Fermentative hydrogen production using organic substrates in batch and continuous conditions

Transcription

1 Fermentative hydrogen production using organic substrates in batch and continuous conditions Gustavo Davila-Vazquez, Elías Razo-Flores* División de Ciencias Ambientales. Instituto Potosino de Investigación Científica y Tecnológica, Camino a la Presa San José 2055, Lomas 4ª sección, San Luis Potosi, SLP, México, * erazo@ipicyt.edu.mx

2 Why biological hydrogen production?! A wide variety of microorganisms can evolve H 2 under anaerobic environments.! Bio-H 2 can be produced from organic wastes or cheap substrates.! Using biomass as a substrate, there is no net emission of CO 2.! Fermentative Bio-H 2 production using mixed cultures has a big potential for full-scale practical application.

3 Anaerobic digestion and acidogenesis Angenent LT, Karim K, Al-Dahhan MH, Wrenn BA, Domiguez-Espinosa R Production of bioenergy and biochemicals from industrial and agricultural wastewater. Trends Biotechnol 22(9):

4 Catabolic pathways of mixed-acid fermentation from glucose Lee H-S, Salerno MB, Rittmann B Thermodynamic evaluation on H 2 production in glucose fermentation. Environ Sci Technol 42(7):

5 Effect of hydrogen partial pressure (P H2 ) on hydrogen yield Angenent LT, Karim K, Al-Dahhan MH, Wrenn BA, Domiguez-Espinosa R Production of bioenergy and biochemicals from industrial and agricultural wastewater. Trends Biotechnol 22(9):

6 Davila-Vazquez G, Arriaga S, Alatriste-Mondragón F, de León-Rodríguez A, Rosales-Colunga LM, Razo-Flores E. Fermentative biohydrogen production: trends and perspectives. Rev Environ Sci Biotechnol 2008;7:27-45.

7 Aim of this work Lettinga, G., et al (1980). Biotechnol Bioeng. 22: To study biohydrogen production using an enriched mixed population from anaerobic granular sludge and lactose or cheese whey (CW) as substrate in batch and continuous fermentations.

8 Experimental strategy: Batch experiments Central composite design Treatments Substrate (g/l) ph Every 3 h Gas sample to GC (TCD) 120 ml serum bottles/headspace of 40 ml. Cultivated at 37ºC/150 rpm. Mineral medium. H Gas liberation from serum bottles. mmol H y = 0,000252x + 0, R 2 = 0, Area CO 2

9 Data analysis: Batch experiments 500 H max H(t) (ml) R Modified Gompertz equation: ÏÔ È H (t) = H max! exp - exp R! e Ô Ì Í (l - t) + 1 ÓÔ Î H max Ô l Time (h) V H, i = V H, i-1 + C H, i V G, i + V H (C H, i - C H, i-1 ) V H,i, V H,i-1 = cumulative H 2 gas volumes at the current (i) and previous (i-1) time intervals (ml) C H, i, C H, i-1 = fraction of H 2 gas in the headspace of the bottle measured by GC, in the current and previous time intervals (%v/v) V G, i = H 2 gas released at current time (i) (ml) V H = Headspace volume (ml) H max (ml H 2 ), R (ml H 2 /h), l (h) Calculate: mol H 2 /mol substrate (HMY) mmol H 2 /L h (VHPR) Analyzed by Surface Response Methodology using Statgraphics 5.0

10 Water displacement device Biocontroller Gas (H 2 + CO 2 ) Continuously stirred tank reactor (CSTR). Applikon,, 3 L. Work volume: : 2 L. Volume produced Stirrer controller Feed NaOH 10M Peristaltic pump for both feeding CW and withdrawn of medium Antifoam Effluent

11 Results

12 Kinetics of Bio-H 2 production from CW

13 Davila-Vazquez G, Alatriste-Mondragón F, de León-Rodríguez A, Razo-Flores E. Fermentative hydrogen production in batch experiments using lactose, cheese whey and glucose: Influence of initial substrate concentration and ph. (In Press) Int J Hydrogen Energy. doi: /j.ijhydene

14 Table 1. Summary of experimental conditions for best HMY and VHPR. Substrate Lactose CWP Highest HMY (mol H 2 /mol substrate) and VHPR (mmol H 2 /L-h) obtained, and conditions at which they were found HMY VHPR 3.6 ± 0.03 ph = 7.5 [S 0 ] = 5 g/l 3.1 ± 0.04 (mol H 2 /mol lactose) ph = 6.0 [S 0 ] = 15 g/l 5.6 ±0.48 ph = 7.5 [S 0 ] = 5 g/l 8.1 ± 1.5 ph = 7.5 [S 0 ] = 25 g/l Davila-Vazquez G, Alatriste-Mondragón F, de León-Rodríguez A, Razo-Flores E. Fermentative hydrogen production in batch experiments using lactose, cheese whey and glucose: Influence of initial substrate concentration and ph. (In Press) Int J Hydrogen Energy. doi: /j.ijhydene

15 VFA detected at the end of CW fermentation 70 Acid concentration (mm) g/l,4.5 5 g/l, g/l, g/l, g/l, g/l,8.12 0,86 g/l, g/l, g/l, g/l,6.0 Acetic Butyric Propionic

16 VFA detected at the end of lactose fermentation [Acid] (mm) g/l,4.5 5 g/l, g/l, g/l, g/l, g/l,8.12 0,86 g/l, g/l, g/l,6.0 Acetic Propionic Butyric

17 Carbonate vs. Phosphate mineral medium using CW 6383 ml H mol H 2 /mol lactose 960 ml H 2 /h 400 ml H 2 /L h 3944 ml H mol H 2 /mol lactose 566 ml H 2 /h 235 ml H 2 /l h Time " Cumulative H 2 production using carbonate-based medium (A),! Cumulative H 2 production using phosphate-based medium (B),! ph:buffer A, " ph: buffer B.

