Fundamental of Nanotechnology PROF. ABSAR AHMAD director.inc@amu.ac.in DIRECTOR- INTERDISCIPLINARY NANOTECHNOLOGY CENTRE (INC),INTERDISCIPLINARY NANOTECHNOLOGY CENTRE (INC) Recent Establishment of INC: Aligarh Muslim University, Aligarh, decided in 2016 to establish a state of the art Nanotechnology Centre with,regard to the founder- Sir Syed's vision to harbor all new and novel fields of knowledge and research at AMU. Having spent 23+ years at CSIR-National Chemical Laboratory, Pune, inclusive of 18+ years of pioneering research work related to nanoscience and nanotechnology, | was invited by the Executive Council of AMU to help in establishing a world class centre of excellence in nanotechnology at the esteemed university. In view of student welfare and knowledge propagation, | accepted the offer and have joined AMU in December 2016 as the Founder-Director of the Interdisciplinary Nanotechnology Center (INC). Since then, we have begun harboring a decent number of positions for Asst. Professors, PhD scholars, etc. and have also undertaken the teaching responsibilities of M.Tech Nanotechnology » v-.09::on's20i cuscen JEREEI u.. ology in full swing.The opportunities in the fields of nanoscience and nanotechnology from biological routes are enormous and provide a platform for young thinkers to have brilliant careers in science.uy 20 Years of Biosynthesis of Nanoparticles and Drugs from fungal Endophytes - Prof. Absar Ahmad Biosynthesis of nanomaterials: Extracellular AgNPS Technical know- how transferred to Tata Chemicals, ‘Mumbsi. DSM (Holland), CIPLA, Gatewell Pharma, Ipca Laboratories, Ross LifeSciences, Cooper Pharma, Or. G Wellness, Genneva Biotech, External Cash Flow: (ECF) Genencor (USA), Torrent Pharmaceuticals, Asian Paints, Tata Chemicals 2] Drugs from fungal endophytes: Taxol & 10 DAB producing endophytic fungus emerging from Taxus baccata. Awards \VASVIK Industrial Research Award: 2006 DBT-Tata Innovation Fellowship: 2007-8 NCL-Sciantist of the Year: 2008 MRS! Medal: 2009 Interested Companies: We are the pione work: “24,500 ; Scholar. Biologically fabricated NPs by our group Ag alloy, bimetallic core shells care. Oxides: 70, Fo,0,, TIO, Si0,, BaTiO., Ce0,, 60405, C10;, MnO, AlOs, Biominer Total funding received: + Rs. 10 Crores 3] Nanoparticle-Dri ‘Quantum dots: CdS, Zn, Nis, MnS, PBS, BS, n: Porous carbon, porous graphene, GOs CACO, SCO, BaCDs, PHCDs, CACO, ton: Biocon) Lee & le’ Fluorescence spectrum: Drug + Nano conjugate ering contributors to the field and have received high international acclaim amongst peers. Total citation of the published heindex = 51, source Google ‘Our Key Publications: (Citations ‘atare Matera, 3(2008)-3738 4.amm.Cham Soc: Papers 890 4. Colloid iearface Se, 2752008): 2512 Biotechnol Prog, 222006): 2005 Colloids Surf 6 Bisinterfces, 222003): 2847 ‘Nan Letters, (2001); 1260 Biotechnol Prog, 192003): 1291 ‘Currant Seenee, 85,2003) 142 4. Maters, Cham, 13,2003): 902 ‘Angew. Chem. nt 10,2001): 866 Langmuie 19,(2003) 683 |LNaposcience:Nanotach, 5 (2005): 618 ‘ChemBioChem, 5 (2002) $80 Chemistry of Materials, 17 (2005): $87Teaching & Research Roadmap: Inter: iplinary Nanotechnology Centre (INC), AMU. Scaling-up techniques for mass iuction of bioinspired nanomaterials Metals, metal oxides Metal sulfides , quantum dots *Biominerals Nan Desalinization of Water *One-step water filtration process to produce potable water from sea water ‘Removes salt ions, heavy metal, fungi, bacteria and viruses, ‘Study of nanomaterials in Energy ‘Energy generation Energy storage Catalysis & Water splitting ‘Study of nanomaterials in Environment ‘Water, soil & ar pollution control ‘Study of nenometerials in Healtheare *HIV-AIDS, Diabetes, Malaria ‘Cancer (Liver, Breast, Lung, etc) *Neuradegenerative diseases ‘Eye Diseases and Retina Regeneration Animal diseases sTheranostics Tailor-made Nanoparticles screened ‘Au, Y20, CeOz, ZnO, Pt, AlsOa, AunS, AgaS, Bi2Ss, MnsOs, Ag, TiO, SiOz, Gda02, FeaOs, et ‘Enhancing Plant Growth and Productivity ‘+Antisnsect activity ‘Safety/Toxicity Assesement ‘Cytotoxicity sGenotoxicity ‘Biodistribution & Clearance time Metal accumulating plant for nano- ‘material leaching '*Prosopis juliflora: tor copper oxide NPs ‘sHoloptelia integrifolia: MnO2 NPs Carbon supported metal/oxide/ sufide/ ‘quentum dors *Hlectrocataysts, energy generation & storage, water purification, etc Novel compounds from biological ‘Low-volume, high-value drugs ike Artemisini, Vinblastine, Vneristine, Calanolide, Camptothecin, Azadirachtin, Picrosides, Gingoldes, etc ‘Flavor compounds: Fructose, Citric acd, Glutamic acid, ete. «Fragrance compounds: Vanilin, Santali, Macrocyctic musks, ec. “Antifungal/bacterial/vira/algal/insect compounds Prof, Absor Ahmed’s Grouproorganisms used in nanomaterials synthesis. Fungal spores are everywhere. Every breath we take is full of fungal spores. Most humans only succumb to fungal pathogens when immunocompromissed. Fungi are universally distributed in a variety of habitats where organic matter is present. Some fungi are aquatic, some are terrestrial while a variety of fungi infect plants, animals, including man. Some fungi show mutualistic association with algae and higher plants forming Lichen and mycorrhiza respectively Endophytic fungi present in terrestrial parts of vascular plants and anaerobic fungi live inside the rumen of herbivorous animals. Endophytic Fungi Actinomycetes Saprophytic fungi Plants are not $0 lucky Plant pathogenic fungi nays (corramut)Occurrence (Contd). Gilled Fungi Chanterelle with wrinkles Coral FungiMicroorganisms used in nanomaterials synthesis (contd). Underground fungi Trained Pigs dig them up Sell for hundreds of Dollars/IB ‘SMALL PUFFBALLS SPORULATINGDictyophora indusiata : Stinkhorn mushroom This very beautiful mushroom, has an unpleasant, decaying flesh odour which attract large number of flies. The spores of the mushroom stick on the legs and bodies of the fliesiand are widely dispersed. Meixner test: Blue color produced by extract from mushroom with concentrated hydrochloric acid, indicative of amatoxins (cause of mushroom poisoning)What ts ENDopHYTE ? te Endophytic fungi reside in a symbiotic fashion inside their host plants, mimic the chemistry of their respective host and interestingly, produce the same bioactive natural products or drugs and derivatives as their host plants; meanwhile causing no damage to them. In addition to this, endophytes provide protection from insect pests and from overgrazing by animals. Vinblastine & Vincristine producing endophytic fungus emerging from C. roseus leaves in between plant cells. (Magnification 400 times) Endophyte strands stained blue(cotton blue stain), growingIsolation of Endophytes from Catharanthus roseus Catharanthus roseus plant : leaves, stem and root Samples | Cut into small pieces(0.5x0.5 cm) Surface sterilized with 0.1% HgCl, solution (for 1 min.) Washed with sterile distilled water Small pieces put on PDA plate After 10-15 days, fungi were observed growing from pieces of leaves etc. Individual hypal tips of the various fungi were removed from PDA plate Fungal culture was checked for purity and transferred to PDA slants 52 endophytic fungal strains Isolated Screened for vinblastine and vincristineIsolation of Endophytes from Catharanthus roseus Catharanthus roseus Leaf pieces of Catharanthus roseus showing the emergence of endophytic fungi after 3 weeks.What is nanoscience / nanotechnology Greek word ‘nanos’, which means dwarf. 