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Hydrogen project


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As the remedy to overcome the crisis following depleting fossil fuels and global climate change, a variety of alternative fuels emerged. Among all the alternative fuels or energy, hydrogen attracted more and more attention due to its being clean, efficient and renewable nature. This study evaluates the potential of employing food and temple waste for fermentative hydrogen production.

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Hydrogen project

  1. 1. FERMENTATIVE HYDROGEN PRODUCTION FROM TEMPLE AND FOOD WASTE BY BATCH MODE USING Enterobacter aerogenes Submitted by- Ingole Charan Uttamrao Supervisor Prof. Subir Kundu School of Biochemical engineering Indian Institute of Technology (Banaras Hindu University) Varanasi -221005 1
  2. 2. Introduction 2 • As the remedy to overcome the crisis following depleting fossil fuels and global climate change, a variety of alternative fuels emerged. Among all the alternative fuels or energy, hydrogen attracted more and more attention due to its being clean, efficient and renewable nature. • Hydrogen is harmless to humans and the environment and is the only carbon-free fuel, on the oxidation produces water alone. H2+1/2 O2 H2O (1) • It gives secure elasticity for a sustainable energy system, in nature only, bacteria and algae have the gift to produce hydrogen, considering the present energy crisis and environmental tribulations.
  3. 3. Introduction cont. • Biological processes, unlike their chemical or electrochemical counterparts, are catalyzed by microorganisms in an aqueous environment at ambient temperature and atmospheric pressure. • These techniques are well suited for decentralized energy production in small-scale installations in locations where biomass or wastes are available, thus avoiding energy expenditure and costs for transport • Intensive research on Biohydrogen is underway, and in the last few years several novel approaches have been proposed and studied to surpass these drawbacks. Among the available biological methods, microbial fermentation of organic carbon sources has been comprehensively studied (Ilgi Karapinar and Kargi 2006). • Glucose is a monomer of cellulose, the most abundant biomass (Lattin and Utgikar 2007). 3
  4. 4. Introduction cont. • This study evaluates the potential of employing food and temple waste for fermentative hydrogen production. For the optimum hydrolysis of these waste materials to release reducing sugars, different concentrations (1-5%, v/v) of HCl were used. • Released reducing sugar is further used as substrate for biohydrogen production by using Enterobacter aerogenes. • The fermentative H2 production mainly depends on temperature, pH and substrate concentration. • In the world a good source for H2 production is considered as microorganisms. 4
  5. 5. Introduction cont. Enzymes involved in hydrogen production • There are four fundamentally different hydrogen producing and metabolizing enzymes found in Microorganism: (1)The reversible or classical hydrogenase (2) The membrane-bound uptake hydrogenase (3) The nitrogenase enzymes and (4) Hydrogenase 5
  6. 6. Introduction cont. • General reaction implicated in the microbial conversion of Biomass to Biohydrogen 6 Process General reaction Microorganism Direct Biophotolysis 2H2O+Light2H2+O2 Algae Photo fermentation CH3COOH+2H2O+Light4H2+2CO2 Purple bacteria, Microalgae Indirect Biophotolysis a. 6H2O+6CO2+LightC6H12O6+6O2 b. C6H12O6+2H2O4H2+2CH3COOH+2CO2 c. 2CH3COOH+4H2O+Light8H2+6O2 Overall reaction:12H2O+Light12H2+6O2 Microalgae,Cynobacteria Water gas shift reaction CO+H2OCO2+H2 Fermentative bacteria +Methanogenic bacteria Two phase H2+CH4 Fermentation a. C6H12O6+2H2O4H2+2CH3COOH+2CO2 b. 2CH3COOH=2CH4+2CO2 Fermentative bacteria High yield dark fermentation C6H12O6+6H2O12H2+6CO2 Fermentative bacteria
  7. 7. Introduction cont. Anaerobic decomposition of organic matter (Zehnder et al. 1982) 7
