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SECOND GENERATION ETHANOL PRODUCTION FROM AGRICULTURAL RESIDUES
Prof. Rintu Banerjee, FNAAS, FBRS, FBRIAT Professor, Agricultural & Food Engineering Department Indian Institute of Technology Kharagpur India
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Sector wise global CO2 emission
Carbon dioxide equivalent emissions (CO2 eq) from transportation sector in India Source: IPCC, 2014)
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World market for Biofuel production: 2013- 2023
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Indian scenario in biofuel production
Bioethanol demand with blending targets (%) in India Demand for ethanol for blending and share in blending
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Indian scenario in biofuel production
Government of India, is planning to set up twelve (12) 2G Ethanol Bio-refineries across 11 States viz. Punjab, Haryana, U.P., M.P, Bihar, Assam, Odisha, Gujarat, Maharashtra, Karnataka and A.P. The estimated investment for the 12 Bio-refineries is Rs 10,000 crores. These Bio-refineries shall produce around crore litres of Ethanol annually, thus contributing significantly towards the Ethanol Blended Petrol Programme. Recently GoI has proposed a new 2G biorefinery in Paradip (Odisha)
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India to commission world’s largest green field refinery by 2022
Signed a Joint Venture (JV) agreement, June -2017
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Second Generation (2G) ethanol production
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Potential source of biomass for 2G Ethanol
Agricultural residues (wheat straw, corn stover) Energy crops (switchgrass, poplar) Wood and paper waste Municipal solid waste, organic Low cost renewable organic resources
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Global biofuels market forecast (2017-2014)
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Gross residue availability from crop production in India
The above scenario illustrates the potential of some of the crop residues which can be utilized for biofuel and biochemical generation
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Biomass Selection Lignocellulosics Grazing Non- Grazing Rice straw
Maize straw Ricinus communis Lantana camara Wheat straw Rice husk Jatropha curcas Pineapple leaf wastes Jute waste Oil seed waste Bambusa bambos Datura stramonium Grazing Non- Grazing
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2G-Ethanol production from Non-conventional and Renewable sources
Lignocellulosic biomass Bambusa bambos Lantana camara Kans Grass Ricinus communis Cotton Stalk Sugarcane Tops 2G-Ethanol production from Non-conventional and Renewable sources CELLULOSE HEMICELLULOSE LIGNIN Enzymatic Pretreatment -Promising Green Technology Rice straw
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Biochemical Characterization of lignocellulosics
composition Cellulose (%, w/w) Hemicellulose Lignin Lantana camara 47.25 16.4 17.26 Ricinus communis 42.00 18.02 19.88 Bambusa bambos 45.00 17.00 19.20 Sugarcane Tops 33.00 22.76 13.45 Kans Grass 38.70 29.00 17.46 Pineapple leaf waste 25.00 13.00 Rice straw 33 16 14 Mixture 43.02 24 14.57
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Biochemistry of Lignocellulosics
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Steps involved in 2G ethanol generation: soil to soil technology
Biomass pretreatment biomass pretreatment Cellulose and Hemicellulose hydrolysis Hexose and Pentose fermentation Ethanol recovery Enzymatic degradation of recalcitrant lignin layer Hydrolysis of cellulose & hemicellulose to hexose and pentose sugar Utilization of hexose and pentose sugar to ethanol Distillation to recover maximum ethanol Residual solid biomass Biomethane Anaerobic Digestion Biomanure Cyanobacterial treatment
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Different Methods of Pretreatment
Acid Hydrolysis Alkali Hydrolysis Physical Treatment Physico-chemical Treatment Biological/ Enzymatic Treatment Costly, Hazardous, energy and labor intensive, loss of target biomolecules
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Major Bottlenecks in Lignocellulosic Bioethanol Production
