feature article feature article

Repair and servicing facility north-eastern states, Sikkim, Jammu and central subsidy is also provided for repair The government pays a job fee of Rs 800 Kashmir, Himachal Pradesh, Uttaranchal, and revival of family-type biogas plants per biogas plant constructed on turnkey hilly districts, and islands. In other that are at least five years old and have basis with three years’ warranty for regions, it is Rs 700 per plant. Financial developed structural defects thereafter. trouble-free functioning of the plant in support limited to 50% of the rate of Courtesy: MNRE, Government of Bioethanol production

Manufacturers of biogas M/s Sweet Home Appliances Pvt. Ltd M/s Malhotra Engineering Company (P) Ltd from 3-E/16, BPNIT 572-B, Nangloi burners Faridabad – 121 001 New Delhi – 110 041

M/s Sunflame Industries (P) Ltd M/s Tulsi Domestic Appliances M/s Rupak Enterprises Shed No. 2, Plot No. 58 30-A, Old Industrial Area 1/46 Vishwas Nagar, Shahdra P O Amar Nagar, Faridabad – 121 003 Alwar - 301 001 New Delhi –110 032 biomass

M/s Batra Investments Pvt. Ltd M/s Associated Engineering Works Bhawna Industries 14/1, Mathura Road Tanuku - 534 211 8-A, Industrial Development Colony P O Amarnagar Andhra Pradesh Kunjpura Road, Karnal – 132 001, Haryana Faridabad – 121 003 M/s Mitaso Appliances Ltd M/s Mech-Ci-Co. M/s Gas and Chemical Industries (P) Ltd Plot No. 63, Sec. 6 1-7, GIDC Industrial Township 14/1 Mathura Road Faridabad – 121 006 Vatwa, Ahmedabad – 382 445 Faridabad - 121 003 M/s Agriculture Associates Inter Gas Appliances Pvt. Ltd M/s Baroda Appliances Station Road C-113, Sector-2 886/A, GIDC Makarpura Alwar (Rajasthan) Noida – 201 301, Uttar Pradesh Baroda - 10

Akshay Urja bids adieu to Mr Vilas Muttemwar, former Inviting articles for Minister for New and Renewable Energy Akshay Urja Shri Vilas Muttemwar represents Akshay Urja publishes news, articles, research Nagpur Lokshabha constituency Sachin Kumar1,2, Pratibha Dheeran1 and papers, case studies, success stories, and write- of and has been re- Dilip K Adhikari1 elected 7th time as a Member of ups on RE. Readers are invited to send material with original photographs and statistical data. The Parliament of the tropical regions such as Brazil, India, (35%–50%), hemicellulose (20%– in May 2009. Shri Vilas Muttemwar photographs should be provided on hard copy ignocellulosic biomass Thailand, Hawaii, and the southern 35%), polyphenolic lignin (10%–25%) completed his full tenure of five or as high resolution (minimum 300 DPI) files on is the most abundant USA. Other lignocellulosic feedstocks and other extractable components. years as Minister for New and a CD. Akshay Urja will pay suitable honorarium for renewable resource on earth. include agricultural residues such The utilization of both cellulose and Renewable Energy. During his each published article of about 1500 words and Lignocellulosic materials, as corncob, corn stover, wheat, hemicellulosic monosachharides like tenure the renewable energy sector above to the authors. The publication material in including wood, grass, forest and rice straw; industrial residue hexose and pentose present in a typical took a leap with installation of over two copies, along with a soft copy on CD/floppy/ Lresidues, agricultural residues, pulp such as pulp and paper processing lignocellulosic biomass hydrolysate 9500 MW of renewable powers in e-mail may be sent to and paper mill wastes, and municipal waste; and energy crops such as switch (composition of various lignocellulosic the country, and the massive installation of solar water heating solid wastes can be used for bioethanol grass. biomass is shown in Table 1) is essential systems, solar lighting systems, family-size biogas plants, and a Editor, Akshay Urja production. Among these resources, Lignocellulosic materials are for the economical production of number of other renewable energy systems. The major initiatives agricultural residues, such as sugarcane comprised of lignin, hemicellulose, ethanol. Therefore, microorganisms Ministry of New and Renewable Energy taken by him for popularizing renewable energy among the bagasse, dominate in terms of tonnage and cellulose in varying proportions. that are able to ferment both glucose Block – 14, CGO Complex, common people include the Akshay Urja magazine, Rajiv Gandhi and can serve as feedstock. Sugarcane The general composition of and xylose are most desirable for an Akshay Urja Diwas, Renewable Energy Clubs, District Advisory Lodhi Road, New Delhi – 110 003 bagasse is plentiful in tropical and sub- lignocellulosic biomass is cellulose efficient bioconversion of biomass to Committees, and so on. Tel. +91 11 2436 3035 Akshay Urja Team wishes him success in all his future Fax +91 11 2436 3035 1Biotechnology Area, Indian Institute of Petroleum, Dehradun (India) endeavours. E-mail [email protected] 2Department of Chemical Engineering, Indian Institute of Technology, Roorkee (India) Tel. +91 135 252 5763; Fax +91 135 266 0202; E-mail [email protected]