0 O00 0, pave One nanometer is thus one billionth of metre (or one millionth of millimetre, etc.). It can be expressed as 10°9 metres and shortened to nm.NANOSCIENCE AND TECHNOLOGY DEFINITIONS Nanoscience The study of phenomena and manipulation of materials at atomic, molecular and macromolecular scale, where properties differ significantly from those at a larger scale. Nanotechnology The design, characterization and production and application of structures, devices and systems by controlling shape and size at nanometer scale.Pioneers of Nanotechnology Richard Feynman 1969 — Delivered a talk “ There’s Plenty of Room at the Bottom” (Nobel Prize 1965). Eric. Drexler 1986- Wrote a book “ Engines of Creation” (Guru). Norio Taniguchi 1974- “ Used the term Nanotechnology”How small is a nanometer ? (and other small sizes) Start with a centimeter. Now divide it into 10 equal parts. ‘Now divide that into 10 equal parts. Now divide that into 100 equal parts. Now divide that into 10 equal parts. Finally divide that into 100 equal parts. Accentimeter is about the size of a bean. Fach partis a millimeter long, About the size ofa flea, Each part is 100 micrometers long. About the size (width) of a human hair, Each part is a micrometer long. About the size of a bacterin Each part is a 100 nanometers long. About the size of a virus. & Fach part is a nanometer. About the size of a few atoms or a small molecule.How Small Are Nanoscale Particles ? ei ny re eens ray es) Nee ac or - Na i) Nes coo ee ear ees Teele ee acd Buckminister or) x4 av ato curren) \Oxide surface h C.N. R. Rao, P. P. Edwards et al. Chem. Eur. J., 8 (1) 2002, 28Nano-sizing causes changes in :- Color Crystal shape eoble alelrie inti tidal Conductivity Magnetism Melting Points Chemical Reactivity Light Absorption | \y W gy 3S Silver nanoparticles of different sizes EASES Gold nanoparticles of different sizes Fluorescence from CdSe nanoparticles of different sizes (bandgap engineering)Nanostructures Dimension: a 5 5 : a fi ° Z g 5 oO n a ° oGOD’S OWN CREATION OF NANO MATERIALSAPPLICATIONS OF INORGANIC NANOPARTICLES @© = Functionalized NPs © = Dug molecule aoe As Targeting molecule Y= Receptor in tumor @ = Tumor in mice Fabrication of nanoparticles with drug molecules and targeting agent and applying in mice model.APPLICATIONS OF INORGANIC NANOPARTICLES » CATALYSIS (HOMOGENEOUS AND HETEROGENEOUS) » AS PRECURSORS FOR THE SYNTHESIS OF COATING FOR ELECTRONIC APPLICATIONS >» HYPERTHERMIA (TUMOUR DESTRUCTION VIA HEATING) >» SENSORS (BIOSENSORS, ENVIRONMENTAL SENSORS AND DETECTION OF POISONOUS SUBSTANCE IN SOIL) >» DRUG DELIVERY » DIAGNOSTICS >» QUANTUM DOTS AS LUMINESCENT PROBES IN BIOLOGICAL SYSTEMS » SEMICONDUCTOR TECHNOLOGY »CdS NANOPARTICLE AS A _ FLUOROSCENT LABEL IN FLUOROSCENCE MICROSCOPY > USED AS A SPECIFIC STAINING AGENT IN BIOLOGICAL ELECTRON MICROSCOPY SUCH AS SEM AND TEM.Bioinspired nanomaterials in Agriculture. Insact-repellent activity observed in Pulse beetle with our Gd,0, NPs NPs may increase plant growth and vigor # Insect Nanoparticles tested | Best Positive Lea 1. Mosquito larvae | Au, Ag, Gd,0,, TiO, Au, Ae 2. | Rice weevil ‘Au, Ag, Gd,0,, TIO, Gd,0,, TIO, 3, | Pulse beetle ‘Au, Ag, Gd;0y TiO; | 64,05 Ag 2, [Cotton bog Ag. G0, TIO, 64,0, Ti0, isles 5 TiO, S10, | Ag 64,04 TO, 6. Drosophila larvae Au, Ag, Gd0, TiO, ‘Ag G,0,,T10, Insect-repellent activity 7. Ticks ‘Au, Ag, Gd;0s, TiO, ‘Au, Ag, Gd,0s, TIO, ‘observed in Drosophila 8. Aphids Au, Ag, Gd,05, TiO; Au, Ag, Gd0s, TiO larvae with our TiO2 NPs List of some of our biofabricated NPs tested in insects contrUses