BIOPRODUCTS BIOMATERIALS Prof Giovanni Sannia Evolution IndustrialWhite Biotechnology
BIOPRODUCTS & BIOMATERIALS Prof. Giovanni Sannia
Evolution
Industrial/White Biotechnology uses enzymes and/or microorganisms 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 H 2 O consumption, waste and CO 2 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.
What are the benefits of industrial Biotechnology? Drivers Profit Economically Viable • Cost reduction • Novel products Hurdles • Regulation • Further technology development • Feedstock prices • Investments Drivers • Less energy • Less waste Drivers • Knowledge-based quality jobs • Responsibility SUSTAINABILITY Hurdles • Unawareness • Acceptance Hurdles • Further technology development • Waste management People Planet Socially Responsible Environmentally Sound The Triple-P Bottom Line
RESOURCES PROCESS Starch, colza, cereals, maize, forestry wastes, co-products of wood industry, beetroots, potatoes, breeding effluents, flax, hemp, miscanthus, other renewable materials… PRODUCTS BIOENERGIES Bioplastics Green heat Wood pellets stoves and central Cash bags in maize starch, disposable heating forks, spoons and knives made systems, wood chips central heating of beetroot, food recipients… systems, heat produced by biomethanation Biolubricants of breeding effluents, cereals stoves Chain oil for chainsaw, engine and lubricants, hydraulic fluids… central heating systems… Vegetal ink Biomaterials Insulation materials, gardening pots and miscanthus… Chemical products Vegetal solvents, paints, additives… 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… Biodetergent Washing-up liquid, shampoo, washing powder, floors and surfaces cleaning detergent… 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…
Bioproducts Terminology Renewable Bioresource Feedstock • Plants crops trees algae Bioprocess Technology Biocatalysis (Enzymes) • Organic residues municipal industrial agricultural forestry aquaculture • Bioenergy and Biofuels Fermentation (Microorganisms) • • Animals, fish • Microorganisms Industrial Bioproducts Physico – Chemical Process Technology Extraction Pyrolysis Gasification Manufactured products: – biochemicals – biosolvents – bioplastics – ‘smart’ biomaterials – biolubricants – biosurfactants – bioadhesives – biocatalysts – biosensors
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.
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.
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!
Some of the disciplines required…. . Chemistry Bioinformatics Proteomics Metabolic engineering Microbiology Molecular biology Evolution & screening Advanced analysis Industrial fermentation Biochemistry/enzymology Bioprocess development Down stream processing
Integrated Research Team • Bio. Material 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
Renewable Bioproducts Vision Goals: Achieve 10% of basic chemical building blocks from plant-derived sources, with concepts in place to achieve 50% by 2050
Classes of Bioproducts The thousands of different industrial bioproducts produced today can be categorized into four major areas: 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.
Life cicle of bioproducts Source: A. J. Ragauskas et al. , Science 311, 484 (2006); CO 2 balance Source: A. K: Mohanty et al. , Natural Fibers, Biopolymer and Biocomposites, CRC Press (2006) The cycle of CO 2 is closed!
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
Bio-plastic Bioplastics Innovation and Commercialization Drive Biotechnologically produced high volume commercial or potentially commercial polymers: • Polylactide (PLA) • 1, 3 --propanediol (PDO) Dupont Sorona • Polyhydroxy alkonates/PHAs • Thermoplastics starch • Soy Polyol-PU/Soy resin
Polylactide (PLA) PLA
Polylactide (PLA) PLA in packaging Corn-based plastic polylactic acid (PLA) is used in several kinds of packaging, including these beverage containers from Nature. Works.
