1 Nano Bio Tech?

2 Environmental NanobiotechnologyG. Sannia

3 A Brief History of NanotechnologyFeynman In 1959, Richard Feynman ( ) of CalTech gave a talk entitled "There's Plenty of Room at the Bottom.“ "Why can’t we compress 24 volumes of Encyclopedia Britannica on a pin head ?“ " The biological example of writing information on a small scale has inspired me to think of something that should be possible " In the 1980s/90s, a team led by Richard Smalley ( ) manipulated Carbon atoms to create Fullerenes (“Buckyballs”) In 1990, IBM scientists wrote the logo IBM using 35 xenon atoms on nickel. Smalley

4 What is Nano?....... From the Greek word for “dwarf”A nanometre is 1/1,000,000,000 (1 billionth) of a metre (10-9 m), which is around 1/50,000 of the diameter of a human hair or the space occupied by 3-4 atoms placed end-to-end. Term “nanotechnology” was first used in 1974 by Norio Taniguchi

5 The Scale of “things”

6 What is Nanotechnology?(Definition from the National Nanotechnology Initiative) Research and technology development aimed to understand and control matter at dimensions of approximately 1 – 100 nanometre – the nanoscale. Ability to understand, create, and use structures, devices and systems that have fundamentally new properties and functions because of their nanoscale structure. Ability to image, measure, model, and manipulate matter on the nanoscale to exploit those properties and functions. Ability to integrate those properties and functions into systems spanning from nano- to macro-scopic scales.

7

8 Alice follows the White Rabbit down the rabbit hole…and falls down a very long chamber full of strange things …….Alice then finds a small bottle labeled "DRINK ME," and drinks it. The drink causes her to shrink. Alice accidentally leaves the key on the table, and with her diminished stature can no longer reach it and becomes very scared….. …..a strange new world for Alice!

9 Like Alice, when Matter Falls Down the “rabbit hole” to Nano- Scale dimensions- strange things happen…. opaque substances become transparent (copper); inert materials become catalysts (platinum); stable materials turn combustible (aluminum); solids turn into liquids at room temperature (gold); insulators become conductors (silicon). A material such as gold, which is chemically inert at normal scales, can serve as a potent chemical catalyst. These behavioral changes –nanotechnology - stem from unique quantum and surface phenomena that matter exhibits at the nanoscale.

10 Nanotechnology approaches...Top-down approach reducing the size of the smallest structures to the nanoscale Cutting, carving and molding This approach starts with a bulk or thin film material and removes selective regions to fabricate nanostructures (similar to micromachining techniques). This top-down approach is an offshoot of standard lithography and micromachining techniques. However, this strategy becomes increasingly challenging, as the dimensions of the target structures approach the nanoscale.

11 Nanotechnology approaches...Bottom-up approach Manipulating individual atoms and molecules Components made of single molecules This method relies on molecular recognition and self-assembly to fabricate nanostructures out of smaller building blocks (molecules, colloids, and clusters). Nature efficiently builds nanostructures by relying on chemical approaches. Molecular building blocks are assembled with a remarkable degree of structural control in a variety of nanoscaled materials with defined shapes, properties, and functions, e.g. DNA.

12 What is nanotechnology?Not just about miniaturisation…

13 What is nanotechnology?Not just about miniaturisation… Emergent phenomena Convergence Self-assembly Self-replication Bio-nanotechnology

14 What is nanotechnology?Not just about miniaturisation… Emergent phenomena As the size of a crystal shrinks towards the size of molecules and atoms, its electrons start to follow the laws of quantum mechanics rather than classical mechanics, and behave more like waves than like particles. The electrical and optical properties can be dramatically changed just by changing the size. Convergence Self-assembly Self-replication Bio-nanotechnology

15 What is nanotechnology?Not just about miniaturisation… Emergent phenomena Convergence Progress in nanotechnology is dependent on understanding ideas from physics, engineering, chemistry and biology: it is a multi-disciplinary or convergent topic. This has implications for how we teach science and how we structure research. Self-assembly Self-replication Bio-nanotechnology

16 What is nanotechnology?Not just about miniaturisation… Emergent phenomena Convergence Self-assembly Although most miniature devices are made by making small patterns on large objects (“top down”), it may be possible to get better control by assembling individual atoms or molecules into larger clusters (“bottom up”). Self-limiting chemical or biological synthesis can be used. Self-replication Bio-nanotechnology

17 What is nanotechnology?Not just about miniaturisation… Emergent phenomena Convergence: Self-assembly Self-replication Borrowing ideas from biology, we can imagine machines which reproduce themselves. This has given rise to some newspaper stories about nanorobots taking over the world. Is this science fact or science fiction? Bio-nanotechnology

18 What is nanotechnology?Not just about miniaturisation… Emergent phenomena Convergence Self-assembly Self-replication Bio-nanotechnology By making structures which are the same size as the components of cells, we can start to manipulate biological processes, or make sensors which are sensitive to a single molecule. Or, we can exploit Nature’s nanotechnology, using DNA to assemble scaffolds or to make biomolecular motors.

