DOI QR코드

DOI QR Code

Microbial Diversity in Three-Stage Methane Production Process Using Food Waste

음식물 쓰레기를 이용한 3단계 메탄생산 공정의 미생물 다양성

  • Nam, Ji-Hyun (Energy and Environment Fusion Technology Center, Myongji University) ;
  • Kim, Si-Wouk (Department of Environmental Engineering, Chosun University) ;
  • Lee, Dong-Hun (Department of Microbiology, Chungbuk National University)
  • 남지현 (명지대학교 에너지환경융합기술연구소) ;
  • 김시욱 (조선대학교 환경공학과) ;
  • 이동훈 (충북대학교 미생물학과)
  • Received : 2012.06.11
  • Accepted : 2012.06.26
  • Published : 2012.06.30

Abstract

Anaerobic digestion is an alternative method to digest food wastes and to produce methane that can be used as a renewable energy source. We investigated bacterial and archaeal community structures in a three-stage methane production process using food wastes with concomitant wastewater treatment. The three-stage methane process is composed of semianaerobic hydrolysis/acidogenic, anaerobic acidogenic, and strictly anaerobic methane production steps in which food wastes are converted methane and carbon dioxide. The microbial diversity was determined by the nucleotide sequences of 16S rRNA gene library and quantitative real-time PCR. The major eubacterial population of the three-stage methane process was belonging to VFA-oxidizing bacteria. The archaeal community consisted mainly of two species of hydrogenotrophic methanogen (Methanoculleus). Family Picrophilaceae (Order Thermoplasmatales) was also observed as a minor population. The predominance of hydrogenotrophic methanogen suggests that the main degradation pathway of this process is different from the classical methane production systems that have the pathway based on acetogenesis. The domination of hydrogenotrophic methanogen (Methanoculleus) may be caused by mesophilic digestion, neutral pH, high concentration of ammonia, short HRT, and interaction with VFA-oxidizing bacteria (Tepidanaerobacter etc.).

혐기성 소화는 음식물 쓰레기와 같은 폐기물로부터 재생 가능한 에너지원으로 메탄을 생성하는 공정이다. 본 연구에서는 음식물 쓰레기와 폐수를 동시에 처리하는 3단계 메탄생산 공정을 이용한 혐기성 소화공정의 bacteria와 archaea 군집 변화를 조사하였다. 3단계 메탄생산 공정은 음식물 쓰레기 및 폐수를 메탄과 이산화탄소로 전환하는 반혐기성 가수분해/산생성, 혐기성 산생성과 혐기성 메탄생성조로 구성되어있으며, 16 rRNA 유전자 라이브러리의 염기서열 분석과 정량 PCR 등의 분자생물학적 방법으로 주요 미생물 군집을 조사하였다. 메탄생산 공정의 주요 미생물 군집은 VFA-산화 박테리아와 Methanoculleus 속에 속하는 hydrogenotrophic methanogen의 두 종(species)이었다. 또한, 소수의 Picrophilaceae 과(Thermoplasmatales 목)의 archaea도 확인하였다. 음식물을 이용한 3단계 메탄생산 공정은 acetogenesis를 기반으로 하는 고전적 메탄생성 공정과 달리 주로 hydrogenotrophic methanogen의 분해 경로에 의해 이루어 짐을 알 수 있다. 이들 균주의 우점은 중온 소화공정, 중성 pH, 높은 암모니아 농도, 짧은 HRT, Tepidanaerobacter 속 등과 같은 VFA 산화세균과의 상호작용 등에서 기인한 것으로 생각된다.