  8. 8. Literature and review • Hydrogen gas is a one of the hopeful and alternate source for reduction of greenhouse effect (Christopher and dimitrios 2012). • Approximately 95% of hydrogen produced is consumed at the site of production, with 1.5 million tons being sold for industrial and chemical uses (Lattin and Utgikar 2007). • Carbohydrate rich, nitrogen deficient solid wastes such as cellulose and starch containing agricultural and food industry wastes and some food industry wastewaters such as cheese whey, olive mill and baker’s yeast industry wastewaters can be used for hydrogen production by using suitable bio-process technologies. • Hydrogen was produced at an average rate of 6 ml/h per g (dry weight) of cells with whey as a hydrogen donor. In continuous cultures with glutamate as a growth-limiting nitrogen source and lactate as a hydrogen donor, hydrogen was evolved at a rate of 20 ml/h per g (dry weight) (Hans and Reinhard 1979). 8
  9. 9. Literature and review cont. • Despite being a clean and high energy fuel, currently only 50 million tons of Hydrogen is traded every year with a growth rate of about 10% (Winter 2005). • The majority of this hydrogen is used to produce ammonia fertilizer, as feedstock for chemical and petroleum refining areas, plastics, solvents and other commodities (Dunn 2002).The technology currently used to make hydrogen has been well established, but the majority of hydrogen produced uses fossil fuels in the production process. • Project to turn flower waste into organic fertilizers – showing the technology for the waste would be amassed and dumped into this dip places to produce cheap vermicompost fertilisers. • During the time of special occasions or pujas the amount of wastes increases leading to major difficulties for us in terms of clearing them. But in this project all the wastes would be cleared and stored (Kenya flower council 2013). 9
  10. 10. Literature and review cont. • The food processing industry in the United States is composed of more than 20,000 companies (Elitzak 2000). • It is estimated that the average large food processing industry annually produces about 1.4 billion liters of wastewater (Van Ginkel et al. 2005). • Wastes from these industries are usually high in organic matter and normally contain sufficient nitrogen, phosphorus, and trace elements for biological growth (Gray 2004). • The biodegradability of food waste is mainly related to carbohydrate materials, which are the main source of hydrogen production (Noike and Mizuno 2000). • India wastes Rs 44,000 cr worth food by food processing industry and others every year. (Deccan herland Aug. 2013). • These waste streams usually require treatment practices before being discharged into local sewer districts. 10
  11. 11. Literature and review cont. • Approximately 50% of hydrogen production globally comes from natural gas, 30% oil, and 20% coal; see Figure.03 (Romm 2005). • Current research has studied many different types of substrates for the use of hydrogen production. The major criteria for substrate selection are the availability, cost, carbohydrate content, and biodegradability (Kapdan and Kargi 2006; Rai et al 2012). 11
  12. 12. Literature and review cont. • Hydrogen production capability of anaerobic facultative bacterial culture Enterobacter aerogenes has been widely studied (Yoko and Saitsu 2001) • In dark fermentation of hydrogen the enzyme hydrogenase present in anaerobic organisms oxidizes reduced ferrodoxin to produce molecular hydrogen, external iron addition is required for hydrogen production. • Hydrogen producing microorganisms • Diverse microbes capable of H2 production by dark fermentation are distributed across a wide variety of bacterial groups (Lee et al., 2011). • The organisms used in dark fermentation studies include anaerobes, facultative anaerobes and aerobes in a wide Temperature range (mesophiles, thermophiles and hyperthermophiles). 12
  13. 13. Literature and review cont. • Mesophiles are mainly affiliated with two genera: facultative Enterobacteriaceae (Kumar and Das, 2000) and strictly anaerobic Clostridiaceae (Collet et al., 2004; Evvyernie et al., 2001; Wang et al., 2003), • Most thermophiles belong to genus Thermoanaerobacterium (Ahn et al., 2005; Ueno et al., 2001; Zhang et al., 2003). Also few aerobes, such as Bacillus (Kalia et al., 1994; Kumar et al., 1995; Shin et al., 2004), • Aeromonons spp., Pseudomonos spp. and Vibrio spp. (Oh et al., 2003b) have been characterized for H2 production under anaerobic conditions but they show H2 yields less than 1.2 mol H2/mol-glucose. 13