16 VOLUME 2 • ISSUE 6 JUNE 2009 VOLUME 2 • ISSUE 6 JUNE 2009 17 feature article feature article

di-hydrogen ortho phosphate, 1.0 g/l Analytical methods Table 1 Composition of various lignocellulosic raw materials ammonium sulphate, and 1.0 g/l yeast Table 2 Sugars and furfural (percentage of bagasse) recovered in first stage hydrolysis at different acid concentrations Reducing sugars in media and Raw materials Glucose Mannose Galactose Xylose Arabinose extract with pH 5.0 at 45 to 60 °C in fermented broth were determined by batch process and continuous process Acid concentration Solid to DNS (di-nitrosalicylic acid) method. Corn stover 39 0.3 0.8 14.8 3.2 (%) (w/w) liquid ratio Xylose(%) Glucose (%) Furfural (%) with recycling the cells. Ethanol was determined by using gas Wheat straw 36.6 0.8 2.4 19.2 2.4 2 1:10 9.2 0.6 0.12 chromatography using a Chemito Rice straw 41 1.8 0.4 14.8 4.5 Fermentation conditions 4 1:8 13.6 0.9 0.16 8600 Refinery Gas Analyser with a Sugarcane bagasse 38.1 -- 1.1 23.3 2.5 Batch fermentation process 6 1:8.8 19.6 1.4 0.23 4-m-long and 1/8 diameter Porapack Rice hulls 38.1 3.0 0.1 14.0 2.6 Batch fermentation of sugarcane 8 1:8.3 23.1 4.4 0.31 column with Chemosorb 80/60. Sample bagasse hydrolysate and cassava starch 1:5.2 25 4.6 0.41 was injected at 120 °C, and the oven hydrolysate was performed in 2 litre ethanol. The bioethanol production 1:4.2 26.14 4.6 0.63 temperature and flame ionization Materials and methods Bioflow-110 bioreactor by free cells from lignocellulosic biomass requires 10 1:8.4 25 5.8 0.95 detector temperature was 150 °C and Microorganisms and culture of Kluyveromyces sp. IIPE453 in batch two essential steps: saccharification of 200 °C, respectively, using helium as a conditions mode. The temperature and agitation lignocellulosic biomass to fermentable carrier gas. Ethanol was also determined The strain used for ethanol production, were controlled at 50 °C and 200 rpm grown cells of Kluyveromyces sp. IIPE453. Hydrolysis of starch biomass sugars and fermentation of sugars by colorimetry method. Furfural was Kluyveromyces sp. IIPE453, was grown respectively. The air/N flow in proper ratio was Hydrolysis of different starch biomass like to ethanol. 2 measured by Double Beam UV-VIS in salt medium containing 0.15 g/l controlled in the bioreactor for in situ soluble starch, cassava, tapioca, sweet Worldwide, bioethanol is currently Spectrophotometer 2600 at 277nm. (gram per litre) di-sodium hydrogen Continuous fermentation process recovery of ethanol. The process was sorghum, and maize was performed in 1 produced by fermentation of monomeric ortho phosphate, 0.15 g/l potassium with recycling the cells performed at different dilution rates, cell litre flasks containing 5% starch biomass sugars by mesophiles, which ferment Results and discussion di-hydrogen ortho phosphate, Continuous fermentation was mass concentration, and temperatures. in 200 ml 0.05 M acetate buffer (pH 5.0) the sugars at 25–37 °C. These mesophiles Hydrolysis of sugarcane bagasse 2.0 g/l ammonium sulphate, 1.0 g/l yeast performed in 2 litre Bioflow-110 The pH and agitation were controlled and 100 ml crude enzyme (5.29 mg/ml). have certain limitations in fermenting In first stage hydrolysis, sulphuric acid extract, and 10 g/l glucose with pH 5.5 bioreactor by using sugarcane bagasse at 5 and 250 rpm, respectively. All the flasks were incubated at 80 °C and pentose sugars produced from was used at different concentrations at 45 °C. Fermentation was carried out hydrolysate solution containing glucose monitored at an interval of 2 hours until lignocellulosic biomass. However, the (2% w/w to 10% w/w) and different in a medium prepared in hydrolysate and xylose sugars. The hydolysate