Keywords

References

  1. Angenent, L.T., Sung, S.W., and Raskin, L. 2002. Methanogenic population dynamics during startup of a full-scale anaerobic sequencing batch reactor treating swine waste. Water Res. 36, 4648-4654. https://doi.org/10.1016/S0043-1354(02)00199-9
  2. APHA. 1998. Standard Methods for the Examination of Water and Wastewater. American Public Health Association/American Water Works Association/Water Pollution Control Federation, Washington, D.C., USA.
  3. Bialek, K., Kim, J., Lee, C., Collins, G., Mahony, T., and O'Flaherty, V. 2011. Quantitative and qualitative analyses of methanogenic community development in high-rate anaerobic bioreactors. Water Res. 45, 1298-1308. https://doi.org/10.1016/j.watres.2010.10.010
  4. Briones, A.M., Daugherty, B.J., Angenent, L.T., Rausch, K., Tumbleson, M., and Raskin, L. 2009. Characterization of microbial trophic structures of two anaerobic bioreactors processing sulfate-rich waste streams. Water Res. 43, 4451-4460. https://doi.org/10.1016/j.watres.2009.07.003
  5. Chen, S. and Dong, X. 2005. Proteiniphilum acetatigenes gen. nov., sp. nov., from a UASB reactor treating brewery wastewater. Int. J. Syst. Evol. Microbiol. 55, 2257-2261. https://doi.org/10.1099/ijs.0.63807-0
  6. Chun, J., Huq, A., and Colwell, R.R. 1999. Analysis of 16S-23S rRNA intergenic Spacer Regions of Vibrio cholerae and Vibrio mimicus. Appl. Environ. Microbiol. 65, 2202-2208.
  7. DeLong, E.F. 1992. Archaea in coastal marine environments. Proc. Natl. Acad. Sci. USA 89, 5685-5689. https://doi.org/10.1073/pnas.89.12.5685
  8. Demirel, B. and Yenigün, O. 2002. Article first published online: 18 Two-phase anaerobic digestion processes: a review. J. Chem. Technol. Biotechnol. 77, 743-755. https://doi.org/10.1002/jctb.630
  9. Felsenstein, J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39, 783-791. https://doi.org/10.2307/2408678
  10. Felsenstein, J. 2002. PHYLIP (Phylogeny Inference Package) version 3.6a3. Distributed by the author, Department of Genome Sciences, University of Washington, Seattle, USA.
  11. Hansen, K.H., Ahring, B.K., and Raskin, L. 1999. Quantification of syntrophic fatty acid-$\beta$-oxidizing bacteria in a mesophilic biogas by oligonucleotide probe hybridization. Appl. Environ. Microbiol. 65, 4767-4774.
  12. Hattori, S. 2008. Syntrophic acetate-oxidizing microbes in methanogenic environments. Microbes Environ. 23, 118-127. https://doi.org/10.1264/jsme2.23.118
  13. Jukes, T.H. and Cantor, C.R. 1969. Evolution of protein molecules, pp. 21-132. In Munro, H.N. (ed.), Mammalian Protein Metabolism. Academic Press, New York, N.Y., USA.
  14. Karakashev, D., Bastone, D.J., and Angelidaki, I. 2005. Influence of environmental conditions on methanogenic compositions in anaerobic biogas reactors. Appl. Environ. Microbiol. 71, 331-338. https://doi.org/10.1128/AEM.71.1.331-338.2005
  15. Kim, J.K., Cho, J.H., Lee, J.S., Hahm, K.S., Park, D.H., and Kim, S.W. 2002. Mass production of methane from food wastes with concomitant wastewater treatment. Appl. Biochem. Biotechnol. 98-100, 753-764. https://doi.org/10.1385/ABAB:98-100:1-9:753
  16. Kim, J.K., Han, G.H., Oh, B.R., Chun, Y.N., Eom, C.Y., and Kim, S.W. 2008. Volumetric scale-up of a three stage fermentation system for food waste treatment. Bioresour. Technol. 99, 4394-4399. https://doi.org/10.1016/j.biortech.2007.08.031
  17. Kongjan, P., O-Thong, S., and Angelidaki, I. 2011. Performance and microbial community analysis of two-stage process with extreme thermophilic hydrogen and thermophilic methane production from hydrolysate in UASB reactors. Bioresour. Technol. 102, 4028-4035. https://doi.org/10.1016/j.biortech.2010.12.009
  18. Lane, D.J. 1991. 16S/23S rRNA sequencing, pp. 115-175. In Stackebrandt, E. and Goodfellow, M. (eds.), Nucleic acid techniques in bacterial systematics. John Wiley & Sons, Chichester, England.
  19. Lee, C., Kim, J., Shin, S.G., O'Flaherty, V., and Hwang, S. 2010. Quantitative and qualitative transition of methanogen community structure during the batch anaerobic digestion of cheese-processing wastewater. Appl. Microbiol. Biotechnol. 87, 1963-1973. https://doi.org/10.1007/s00253-010-2685-1
  20. Maestrojuan, G.M., Boone, D.R., Xun, L., Mah, R.A., and Zhang, L. 1990. Transfer of Methanogenium bourgense, Methanogenium marisnigri, Methanogenium olentangyi, and Methanogenium thermophilicum to the Genus Methanoculleus gen. nov., emendation of Methanoculleus marisnigri and Methanogenium, and description of new strains of Methanoculleus bourgense and Methanoculleus marisnigri. Int. J. Syst. Bacteriol. 40, 117-122. https://doi.org/10.1099/00207713-40-2-117
