Home Page Researchers Oded Shoseyov
Institute of Plant Science and Genetics in Agriculture
The Faculty of Agriculture, Food and Environmental Science
The Hebrew University, Rehovot, Israel
Tel: +972-8-9489084; Fax: +972-8-9462283
Genetic engineering of self-assembled proteins
Oded Shoseyov research will focus on two areas. The first one is using biopolymers, such as cellulose and silks for textile, construction and medical applications. In recent years, due to climate changes and declining oil supply, synthetic materials are becoming more and more unfavorable, thereby enhancing the need to find green alternatives. In this research, we describe the fabrication of Nano Crystalline cellulose (NCC), cellulose-silk bio-nanocomposite comprised of NCC and recombinant spider silk protein fused to a cellulose binding domain (CBD). Silk-CBD successfully bound cellulose and unlike recombinant silk alone, self-assembled to form micro-fibers even in the absence of NCC.
The second area of our research related to the Carbon Nanotubes (CNT)-SP1 composite. CNT have stimulated research due to their wide range of applications. However, their existence as aggregates and the difficulty in debundling and dispersion makes them poorly soluble and limit the improvement of properties when used as fillers. Many techniques have been employed to obtain such dispersions including mechanical, ultrasonic and solution mixing resulting in limited effect. Attaching a protein moiety such as SP1 showed promising results. SP1 is a thermally stable protein, originally isolated from Poplar trees, which self assembles to an extremely stable 11 nm ring-shape dodecamer. SP1 is stable under extreme conditions (high temp, high pH and detergents), thus allowing inexpensive production for a variety of industrial application. Linkage of CNT to specific peptides on SP1 N-terminus by genetic engineering resulted in 12 CNT binding sites per ring. This enabled the creation of SP1 variants which tightly bind to carbon nanotubes to form a stable SP1/CNT complex.
Production of Nano Crystalline Cellulose (NCC) nano-structured foams
NCC was produced by H2SO4 hydrolysis of 200?m Micro Crystalline Cellulose (MCC). The process involved suspension of the MCC powder in water, hydrolysis in controlled temperatures and acid concentration, washing cycles in water followed by sonication until a clear liquid crystal suspension was achieved (Figure 1A). A sample of the final product was mounted onto thin carbon supports and examined by TEM (Figure 1B). NCC foams were produced by casting the suspensions into aluminum or Teflon molds followed by freezing and lyophilization yielding highly porous foams (Figure 1C). SEMs of the NCC foams revealed networks of smooth nanopaper sheets stacked upon each other with high order (Figure 1D).
Figure 1: Nano structured foams produced from NCC liquid crystal suspensions. (A) clear NCC 2.5% suspension. (B) TEM image the suspension showing nano rods at dimensions of 10-20 nm width and 200-400 nm length. (C) lightweight porous foam produced by freezing and lyophilization of the suspension. (D) SEM image of the foam internal structure showing its arrangement in ?puffed pastry? laminated sheets nano structures.
Production of composite silk/silk-CBD and NCC sponges
Recently we produced silk/silk-CBD and NCC sponges. First, purified, concentrated spider silk protein was mixed with NCC solution and then sonicated. This procedure has a double effect; in addition to homogeneous dispersion of NCC within the polymeric matrix, the sonication process induces rapid gelation of spider silk proteins by accelerating formation of physical cross-links, such as initial chain interactions related to ?-sheet formation. After sonication, three dimensional cellulose NCC-spider silk porous matrices were generated using the freeze drying method. SEM pictures of the resulting sponges clearly showed that pore architecture and alignment differed between the silk-CBD and control silk sponges (Figure 2). Silk sponges included nonoriented, 30-100 µm pores (Figure 2C) of irregular shape, much like the NCC-silk composites (Figure 2D). Silk-CBD sponges featured 300-500 µm leaf-shaped pores arranged in a relatively consistent direction (Figure 2E). Similar characteristics were observed in sponges formed of native silkworm silk produced under the conditions applied here and were said to result from parallel arrangement of silk fibroin crystal flakes. The composite NCC-silk-CBD sponge bore ~100 µm structurally aligned pores (Figure 2F, G, H).
Figure 2: SEM pictures of silk, silk-CBD, or composite silk-NCC sponges. A-B: NCC sponge, C-100% silk sponge, D- silk-NCC composite sponge, E-100% silk-CBD sponge, F-H- silk-CBD-NCC composite sponge on an expanded scale. Pictures scale bar: A,C, E 1 mm; B,D,E 200 µm; G,H 100 µm
Mechanical and Thermal Properties of CNT / SP1 Nano Fillers
Our lab and Fulcrum Ltd. linkage carbon nanotube (CNT) to specific peptides on SP1 N-terminus by genetic engineering resulted in 12 CNT binding sites per ring. This enabled the creation of SP1 variants which tightly bind to carbon nanotubes to form a stable SP1/CNT complex.
