WO2017109794A1 - Dispositif photovoltaïque à base de protéines - Google Patents
Dispositif photovoltaïque à base de protéines Download PDFInfo
- Publication number
- WO2017109794A1 WO2017109794A1 PCT/IN2016/050450 IN2016050450W WO2017109794A1 WO 2017109794 A1 WO2017109794 A1 WO 2017109794A1 IN 2016050450 W IN2016050450 W IN 2016050450W WO 2017109794 A1 WO2017109794 A1 WO 2017109794A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- photosynthetic
- protein
- photovoltaic device
- based photovoltaic
- protein based
- Prior art date
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Classifications
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- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/24—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- C07K14/265—Enterobacter (G)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12N1/205—Bacterial isolates
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- H—ELECTRICITY
- H10—SEMICONDUCTOR DEVICES; ELECTRIC SOLID-STATE DEVICES NOT OTHERWISE PROVIDED FOR
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- H10K30/00—Organic devices sensitive to infrared radiation, light, electromagnetic radiation of shorter wavelength or corpuscular radiation
- H10K30/451—Organic devices sensitive to infrared radiation, light, electromagnetic radiation of shorter wavelength or corpuscular radiation comprising a metal-semiconductor-metal [m-s-m] structure
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Definitions
- the present invention provides protein based photovoltaic device.
- the invention describes embedding a microbial photosynthetic protein pigment complex between the two electrodes.
- the said photosynthetic protein pigment complex is isolated from a novel photosynthetic bacterium Enterobacter sp. MTCC 5804.
- the said device produces high photocurrent when the photosynthetic protein pigment complex is layered on a semiconducting material comprising of titanium oxide and zinc oxide in the said device set up coated on transparent conductive oxide ( ⁇ /FTO) and platinum as cathode.
- RC photosynthetic reaction centre
- the protein subunits organize the redox cofactors which include primary electron donor comprising of a special pair or dimer of bacteriochlorophyll termed as P or B 8 75 or B 8 9o (B refers to "Bulk pigment” and the number refers to the wavelength of absorption peak in the near infrared region), two accessory monomer bacteriochlorophylls, two bacteriopheophytins, and two quinones electron acceptors.
- P or B 8 75 or B 8 9o B refers to "Bulk pigment” and the number refers to the wavelength of absorption peak in the near infrared region
- B refers to "Bulk pigment” and the number refers to the wavelength of absorption peak in the near infrared region
- two accessory monomer bacteriochlorophylls two bacteriopheophytins
- two quinones electron acceptors two quinones electron acceptors.
- the critical aspect of the photochemistry of reaction centers is their ability to perform
- Bacterial RC such as Langmuir Blodgett films, Adsorption, Reconstitution into Liposomal Lipid Bilayers, Bilayer Lipid Membranes (BLMs) for conceivable applications such as photoelectronic molecular devices, wet- type photo cells, herbicide sensors etc.
- FTO fluorine-doped tin-oxide
- a electrochemical solar cell comprising of reaction center from R. sphaeroids along with redox species such as cytochrome, ferrocene, Diaminodurene, PMS, DCPIP, ferrocyanide, quinones, MV, BV and the cathode and anode comprising of platinum, silver, gold, indium tin oxide, tungsten oxide, tin oxide and a variety of semiconductor organic and inorganic materials.
- redox species such as cytochrome, ferrocene, Diaminodurene, PMS, DCPIP, ferrocyanide, quinones, MV, BV and the cathode and anode comprising of platinum, silver, gold, indium tin oxide, tungsten oxide, tin oxide and a variety of semiconductor organic and inorganic materials.
- redox species such as cytochrome, ferrocene, Diaminodurene, PMS, DCPIP, ferrocyanide, quinones, MV,
- Pizziconi have claimed a nanoengineered biophotonic hybrid device using chlorosomes of Chloroflexus aurantiacus ATCC 29366 in their patent application US 2009/0090410 Al .
- Birge et al have claimed a method of preparing a protein based photovoltaic device in which the genetic modification of the bacteriorhodopsin isolated from Halobacterium salinarum was layered via electrodeposition or by electrostatic layer by layer placement.
- the technical advancement that the present invention offers over the existing state of art is an efficient, simple and inexpensive wet or dry protein based photovoltaic device.
- the present invention offer the following advantages: - a) A simple low cost process for cultivation is involved in growth of Enterobacter sp. MTCC 5804 and production of photosynthetic protein or RC.
- the isolated RC from Enterobacter sp. MTCC 5804 is capable of physically absorbing and orienting on the surface in a way that its natural functionality and spatial organization of harvesting the light is preserved with efficient electron transfer.