of nanotechnology in Plant Sciences | & w Ag NPS Control ‘Nanotechnology ad te seruscaownto ven aos, ENHANCEMENT OF TILLERING GROWTH ra pore ‘rau acu = smaeeracam se fanaa ee ‘arama cy fans i Sanu ore eae ee Aino aut put re So Succ ortaoec oo archaeon aa ee ia cy Slide cout fr. Hara Sac, VT randMesoporous silica nanoparticles deliver DNA & chemicals into plants Surfacefunctionalized mesoporous silica nanoparticles (MSNs) are successfully used for the intracellular controlled release of genes and chemicals into plant cells. This will help in the farure investigations of plant genomics and gene function as well as improvement of crops Delivering DNA and their effector molecules into intact plant cells using mesoporous silica nanoparticles. a, A typical plant cell, illustrating the cell wall (cw), large central vacuole (v), got rei apo rxles (0h ‘mitochondria (m), chloroplasts (cp), endoplasmic reticulum (er), golgi (g)) within the cytoplasm (white region). b, After action of the gene gun, MSNs (small circles), carrying the small effector molecule (B-estradiol) within the gold-capped structure and externally coated with plasmid DNA, penetrate the cell wall and, in some cases, enter the cytoplasm. c, Within the cytoplasm, the plasmid DNA dissociates from the MSN surface and enters the nucleus (not shown). The MSNs are uncapped by incubating the plant cells with dithiothreitol (DTT). This releases the B-oestradiol effector molecules (orange hexagons) and activates expression of the plasmid DNA in the nucleus. Nature Nanotechnology,2,2007,272 David W. GalbraithComplex genetic, photothermal, and photoacoustic analysis of nanoparticle-plant interactions Nanotechnology ‘Tomato plants Detection impact Phaotermal — Meroar Regime PCR Singlewall carbon nanotubes Multiwall carbon nanotubes Physiological response Graphene eo ‘onto! medium graphene biomass, my Additions to medium Schematics of integrated genomic and photothermal-based analysis of nanoparticle-plant interaction. The right bottom shows the effect of carbon materials (activated carbon, few- layer graphene structures, and single- and multiwall CNTs) on biomass accumulation of tomato plants. Pseudocolor in example of photothermal leaf map on right top indicates signals from small CNT clusters (red- maximum signal, green low signals). Mariya et al. 2010Smart delivery of nanoencapsulated herbicide in the crop-weed environment Target site of underground parts of weed Encapsulated nanoherbicide Target plant root/stolon/ thizome/seed Nanoparticle targeting the specific receptors of weed plantsWhich sort of teeth appeal to you ? These ?? Or these ?? Cc» > |Applications of Nanotechnology in Dentistry ‘The main cause of endodontic disease is bacteria, Disinfection is presently achieved by cleaning the root canal system priog to obturation. Following setting, root canal filing is devoid of any antibacterial effect. Endodontic sealers (AH Plus, Epiphany, end GuttaFlow) with antimicrobial properties yet biocompatible may enhance root canal therapy. For this purpose, a range of Inorganic ianomaterials of different sizes, shapes and chemical compositions such as metals, ‘metal oxides, metal sulphides, quantum dots, bo minerals etc. are being used. The potential of ‘nanoparticles to control the formation of biofilms within the oral cavity, as a function of their biocidal, anti-adhesive and delivery capabilities, is now being further explored. Steps of biofilm formation: 1. Conditioning film formation: (0: adhesive; (B): bracket; (Or ligating with 2. Reversible adhesion; slastomerie ring: 3. Transition to irreversible (Dy: ligating with ste igature; (E):selfigating adhesion; ‘racket with the lip open 4. Corathesion of microorganisms (2 selfdigating brackot with a dosed dip; (G): to already adhering: arch wire; and (H): bonded retainer. 