Design and start-up of a fluidized bed bioreactor for ethanol fermentation

Sinsupha Chuichulcherm
Department of Chemical Engineering, Srinakharinwirot University, Thailand

Due to an increase in fuel price, ethanol is an interesting alternative fuel since it can be fermented from agricultural waste. A system allowed higher ethanol production and shorter fermentation period compared to a conventional system is preferred. A fluidized bed reactor was investigated. A 20 L anaerobic fluidized bed reactor was constructed for ethanol fermentation. PVC pellets were used as the solid support for biomass. The reactor was designed and constructed from material available in Thailand. 16L pineapple juice was fermented by saccharomyces cerevisiea in the fluidized bed reactor with a circulation flow rate of 16L/min. A final concentration of 17vol% ethanol production was obtained from the system within 200 hours in a batch mode during the start-up period. A conventional method for ethanol fermentation in 20L tank without circulation was also performed. After the fermentation was completed, the ethanol concentration in the fluidized bed reactor was 132.72 g/L which is 30%higher than the ethanol concentration in the fermentation batch tank and also contained less residual sugar concentration. It could be concluded that the fluidized bed reactor can be used in ethanol fermentation process with a higher efficiency compared to a conventional one.
Keywords: fluidization, ethanol, reactor design

Introduction
Ethanol is a product of sugar fermentation under anaerobic conditions. In order to get a high ethanol concentration, an immobilization of microorganisms either inside or at the surface of a support is needed since the immobilized microbes can tolerate more severe conditions than individual cells especially when the product is a growth inhibitor (Nagpal et al.,2000). Moreover, advantages of using an immobilized bioreactor over a continuous stirred tank bioreactor are (i)the achievable biomass concentration, (ii) the longer hydraulic resident time or a higher dilution rate (Marin et al., 1999). Biofilm formation onto the surface of the support is the result of a number of processes, including growth of microbes, attachment and adhesion, decay of microbes and detachment from the support surface (Nioella et.al, 1999). When the biofilm grows, it alters size, shape, density and surface roughness and bulk density of the support particle. It can also increase the surface area so that the heat and mass transfer can be obtained easily. A fluidized bed bioreactor is a fermenter that uses the advantages of immobilized biomass. Moreover, the temperature of the liquid in the reactor is almost constant since the immobilized biomass particles behave like a fluid resulted in a good mixing. The use of fluidized bed technology for wastewater treatment has been studied at the laboratory scale and pilot plant level. It is also applied to ethanol fermentation. Nevertheless, the fluidized bed reactor used in Thailand is usually bought from aboard. Due to a large number of variables involve in a fluidized bed bioreactor

design, empirical values are more useful than the theoretical one. The design strategy will be based on experimental result from published work and it is adapted for the material that can be found in Thailand.

Design considerations of a fluidized bed reactor

a) Support material: material used should have a high surface area to volume ratio, a rough surface or high porosity with a slightly high density compared to the fermentation broth. Porous support such as clay and activated carbon is suitable for biofilm attachment since it has a high adsorptive power but its strength is not good enough when to be used in a fluidized bed reactor. Sand is always reported to be used as it has a rough surface and the biomass can attach onto the surface but not inside the sand particle. Other physical characteristics which have to be considered are size, shape, and hardness. It is also have to consider the cost of the material and its poisons to the microbes. b) Support particle diameter and the fluidized bed column diameter: according to Di Filice (1996), column diameter should be 30 times larger than average diameter of a support particle to avoid hinder settling velocity effect. Also, Marin et al., 1999 recommended the height of the column should be around ten times of the column diameter. However, the surface velocity in the fluidized-bed system needs to achieve 20% to 40% expansion to get a better mix and higher heat and mass transfer rate. c) Alkalinity and recirculation: due to an increase of a fermentation product, CO2, pH of the fermentation broth seem to shift to (more than pH 7) or (a range of slightly alkaline). Also, a need of high surface velocity in the fluidized column, sometimes higher than the dilution rate, makes a recirculation and a recycle port necessary. The recycle port is a small box where the feed inlet port, the effluent port, pH probe, are situate and is attached to the fluidized bed column. A recirculation pump can be used to circulate fermentation broth form the recycle port to and from the column with a flow rate higher than the dilution rate Nagpal et al, 2000 used a recirculation flow rate 50 times higher than the dilution rate in an anaerobic system to achieve a better yield. d) Velocity in fluidization operation: two velocities which have to be considered are VOM, a minimum fluidization velocity and Ut, a terminal fluidization velocity. The two velocities can be calculated using Ergun equation. However, if the particle size is larger than 1 mm, the laminar flow term in the Ergun equation becomes negligible, and VOM can be written as shown in equation (1) (McCabe et al., 1993).
3 D p ( S l ) M (1) VOM = 1.75 l The terminal velocity is proportional to VOM and void fraction as can be seen in equation (2). 2.32 (2) Ut = 3 / 2 VOM M The two equations can be used when the particles Reynolds number is higher than 1000 (a turbulent regime). If the velocity used in the operation is lower than the VOM, the bed would not expand. On the other hand, when the operating velocity is higher than the Ut, all the support particles would be carried out of the column with the 0.5