- MTCC 5804 is capable of generating an open circuit photo voltage, a photocurrent density and an efficiency of not less than 500 mV, 500 ⁇ /cm 2 and 0.1% respectively.
- the main objective of the invention is to provide protein based photovoltaic device.
- Still yet another objective of the present invention is the incorporation of the photosynthetic protein pigment complex in the said device, which is isolated from a novel Enterobacter sp. MTCC 5804 without any genetic modification and using low cost cultivation, extraction methods that reduces the overall cost of the device.
- Still another objective of the invention is configurating device using photosynthetic RC Enterobacter sp. MTCC 5804 capable of generating an open circuit photo voltage, a photocurrent density and an efficiency of not less than 500 mV, 500 ⁇ /cm 2 and 0.1% respectively.
- the present invention provides an efficient, simple and inexpensive wet or dry protein based photovoltaic device employing photosynthetic protein pigment complexes isolated from anovel photosynthetic bacteria namely Enterobacter sp. MTCC 5804.
- the said invention uses a photosynthetic RC which exhibits high photovoltaic efficiency in a photovoltaic device set up without any genetic modification.
- the photosynthetic RC self-orients on the semiconductor layer in such a way that its functionality is preserved resulting in high efficiency.
- the present invention provides a Photovoltaic device comprising of extracted, stabilized and partially purified photosynthetic protein pigment complex isolated from a novel photosynthetic bacterium Enterobacter sp. MTCC 5804 deposited on a semiconductor material, a cathode; an anode, an electrolyte system which may be solid/liquid or inorganic/organic in nature and a support.
- the photosynthetic protein pigment complex may be extracted from Enterobacter sp. MTCC 5804 by sodium dodecyl sulphate, sodium cholate, sodium deoxycholate, CHAPS, Triton X-100, Tween 20, Tween 80, Lauryl dimethyl amine oxide (LDAO).
- the photosynthetic protein pigment complex is extracted from Enterobacter sp. MTCC 5804 using 0.08 % LDAO, wherein the ratio of A280: Asoo during the extraction never exceeds beyond 3.5.
- the photosynthetic protein pigment complex is specifically extracted with amine oxide surfactant lauryl dimethyl amine oxide.
- the partial purification of photosynthetic protein pigment complex from Enterobacter sp. MTCC 5804 is done with 50 KDa membrane filters and the ratio of A 2 so: Asoo lies in the range of 1.2-1.3.
- the photosynthetic protein pigment complex is stabilized during the layering step.
- the stabilization of the photosynthetic protein pigment complex is done using 5% PEG-6000 or 5% PEG-4000.
- the photosynthetic protein pigment complex is layered on a semi-conducting material.
- the semi conducting materials are T1O2 and ZnO.
- the semi-conducting material is seeded individually or in combination on transparent conductive oxides ( ⁇ /FTO).
- the semiconducting material is seeded on the transparent conductive oxides ( ⁇ /FTO) by taking T1O2 and ZnO in varying proportions (1:3-3: 1).
- the anode is fluorine doped tin oxide or indium tin oxide.
- the cathode is selected from platinum, aluminium, silver, gold, calcium and magnesium.
- the anode and cathode is FTO/ITO and platinum respectively.
- the support is a glass slide.
- the electrolyte may be solid or liquid, organic or inorganic in nature.
- the electrolyte may be Iodolyte, potassium chloride, ionic liquids, Tris-HCl containing various types of redox mediators such as methyl viologen, soluble cytochromes and quinones, cobalt(II)/(III) and manganese(iii)/(iv)-based redox mediators.
- the electrolyte is Iodolyte.
- the photovoltaic device under optimum conditions can produce an open circuit photo voltage, a photocurrent density and an efficiency of not less than 500 mV, 500 ⁇ /cm 2 and 0.1% respectively.
- Fig. 1 shows UV- Visible spectra of the photosynthetic RC in crude form and partially purified form, isolated from Enterobacter sp. MTCC 5804. List of abbreviations
- the present invention offers an efficient, simple and inexpensive wet or dry protein based photovoltaic device embedding photosynthetic RC isolated from a novel photosynthetic bacterium namely Enterobacter sp. MTCC 5804 having been deposited at MTCC, Institute of Microbial Technology, Chandigarh, India (MTCC 5804) on 06.02.2014.
- the said organism was isolated using Winogradsky column, from the mud and water samples collected from Renuka Lake (30°36'36"N 77°27'30"E), Himachal Pradesh, India. The sample was then enriched and purified using photosynthetic media popularly known as Pfennig media. The said organism showed orange pigmentation on the solid media and red pigmentation in the liquid media with absorption maxima at 800 and 850 nm, indicating presence of photosynthetic reaction centre (RC) and Light harvesting complex (LHC).