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
Polylactide (PLA) PLA in Automotive Poly Lactic Acid in Automotive Existing Technology: Limited Scope ØA biodegradable thermoplastic Corn Toyota Tire Cover Ø Functional properties match petroleum plastics Ø High Price Ø Poor durability APPLICATIONS: Tire Cover, Carpet, Antenna Cargill-USA, - Apack Germany/Fortum Oyj. Finland, Mitsui Chemicals. Japan, Brimingham Polymers USA/Phusi France
Polyhydroxy alkonates/PHAs in Automotive Outlook - More Promising than PLA q Exceeds PP Properties q Price Claimed < $1. 00/lb q Projected Production 2008 q No Commercial Availability q Small Player q (R&D + Investment Risk) Corn & Diverse Grass Metabolix to ADM, (USA); Procter & Gamble Tech to Kaneka Corporation (Japan), Biomatera Inc. , Quebec
Polyhydroxy alkonates/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
Polyhydroxy alkonates/PHAs 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 General structure of polyhydroxyalkanoates; R 1/R 2= alkyl groups C 1 -C 13; x=1 -4; n=100 -30000 n. Cmon≤ 5, short chain length (scl-PHA); n. Cmon≥ 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 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 Poor Good Biodegradability Good None Density (g/cm-3)
Polyhydroxy alkonates/PHAs composition is influenced by the type of supplied Carbon source: ØSugars (unrelated) ØFatty acids (related) according to the biosynthetic pathways shown in figure Poly 3 -hydroxybutyrate Poly 3 -hydroxyalkanoates
Recipient E. coli Polyhydroxy alkonates/PHAs Ø Isolation of pha. RBC biosynthetic operon Pha. R Pha. B PHA synthases subunit/ ketoacyl-Co. A reductase Transcriptional regulator? Pha. C PHA synthases subunit Ø Construction of recombinant E. coli biopolymer producing systems Case study
Polyhydroxy alkonates/PHAs Growth medium Strain Additional carbon source CDW (mg/L) PHA (%) Composition (%) 3 HB LB 0. 2% Octanoate 380. 0 MM 0. 2% Octanoate 156. 2 MM MM MM 0. 3 0. 2 9. 9 0. 05 tial n e t o p e h pt” of t e c n ion of t o a c s f i o r f o l o a o 0. 1% Octanoate 128. 0 7. 0 0. 1 r “p for v a x) H e m d e H i t 3 v s ( o y r P s p r t a n s t e l a Resu combin h as a n e c r u a s f , o s t n c o Decanoate 5. 7 0. 1 e produ 185. 7 l b exploitati 0. 1% a u l a v ls into waste oi 0. 05% 334. 9 0. 1 lymer. Decanoate o p o m o h 3 HHx 99. 8 99. 95 99. 9 MM 0. 05% Dodecanoate 344. 7 6. 65 0. 05 99. 95 MM MM 6% Corn oil 6% Coconut oil 553. 6 365. 5 0. 9 0. 4 0. 6 0. 5 99. 4 99. 5 Case study
Polyhydroxy alkonates/PHAs Sustainable mcl-PHAs production from Waste Frying Oils (WFOs) Medium MM MM Additional Extraction water Carbon Source Frying oil A Frying oil B Prefered C-sources Case study D. C. W. [mg/L] PHA amount [mg/L] Tap water 709, 75 359, 00 0, 6 0, 0% 0, 2% Demineralised W. 377, 63 0, 6 0, 2% Elix 362, 75 1, 4 0, 4% - 466, 25 0, 0% Tap water 409, 87 3, 5 0, 8% Demineralised W. 437, 50 4, 5 1, 0% Elix 495, 63 3, 7 0, 8% Aqueous extraction 0, 0 PHA % “Waste water valorization”
Polyhydroxy alkonates/PHAs production from glucidic carbon sources pha. RBC Isolation of Pha. A coding gene β-ketothiolase • Implementation of biosynthetic gene(s) to complete pathways in order to obtain more flexible E. coli PHAs producer pha. ARBC Case study To this aim, a B. cereus Pha. A already characterized for the production of P(3 HB) when expressed in E. coli (Davis et al. 2008) has been chosen. Reported two promoters vectors has been constructed.