18 CSTR: Operation parameters Parameters Experimental period A B C D E F Duration [d] Hydraulic retention time (HRT) [h] Organic loading rate [g lactose L -1 d -1 ] Phosphate buffer was used and ph was controlled at 5.9 using NaOH 10M Davila-Vazquez G, Alatriste-Mondragón F, de León-Rodríguez A, Razo-Flores E. Continuous biohydrogen production using cheese whey: Improving volumetric hydrogen production rate. (In preparation).

19 F E A B C D

20 92.4 g lactose L -1 d g lactose L -1 d mmol H 2 L -1 d L H 2 L -1 d g lactose L -1 d -1

21 CSTR operation and performance Parameters Experimental period A B C D E F Duration [d] Hydraulic retention time (HRT) [h] Organic loading rate [g lactose L -1 d -1 ] Volumetric hydrogen production rate* [mmol H 2 L -1 d -1 ] Hydrogen molar yield* [mol H 2 mol -1 lactose] Total volatile fatty acids* [mg L -1 ] Davila-Vazquez G, Alatriste-Mondragón F, de León-Rodríguez A, Razo-Flores E. Continuous biohydrogen production using cheese whey: Improving volumetric hydrogen production rate. (In preparation).

22 Culture Inoculum Carbon substrate Volumetric H 2 production rate (mmol H 2 L -1 d -1 ) H 2 yield (mol H 2 mol -1 lactose) conditions [HRT (h), ph, Temperature (!C),OLR (g L -1 Reference d -1 )] Clostridium thermolacticum Lactose (10 g L -1 ) , 7, 58, Collet et al Anaerobic sludge Cheese whey powder solution L H 2 L -1 d mm g COD 24, 4-5, 35-38, 10 Yang et al Compost (Fed-batch process) Lactose (2 g L -1 ) , 5, 55, , 5.3, 55, 2.2 Calli et al Sewage sludge Sucrose (20 g COD L -1 ) , 6.8, 35, 40 g COD L -1 d -1 Lin and Lay 2005 Mixed culture immobilized in silicone gel Sucrose (30 g L -1 ) , 6.5, 40, 1440 Wu et al Anaerobic granular sludge Cheese whey powder solution , 5.9, 37, , 5.9, 37, , 5.9, 37, This study

23 Conclusions 1. In this work we demonstrated that undefined microflora from anaerobic granular sludge was able to produce Bio-H 2 using CW powder solution as sole carbon source. 2. Significant effect of initial CW concentration and initial ph was observed and must be considered in the design of a Bio-H 2 generation process from CW. 3. It is possible to improve VHPR in a continuous process using cheese whey as substrate, by an appropriate selection of the operation parameters such as HRT and OLR. 4. Best operation conditions found were HRT=6h and OLR=138.6 g lactose L -1 d -1. A further increase in OLR (to 184.8, keeping HRT=6h) caused a decrease in VHPR probably due to VFAs toxicity. 5. We found higher VHPR than the first report (Yang et al. 2007) using CW and mixed microflora for continuous hydrogen production.

24 Acknowledgements This work was supported by the Fondo Mixto San Luis Potosí Consejo Nacional de Ciencia y Tecnología a (FMSLP-2005-C01-23).

25 Cited references Angenent LT, Karim K, Al-Dahhan MH, Wrenn BA, Domiguez-Espinosa R Production of bioenergy and biochemicals from industrial and agricultural wastewater. Trends Biotechnol 22(9): Calli B, Schoenmaekers K, Vanbroekhoven K, Diels L Dark fermentative H 2 production from xylose and lactose-effects of on-line ph control. Int J Hydrogen Energy 33(2): Collet C, Adler N, Schwitzguebel JP, Peringer P Hydrogen production by Clostridium thermolacticum during continuous fermentation of lactose. Int J Hydrogen Energy 29(14): Davila-Vazquez G, Arriaga S, Alatriste-Mondragón F, de León-Rodríguez A, Rosales-Colunga LM, Razo- Flores E Fermentative biohydrogen production: trends and perspectives. Rev Environ Sci Biotechnol 7(1): Davila-Vazquez G, Alatriste-Mondragón F, de León-Rodriguez A, Razo-Flores E. In press. Fermentative hydrogen production in batch experiments using lactose, cheese whey and glucose: Influence of initial substrate concentration and ph. Int J Hydrogen Energy: doi: /j.ijhydene Lee H-S, Salerno MB, Rittmann B Thermodynamic evaluation on H 2 production in glucose fermentation. Environ Sci Technol 42(7): Lin CY, Lay CH A nutrient formulation for fermentative hydrogen production using anaerobic sewage sludge microflora. Int J Hydrogen Energy 30(3): Yang P, Zhang R, McGarvey JA, Benemann JR Biohydrogen production from cheese processing wastewater by anaerobic fermentation using mixed microbial communities. Int J Hydrogen Energy 32(18): Wu SY, Hung CH, Lin CN, Chen HW, Lee AS, Chang JS Fermentative hydrogen production and bacterial community structure in high-rate anaerobic bioreactors containing silicone-immobilized and self-flocculated sludge. Biotechnol Bioeng 93(5):