- RC photosynthetic reaction centre
- LHC Light harvesting complex
- the strain was identified as Enterobacter sp. based on its 16S rRNA gene sequence (1,411 bp), which was aligned with sequences available in the NCBI database using ClustalX software, and further named as Enterobacter sp IIPPS850R. It had 55.21% (G+C) content and 44.79% (A+T) content. The phylogenetic tree clearly showed that the isolated strain had more than 99% homology with the strain Enterobacter sp.
- An IDA deposit has been made to MTCC, Institute of Microbial Technology, Chandigarh, India (MTCC 5804).
- the 16S rRNA gene sequence of Enterobacter has been submitted at NCBI Genbank with accession number being JX855136.
- an efficient, simple and inexpensive wet or dry protein based photovoltaic device is prepared using following steps:
- the photosynthetic bacteria namely Enterobacter sp. MTCC 5804 was grown on modified Pfennig media comprising of g/L of trisodium citrate - 2; KH 2 P0 4 - 0.33; MgSO 4 .7H 2 O-0.33; NaCl-0.33; NH 4 Cl-0.5; CaCl 2 .2H 2 O-0.05; yeast extract-0.4 fortified with 0.5 ml of 0.02% FeS0 4 .7H 2 0 and 1 ml of trace metal solution (mg/ml) containing ZnSO .7H 2 O-10; MnCl 2 .4H 2 0-3; H 3 BO 3 -30; CoCl 2 .6H 2 O-20; CuCl 2 .2H 2 0-l; NiCl 2 .6H 2 0-2; Na 2 Mo0 -3.
- the said organism was grown for 3-5 days under illuminated conditions (preferably indirect sunlight) at 32 + 2°C
- the cells were centrifuged down at 8000 rpm for 15 min at 4°C and resuspended in 50mM Tris HC1 (pH-8.0) so as to make 30% slurry (i.e, 30% packed cell volume).
- This slurry was subjected to high pressure homogenizer (HomoGenius, GEA Niro Soavi; Panda Plus) maintained at 4°C with a pressure range of 800-1400 bar for 15 minutes.
- the disrupted cells were centrifuged at 12000 for 30 min at 4°C to remove the lysed cell debris and the photosynthetic RC recovered in the supernatant was stabilized with 0.08% N, N- Dimethyldodecylamine N-oxide (LDAO) and stored at 4°C until use.
- LDAO N- Dimethyldodecylamine N-oxide
- the ITO/FTO were cleaned with acetone in a sonication bath for 30 min followed by washing with deionised water in a sonication bath for 30 min and dried in nitrogen atmosphere.
- a paste of titanium oxide anatase powder (size less than 25 nm; 637254-Sigma Aldrich) was prepared in ethylene glycol with 50 ⁇ L ⁇ of Triton X- 100, by mixing them in the ratio of 1: 1 (w/v) using mortar pestle followed by sonication in ultrasonic bath for an hour.
- a paste of zinc oxide nanopowder (size less than 100 nm; 544906-Sigma Aldrich) was prepared with varying proportions of titanium oxide under similar conditions.
- the said paste of semiconducting material was layered on the cleaned ITO/FTO slides (thickness; 7- 10 ⁇ ) using doctor's blade, followed by drying in an oven at 100°C for 30 min and then annealed at 450°C for 30 min.
- the stabilized photosynthetic RC isolated from Enterobacter sp. MTCC 5804 was partially purified using Amicon Ultra- 15 Centrifugal Filter Units (3()/50KDa cut off membrane) centrifuged at 5000 rpm for 30 min at 4°C. Since the photosynthetic RC of the said bacterium has a molecular weigh of more than lOGKDa, it was recovered in the Reteniate. This resulted in recovery of a concentrated and partially purified photosynthetic RC. After passing through 30/50 KDa membrane, the ratio of the A28o Asoo of the photosynthetic RC solution never exceeded 1.5.
- the typical absorption maxima of the said photosynthetic protein is seen at following wavelengths that is 851nm, 800nm, 588nm, 510nm, 480nm, 397nm and 378nm.
- stabilization of the partially purified photosynthetic RC was done using 5% PEG-6000.
- the unpurified / partially purified photosynthetic RC stabilized with or without 5% PEG-6000 was layered onto semiconductor surface and vacuum dried under dark conditions.