It was demonstrated that the SP1/CNT complex prevents CNT aggregation and allows its homogenous mixing in water. In order to obtain homogenous CNTs in a polymer matrix, the dehydrated complex was complex easily re-dispersed in epoxy resin or polyurethane resin. The SP1/CNT is covalently bound to epoxy groups prior to polymerization with the curing agent. Dispersion and uniformity were improved by using speed-mixer and a 3 wheel roller.
Table 1 summarizes the results of the bulk properties of the adhesive epoxy samples with and without the CNT/SP1 nano-composite. Two references were tested: a neat adhesive and an adhesive with SP1 protein without CNT. The results are presented following two ways of dispersion: Speed-mixing or mill-rolling + speed mixing. Two concentrations were measured: 0.35% and 0.7% wt.
Table 1: Bulk mechanical results with and without CNT/SP1 and effect of different mixing methods.
The table shows slight improvement in elastic modulus and hardness on the expense of tensile strength and elongation. Mill rolling compared to mixing gives better results.
Table 2 summarizes the results of shear, peel and tensile adhesion strength for the epoxy adhesive with and without nano-filler. Three kinds of samples were tested: single lap shear (SLS), T-Peel, and tensile adhesion (Butt-Joint). Each kind of sample had 5 duplicates. All tests were produced at room temperature.
Table 2: Shear, T-peel and tensile adhesion strengths and failure mode of the epoxy adhesive with and without nano-filler on Al adherends.
A second reference used for shear test was epoxy 828/140 with SP1 (without CNT). This reference gave the result of 16.1 ± 1.5 MPa. This shows that the addition of protein SP1 into the epoxy causes some improvement of the adhesion strength, but the major improvement is caused by the CNTs.
Specific research topics related to Nanoscience and Nanotechnology:
- SP1 as A Novel Scaffold Building Block for Self-Assembly Nanofabrication
- Novel nano composites of polymeric proteins-cellulose whiskers for stem cell-based spine therapy.
- Recombinant Exon-Encoded Resilins for Elastomeric Biomaterials
- Production of recombinant resilin nanocomposites in transgenic plants.
- The production of spider silk fibers and their uses.
- Core shell biocomposite fibers for medical applications.
List of publications in Nanoscience and Nanotechnology (2011-2012)
- Qin G, Rivkin A, Lapidot S, Hua X, Preis I, Arinus S.B, Dgany O, Shoseyov O and Kaplan D L. (2011). Recombinant exon-encoded resilins for elastomeric biomaterials. Biomaterials, 32, 35, 9231-9243.
- Shani N, Shani Z, Shoseyov O, Mruwat R and Shoseyov D. (2011). Oxidized Cellulose Binding to Allergens with a Carbohydrate-Binding Module Attenuates Allergic Reactions. J. Immunol. 86, 1240-1247.
- Qin LX , Yang L, Li DW , Jing C, Chen BQ, Ma W, Heyman A, Shoseyov O, Willner I, Tian H, and Long TL. (2011). Electrodeposition of Single-Metal Nanoparticles on Stable Protein 1Membranes: Application of Plasmonic Sensing by Single Nanoparticles. Angew. Chem. Int. Ed.; 50, 1? 6.
- Lapidot S, Meirovitch S, Sharon S, Heyman A, Kaplan D L, and Shoseyov O. (2012). Clues for biomimetics from natural composite materials. Nanomedicine. 7(9), 1-15.
Published articles resulting from cooperation between the universities:
- Frasconi M, Heyman A, Medalsy I, Porath D, Mazzei F, and Shoseyov O. (2011). Wiring of Redox Enzymes on Three Dimensional Self-Assembled Molecular Scaffold. Langmuir, 27, 20, 12606?12613.
- 2. Khoutorsky A, Heyman A, Shoseyov O and Spira M.E. (2011). Formation of Hydrophilic Nanochannels in the Membrane of Living Cells by the Ringlike Stable Protein-SP1. NanoLetters, 11, 2901-2904.
- Medalsy I, Klein M, Heyman A, Shoseyov O, Remacle F, Levine R D, Porath D. Logicimplementations using a single nanoparticle-protein hybrid. (2010). Nature Nanotechnology. 5, 451-457. Impact factor of 26.31(from Google Scholar).
- Heyman A, Medalsy I, Bet Or O, Dgany O, Gottleib M ,Porath D and Shoseyov O. (2009). Protein Scaffold Engineering Towards Tunable Surface Attachment. AngewandteChemie- International Edition. 121(49), 9454? 9458. Impact factor of 12.730 (from Google Scholar).