5 Growth into mature biofilm. Orthodontic biofilm, visualized by staining with GUM red cote, before (lower dentition) and after (opper dentition) removal of brackets. Stained areas can be clearly seen on the tooth surfices around the area where the brackets have been ‘bonded or around the brackets stl present.Role of Nanomaterials in Treatment of Eye Disorders Anatomy of the Eye Owing to their minute size (20-30 nm), these inorganic nanomaterials may easily penetrate into the corneal epithelium and effectively inhibit VEGF-induced angiogenesis in evi segment of the eye including anterior, middle (ciliary body and trabecular mesh work) and posterior segments and help in the treatment of corneal NV and various other eye disorders. Squint CataractNanodevices for delivery applications Protein coat Schematic representation of different nanodevices for delivery of pesticides, fertilizers or nucleic acids (a) adsorption on nanoparticle; (b) attachment on nanoparticle mediated by different ligands; (c) encapsulation in nanoparticulate polymeric shell; (4) entrapment in polymeric nanoparticle. (e) viral capsid Se Se droplet: oe os Nanosized pesticide Schematic representation of incorporation of nanoformulations in spray droplets to maximize efficient spraying. (a) spray droplets; (b) droplets containing nanoparticles loaded with pesticide; (c) droplets containing nanosized pesticides.Corporations active in nanotechnology BASF ASML Raytheon ExonMobil GD —_degussa. @ BREE Nec fmt SUMITOMO CHEMICAL <> CATERPILLAR” ee PHILIPS ea” —-=XEROX Microsoft PRAXAIR fn /~ oO Canon Unikover en ChevronTexaco e NOKIA wD) rufirsu PeG Nestle Fer THEBOCGROUP CERT ae oO sent onsen =e Testctae QSchematic outlining the various approaches for the synthesis of nanoparticles. Nanosynthesis routes — | ™ Physical Biological / / 4 \ \ x ~ Chemical Vapour condensation Biosynthesis/Biomimetic Biotemplates Sol-Gel Laser ablation Algae as DNA/RNA Co precipitation Photoirradation Diatoms Proteins Chemical etc Bacteria reduction Viruses Fungi Chemical pollens Oxidation Sponges S-Layers Amino acids PeptidesCHARACTERIZATION OF NANOMATERIALS UV-visible absorption spectroscopy (UV-vis) Fourier transform infrared spectroscopy (FTIR) X-ray diffraction (XRD) X-ray photoemission spectroscopy (XPS) Energy dispersive analysis of X-rays (EDAX) Transmission electron microscopy (TEM) Scanning electron microscopy (SEM) Selected area electron diffraction (SAED)Transmission Electron Microscopy for Nanotechnology Transmission electron microscopy (TEM) is one of the most powerful tools used for characterizing nanomaterials, and it is indispensable for nanotechnology. In recent years, a number of new and novel developments have been made in TEM for nanotechnology, such as; In situ microscopy for observing dynamic processes at nanoscale. . Nano measurement techniques that directly correlate physical properties with structures. . Holographic imaging of electric and magnetic fields. . Quantitative chemical mapping at sub-nanometer resolution Ultra-high resolution —_—imaging techniques. >D.BWilliams, C.B.Carter, Transmission Electron Microscopy, Plenum Press, New York 1996. >Z.L.Wang, Elastic and inelastic Scattering in Electron Diffraction and Imaging, Plenum Press, New York 1995 >L.Reimer, Transmission Electron Miscoscopy, 3" ed., Springer-Verlag, New York 1033Folvexsatal AnaelavesCharacterization by A] SAED and B] XRD A D= Diameter of the ring d= 2X3.59 2 Camera constant = A = 3.59 Sin 30° = 0.5 Sin 0°=0 Sin 15°= 0.2588 6 =30°0=15° Bragg’s Law: n ) = 2d