Maximum lignin degradation with minimum loss of cellulose and hemicellulose Efficient depolymerization of lignin without the production of furfurals and hydroxymethyl furfurals Simultaneous utilization of Pentose and Hexose sugars Less tolerance of yeast towards higher concentration of ethanol
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Water requirement during biomass pretreatment
Biological pretreatment requires litre of water for per litre of ethanol production compared to more than 1 or 5 litre of water requirement in physical, chemical and physico-chemical pretreatment methods
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Leading Pretreatment technologies
Partial hydrolysis of hemicellulose Ammonia recovery during pilot scale operation is difficult Lignin is deposited on the surfaces of the material possibly causing blockage of cellulases to cellulose Expensive Needs improved method for ILs recovery and reuse
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Enzymes/microbes involved in second generation bioethanol production
Lignolytic enzymes Lignin peroxidase Manganese peroxidase Laccase Versatile peroxidase Cellulolytic enzymes Endo β-glucanase β-glucosidase Cellobiohydrolase Oxidative cellulases Cellulose phosphorylase Xylanase Swollenin Expansin Yeast Saccharomyces cerevisae Saccharomyces pastorianus Pichia pastoris Saccharomyces fermentati Saccharomyces paradoxus
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IIT Kharagpur contribution in biofuel production
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Enzymatic Delignification of Lignocellulosics
Microbial Biotechnology and DSP Laboratory, Department of AgFE, IIT Kharagpur Blue laccase from a hyperactive strain of Pleurotus djamor Yellow laccase from Lentinus squarrosulus
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Result of Delignification
Biomass Condition Solid loading (% dry weight/vol.) % Delignification Ricinus communis Initial mixing followed by static 30 75-80 Lantana camara Kans grass Bamubusa bambos Cotton stalk 25 77-81 Sugarcane top Banana waste 20 70-72 Pineapple leaf waste Rice straw 78-82 Mixture
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SEM analysis Control Lantana camara Pretreated Control
Ricinus communis Pretreated Control Bambusa Bambos Pretreated Control Pineapple leaf Pretreated
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SEM analysis cont… Control Kans grass Pretreated Control Sugarcane top
Mixture Pretreated
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Porosity Analysis by BET/BGH Analyzer
Biomass Pore size (Before (b); After (a)) Angstrom Pore volume (Before (b); After (a)) cm3/g % Delignification Ricinus communis (b); (a) 13.01 x 10-3 (b); 20.50 x 10-3 (a) 75-80 Lantana camara 86.74 (b); (a) 5.968 x 10-3 (b); 7.496 x 10-3 (a) Kans grass 61.9 (b); 128.5 (a) 4.259 x 10-3 (b); 5.365 x 10-3 (a) Pineapple leaf waste 120 (b); 134 (a) 4.26 x 10-3 (b); 6.57 x 10-3 (a)
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C–H methyl and methylene groups
FTIR analysis Ricinus communis Lantana camara Bambusa bambos Kans grass Pineapple leaf Sugarcane top Mixture Wavenumber (cm-1 ) Particular OH Vibration C–H methyl and methylene groups O–H phenolic lignin
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XRD Analysis Control Lantana camara Pretreated Control Sugarcane top
Kans grass Pretreated
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XRD Analysis cont… Pineapple leaf Mixture Lignocellulosic biomass
Increase in crystallinity (%) Ricinus communis 6.82 Lantana camara 7.46 Kans grass 8.54 Bambusa bambos 4.62 Pineapple leaf waste 6.25 Sugarcane top 4.30 Mixture 3.66
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HPLC chromatogram of Lignin degradation Products
With time peak area of the individual peaks increased Lignin degraded products released over time
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GC-MS Analysis of lignin degraded compounds
Identified compounds 0 h 1st h 2nd h 3rd h 4th h 5th h 6th h 7th h Retention time Propanoic acid - + 4.748 Butanoic acid 5.731 Acetic acid 5.974 Chromone 7.167 Methanone 7.544 Pentacene 8.007 Butanedioic acid 8.194 2-Butenedioic acid 8.747 2(3H)-Furanone 9.186 3,4-Methylenedioxyphenylacetic acid 10.876 3,5-di-tert-Butyl-4-hydroxyphenyl propionic acid 11.282 3-Bromo-5-ethoxy-4-hydroxybenzaldehyde 12.269 2',6'-Dihydroxyacetophenone 15.882 11H-Dibenzo[b,e][1,4]diazepin-11-one 15.863 5(2-Dimethylamino-1-phenyl)-vinyl-thiadiazol 16.264 Trimethylsilyl-3-methoxy-4-(trimethylsilyloxy)cinnamate 17.023 Ferulic acid 17.013 Trimethylsilyl-3,4-bis(trimethylsiloxy)cinnamate 17.452 Ethyl-2-ethoxyquinoline-4-carboxylate 21.628 Salbutamol 23.876 2-(p(Dimethylamino)phenyl) benzimidazole 25.070 3,4-Dimethylbenzamide 25.375 Benzoic acid 26.664 1,2-Benzenedicarboxylic acid 26.425
