Applied bioethanol technology in Brazil* Angewandte Bioethanoltechnologie in Brasi Josef Dérfler and Henrique V. Amorim The Brazilian bioethanol industry has plenty of experience. As result of the international oil erisis in 1973 and. 1979 the govern ‘ment supported ethanol technology and production by the n development program “Proalcool”. Actually 329 processing plants with an average production quantity between 600 and 800 md fare in operation resulting in 17.6 bn L of bioethanol in 2006. Until the year 2012, 7 new mills will be established inereasing the an- ‘ual production volume up to 28 bn L. Bioethanol production as an integrated part of sugarcane processing can be based on raw molasses or a mixture of both, On average a yield of 85 L of etha- nol per tonne of sugarcane is calculated. Since 1979 ongoing improvements, developments and experiences have been mastering the applied bioethanol technology thus result- ing in highly efficient processes. Intelligent fermentation concepts ‘such as “high density processes” through yeast treatment and re- cycling are generating fast batch cycles of less than 12 hand high a yields of up to 92% (based on the theoretical equation from sugar to ethanol). Using bagasse as the energy source also provides a high-yield operating energy balance. As fermentation concepts batch and continuous processes are applied. Due to higher production stability and lower contamination risk the batch proce ess concept is favored. Approximately 75% of all existing plants ‘work with batch processes, Scientific knowledge related to genetic stability and viability of yeast strains as well as microbiological in- process controls are the main success factors for the ongoing high productivity within the bioethanol process. Therefore the applied yeast strains and fermentation parameters have a significant impact ‘on the profitability of ethanol plants Key words: bioethanol, technology, processing, y Brazil st reeycling, 1 Actual ethanol technology in Brazil 1.1 Introduction and general remarks for high yield processing Based on long term experience and practice the Brazilian sugar tnd alcohol industry was able to develop highly efficient ethanol processes with robust and best technology om the lange scale. Those ‘developments and improvements were originally supported and initiated through the national government program “Proalcool”, launched in 1975, with the goal of using ethanol as a fuel to reduce oil imports Besides sugarcane productivity and sugar extraction an important impact is contributed by the design and process performance of the lien Die brasilianische Alkoholindustrie kann auf eine langjihrige Er- fahrung verweisen, Die internationalen Erdolkrisen der Jahre 1973 ‘und 1979 waren der Anlass 2um Star des nationalen Forschungs- programms ,Proalcoo!” zur Entwicklung der Alkoholtechnolo- ‘gic und -produktion, Im Jahr 2006 waren 329 Fabriken mit einer ‘durchschnittlichen tglichen Produktionsmenge von 600 bis $00 1m’ in Betrieb, die insgesamt 17,6 Mrd. L Bioethanol produzierten Bis zum Jahr 2012 sollen weitere 77 Fabriken errichtet werden, um, je jdhrliche Menge auf 28 Mrd. L zu ethohen. Die Bioethanol- produktion witd als integraler Prozess bei der Rohrzuckergewin- ‘nung beirieben, wobei als Fermentationsrohstoff Rohsaft, Ablaut oder eine Mischung aus beiden Komponenten verwendet wird. Ein ddurehschniitlicher Ertrag von 85 L Ethanol pro Tonne Zuckerrobr ‘wird erreicht. Seit dem Jahr 1979 werden permanent Verbesserungen und Ent- wicklungsarbeiten beim industrellen Bioethanolprozess vor: ‘genommen, Durch hohe Biomassekonzentrationen _wihrend des Fermentationsprozesses sind sehr kurze Zykluszeiten. von < 12h fireinen Batchansatz miglich, Durch Hefertickfuhrung und -aufbercitung werden hohe Transformationsraten des vorliegenden Zauckers 70 Ethanol von bis zu 92 % erreicht, Ebenso fibre die ther- rmische Nutzung der Bagasse 7u einer energieautarken Bilanz des Gesamtprozesses der Zucker- und Ethanolproduktion. Als Fermen- tationsverfalren werden sowohl Batch- als auch kontinuierliche Prozesse angewandl. Allerdings wird dem Batchkonzept, mit dem twa 75 % aller Alkoholfabriken arbeiten, aufgrund seiner hoheren Produktionsstabilitit und geringerer Infektionsrisiken der Vorzug egeben, Stichwiirter: Bioethanol, Technologie, Produktion, Heferickith rung, Brasilien alcoholic fermentation, General basic knowledge of biotechnoloy cal processes such as cleaning and sanitation of equipment, st lization and pasteurization, nutrient and growth requirements for ‘yeast, possible biochemical pathways of yeast and bacteria under ‘aerobic and anaerobic condition is essential for efficient manage~ ‘ment of an ethanol plant. ‘Therefore many industrial parameters related to heat treatment [processes of equipment and raw juice for reducing bacterial con tamination, yeast viability and optimal nutrient composition have to be carefully controlled within ethanol processes. Taking only account of engineering aspects and simple measurable on-line pa- * Piper rset tthe 3 Bioethanol ecology Metin in Peto. Gemmany, 24-25 A207Generally the energy balance of a liquid-liquid distillation process as applied in Brazil is better, because the beer sent to the column is free of solids and reduced by volume through previous yeast separation and recycling. 