Thermoplastic Starch in Automotive Rapidly Replacing PS Foam in Auto-part Packaging ü Fastest Growing ü Least expensive biopolymer ü Stable price vs Oil (low cost) ü Proven technology and Process versatility ü Poor performance Potato, Corn. BIOTECH®-Germany. , COHPOL™-Finland, ECOPLAST®Holland, VEGEMAT®France; US, Mich. State; Canada, Uof. TInnovation
Soy-Polyol Foam in Automotive Promise with Competitive Technologies Ø Price Neutral Ø Improved properties over synthetic polyol Ø Adaptable with minimum adjustment of existing process Ø 100% Polyol replacement possible Brookestone Cargill, Soy Polyurethane Systems, Dow and 6 others; BASF?
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
New Daimler. Chrysler 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
Toyota’s Green Car Drive Toyota–A Global Model (Sweet Potato)
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.
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.
Bio-Materials 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 BIOMATERIALS ENGINEERING SCIENCES FORENSIS SCIENCES MEDICAL SCIENCES
Commercially, biomaterials are of enormous importance. It is thought that biomaterials based devices cost some $AUS 400 billion dollars per year which constitutes roughly 8% of money spent on health-related issues. Under FP 5 and FP 6 the EU has funded biomaterials research by a total of € 173. 7 million.
A Little History on Biomaterials v Romans, Chinese, and Aztecs used gold in dentistry over 2000 years ago, Cu not good. v Ivory & wood teeth v Aseptic surgery 1860 (Lister) v Bone plates 1900, joints 1930 v 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 v 1960 - Polyethylene and stainless steel being used for hip implants Kolff Artificial Kidney, 1943
Applications include: üimplants eg. titanium hip joints, artificial lenses ütissue engineering for the regeneration of damaged or diseased tissues eg. nerve regeneration for spinal cord injuries ü‘smart’ surfaces for the culture of embryonic stem cells üartificial muscles using electroactive polymers üusing peptides and DNA molecules as building blocks to build new nanostructures ü‘bionanotech’ engineering approaches to manipulate cell function üsite specific drug delivery using nanostructured materials üimproved dressings for chronic wounds
Evolution of Biomaterials Structural Soft Tissue Replacements Functional Tissue Engineering Constructs
Advances in Biomaterials Technology üCell matrices for 3 -D growth and tissue reconstruction üBiosensors, Biomimetic , and smart devices üControlled Drug Delivery/ Targeted delivery üBiohybrid organs and Cell immunoisolation • New biomaterials - bioactive, biodegradable, inorganic • New processing techniques
Classes of Synthetic Biomaterials q Metals stainless steel, cobalt alloys, titanium alloys q. Ceramics aluminum oxide, zirconia, calcium phosphates q. Polymers silicones, poly(ethylene), poly(vinyl chloride), polyurethanes, polylactides q. Natural polymers collagen, gelatin, elastin, silk, polysaccharides q. Semiconductor Materials
Application of Synthetic Biomaterials q Metals Dental Implants, Orthopedic screws/fixation q. Ceramics Bone replacements, Heart valves, Dental Implants q. Polymers Drug Delivery Devices, Skin/cartilage, Ocular implants q. Semiconductor Materials Implantable Microelectrodes, Biosensors
Artificial Hip Joints Basic Material: Stainless Steel, titanium and its alloys, and UHMWPE. Challenges: Prevention of wear & loosening over extended periods (10 -15 yrs. ).
Vascular Grafts Basic Material: Polyurethane, Teflon & Dacron Challenges: Maintenance of mechanical integrity Long term blood compatibility (avoidance of blood clotting).
Substitute Heart Valves
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.
Block Diagram of a Biosensor Olfactory Membrane Sample (Analyte or Substrate) Biorecognition Element Olfactory Nerve Cell Transducer Signal Processing Device
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
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
Optically based strips Source Reflected Rays Spreading Element Analytical Element Reflective Element Support Element Commercial Biosensors – Ramsay - John Wiley & Sons © 1998
Support 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 Ti. O 2, Ba. SO 4, Mg. O, and Zn. O 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
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 backto-back with a sensing element, thus making a very small sensor
- Slides: 54