- the counter electrode was prepared by depositing a thin layer of platisol T/SP (Solaronix make), which can be squeegee -printed by using a polyester mesh, and heat-treated in air for 30 min at 450°C. A drop of electrolyte was added to the photosynthetic RC coated on semiconductor photoanode.
- a Pt-coated ITO/FTO electrode was then clipped onto the top of the photosynthetic protein-TiC>2 working electrode to form the complete solar cell.
- the active area of the said device was 0.25 cm "2 + 0.02.
- the solar conversion efficiency was calculated by using the formula:
- Example 1 Effect of various detergents on the extraction and stabilization of photosynthetic protein from Enterobacter sp. MTCC 5804
- the bacteria were grown in 5L bottles in modified Pfennig media comprising of g/L of Trisodium citrate - 2; KH 2 P0 4 - 0.33; MgSO 4 .7H 2 O-0.33; NaCl-0.33; NH 4 Cl-0.5; CaCl 2 .2H 2 O-0.05; yeast extract-0.4 fortified with 0.5 ml of 0.02% FeS04.7H 2 0 and 1 ml of trace metal solution (mg/ml) containing ZnSO4.7H 2 O-10; MnCl 2 .4H 2 0-3 ; H3BO3-3O; C0CI2.6H2O-2O; CuCl 2 .2H 2 0-l; NiCl 2 .6H 2 0-2; Na 2 Mo0 4 -3.
- modified Pfennig media comprising of g/L of
- EXAMPLE 2 Effect of semiconductor material and protein stabilizer on photocurrent efficiency of photosynthetic protein isolated from Enterobacter sp. MTCC 5804
- Table 3 Effect of semi-conductor material and protein stabilizer on photocurrent efficiency of photosynthetic protein isolated from Enterobacter sp. MTCC 5804.
- V oc pen c rcu t p otovo tage
- I sc ort c rcu t current
- J sc p oto current ens ty, V
- EXAMPLE 2 Photovoltaic device preparations with homogenised cell free extract of Enterobacter sp. MTCC 5804 and partially purified protein.
- EXAMPLE 3 Effect of different semi conductor material on the photocurrent efficiency of the photosynthetic protein isolated from Enterobacter sp. MTCC 5804
- Voc Open circuit photovoltage
- Jsc photo current density
- Vmax Maximum voltage
- Pmax Maximum Power output.
- EXAMPLE 4 Effect of PEG-4000 addition during layering of photosynthetic protein isolated from Enterobacter sp. MTCC 5804
- Voc Open circuit photovoltage
- Isc Short circuit current
- Jsc photo current density
- Imax maximum current
- Vmax Maximum voltage
- Pmax Maximum Power output.
- EXAMPLE 5 Effect of incorporating HTL layer on photocurrent efficiency of protein-pigment photovoltaic device from IIPPS850R strain
- Photocurrent efficiency of partially purified photosynthetic protein (using 50KD membrane) from Enterobacter sp. MTCC 5804 stabilized with 5% PEG-6000 was tested with different hole transport materials. The experiment was conducted by coating them onto ITO Slides pre-coated with metal oxides and were subjected to solar stimulator with an illumination of light intensity of 1000W/m 2 . Following results were obtained as shown in Table 7.
- this invention claims protein based photovoltaic device without any genetic modification of the photosynthetic protein with high photocurrent efficiency, this set up is inexpensive as compared to existing protein-based photovoltaic devices.
- the said invention is novel, since this invention claims the incorporation of a photosynthetic RC from a novel photosynthetic bacterium Enterobacter sp. MTCC 5804 which can easily be extracted and partially purified and retains the natural functionality during the physisorption on photo anode. Moreover, a simple, low cost process of cultivation of novel photo synthetic bacterium Enterobacter sp. MTCC 5804 gives an added advantage to device preparation.
- the claimed protein based photovoltaic device in an optimum step up can produce an open circuit photo voltage, a photocurrent density and an efficiency of not less than 500 mV, 500 ⁇ /cm 2 and 0.1% respectively.
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Abstract
La présente invention concerne un dispositif photovoltaïque à base de protéines. Plus précisément, l'invention concerne un dispositif photovoltaïque à base de protéines incorporant un complexe de pigment protéique photosynthétique, isolé à partir d'une nouvelle bactérie photosynthétique Enterobacter sp. MTCC 5804, entre deux électrodes. Le dispositif produit un photocourant élevé quand le complexe de pigment protéine photosynthétique est déposé en couche sur un matériau semi-conducteur à base d'oxyde de titane et d'oxyde de zinc et en prenant de l'iodolyte comme électrolyte, le mélange ITO/FTO et le platine jouant respectivement le rôle d'anode et de cathode dans ledit dispositif.
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