19 The First NanotechnologyAncient stained-glass makers knew that by putting varying, tiny amounts of gold and silver in the glass, the could produce the red and yellow found in stained-glass windows. Similarly, today’s scientists and engineers have found that it takes only small amounts of a nanoparticle, precisely placed, to change a material’s physical properties.

20 Nanotechnology Facts Nanotechnology is expected to be one of the most important emerging technologies for the 21st century.

21 Impact on Global EconomyNational Nanotechnology Initiative (NNI) was started in 2000 by President Clinton. Since 2000, the federal government has allocated over $2 billion for nanotechnology research. $480 million of venture capital went into nanotechnology startups in 2005. (2001) European Union approved budget > €16B ($20B) for R&D under EU Framework Programme. Nanotechnology, a major theme and priority, was slated to receive nearly 10% of this funding allocation. Japan, Taiwan, Singapore, China, Israel and Switzerland have all begun similar measures.

22 Nanotechnology Predicted Growth$15 billion annual investment predicted within 10 years. 50% of all products produced will be influenced by nano within 10 years. Employment in the nanotechnology sector is expected to grow to 2 million workers within the next decade (US Department of Labor). Federal govt. spent $3.7 billion for nanotechnology R&D from FY EU expects to spend an equal amount on nano R&D.

23 Nanotechnology Predicted Growth

24 Nano-Biotechnology Nanotech that looks nature for its start “At present, nanobiotechnology is a field that concerns the utilization of biological systems optimized through evolution, such as cells, cellular components, nucleic acids, and proteins, for the development of new biomaterials and analytical toolkits as well as for understanding biological phenomena in more detail.”

25 Nano-Biotechnology

26 Nano-Biotechnology

27 Nano-Biotechnology Biological Systems Modular and Replaceable PartsMolecular Motors with Specific Targeting Durable Catalytic at Ambient Temperatures “Bottom-Up” Manufacturing Self-Assembly Genetically Re-Engineered “Functional biomolecules possess a wide range of highly desirable, intrinsic properties (e.g., thermostability, energy conversion, actuation, etc). Such proteins can be isolated, engineered and mass-produced for integration as structural and functional components of nanoscale materials and systems.”

28 Nano-Biotechnology Biological Systems Modular and Replaceable PartsBiological Systems, Continued: Modular and Replaceable Parts: Biological parts are replaceable, durable, work at ambient temperatures, and some are catalytic (e.g. enzymes). Examples are molecular motors (circular, linear, and antibodies). Nature provides examples of nanoscale actuators and pumps that can efficiently convert chemical and light energy into mechanical work. These classes of functional proteins may be exploited to incorporate bio-mimetic and inspired functionality into integrated systems “Functional biomolecules such as motor proteins possess a wide range highly desirable, intrinsic properties (e.g., thermostability, energy conversion, actuation, etc). Such proteins be isolated and engineered for integration as structural and functional components of nanoscale materials and systems.” References: Engineering Active Biomolecules for Integrated Nanomaterials and Nanodevices. Center for Integrated Nanotechnologies. Department of Energy Fig. 9.7 – Nature provides examples of nanoscale actuators and pumps. Modular and Replaceable Parts 28