sin 8 d I bkl 2.97 12 311 n=l deh = 1.54 2sin® 5x sin (15°) = 154 2X 0.2588 d= 2.97 A®PARTICLE SIZE CALCULATION BY XRD CENTER = 39.158 WIDTH = 0.99207 PARTICLE SIZE BY DEBYE-SCHERRER FORMULA = ni/B cosé B = WIDTH / 57.29 @ =CENTRE / 2 n=09 R= 154A PARTICLE SIZE = 0.9 X 1.54 / ( 0.99207/57.29) / cos( 39.158/2) = 0.9 X 1.54 X 57.29 / 0.99207 / cos(19.579) = 0.9 X 1.54 X 57.29 / 0.99207 / 0.745545 = 107.3559 A = 10.73559 nmCharacterization by XPS The photoelectron spectroscopy of the CdCI2 and SeCl4 reaction mixture was carried over a wide range of binding energies, i.e., 0 to 1400 eV, as shown in the Figure 6. 600 800 1000 1200 a=) Binding energy (eV) Core level binding energies and atomic percentages of various elements detected in the CdCl2 and SeCl4 reaction mixture. It is evident that Cd, Se, C, O, N, and Na are the Element Cis Ols Nis Cd 3dgy Se 2P. 1 rf 1 Atomic % WA 225 46 13 12 only prominent elements Binding energy [eV] 285.0 $31.1 309.7 4055 = 64g__—~Present in the reaction mixture (Fig.).Characterization by XPS...contd “ int (2.4) 3 ew ° Intent (a) ‘ag onary (¥) Being nery(¥) The core level binding energies and atomic percentages of various elements detected in the reaction mixture is given in Table I. The Cd 4f spectrum could be decomposed into two spin-orbit components | and 2 (spin-orbit splitting ~6 eV). The Cd 3d5/2 and 343/2 peaks occurred at a binding energy of 406 eV and 412 eV, respectively, agree with the core level binding energies reported earlierl8 and are characteristic of metallic Cd (Fig A.). The Se 3d peak occurred at a binding energy of 54.8 eV, agrees with the core level binding energies reported earlier18 and is characteristic of metallic Se (Fig.B).WHY BIOSYNTHETIC APPROACH TO NANOMATERIAL SYNTHESIS ? > Physical (vapour deposition, lithography, radiolysis) and Chemical (reduction by borohydrate or citrate) methods of nanoparticles synthesis are Eco-unfriendly, Cumbersome and yield big particles. > Need for reliable Ecofriendly and Economic synthesis of nanomaterials. hk Strategy > Screening of fungi and actinomycetes (more than 200 cultures for metal salt reduction). > Fungi and actinomycetes have also been found to carry out oxidation reduction reactions. Care | : a fi mate J. Molecular Catalysis A: Chemical 181: 2003, 237 cL J, Molecular Catalysis B: Enzymatic 27,2004,61 . 4 = U. S. Patent No.6,559,322, 2003 Verticlium sp. F. oxysporum Indian Patent application no 1304/Del/2001Vv Vv Vv Advantages Of Biological Synthesis Of Nanoparticles Protein capped water dispersible nanoparticles Functionalization Biomilling Selective Bioleaching Monodispersity Size and Shape VariationBenefits of Biological Synthesis over Chemical and Physical protocols. India has a range of rich untapped areas which are home to a vast variety of rare and exotic medicinal and aromatic plants, endophytic fungi, mushrooms, algae, etc. These are an excellent source to work upon and should be explored for the bio-synthesis of a range of bio-compatible inorganic nanomaterials, the fabrication of which is very difficult via chemical and physical methods. The bio- compatible nanomaterials will find use in a number of applications listed as follows: a] Energy storage, production and conversion b] Agricultural productivity enhancement c] Food processing and storage d] Vector and pest - detection and control e] Conversion of toxic environmental contaminants into their non-toxic forms f] Nutritional supplements, foods and drinks. g] Delivery of DNA and chemicals into plants h] Anti-microbial and anti-insect activity i] Smart Treatment - delivery system in plants j] Smart Delivery of herbicides and fungicides, ete.Intracellular biosynthesis