fermentation broth. Then, the volume flow rate of the fermentation broth can be determined by multiply the velocities calculated with a cross-sectional area of the column. Once the volume flow rate known, a suitable recirculating pump and rotameter can be selected.

Material and methods

a) Fluidized bed bioreactor: fluidized bed column was made of Perspex with a 0.08m inside diameter and a 1.0m height. The Perspex was bought from Wongwaien 22, Bangkok. A round stainless steel sheet with a diameter equal to the diameter of the column was fixed at the top of the column or at the fluid exit to prevent loss of the support particles. Liquid was pumped from the bottom of the recycle port, made from a plastic box, to the bottom of the fluidized bed column using a centrifugal pump. Recirculation flow rates in a range of 15-16 L/min were used. Figure 1 shows the apparatus without the support particle filling. Support particles used in the experiment were PVC pellets with an average diameter of 2.35x10-3m. They were rubbed with sand paper to give a rough surface. PVC pellets were selected since they are inert to alcohol, easy to buy, cheap and non-toxic to bacteria. Density of the PVC pellets was 2053 kg/m3. Figure 2 shows a closed-up picture of PVC pellets before and after use in the bioreactor.

Figure 1 the20L fluidized bed bioreactor.

b) Microorganism and fermentation: Saccharomyces cerevisiea were cultivated in pineapple juice to be used as a starter. 16L boiled pineapple juice was fermented in the fluidized bed bioreactor in a batch mode during a start-up period. Sugar and ethanol concentrations in the fermentation broth were measure using phenol-sulfuric method and gas chromatography method respectively.

Figure 2 pictures of PVC pellets before and after use in the bioreactor

Results and discussion

After the fluidized-bed reactor was assembled, an experiment on fluidization behavior was conducted by altering the liquid flow rate and measuring the bed height. The results show in figure 3. It can be seen that when liquid (tap water) flowed upward through the bed at low velocity range (at flow rates of 0-6L/min), the particles remained stationary. Further increase in water flow rate, the particles began to move. It is because the force of the pressure times the cross-sectional area is equal to the gravitational force. At this point it is called the minimum fluidization (Geanckoplis, 1993).Upon further increase in the water flow rate, the bed expanded and the void volume increased. The particles flowed upward to the top of the column and would be carried out unless there was a screen to prevent the loss. At that time, the water flow rate was 20L/min, which were higher than the terminal velocity. A plot between pressure drops and water flow rates is also shown in figure 3. The results supported the results discussed earlier. The pressure drop increased as the water flow rate was increased until the minimum fluidization was observed. Then at a flow rate of 8L/min the pressure drop decreased dramatically and remained unchanged as the bed continue to expand. However, when using pineapple as fermentation broth, a bed height was slightly higher than a height when using pure water. It was probably due to a change in fluid density and viscosity.