- Medalsy I, Dgany O, Sowwan M, Cohen H, Yukashevska A, Wolf SG, Wolf A, Koster A, Almog O, Marton I, Pouny Y, Altman A, Shoseyov O, Porath D. (2008) SP1 protein-based nanostructures and arrays. Nano Lett. 8(2): 473-477. Impact factor of 9.99 (from Google Scholar).
- Heyman A, Levy I S, Altman A C and Shoseyov O. (2007). SP1 as a novel scaffold building block for self-assembly nanofabrication of submicron enzymatic structures. Nano Lett. 7(6), 1575-1579. Impact factor of 9.99 (from Google Scholar).
- Shoseyov O, Shani Z. and Levy I. Carbohydrate Binding Modules: Biochemical Properties and Novel Applications. Microbiol. Mol. Biol. Rev. (2006) 70(2), 283?295. Impact factor of 15.864 (from Google Scholar).
New patents and patents utilization (2011-2012):
Applied Patents: (represented patents families)
- Shoseyov O and Spira M. SP1 to electrically couple cells and sensing pads and thereby facilitate the assembly of neuroelectronic hybrid systems. PCT/IL2012/050188.
- Lapidot S, Meirovitch S, and Shoseyov O. CBD-spider silk protein. PCT/IL2012/000058.
- Lapidot S, Rivkin A, and Shoseyov O. Modified resilline as a bioadhesive. US patent 61/530,167.
- Paltiel Y and Shoseyov O. Green House - Use bio materials made from cellulose as a building material for biodegradable light spectrum tunable plastic sheets for walk-in tunnels and greenhouse. US patent 61/607,185.
- Lapidot S and Shoseyov O. Utilization of Nano Crystalline Cellulose for fuel and explosives. Israel patent 219814.
- Shoseyov O. Production of recombinant glycoproteins with reduced immunogenicity in plants. US patent 61/584,920.
- Carmel Goren L, Gustafsson T, Lapidot S, and Shoseyov O. Nano Crystalline Cellulose as additive to cement and gypsum mixtures. US patent 61/643,696.
- Shani N, Shani Z, and Shoseyov O. Oxidized cellulose based protection against airborne pathogens. US patent 61/655,038.
Cooperation with industries and defense projects (2011-2012):
- 2007-2012: DFG. Assembly of SP1-metal/semiconductor nano particles arrays. Oded Shoseyov (projects performer). SP1 is a boiling stable stress induced protein that self-assemble to an extremely stable dodecamer isolated from poplar tree. The biological role of SP1 is still under investigation. Nevertheless, the unique physical characteristics of this self-assembled nano-ring shaped particle enable us to utilize it in the assembly of nano-bioreactors, electronic devices and incorporation of carbon-nanotubes into epoxy resins, carbon and Kevlar fabrics.
- 2009-2012: "TASHTIOT" Ministry of Science. Bio-composites for regenerative medicine. Oded Shoseyov (projects performer). In this project we propose to design unique injectable gels for stem cell-based spine therapy, which will be composed of NCC, resilin and collagen. The nano structure of the cellulose whiskers will allow us to produce injectable gels with controllable biomechanical properties that can sustain the different loads in the spine, according to the therapeutic application.
- 2010-2013: MATERA, An ERA-NET+ financed by FP7. COLCOMP: core shell biocomposite fibers for medical applications. Oded Shoseyov (projects coordinator). This project aim is to design novel composite surgical hernia meshes with a special designed biocompatible formulation and surface layer composed of: biopolymer material like PLA polymer, Nano Cellulose Crystals (NCC) and human recombinant collagen (rhCollagen). This innovative approach will answer the need for a biocompatibilised and bioresorbable textile monofilament product with high mechanical strength that can be used to develop novel hernia mesh structures with high regenerative capability which have become a real need due to the long time failure and complications that are associated with the synthetic meshes that are currently in use.
- 2011-2014: NMP FP7. Green nanomesh: Targeting Hernia Operation Using Sustainable Resources and Green Nanotechnologies. Oded Shoseyov (projects performer). Herein, we propose a novel approach that employs sustainable raw materials and recent advances in green nanotechnology to fabricate a Green Nano Mesh prototype for hernia repair that not only will eliminate toxic chemicals from the manufacturing processes, but will also enhance functional repair of injured or degenerated tissues due to superior inherent biological properties. Specifically, we aim to fabricate a green nano-fibrous mesh with well-defined nano-topography using: (a) cellulose nano-crystals as a reinforcement; (b) human recombinant collagen, derived from transgenic tobacco plants as opposed to acid extracted animal collagen that harbours risks of interspecies transmission of disease; and (c) biodegradable polyesters (e.g. polylactic/polyglycolic acid or poly-?-caprolactone) as opposed to non-sustainable and non-degradable plastics such as polypropylene, polytetrafluoroethylene and nylon.The green credentials of this innovative approach lie in the use of sustainable eco-friendly raw materials that will produce biodegradable waste products, therefore replacing hazardous chemicals currently in use. Thus, this proposal directly aligns with the European Commission commitment for a low carbon economy and fits the call for the substitution of hazardous materials, components and processes using green nano-technology.