29 Nano-Biotechnology Biological SystemsBiological Systems, Continued: Molecular Motors with Specific Targeting: How can motors be used in nanobiotechnology? The genes for these motors could be cloned, mass-produced, and then put on nanodevices for moving cargo, fluids or generating or converting energy. In nature, there are motors that convert light energy into chemical energy and utilize the movement of ions to generate chemical energy; many of these motors are reversible. These motors have been described as being more efficient than man-made motors and cheap in terms of the amount of energy they require compared to how much work they put out. And of course, they are very small (nm range), so they are perfect for nanotechnology. Chemists at Italy's University of Bologna, UCLA and the California NanoSystems Institute (CNSI) have designed and constructed a molecular motor of nanometer size that does not consume fuels; their nano motor is powered only by sunlight. It could be used in drug delivery (Nature 2005, 437, 1337), and since it runs on light energy, it is considered eco-friendly (http://advancednano.blogsport.com) and (www.nanoform.org) and (http://www.physicalsciences.ucla.edu/research/nano_motor.asp). An example of a linear motor is a DNA polymerase (e.g. Taq polymerase used in polymerase chain reactions). To watch a molecular motor work go to the movie, “The Inner Cell,” at YOUTUBE. There are proteins (in this case antibodies) that can be used as probes for targeting specific proteins. The “targeting part” (the variable region of the protein) of the antibody can be selected and even changed (http://library.lanl.gov/cgi-bin/getfile?28-25.pdf). Disadvantages of proteins isolated from biological systems: Many of them only work in a water world under specific solvent conditions! Reference: Andrew M. Bradbury, Geoffrey S. Waldo, and Ahmet Zeytun, Fluorobodies, Los Alamos Science, Number Fig. 9.8a - Fig. 9.8b - Molecular Motors with Specific Targeting

30 “Bottom-Up” ManufacturingNano-Biotechnology Biological Systems “Bottom-Up” Manufacturing DNA RNA protein Eukaryotes DNA (exons and introns) Final mRNA is composed of exons. cDNA is complementary to mRNA. Biological Systems, Continued: Bottom-Up Manufacturing: Biological systems are also manufactured from “bottom up,” can self-assemble, be re-engineered and mass produced. Francis Crick (http://www.ncc.gmu.edu/dna/base.htm) proposed that DNA was the template from which a messenger RNA (mRNA) was produced. This process is known as transcription. Then the mRNA is decoded (translated) into protein. Crick named the transcription of DNA to RNA, and the translation of RNA to protein the Central Dogma of biology, and he felt, at the time, that all life must follow this blueprint. Of course this was before RNA viruses, such as HIV, were discovered. Anyway, these two processes can occur simultaneously in bacterial cells (prokaryotes) because a bacterium does not have a nucleus. In Eukaryotes, such as human cells, transcription occurs in the nucleus. The mRNA migrates out of the nucleus and is translated into a protein. Translation occurs in the cytoplasm. Note that both DNA and RNA are acidic compounds just like lemon juice or vinegar is acidic. In Eukaryotes, DNA replication (i.e. the duplication of DNA) occurs in the nucleus. Gene expression, that is the production and action of proteins, is what actually leads to the traits that people show (e.g. eye color, personality, athletic abilities, likes, dislikes). Of course how a person interacts with their environment, from the moment they are aware of their surroundings, also contributes to personal traits. Genes in eukaryotes, such as humans, are made-up of both coding sequences known as exons and noncoding regions known as introns. The final mRNA (messenger RNA) for a protein usually just represents the exons. If DNA is artificially made from the exon-only-containing mRNA, it is known as complementary DNA (cDNA). The enzyme that catalyzes this reaction was cloned from a group of RNA-containing viruses known as retroviruses (HIV is a retrovirus). Transcription of cDNAs typically results in the production of the “correct” protein, barring any other post-translational modifications that needs to be done on the translated protein. Fig

31 Nano-Biotechnology Biological Systems Self-AssemblySelf-assembly is assembly of molecules without use of outside source. Nature provides brilliant examples of multiscale self- assembly of energy-dissipative materials, which together exhibit a range of emergent phenomena. The overarching goal of scientist work is to understand, exploit, and engineer structural and functional biomolecules to assemble integrated nanomaterials and nanodevices with unique properties” Biological Systems, Continued: Self-Assembly: Molecular self-assembly is the assembly of molecules without guidance or management from an outside source. It is important to note that insulin, like many other proteins and other cellular materials like membranes, self-assemble in the cell as it is being synthesized. Man-made products generally do not self-assemble; they must be assembled. This ability to self-assemble is something that nanotechnologists would like to exploit and/or build into some of their structures. Some proteins (e.g. SOD), especially small ones, can reassemble if they have become denatured, once normal conditions are reinstituted (e.g. lower temperature, lower salt). One has to remember this self-assembly happens in a very specific cellular environment (watery, defined ionic strength and composition, and pH) and has been perfected through evolution. Of course, not all cellular proteins self-assemble; some require help from other proteins to assemble correctly. Plus insulin, like many other proteins, to work correctly requires some post-translational modifications. In eukaryotic cells, these modifications are taken care of by other enzymes in other parts of the cell. “Nature provides brilliant examples of multiscale self-assembly of energy-dissipative materials, which together exhibit a range of emergent phenomena. The overarching goal of our work is to understand, exploit, and engineer structural and functional biomolecules to assemble integrated nanomaterials and nanodevices with unique properties” References: Engineering Active Biomolecules for Integrated Nanomaterials and Nanodevices. Center for Integrated Nanotechnologies. Department of Energy Water Micelles Self assembly 31