of inorganic nanoparticles (Au and Ag) using Endophytic fungus, Verticillium sp. and Alkalotolerant Actinomycete, Rhodococcus sp. > Mycelial (cell) mass, separated from culture broth, is suspended in sterile water along with aq. Solution of metal salt ( HAuCl, and AgNO, ) under shaking at ambient conditions. > After few hours, the fungal mycelial (cell) mass develops distinct characteristic colour & solution remains colourless after filtration. > Biomass-nanoparticle composite is characterized by XRD, UV-Vis, SEM, EDAX, TEM and SAED. > These nanoparticles are suitable for bio-inorganic composite film, in catalysis and other electronic applications. J. Nanoscience and Nanotechnology, 17, 2017, 1787. J. Nanoscience and Nanotechnology, 14, 2014, 1 Materials Letters 79,2012, 116. Angew. Chem. Int. Ed. Engl.40, 2001, 3585 Nano. Lett. 1, 2001, 515 Nanotechnology, 14, 2003, 824 J. Biomed, Nanotechnol 1,2005, 47 U.S. Patent No. 6,537, 344, 2003Green Synthesis of ‘Metal’ Nanoparticles: Intracellular Gold NPs SEM analysis of Verticillium- ‘TEM analysis of ‘carbon ‘Au Nanoparticles supported’ Nanoparticles Yeawdergth (rf 200) ‘Materiale Letters 129, 2014, 159, Materials Letters 79, 2012, 136, Angew. Chem. int. Ed. Engl.0, 2001, 3585 hon ba.5, 2, 5 UV-Vis / XRD analysis of Verticillium - Au nanoparticlesTransmission electron microscopy (TEM) studies of ultra thin section of Au nano Verticillium mycelial mass Asmall portion of the mycelial mass which showed the formation of gold nano particles was taken and fixed in 2.5 % gluteraldehyde for 24 h at R. T. After fixation, mycelial mass was centrifuged, washed several times in D. W. Mycelial mass was dehydrated with 30, 50, 70, 90% and finally with absolute ethanol After dehydration mycelial mass was kept in propylene oxide and finally kept in 1:1 mixture of propylene oxide and Epon 812 and kept over night at R. T. Then kept for embedding in Epon 812 + doddecynyl succunie anhydride (DDSA) + methyl nadie anhydride (MNA) in the ratio of 1:1:5 and added two drops of tridimethyl amino methyl phenol (DMP3o0) to accelerate polymerization process and kept three days at 60 °C Ultrathin sections were cut on utramicrotome and were taken on copper TEM grids | Sections were stained with uranyl acetate and lead citrate prior to TEM analysisTEM OF THIN SECTIONS OF VERTICILLIUM - Au AND RHODOCOCCUS- Au NANOPARTICLES Verticillium sp. Rhodococcus sp. PROBABLE MECHANISM OF INTRACELLULAR BIOSYNTHESIS > Nanoparticles are formed on the inner surface of the fungal /actinomycete cell and not in solution. > Trapping of ions on the surface of fungal/ actinomycete cell. » lons are reduced by enzyme (reductases) present in the cell wall or in the cytoplasmic membrane.Extracellular Biosynthesis of Inorganic(Au,Ag and CdS) Nanoparticles Using Plant pathogenic fungus Fusarium oxysporum and Extremophilic actinomycete, Thermomonospora sp. = Mycelial (cell) mass, separated from culture broth, is suspended in sterile water along with aq. Solution of metal salt (HAuCI,, AgNO, Cdso,) under shaking at ambient conditions. . + After few hours, the fungal mycellial (cell) mass develops distinct characteristic colour. Biomass remains colourless after filtration Solution containing metal nanoparticle is then characterized by UV-Vis, FT-IR, TEM, SAED. These nanoparticles are suitable for homogenous catalysis and can be immobilized in different matrices or in thin film form for optoelectronic and other electronic application. Curr. Top. Med. Chem. , 16, 2016, 2036. Langmuir, 19, 2003, 3550. Chem Bio Chem, 3, 2002, 461 (Au nanoparticles).Green Synthesis of ‘Metal’ Nanoparticles: Extracellular Gold NPs Before Upon filtration, aqueous solution