  21. Menes, R.J. and Muxi, L. 2002. Anaerobaculum mobile sp. nov., a novel anaerobic, moderately thermophilic, peptide-fermenting bacterium that uses crotonate as an electron acceptor, and emended description of the genus Anaerobaculum. Int. J. Syst. Evol. Microbiol. 52, 157-164. https://doi.org/10.1099/00207713-52-1-157
  22. Miller, D.N., Bryant, J.E., Madsen, E.L., and Ghiorse, W.C. 1999. Evaluation and optimization of DNA extraction and purification procedures for soil and sediment samples. Appl. Environ. Microbiol. 65, 4715-4724.
  23. Muyzer, G., De Waal, E.C., and Uitterlinden, A.G. 1993. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl. Environ. Microbiol. 59, 695-700.
  24. Ollivier, B.M., Mah, R.A., Garcia, J.L., and Boone, D.R. 1986. Isolation and characterization of Methanogenium bourgense sp. nov. Int. J. Syst. Bacteriol. 36, 297-301. https://doi.org/10.1099/00207713-36-2-297
  25. Ovreas, L., Forney, L., Daae, F.L., and Torsvik, V. 1997. Distribution of bacterioplankton in meromictic Lake Saelenvannet, as determined by denaturing gradient gel electrophoresis of PCR-amplified gene fragments coding for 16S rRNA. Appl. Environ. Microbiol. 63, 3367-3373.
  26. Qiu, Y.L., Sekiguchi, Y., Hanada, S., Imachi, H., Tseng, I.C., Cheng, S.S., Ohashi, A., Harada, H., and Kamagata, Y. 2006. Pelotomaculum terephthalicum sp. nov. and Pelotomaculum isophthalicum sp. nov.: two anaerobic bacteria that degrade phthalate isomers in syntrophic association with hydrogenotrophic methanogens. Arch. Microbiol. 185, 172-182. https://doi.org/10.1007/s00203-005-0081-5
  27. Rees, G.N., Patel, B.K., Grassia, G.S., and Sheehy, A.J. 1997. Anaerobaculum thermoterrenum gen. nov., sp. nov., a novel, thermophilic bacterium which ferments citrate. Int. J. Syst. Bacteriol. 47, 150-154. https://doi.org/10.1099/00207713-47-1-150
  28. Saitou, N. and Nei, M. 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4, 406-425.
  29. Sawayama, S., Tada, C., Tsukahara, K., and Yagishita, T. 2004. Effect of ammonium addition on methanogenic community in a fluidized bed anaerobic digestion. J. Biosci. Bioeng. 97, 65-70. https://doi.org/10.1016/S1389-1723(04)70167-X
  30. Schnürer, A., Schink, B.G., and Svensson, B.H. 1996. Clostridium ultunense sp. nov., a mesophilic bacterium oxidizing acetate in syntrophic association with a hydrogenotrophic methanogenic bacterium. Int. J. Syst. Evol. Microbiol. 46, 1145-1152.
  31. Schnurer, A., Zellner, G., and Svensson, B.H. 1999. Mesophilic syntrophic acetate oxidation during methane formation in biogas reactors. FEMS Microbiol. Ecol. 29, 249-261. https://doi.org/10.1111/j.1574-6941.1999.tb00616.x
  32. Sekiguchi, Y., Imachi, H., Susilorukmi, A., Muramatsu, M., Ohashi, A., Harada, H., Hanada, S., and Kamagata, Y. 2006. Tepidanaerobacter syntrophicus gen. nov., sp. nov., an anaerobic, moderately thermophilic, syntrophic alcohol- and lactate-degrading bacterium isolated from thermophilic digested sludges. Int. J. Syst. Evol. Microbiol. 56, 1621-1629. https://doi.org/10.1099/ijs.0.64112-0
  33. Song, M., Shin, S.G., and Hwang, S. 2010. Methanogenic population dynamics assessed by real-timequantitativePCR in sludge granule in upflow anaerobic sludge blanket treating swine wastewater. Bioresour. Technol. 101, S23-S28. https://doi.org/10.1016/j.biortech.2009.03.054
  34. Steinhaus, B., Garcia, M.L., Shen, A.Q., and Angenent, L.T. 2007. A portable anaerobic microbioreactor reveals optimum growth conditions for the methanogen Methanosaeta concilii. Appl. Environ. Microbiol. 73, 1653-1658. https://doi.org/10.1128/AEM.01827-06
  35. Thompson, J.D., Gilson, T.J., Plewniak, F., Jeanmougin, F., and Higgins, D.G. 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 25, 4876-4882. https://doi.org/10.1093/nar/25.24.4876
  36. Tian, J., Wang, Y., and Dong, X. 2010. Methanoculleus hydrogenitrophicus sp. nov., a methanogenic archaeon isolated from wetland soil. Int. J. Syst. Evol. Microbiol. 60, 2165-2169. https://doi.org/10.1099/ijs.0.019273-0
  37. Westerholm, M., Roos, S., and Schnürer, A. 2011. Tepidanaerobacter acetatoxydans sp. nov., an anaerobic, syntrophic acetate-oxidizing bacterium isolated from two ammonium-enriched mesophilic methanogenic processes. Syst. Appl. Microbiol. 34, 260-266. https://doi.org/10.1016/j.syapm.2010.11.018
  38. Zellner, G., Messner, P., Winter, J., and Stackebrandt, E. 1998. Methanoculleus palmolei sp. nov., an irregularly coccoid methanogen from an anaerobic digester treating wastewater of a palm oil plant in north-Sumatra, Indonesia. Int. J. Syst. Bacteriol. 48, 1111-1117. https://doi.org/10.1099/00207713-48-4-1111

Cited by

  1. Evaluating Feeding of Organic Waste and Stirring Interval to Optimize Anaerobic Digestion vol.39, pp.4, 2014, https://doi.org/10.5307/JBE.2014.39.4.366
  2. A Study on Improvement of Distribution Facility in Wholesale Agricultural Products Market vol.16, pp.2, 2012, https://doi.org/10.15722/jds.16.2.201802.53