Hydrolysis of sugarcane bagasse the total starch was hydrolysed. thermophiles have certain advantages solid-to-liquid ratios. The temperature containing 0.15 g/l di-sodium hydrogen solution, supplemented with inorganic The sugarcane bagasse powder was over mesophiles, which could be was maintained at 100 ºC for 1 hour. ortho phosphate, 0.15 g/l potassium salts, was fermented using previously collected from sugar mill. It was Recovery of sugars from exploited for ethanol production. The hydrolysate was separated from hydrolysed by sulphuric acid treatment hydrolysate Solvent tolerance, energy savings the residual bagasse and sugars and in two stages. In the first stage hydrolysis, The sugars from the bagasse hydrolysate through reduced cooling costs, higher furfural in hydrolysate were estimated. the sugarcane bagasse was soaked were recovered by ion exchange saccharification and fermentation The maximum 26.14% xylose and 4.6% into 2%–10% w/w sulphuric acid with chromatography using strong anion and rates, continuous ethanol removal, glucose (47% sugars of total cellulose a solid-to-liquid ratio of 1:10 to 1:4.2. weak anion resins in the ratio of 5:1 to and the reduced risk of contamination and hemicellulose present in the The temperature in the digester was 1:1. A glass column with 100 cm length have stimulated a search for sugarcane bagasse) was obtained at maintained at 100 °C for an hour and and 3 cm diameter was packed with thermophilic or thermotolerant yeasts. 8% w/w acid concentration and at 1:4.2 agitation in the reactor was maintained 700 g resins. The bagasse hydrolysates Less energy is required for mixing solid-to-liquid ratio, as shown in Table 2. 1000 rpm. The aqueous phase was obtained in the first stage of hydrolysis and product recovery in thermophilic In second stage hydrolysis separated from the residual bagasse contained sugar concentration of fermentations because of lower 6%–18% glucose and 2.5%–4.64% followed by its washing to collect 35 g/l and sulphuric acid concentration of viscosity, surface tension, higher vapour Screening xylose were obtained, as shown in test for xylose-rich hydrolysate-rich stream. 60 g/l. In the second stage of hydrolysis, pressure, and increased solubility of Table 3. The maximum 33% sugars ther- In second stage hydrolysis 18%– it contained sugar concentration of 80 organic compounds. moamylase of total cellulose and hemicellulose 65% w/w sulphuric acid was added to g/l and sulphuric acid concentration This article describes the present in sugarcane bagasse was residual bagasse taken from first stage of 200 g/l. The hydrolysate was passed saccharification of sugarcane bagasse achieved at 65% w/w sulphuric acid hydrolysis. The temperature in the through the column with flow rate by acid treatment followed by sugar concentration when the digester digester was maintained at 80 °C for 1 ranging from 4 to 17 ml/min. The acid recovery from the hydrolysate by temperature was maintained at 80 °C hour and agitation was maintained 1000 was retained in the column and sugars ion exchange chromatography and for 1 hour followed by diluting the acid rpm. The aqueous phase was separated were eluted through the column. The fermentation of sugars in batch and to 20% w/w and again maintained the from the residual bagasse followed column was regenerated with water to continuous mode with recycle of temperature 100 °C for 30 minutes and by its washing to collect glucose rich recover the acid and the acid solution thermophilic yeast, Klyuveromyces agitation was maintained 1000 rpm. The hydrolysate rich stream. was recycled back for hydrolysis of fresh sp. IIPE453 in the temperature range furfural concentration was negligible. sugarcane bagasse. of 50 °C.