of the control flask was colorless. After Upon ftration, ‘aqueous solution of treated flask showed ruby red color. TEM micrograph of a drop coated film of Au nanoparticles synthesized extracellularly UV-Vis spectra recorded as a function of time of reaction of a 10-M aqueous solution of HAUCI, Selected area _ electron diffraction pattern (SAED) of the nanoparticles shown in TEM micrograph Langmuir, 19, 2003, 3550 ChemBioChem 3, 2002, 461 Colloids and Surfaces 8 97,2012, 27Green Synthesis and Biodistribution of ‘Metal’ Nanoparticles: Extracellular Gold NPs, continued. “econ meee Cell viability assay of gold nanoparticles against NIH3T3 mouse embryonic fibroblast cell line and MDA-MB-231 human breast carcinoma cell line. The data represented in the form of a bar graph and plotted using means + SE. of triplicate determinations. The values were analyzed by Student's t-test (p < 0.005). Statistical analysis, P values for significantly different means, *P< 0.005 and **P> 0.05 vs control. Gamma scintigraphic image of biodistribution of Gold ‘nanoparticles in rat. Intensity soo 30 ~ * 2 M 2 a 2 a Mallat, ~ = 200 360 400 500 600 700 800 4 50 «60D Wavelength (nm) 26 (Degree) TEM micrograph SAED pattern o eae paok 4 yg ne q B os 2 5 \ i Foal \ aod s NN = 09 Sto —xa0 65000 ae ae Wavelength (am) 20 (Degree) Alga 2 TEM micrographs UV-vis spectra of Au 10 Tatensiy 200-40 Com) Wavelength (nm)Green Synthesis of ‘Bi-Metallic’ Nanoparticles: Extracellular Au-Ag Alloy NPs 1mM HAuCl, 1mM AgNO, A BP ce The gradual change of color from purple to deep a pues u brown jis the clear indicative of the formation of . bimetallic Au-Ag alloy nanoparticles | 443nm i 470nm ~ |Ioee 500nm st 543nm A ixm ——+ UV-Vis spectra of Au-Ag alloy nanopartilces with respect to time TEM micrograph of Au- Ag all nanoparticles of 6-14 nm Small 1:2005,517Green Synthesis of ‘Metal’ Nanoparticles: Extracellular Platinum NPs A ® Betoe Aer ‘Wavelength (wm) Chloroplatinic eilieed UV-vis spe TEM micrograph/SAED pattern & Particle size distribution Platinum NPs z 3 a 2 3| Zi s & 2 z a s 4 a o 18001600. 17001800 20 (Degree) ‘Wave number (em) XRD pattern X-ray photoelectron spectra FTIR spectrum Colloids and Surfaces 8: Biointerfaces 97 (2012) 27-31Green Synthesis of ‘Semi-conductor’ Nanoparticles: Extracellular CdS NPs Uponfiltration, Upon filtration, ‘aqueous solution of aqueous solution of ‘the control flask was treated flask Slee onl le TEM mi XRD of a drop coated film of CdS nanoparticles a synthesized extracellularly UV-Vis _spectra_recorded from the ‘aqueous 10-3 M CdSO4 solution as a function of time (in days) of addition of the fungal biomass. Absorbance (a.u) Current Nanoscience, 10, 2014, 588. J. Am. Chem. Soe. 124 ,2002, 12108. Journal of Biomedical 200 300400600600 700 Nanotechnology 3: 2007,190. Wavelength (nm) {Us Patent No.6, 783,963, 2004) Indian patent application no. 412/Del/2002,Green Synthesis of ‘Quantum Dots’ Nanoparticles: Extracellular highly fluorescent CdTe NPs Babi 2065 Powerbar tas) —— iiereyiau) of CdTe inhibition zones on nanoparticle exhibited in term: exposure to various bacteria. P a1n Synthesis of ‘Oxide’ Nanoparticles: Extracellular fluorescent CuAlO, Nanoparticles Fungus does what chemistry finds hard to do !!! Humicola sp. 208 AmM Cu(NO,),.3H,0 + ImM Al(NO3),.9H,0 Cualo, To synthesize CuAlO,, controlling the valence state of Cu to 1+ is the key ‘TEM image of fun CuAlo, (CUAL, NP in water . o 7o| RF one 80 Wats) o. ©. ee eee) oe a0 982 oda ade ote Tine (Sec) Binding Energy (@V) X-ray photoelectron spectra showing ——— ©uitation and luminescence _the presence of the Cu state in the (ual Wes BE spectra for CuAlO2 NPs. sample ‘Adv. Mater. 19: 2007, 3295