  14. 14. Literature and review cont. • Various wastewaters viz., paper mill wastewater (Idania, et al., 2005), starch effluent (Zhang, et al., 2003), food processing wastewater (Shin et al., 2004, van Ginkel, et al., 2005), • Domestic wastewater (Shin, et al., 2004, 2008), rice winery wastewater (Yu et al., 2002), distillery and molasses based wastewater (Ren, et al., 2007, Venkata Mohan, et al., 2008), • wheat straw wastes (Fan, et al., 2006) and palm oil mill wastewater (Vijayaraghavan and Ahmed, 2006) have been studied as fermentable substrates for H2 production along with wastewater treatment. 14
  15. 15. Literature and review cont. • Hydrogen production capability of anaerobic facultative bacterial culture Enterobacter aerogenes has been widely studied (Yoko and Saitsu 2001),in dark • Fermentation of hydrogen the enzyme hydrogenase present in anaerobic organisms oxidizes reduced ferrodoxin to produce molecular hydrogen, external iron addition is required for hydrogen production. • Ten milligram per liter iron concentration was determined to be the optimum in batch hydrogen production by C. Pasteurianum from starch (Liu Gand Shen J. 2004) 15
  16. 16. Biochemical Processes • There are two main stages in the biochemical process of AD for hydrogen production: hydrolysis, acidogenesis, which are shown in Fig. Schematic diagramme for biochemical process modified (Metcalf and Eddy, 1985). • Hydrolysis , Acetogenesis 16
  17. 17. Literature and review cont. • Schematic diagramme (mechanism) for Enteric-type mixed- acid fermentation (Modified from Turcot et al. 2008) 17
  18. 18. Objectives • To characterize the physical properties of the food and temple waste that is collected from Hostel mess (BHU) Varanasi and different temples from Varanasi (UP). • To determine the potential of this food waste and temple waste as a feedstock for fermentative H2 production. • The overall objective of this work will be to develop Processing strategy for the production of hydrogen from flower waste (TW), and food waste generated during food processing and production. • Comparative studies of Hydrogen production from different wastes. 18
  19. 19. Plan of work • Procurement of suitable microorganism for the production of Hydrogen. • Collection of different waste materials. • Pretreatment of the waste materials for the bioconversion. • Formulation of suitable medium using waste for the production of hydrogen. • Determine the experimental setup for anaerobic fermentation system. • Testing for the existence of Hydrogen gas 19
  20. 20. Materials and methods  Hydrogen-producing microbial strain and medium.  Enterobacter aerogenes (MTCC 2822) • Microorganism and culture condition • The strains were procured from Microbial type culture collection and Gene Bank Chandigarh (IMTECH) Enterobacter aerogenes( MTCC 2822). • These strains belong to the family of Enterobacteriaceae which are known for their H2 production ability. • They are obligate anaerobes capable of producing endospores Individual cells are rod-shaped, these characteristics traditionally defined the genus. 20
  21. 21. Materials and methods • The Enterobacter sp. was reactivated in pre-culture basal medium with the following composition (g/l): • The basic culture medium for E.aerogens • Glucose 10.0 g • Tryptone 5.0 g • (NH4)2SO4 2.0 g and MgSO4 0.2 g.100mM of monophosphate (NaH2PO4, Pi) as phosphate sources was added into the basic culture medium. • All chemicals used were of analytical grade, and sterilized individually by autoclave. The initial pH value of medium was controlled at 6.0 by addition of NaOH or HCl. 21
  22. 22. Materials and methods • Substrate and raw materials • Temple Waste includes- • 1. New Vishwanath Temple waste include marigold (Genda) flower with rose. • 2. Durga Temple, Durgakund Varanasi the waste of flower include Hibiscus Rosa sinesis (Gudhal) • 3. Shiv-Temple (Mrityujaya Mahadev), Visheshwarganj, Varanasi the waste flower include Calotropis gigantea (Madar) • 4. Waste food material Food waste from Different Hostel Mess of Banaras Hindu University (BHU), Varanasi 22
  23. 23. Experimental setup • For biohydrogen production from food waste and temple waste the modified laboratory setup made as shown in Fig. the batch reactor with working volume is 120 ml was operated at 32 to 35 °C for hydrogen production the reactor operated till the production end. The generated biohydrogen measured by the water disceplacement method. 23