50 45 40

800

600 Pressure drop (Pa)

Height of bed (cm)

35 30 25 20 15 10 0 5 10 15 20 25 200 400

Bed height Pressure drop 0

Liquid flow rate (L/min)

Figure 3 a plot between bed height and pressure drop when altering liquid flow rate. Results obtained during the experiment on fluidization behavior using tap water as liquid. Ethanol fermentation After circulating a mix of pineapple juice and starter for a week and the yeast was attached onto the support particles, freshly boiled pineapple juice was substituted to the previous one to start a new fermentation batch. Sugar and ethanol concentrations were measured at a time interval. The results were compared with the results from pineapple fermentation using conventional method without immobilized biomass. Initial sugar concentration in each fermentation broth was 350g/L. While the fermentation proceeded, the sugar concentrations decreased and ethanol concentration increased as can be seen in figure 5. After the first 3 hours, the sugar concentrations decreased gradually due to a use in cell growth, maintenance and ethanol formation. However, the sugar concentration in the fluidized bed bioreactor was lower than that in the conventional fermenter after 30 hours. The sugar concentration in the conventional fermenter still decreased until it was constant at 31.67g/L within 200 hours while the residual sugar concentration in the fluidized bed bioreactor was constant at 8.33g/L after 170 hours and it is lower than that reported by Posuwan et al., (1996). Posuwan et al. fermented pineapple juice in a batch process with suspend culture and found the sugar left in the fermentation broth after the process had completed was 24.6g/L. A reason why the sugar concentration in the fluidized bed bioreactor was lower may be because an accumulation of biomass layers onto the support particles made the microorganism more tolerate to ethanol, which is a product inhibitor, than the suspend

microorganism. 97.41% conversion of the sugar in the fluidized bed bioreactor was obtained while 90%conversion was obtained in the conventional fermenter. Also, the ethanol concentration in the fluidized bed bioreactor increased until it reached 132g/L with a yield of 76.72% after 168 hours while the final ethanol concentration in the conventional fermenter was 102.42g/L at 200 hours.

500 ethanol in the fluidised bed reactor ethanol in the conventional reactor sugar in the fluidised bed reactor sugar in the conventional reactor

500

400

400

ethanol concentration (g/L)

300

300

200

200

100

100

0 0 50 100 150 200

time (hour)

Figure 4 residual sugar and ethanol concentration during the fermentation process.

Conclusion
Ethanol can be successfully produced using a fluidized bed bioreactor assembled from materials available in Thailand. PVC pellets were used as support particles. Higher percentage of sugar conversion was achieved when using a fluidized bed bioreactor compared to that using a conventional fermenter.

Acknowledgement
The author would like to thank Miss Kanjana Sudjit, Miss Piyapa Boonsang and Mr. Sutus Sonkami for their help in assembling the experimental apparatus and doing some experiments.

Symbols
Dp Ut VOM M particle diameter terminal velocity minimum superficial velocity for fluidization minimum porosity for fluidization or void fraction sphericity

sugar concentration (g/L)

density; p,particle density; l , liquid density

References
Di Filice R (1996) A relationship for the wall effect on settling velocity of a sphere at any flow regime, Int. J. Multiphase Flow, 22, p527-533 Geanckoplis, Christie J (1993) Transport Processes and Unit Operations, 3rd edit, Prentice Hall PTR, NewJersey, p 123 Marin P., Alklay D., Guerrero L., Chamy R., and Schiappacasse M C. (1999) Design and startup of an anaerobic fluidized bed reactor, Wat.SciTech., 40(8), p 63-70. McCabe W L., Smith J C., and Harriott P., (1993) Unit Operations of Chemical Engineering, 5th edit, McGrawHill, Singapore, p 173-174 Nagpal S., Chuichulcherm S., Peeva L., and Livingston A. (2000) Microbial sulfate reduction in a liquid-solid fluidized bed reactor, Biotechnol Bioeng, 70(4), p370-380 Nicoella C., van Loosdrecht M M C., Di Filice R., and Rovatti M. (1999) Terminal settling velocity and bed expansion characteristics of biofilm-coated particles, Biotechnol Bioeng., 62(1), p 62-70