32 Genetically Re-EngineeredNano-Biotechnology Biological Systems Genetically Re-Engineered Tailor-Made Proteins: Changing Amino Acids Biological Systems, Continued: Genetically Re-Engineered, Continued: With protein design, it has, for example, been possible to improve the stability of an enzyme which is an important component of detergents, by specifically changing an amino acid (orange) close to the catalytic region (yellow). The enzyme can thereby survive the chemicals also needed to make our clothes clean. In this example, the scientists at the biotech company, Genencor International, specifically changed amino acids in the enzyme, subtilisin, a fungal protein degrader, so that it would work better in the washing machine. To do this, fungi were treated with chemicals that causes changes in the their DNA code and then grown under “washing machine” conditions. This procedure resulted in a fungi that contained enzymes that worked well under those specific conditions. Fig

33 Humanity’s top ten problems for next 50 yrsENERGY WATER FOOD ENVIRONMENT POVERTY TERRORISM & WAR DISEASE EDUCATION DEMOCRACY POPULATION

34 Environmental applicationsPollution prevention Treatment remediation Sensors/detection Green manufacturing Energy production and utilization

35 Pollution prevention Synthetic or manufacturing processes which can occur at ambient temperature and pressure. Use of non-toxic catalysts with minimal production of resultant pollutants. Use of aqueous-based reactions. Build molecules as needed --“just in time.” Nanoscale information technologies for product identification and tracking to manage recycling, remanufacture, and end of life disposal of solvents.

36 Laccase catalysed azo bond formationPollution prevention Enzymatic dye synthesis Laccase LAR1 ABu62 Laccase S3 CURIE_22 And during the meeting in Istanbul last december, we selected four dyes with the following selection criteria LAR1 was selected as the red dye, CURIE 22 is yellow, ITU_G is green. These three dyes are made by laccase catalysed oxidations And the natural red 8 is extracted from madder. Laccase catalysed azo bond formation Laccase ITU_G P6 P8

37 Pollution prevention Biomolecular nanolithography5mm Biomimetic methods of organizing metal particles 1.5 nanometers in diameter. Assembling the particles on a biopolymer template or scaffold stretched out on a surface. Nanostructures are organized into well-defined chip architectures, such as lines and grids. Process eliminates the current process chemicals that are harmful to the environment. Nanoscale assemblies have been made that demonstrate stable, room-temperature electrical behavior that may be tolerant of defects and useful in building nanoscale circuits.

38 Surface properties Inversion of wettability by nanometric biofilm of hydrophobins Fungal proteins self-assembling at hydrophobic/idrophilic interfaces Hydrophobins Water contact angle (WCA) Hydrophilic surface Hydrophobic surface

39 Surface properties Creation of chemical nano-patterns on surfacesKOH etching is used in micromachining processes for the realization of microsystems Unetched surface protected by hydrophobin biofim Surface etched by KOH

40 Development of biosensors and microarraySurface properties Immobilization of biological macromolecules on hydrophobin biofilm without loosing activity Immobilization of antibodies Immobilization of enzymes Development of biosensors and microarray

41 Nanotechnology enhancementsTreatment & Remediation Water Purification Nanotechnology enhancements Easier contamination removal: filters made of nanofibers that can remove small contaminants Improved desalination methods: nanoparticle or nanotube membranes that allow only pure water to pass through Lower costs Lower energy use

42 Nanotechnology enhancementsEnergy production and utilization Solar cells Nanotechnology enhancements Improved efficiencies: novel nanomaterials can harness more of the sun’s energy Lower costs: some novel nanomaterials can be made cheaper than alternatives Flexibility: thin film flexible polymers can be manipulated to generate electricity from the sun’s energy

43 Nanotechnology enhancementsEnergy production and utilization Batteries Nanotechnology enhancements Higher energy storage capacity and quicker recharge: nanoparticles or nanotubes on electrodes provide high surface area and allow more current to flow Longer life: nanoparticles on electrodes prevent electrolytes from degrading so batteries can be recharged over and over A safer alternative: novel nano- enhanced electrodes can be less flammable, costly and toxic than conventional electrodes

44 Monitoring BiosensorsAnalytical tools for the analysis of bio-material samples to gain an understanding of their bio-composition, structure and function by converting a biological response into an electrical signal. The analytical devices composed of a biological recognition element directly interfaced to a signal transducer which together relate the concentration of an analyte (or group of related analytes) to a measurable response.