18 VOLUME 2 • ISSUE 6 JUNE 2009 VOLUME 2 • ISSUE 6 JUNE 2009 19 feature article feature article

Continuous fermentation process at 45% on the basis of fermentable Table 3 Sugars and furfural (percentage of bagasse) recovered by second with recycling the cells sugars and the overall ethanol yield on hydrolysis at different acid concentrations Sugar conc. (G/l) The sugarcane bagasse was fed into 35 dry cassava basis was 33%. The dry cell Soluble starch Acid concentration Acid (%) (w/w) the bioreactor with a dilution rate of 30 mass was almost constant through out (%) (w/w) (acid/sugar) Glucose (%) Xylose (%) Furfural (%) Cassava starch 0.075/h and 0.1/h at 45–60 °C. The Tapioca starch the fermentation. 18 50 6 3.5 0.21 highest ethanol yield and productivity 25 26.4 75 9.7 2.5 0.16 of 42% and 2.3 g/l/h, respectively, was 20 Sweet sorghum Conclusion Maize 33.8 100 14.5 4.64 0.14 obtained at 45 °C and 0.1/h. At 50 °C, 15 The overall yield of fermentable sugars 65 125 18 3.34 0.11 air/N was passed through the in acid treatment was 65%–80% 2 10 bioreactor for in situ recovery of ethanol (w/w) of total cellulose and hemicellulose at different conditions as shown in 5 present in sugarcane bagasse under Hydrolysate sugar Ethanol conc. (G/l) Figure 2. The overall ethanol yield on 0 different operating conditions. The conc. (G/l) DCM (g/l) the basis of total fermentable sugars 0 2 4 6 8 10 12 overall ethanol yield on the basis of 35 12 Time (h) present in hydrolysate in continuous total fermentable sugars present in Ethanol 30 10 fermentation with cell recycle at hydrolysate was obtained 35% in batch 50 °C was 35%–38% with ethanol Figure 3 Enzyme hydrolysis of starch base biomass at 80 °C by thermoamylase produced process with ethanol productivity of 25 by Geobacillus sp. IIPTN 8 productivity of 0.216–1.86 g/l/h. 0.52 g/l/h at 50 °C. The overall ethanol 20 DCM Almost 90% of ethanol was recovered yield on the basis of total fermentable 6 during fermentation on stripping by sugars present in hydrolysate in 15 Conc. (G/l) air/N and five times concentrated 30 continuous fermentation with recycling 4 2 10 ethanol was obtained as compared to the cells at 50 °C was 35%–38% Sugar conc. 25 2 ethanol in fermented broth. with ethanol productivity of 5 Hydrolysate sugar 0.216–1.86 g/l/h. Almost 90% of 0 0 Simultaneous liquification and 20 ethanol was recovered during 0 2 4 6 8 10 12 14 16 18 20 Time (h) saccharification of starch biomass fermentation on stripping by Different starchy biomass like soluble 15 air/N and five times concentrated Figure 1 Ethanol production in batch mode by Klyuveromyces sp. IIPE453 in sugarcane Ethanol conc. 2 bagasse hydrolysate at 50 ºC starch, cassava starch, tapioca starch, ethanol was than ethanol in broth. sweet sorghum, and maize were 10 The thermoamylase was found very hydrolysed by thermoamylase isolated effective for hydrolysing different types Recovery of sugars from consumed within 20 hours as shown in 5 from Geobacillus sp. IIPTN at 80 °C as of starch biomass with a considerable hydrolysate Figure 1. The final ethanol concentration shown in Figure 3. The total starch yield at high temperature. The sugarcane hydrolysate, containing in broth was 10.2 g/l with ethanol yield 0 for each substrate was hydrolysed in 0 20 40 60 80 100 120 140 fermentable sugars and sulphuric of 35% and productivity of 0.52 g/l/h. Time (hr) acid, was passed through the column Figure 4 Ethanol production in batch mode by free cells of IIPE453 from cassava containing ion exchange resins. About Conc. (G/l) 70 hydrolysate. 95%–100% acid-free sugars were 60 recovered, with strong anion and weak anion mixture in the ratio of 5:2 and 50 10 hours. The sugar yield on tapioca flow rate 17 ml/min. Ninety five per cent 40 and sweet sorghum were obtained at acid was recovered in regeneration of 30 81.7% and 58%, respectively. the column. The recovered acid was 20 recycled for further hydrolysis of fresh 10 Ethanol production from cassava sugarcane bagasse. 0 hydrolysate 0 2 4 6 8 10 12 14 16 18 20 22

–1 –1 Fermentation was carried out with D=0.1 h D=0.075 h FeCl3 Air sparging cassava hydrolysate in batch mode by Ethanol fermentation from Nitrogen sparging sugarcane bagasse hydrolysate Time (days) free cells of Kluyveromyces sp. IIPE453 Hydrolysate sugar in feed Hydrolysate sugar in recycle Batch fermentation process Hydrolysate sugar in outlet DCM in fermenter as shown in Figure 4. The total sugar The concentration of thermophilic Ethanol in outlet Ethanol in condensate in hydrolysate was consumed in yeast, Kluyveromyces sp. IIPE453, was 138 hours with productivity of Figure 2 Continuous fermentation with cell recycle at 50 °C and air/N2 stripping in sugarcane kept at about 10 g/l. Total sugar was bagasse hydrolysate by Kluyveromyces sp. IIPE453 0.09 g/l/h. The ethanol yield was obtained

20 VOLUME 2 • ISSUE 6 JUNE 2009 VOLUME 2 • ISSUE 6 JUNE 2009 21