2. Future trends and developments Driven by increased competition and cost pressure in the bioeths- ‘ol sector the industry has to look for applicable and well proven ‘technologies with higher ethanol conversion yields and energy Sav- ing concepts. Taking into account the advantage of the Brazilian technology with yeast recycling it wll be a step forward for further process improvement. Transformation of that technology to etha- ‘ol production from starch-based raw materials will leverage the -y including less energy’ costs for drying of the resulting Ingledew (2007) remarked that through grain removal after dex ‘rinization in the dry corn milling process an approximate 11% volume gain in the fermentation vessel can be realized including saving energy for cooling of the insoluble particles in the fermenter ‘and for heating of those in the distillation column. No dry grind fuel ethanol plant is yet known to carry out grain removal prior to fermentation, Technology in wet milling, of course, does lead 10 solid-free mashes, and in this process spent yeast iy easily removed at the end of the fermentation leading to a liquid-liquid distilla- tion It has also to be considered that through higher viable yeast cell ‘concentrations in the fermentation vessel higher alcohol concentra- tions can be achieved more easily. How the fermentation conditions (cg. temperature, feeding of the carbon source) have tobe adjusted should be the program of further research activities in the mid-term ‘un. It will be also a question to find the optimal balance between alcohol concentration, fermentation time and yeast cell viabi lty for achieving that goal. Those parameters will strongly influ- ence the economic results of ethanol plants in the future. References Aunorin, HV (2007): Globalization ofthe Brazilian Bioenergy Program ~ Op: ertunties & Challenges, Presentation athe Andrew Van Hook Symposi, notion and Environment Protection fora Sustanahle Sugar Indu", March 2007, Reims/France Amorim, HV. Lipes LM, 2005): Ethanol Production ina Petroleum Depend ent Word: The Brazilian Experience. Sugar Joural, May, 11-15 Ingledevs M.W (2007) Fermentation: Recent Advances. Presentation at the Sd "European Ettanol Technology Mesting, April 2007, Detmold/Germany ‘Technologie du bioéthanol appliquée au Brésil (Résumé) Lindustie du bioéthanol au Brésil posséde une grande experience. ‘Ala suite des crises internationales du pétrole en 1973 et 1979, le gouyemement a soutenu la technologie et Ia production de éths- ‘nol par le programme de développement national ,Proaleool En fait, 329 usines avec une prxluction moyenne comprise entre 600 et 800 m'fjour sont en activité se tradusant par une produc tion de 17,6 milliards de litres de bioéthanol en 2006. Jusqu'en 2012, 77 nouveaux moulins seront installés aceroissant la produc tion annuelle jusqu’a 28 milliads de lies. La production de bio: ihanol comme partic intéorante du traitement de la eanne & suere ‘au biogthanol se traduisant par des procédés hautement efficaces. Des concepts intelligents de fermentation tels que les “processus & ‘haute densité” grace & un traitement des levures et au recyclage en- ‘trainent des cycles discontinus rapides de moins de 12h et des taux. de conversion élevés allant jusqu’a 92 % (caleulé sur I’équation ‘théorique de Ia transformation du suere en éthanol). L'utilisation de la bagasse comme source énergie procure également un bilan <énergétique de haut rendement, Pour les concepts de fern ‘on applique les processus continus ou discontinus. A. stabilité élevée de la production et du faible risque de contamina tion, Ie concept de processus discontinu est prétéré. Environ 75 % de toutes les usines existantes travaillent avec le processus discon- tin, La connaissance scientifique concernant la stabilité génétique ‘et la viabilité des souches de levute aussi bien que les contrales mi ‘crobiologiques en cours de fabrication sont les principaux facteurs ‘de réussite pour la haute productivité au cours de la fabrication du ‘boéthanol. C'est pourquoi le choix des souches de levure adoptses ‘et les parametres de la fermentation ont un impact significatif sur la rentabilité des usines déthanol, ase d'une La tecnologia del bioetanol aplicada en el Brasil (Resumen) La industria alcoholera det Brasil ha adquirido muchos ats de ex periencia, Las crisis internacionales del petr6leo de los aftos 1973 y 1979 fueron cl motivo para el comienzo del programa de inves tigacién nacional “Proalcoo!” para el desarrollo de la tecnologia alcoholera y la produccién del alcohol. En el ato 2006 