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Some of the detected compounds
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Compounds detected during laccase mediated delignification
Low molecular weight compounds like 2(3H)-Furanone (RT 9.186), 3,4-Methylenedioxyphenylacetic acid (RT ), Ethyl-2-ethoxyquinoline-4-carboxylate (RT ) Aromatic compounds Trimethylsilyl-3-methoxy-4-(trimethylsilyloxy) cinnamate (RT ), 3-Bromo-5-ethoxy-4-hydroxybenzaldehyde (RT ) and Ferulic acid (RT ) Low molecular organic acids Propanoic acid (RT 4.748), Butanoic acid (RT 5.731), Acetic acid (RT 5.974), Butanedioic acid (RT 8.194)
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Microscopic analysis of raw and pretreated biomass
Raw biomass Pretreated biomass
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Established biomass pretreatment technology with different substrates
Blue laccase Substrates: L. camara, R. communis, S. spontaneum, B. bambos, S. Officinarum tops, A. comosus leaf waste, Lignocellulosics Mixture Optimum conditions: ºC, 6-8 h, 15-25% Solid loading, 6-7 pH, IU/mL laccase titre, 75-86% delignification % Ref: Gujjala et al., 2016; Mukhopadhyay et al., 2011; Rajak and Banerjee, 2015; Kulia et al., 2011; Avanthi and Banerjee, 2016. Yellow laccase Substrates: L. camara, R. communis, S. spontaneum, B. bambos, S. Officinarum tops, A. comosus leaf waste, Lignocellulosics Mixture Optimum conditions: ºC, 4-6 h, 15-25% Solid loading, 5-7 pH, IU/mL laccase titre, 65-87% delignification % Ref: Mukhopadhyay et al., 2011; Rajak and Banerjee, 2016.
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Enzymatic Saccharification of Pretreated Lignocellulosic Biomass
Delignification A mixture of carbohydratases produced from T. reesei RUT C30
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Correlation between Delignification, Reducing sugar and Ethanol Production from Lignocellulosic Biomass
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Strategies to improve the ethanol yield from lignocellulosic biomass
Ethanol (%, v/v) SHF SSF CBP PCBP SSCF Ricinus communis 2.78 3.58 6.81 7.12 6.24 Lantana camara 2.51 3.38 6.5 6.9 6.95 Sugarcane top 3.21 6.02 5.91 5.96 7.50 Kans grass 3.50 6.30 4.9 7.80 8.00 Pineapple leaf waste 3.25 6.42 6.78 7.18 Mixture 3.00 5.14 7.52 7.65
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Integrated approach towards soil to soil technology
Processing Processed solid biomass Ethanol, butanol, lactic acid Residual fermented biomass By-products (Biogas) Removal of CO2 by water stripping Left over residual solid biomass Enriched biomethane CO2 as carbon source used in algal cultivation Enrichment with cyanobacteria Biomanure applied back to the soil Fermentation (MAC) Fermentation Microbes / Enzymes Biodiesel Agricultural residue
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Pilot Plant facility for Ethanol Production
(Capacity: 500 kg fresh biomass processing per batch)
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Lignocellulosic Biomass Hot Air Oven Pulverizing unit
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Saccharification tank
Pilot plant Pre-treatment tank Saccharification tank Fermentation tank Tincture press Gantry system Control panel
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Distillation assembly
Boiler assembly Distillation assembly
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Gantry system for lifting the immobilization tray
Biomass collection tank Gantry system for lifting the immobilization tray
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Students’s Contribution
Dr. Mainak Mukhopadhyay Dr. Arindam Kuila Mr. Rajiv Ch. Rajak Mr. Sanjeev Kumar BIOENERGY GROUP AT P.K.SINHA CENTRE FOR BIOENERGY, IIT KHARAGPUR Ms. Althuri Avanthi Ms. Anjani Devi Chintagunta Ms. Knawang Ch. Sherpa Mr. G. Lohit K. Srinivas Mr. Debajyoti Kundu Ms. Tania Dev Ms. Reddhy Mahle
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