- 2011-2014: Marie Curie IAPP. ?Tendon Regeneration?: Targeting Functional Tendon Regeneration Using a Loaded Biomimetic Scaffold. Oded Shoseyov (projects performer).In this project, we anticipate to produce the first bio mimetic three-dimensional fibrous composite of collagen-resilin scaffold to match the properties (e.g. structural, mechanical) of native tendons and to facilitate the incorporation of bioactive or therapeutic molecules (e.g. drugs, genes, growth factors, respectively).
- 2012-2015: Academia Sinica, Taiwan. The Mechanics of Solid Foam with Nanocomposite. Oded Shoseyov (projects performer). In this proposal, we will devise solid foam with well-defined microscopic structures such as foam with body-centered-cubic structure and with face-centered-cubic- structure which can be tested both theoretically and experimentally. The results will give us understanding on the microscopic picture on how the structure deforms under macroscopic force. In addition, we will engineer the constituent materials with nanocomponent such as nanocrystalline cellulose which will enhance its mechanical properties such as elasticity, anisotropy, and so on. Our work covers multi-length scale will provide comprehensive understanding and insight to the important but understudied subject.
- 2012-2016: Proposal for Focal Technology Area (FTA). Hybrid Nanomaterials and Formulations for Functional Coatings and Printed Devices. Oded Shoseyov (projects coordinator). In this proposal we will focus on developing new hybrid nanomaterials (NM) and their formulations and demonstrating their applicability to industrial products in two closely linked areas:
- Functional coatings: Smart paints with self-assembly properties for application on different surfaces offering functionality beyond color, such as light spectrum converters for home and greenhouses, thermo-solar paints for high temperatures, antibacterial and anti-fouling coatings for walls and food packaging, impact resistant reflective coating for mirrors and anti-corrosion coatings.
- Printed devices: Materials and inks for electronic and optical devices printed on paper and plastics offering low cost flexible products such as RFID tags, fluorescent signage, electrodes for plastic solar cells and printed transistors.
- Omer Terapiotics Ltd (2011) is involved in developing of a preventive treatment for allergic disorders. The core technology of the company is based on the research conducted by me. In my studies I identified a polysaccharide, Oxidized Cellulose (OC), with high allergen-binding capacities.
- Paulee Cleantec Ltd (2012) has developed the first device in the world which removes and does away with dog waste in real time at the location where it occurred; all this without human contact or environmental pollution.
Cooperation with other universities in Israel:
Within Hebrew University:
- Proposal for Focal Technology Area (FTA): Uri Banin, Shlomo Magdassi, Danny Porath, Daniel Mandler, David Avnir, Shlomo Yitzchaik, Yossi Paltiel, Itamar Willner, Oded Millo, Meital Reches, Roy Shenhar, and Uri Raviv. Hybrid Nanomaterials and Formulations for Functional Coatings and Printed Devices.
- Itamar Willner. Institute of Chemistry; Electrochemistry of SP1.
- Danny Porath. Institute of Chemistry and Center for Nanoscience and Nanotechnology. Nanoelectronics of SP1 scaffolds.
- Micha E. Spira. Department of Neurobiology, the Life Science Institute, Faculty of Science. SP1 as a nanopore between a damaged neuron and a nursing cell.
- Yossi Paltiel. Nano materials and nano-optics.
- Dan Gazit. Novel nano composites of polymeric proteins-cellulose whiskers for stem cell-based spine therapy.
With other universities:
- Prof. Gil Markovich. Tel-Aviv University. Nanoparticles and SP1 arrays.
- Prof. Gil Navon. Tel-Aviv University. Novel nano composites of polymeric proteins-cellulose whiskers for stem cell-based spine therapy.
- Prof. Yael Hanein. Tel-Aviv University. Development of an artificial retina using SP1 arrays.
Students, postdocs and researchers:
Staff scientists: Rachel Algom, Mara Dekel, Dr. Levava Roiz, Dr. Patricia Smirnof, and Dr. Arnon Heyman.
Junior scientists/post-docs: Dr. Sigal Sharon, Dr. Shaul Lapidot, and Dr. Lena Magrisso.
Ph.D. students: Myron Abramson, Liron Nutman, Dorit Levy, Itan Price, Amit Rivkin, Amit Yaari, Yuval Nevo, and Tsvika Zvirin.
M.Sc students: Daniela Zecharia, Shira Bella Arinos, Tsvika Shtein, and Iftach Birger.
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