  24. 24. Estimation • Carbohydrate (X-PertTM Carbohydrates Estimation Teaching Kit) • Reducing sugar by DNS Method Standard curve for reducing sugar estimation by DNS method 24
  25. 25. Estimation • Carbohydrate (X-PertTM Carbohydrates Estimation Teaching Kit) • Phenol Sulphuric Acid Method: Standard curve for total sugar by PSA method 25
  26. 26. Estimation • C: N Ratio Analysis (Allison, LE in Black, CA et al. 1965) • Carbon analysis: Walkley-Black chromic acid wet oxidation method • Nitrogen analysis 26
  27. 27. Characteristic • Physico-chemical Analysis of Food and the Temple waste • Waste food from Hostel mess of BHU and the different temple waste i.e. Flower waste sample was collected from in zip lock bags and all were dried under sun natural drying and made pieces and with make fine powder after all kept all sample by labeling accordingly in research laboratory biochemical engineering oven for 1 hour under 90 -105°C. 27
  28. 28. . 28
  29. 29. Study for MC, TS, VS, and FS [APHA, 1989] • The food and temple waste sample was finely ground to maintain homogeneity. Specific amounts of food sample were weighed, using a laboratory weight box in three separately prepared crucible dishes and same for all separated sample of temple waste. • The crucible dishes were also weighed individually using the same balance and were then placed in an oven at 105ºC overnight and weighed again the next morning. The crucible dishes were then transferred to a cool muffle furnace heated to 550ºC ± 50ºC and ignited for an hour to remove volatile organics. • Bulk Density Determination (FMP Group (Australia) Ltd. Information QC 125 Issue 3) 29
  30. 30. Theoretical consideration • The biological hydrogen production by anaerobic condition by specific microorganism, usually use the organic substrate as their carbon and energy and hydrogen ion (H+ ) as an electron acceptor, the specified mechanism for biological production of hydrogen by fermentation is generally associated with the presence of an iron-sulphur, protein called “ferredoxin, an electron carrier and low redox potential. • Different type of microorganism including bacteria and archaea domain, have the ability to show the potential of hydrogen production by naturally metabolic process, with that the metabolically observed product formation in microorganism with H2 gas with other co-product get 30
  31. 31. Theoretical consideration 31 Energetic view of anaerobic bacterial hydrogen production. (Biohydrogen by Oskar R.Zaborsky)
  32. 32. Result and Discussion • Hydrogen is an alternative energy source is produced biologically via various methods using different types of materials. As the case of substrate for hydrogen production the important consideration is of the economics of biohydrogen production, there is need to go for cheaper and abundant feedstock for making the process cost effective. (Rai 2014, Bioresource technology 152:140-146). 32
  33. 33. Effects of dilute acids treatment on hydrolysis of flower waste 33 Schematic diagramme for hydrogen production from acid hydrolysate (Rong Chen, et al 2013)
  34. 34. References • Bartlett, GN, Craze, B, Stone, MJ & Crouch, R (Ed) 1994, Guidelines for Analytical Laboratory Safety. Department of Conservation & Land Management, Sydney. • Benemann J (1996) Hydrogen biotechnology: progress and prospects. Nature Biotechnology 14:1101-1103. • Benmeman, J.R 2000. Hydrogen Production by Algae. Journal of Applied Phycology. 12291-300. • Chen CC, Lin CY, Lin MC. Acid-base enrichment enhances anaerobic hydrogen process. Appl Micro Biotechnol 2002. 58:224–228. • Christopher and dimitrios Energy & Environmental Science, Royal Society of Chemistry Publishing, Energy Environ. Sci., 2012, 5, 6640. 34
  35. 35. References • Chun-Mei Pan, Hong-Cui Ma,Yao-Ting Fan,Hong-Wei Hou International journal of hydrogen energy 36 (2011) 4852-4862 • Collet C, Adler N, Schwitzguebel JP, Peringer P. Hydrogen production by Clostridium thermolacticum during continuous fermentation of lactose. Int J Hydrogen Energy 2004. 29: 1479–1485. • Das, D. and T.N. Veziroglu. 2001. Hydrogen production by biological processes: a survey of literature. International Journal of Hydrogen Energy.26 (1):13-28. 35