45 Monitoring BiosensorsBiosensors: Bacteria that can locate biologically active pollutants. Bioluminescence: Reports presence of a pollutant. Current Applications, Continued: Sensors: Annually in the U.S., industrial plants generate 265 million metric tons of hazardous waste, 80% of which make its way into landfills. Burying these chemicals does not remove them from the ecosystem, it just moves it out of sight. Water run-off from both the plants and the landfills can lead to water pollution. Traditional chemical analyses to locate these chemicals are expensive and cannot distinguish between chemicals that affect biological systems from those that lie inert in the environment. In response to this problem, scientists developed biosensors, bacteria that can locate biologically active pollutants. These biosensors do not require costly chemicals or equipment and work within minutes. These bacteria contain a receptor that is activated in the presence of the pollutant and a reporter (in this case, bioluminescence) that is visually seen. There are several examples of this type of system being used all over the world. In South Korea, bacteria containing this system are being used to determine wastewater treatment failures. Before treated water is released into the environment, it is pumped through a bioreactor containing the bacteria. These bacteria will emit light as long as they are healthy but will stop emitting light if they have been killed by toxic pollutants. Fig Omphalotus nidiformis, glowing with the lights off bioluminescence. 45

46 Nanotechnology and the Environment Videos Nanotechnology and the Environment Videos Using Nanotechnology to Purify Water This video examines the work of Michael Wong in using gold and palladium nanoparticles to remove contaminants from water

47 Nanotechnology Providing Clean Water for EveryoneNanotechnology can help provide clean water for NASA astronauts, disaster relief teams, and field clinics. The CEO of a Vermont nanotech start-up company drinks water out of the Charles River to make his point and MOS tests the water filtering device in front of a live NECN audience.

48 Water Purification using Magnetic NanoparticlesTurboBeads magnetic nanoparticles are used to magnetically extract a dye (methylorange) from drinking water

49 Coating Uses Nanotechnology to Keep Building Exteriors CleanRadiant Shield Coating (RSC) by Hyperion Environmental uses the power of light to keep building exteriors clean. RSC destroys organic contaminants before they accumulate, keeping buildings cleaner, longer than ever before.

50 Nanotechnology Offers Solution for Mexico Gulf Oil Spill Clean-upA piece of chemically treated cotton cloth is able to separate crude oil from sea water (both from Mexico Gulf) completely within seconds by using gravity alone. The treated cloth allows water to path through but not oil. The novel surface chemical treatment method is developed by University of Pittsburgh.

51 IBM Makes Water Clean With Smarter, Energy-Efficient PurificationIBM has unveiled a novel nanomembrane technology that stands to alleviate the growing shortage of drinkable water worldwide. New membrane that filters out salts as well as potentially harmful toxins in water such as arsenic while using less energy than other forms of water purification.

52 Smart Ways to Manage and Reuse Water in South AustraliaUniSA and SA Water have extended a research partnership deal that has seen SA Water invest $3.5m of funding into finding smart ways to manage and re-use water in South Australia.

53 Nanotechnology for Removing Arsenic from Drinking WaterIn March 2010, researchers from Rice University in Houston traveled to Guanajuato, Mexico, to conduct field tests on a new nanotechnology for removing arsenic from drinking water. The system uses nanoscale magnetite, or nanorust.

54 The Media and Nanotechnology (USA)Nanotechnology Regulation Needed, Critics Say December 5, 2005 Study Raises Concerns About Carbon Particles March 29, 2004 ASSESSING RISKS; Technology's Future: A Look at the Dark Side May 17, 2006 The promise and perils of the nanotech revolution; Possibilities range from disaster to advances in medicine, space July 26, 2004 Solar Energy Nanotechnology Can Replace Fossil Fuels July 11, 2005

55 ENVIRONMENTAL HEALTH ISSUESSome beleive that nanostructures/nanomaterials have not been adequately studied Unknown toxicity of some nanomaterials Fate of such structures in the environment Bioassimilation of such structures in ecosystems Mobility and persistence of nanomaterials Trasformation/degradation products unknown

56

57