trabajaron 329 fabricas en el pais con una proxluccién diaria media de 600 '800 my una produccién total de 17,6 mil millones L de bioetanol. Hasta el aflo 2012 se construiin otras 77 fabricas y se aumentarii Ja cantidad anual a 28 mil millones L.. La produccidn de bioetanol 5 um proceso integral de la produccin de azsicar de cana, la mate- ria prima de fermentacion es jugo crudo, melaza o una mezcla de ambos componentes. De una tonclada de cafa de aztcar se obtiene ‘un rendimiento medio de 85 L de etanol. El proceso industrial del bioctanol vive continuamente mejoras y desarrollos. Las concen= traciones altas de biomasa durante el proceso de fermentacisn per- :miten tiempos de ciclos muy cortos (< 12 h) para una mezcla bateh. Con la recirculacién y 1a recuperaciGn de Ia levadura se obtiene un alto tanto por ciento de transformacién desde el azdcar hacia el eta nol de hasta 92 %. El aprovechamiento térmico del bagazo conduce dems « un balance autosuficiente en enengfa del proceso total de la producciGn de azsicar y etanol. Como métodos de fermentacién ‘se aplican no sélo procesos batch sino también procesos continuos. Pero, la mayor estabilidad del producto y el menor riesgo de infec~ cin son el motivo para que el 75 % de toxas las fabricas aleohole- 1s trabajen en base del concepte batch, Authors’ addresses: Josef Drfler, Partner & Representative of Fet= ‘mentee Lida. for Europe, Asia & North Africa, PhD. Oberer Gries 22, A-3281 Oberndorf, Austria; e-mail: josef.doerfler@ utanet.at Henrique V. Amorim, PhD, Professor, Founder & President of Fermentee Lida, Fermentee Ltda., Av. Antonia Pizzinato Sturion, 1155, CEP 13420-640 Piracicaba ~ SP; Brazil; www-fermentec. ‘com.br ~ Fermentee Lida is a company established in 1977 and specializing in alcoholic fermentation and laboratory conteol of susar and alcohol production. —i, 2: Batch proo 3s with yeast reeycle applied in Brazil (Fermen. “= aE Fe 3: Continuous process with yeast recycle applied in Brazil mentec, 2007) As fermentation concepts batch and continuous processes are ap- plied (Figures 2 and 3). Due to higher production stability and lower contamination risks approximately 75% of al existing plants work with batch processes, Besides knowledge of the optimal fermentation parameters. the choice of yeast strains is fundamental to obtain high yields and alow consumption of antifoam, Fermentec Ltda. selected the yeast stains PE-2, CAT-1 and VR-1 from industrial processes by qualification through a karyotyping technique. These strains are very resistant to stress conditions as existing through permanent yeast recycling during the whole production campaign of approximately 8 months Figures 4 and 5 show the technical fermentation infrastructure of batch cycle as deseribed above. In Table 1 the processed sugare: Cf the largest Brazilian factories in 2006 are listed. For runni these industrial processes under stable conditions and for imple ‘mentation of further improvements it is required to have permanent access to actual technological and scientific support Table 1: factories in 2006 roduction Figures from the larg Brazilian sugar Factory Sugareane Susur Ethanol mnt mat mal ‘Slo Marinto 738 oes Da Bara 620 0.500 2 Vale do Rosé $359 0375 20 Santa Elsa 0372 4 amar 907 0.286 2st Colorado ash 0.339 193 Bonfin 188 0.356 10 Meera” 4140 282 x2 Colombo 138 032 rn} Da Pee 3910 0.195 203 These plants receive tesnological consulianey from Fermenee Lid ttvough annual comic 14 Distillat The distillation process for bioethanol purification iy performed within 2 stages which can be linked and integrated together for 1 stage for hydrated aleohol of 96% (w/s}: © for anhydrous alcohol of more than 99.5% (v/v. on market requirements (e.g. use as fuel for flex fuel vehicles in Brazil or for customs regulations on export markets) the anhydrous step is cancelled and therefore only hydrated alco- hol of 96% (v/v) is produced. Besides molecular sieve technology purification the Bruzilian mills apply an extraction process with‘well as excessive yeast growth, stresses caused by sugar, alcohol salt, high temperature, acidity and contamination by wild yeast are often troubles arising, and these can be only managed with a background of profound biotechnological expertise. To allow the ‘best management of the industrial process the key parameters and their interactions have to be identified to obtain knowledge of the ‘hole ethanol process; ie. where to invest to improve yields, to obiain high-quality ethanol, to decrease losses and to reduce the consumption of chemicals such as sulfuric acid, nutrient supple- ‘ments, antifoams and others. By measuring, controlling and improving the relevant process pa- rameters the Brazilian ethanol industry has been able to improve the fermentation yield from 75% to more than 92% (based on theoretical maximum production of S11 g ethanol/kg glucose as shown in Fig. 1). Without those improvements it would be impos- sible