wallet sized case that contains the testing equipment
A lancet pierces the skin on the finger
The user places this blood sample on a test strip and inserts it into the reader

50. Commercial Biosensors – Ramsay - John Wiley & Sons © 1998
There are two types of glucose test strips
Reflectance-based strips
Explained in the following slides
Electrochemical-based strips
Monitor chemical reactions that involve electrochemically active molecules
Commercial Biosensors – Ramsay - John Wiley & Sons © 1998

51. Optically based strips
Reflected Rays
Source
Spreading Element
Analytical Element
Reflective Element
Support Element
Commercial Biosensors – Ramsay - John Wiley & Sons © 1998

52. Support Element Reflective Element Analytical Element
Serves as the foundation for the strip
Made of a thin and rigid plastic material that might also contain the reflective function
Reflective Element
Reflects light back through the sample
It may be integrated as part of the support element
Common elements used are TiO2, BaSO4, MgO, and ZnO
Analytical Element
Glucose oxidase catalyzes the oxidation of glucose in the blood by oxygen in the atmosphere and in the blood
This produces gluconic acid and hydrogen peroxide
The peroxidase triggers the reaction of the hydrogen peroxide with 3-methyl-2-benzothiazolinone hydrazone (MBTH) and 3-dimethylaminobenzoic acid (DMAB)
A naphthalenesulfonic acid salt replaces the DMAB in the strip

53. Future Directions of Glucose Biosensors
Noninvasive methods
Glucowatch
Small electric current to pull glucose through the skin
OLED based
organic light-emitting device pixels which serve as the light source are attached back-to-back with a sensing element, thus making a very small sensor