for the distilleries to survive with the low prices on the inter= national market as was the case up to 1999, 1009 ‘SUGAR 209 Kc! (00%) 175 Kea (46%) Kea! 24%) ae Fig. 1: Energy balance of aleo- ALCOHOL 264 Kos! (99%) + holie fermentation (Fermentec, 42.9900, 2007) ‘The success of high yield processing in the aleohol industry has. also resulted from research and scientific activities performed dur- ing the last 30 years in Brazil such as ~ Conditions to improve the fermentation yield ~ Selection of the yeast strains from the industrial environment: ~ Development of analytical methods to measure accurate and rapid sugar concentrations in juices, molasses and beers appli- cable in each industrial laboratory ~ Direct counting of living bacteria by light microscopy in up to 15 minutes — Method to choose an effective bacteriostatic agent (in 6 hours) to control bacteria ~ Karyotyping of yeast cells for identification ~ Identification of contaminating bacteria by DNA sequencing: Accurate and precise determination of alcoholic content by dig- ital densimeters ~ Introduction of NIRS (Near Infrared Spectroscopy) to routine analysis = Reduction of the sugar losses and consumption of chemical. 1.2 Raw material management and sugar extraction, In 2006 sugarcane was grown on 6 mn hectares (0.7%) from the total Bruvilian land area of 851 mn hectares. For comparison soy ‘bean and cotn were cultivated on 34 mn hectares (4.0%) and other crops (e.g. coffee. oranges) on 22 mn hectares (2.6%). For 2006 the following production numbers related to sugarcane were accounted (Amorim, 2007): Ethanol productivity of 7500 Libectre. ‘The main area for sugarcane growing is located inthe north of Sao Paulo state, where the crop is harvested within time period of approximately 8 months starting in March each year. Sugarcane is seeded only onetime within to 7 years and is cut yearly with a certain productivity profile. In Sao Paulo state the typical harvest Yield starts with 120 vectare inthe second year and deereases to 70 uhectae in the final yea. Depending on the harvest time ofeach Year, the sugar content varies inthe range of 13 to 17% In Brazil ethanol is almost always produced in combination with sugar at the same facility. Therefore a special flexibility exists ine the production processes. Wien the ethanol prices the international market are more profitable than for erystallized sugar, more sugarcane is dedicated to ethanol production. Also the ccopreduct molasses is directly sent to the fermentation making an ‘optimal logistical management with higher economic output. Ac- ‘wally around 50% of the processed sigarcane i used for ethanol production in the factories (Amorim and Lopes, 2005). In most mills the cane is crushed ina series of $ large roller mills, ‘The raw juice gushes out and the residual sugar in the fiber is ex- tracted by hot water. In some factories horizontal diffusers are used applying a similar concept as is well known for sugarbeet extrac- tion, The raw juice is clarified, e.g. with Hime and afterwards fil tered through a rotary vacuum filter for eliminating the precipitate and solids. Important forthe liming step is to adjust the applied process parameters onto the nutrient requirements of the ethanol step avoiding any limitation of yeast metabolism. For higher alcohol concentrations in the fermentation stage an in- crease of the sugar concentration is required through an evapora tion treatment of the raw juice or mixing it with molasses. The ‘extracted sugarcane fiber (bagasse) is fed to the boilers for generat ing high pressure steam and electricity. No extemal energy source the surplus of produced energy is sold to the electrical 1.3. Fermentation process and yeast handling 1m 2006, 329 ethanol plants with an average production quantity between 600 and 800 m'/d were in operation resulting in 17.6 ba L. Until the year 2012, 77 new mills will be constructed increasing the annual production volume up to 28 bn L. Typically for the Brazil- jan technology isthe yeast recycling concept which results in high ‘transformation rates fo ethanol, because less carbon source is used for yeast growth (see section 1.1). Fermentations are carried out with very high yeast cell concentra- tions of up to 12% (wet base), alcoholic concentrations of up to 12% (v/v) and very short fermentation times between 8 and 10 h. -Afler the end of each cycle the yeast cells are separated from the fermented medium (beer) and concentrated by centrifugation 10 be ‘used in the next fermentation cycle. The beer goes to distillation ‘while the concentrated yeast cream is treated with diluted sulfuric acid to kill bacteria, After the treatment the yeast cells return to the {ermentation vessels, where they restart a new fermentation cycle, For applying that concept it is required to have a juice and beer with a low concentration of inert solids, otherwise the biological process will fail or will not work property.