Activated zeolite—suitable carriers for microorganisms in anaerobic digestion processes?
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Plant cell wall structures represent a barrier in the biodegradation process to produce biogas for combustion and energy production. Consequently, approaches concerning a more efficient de-polymerisation of cellulose and hemicellulose to monomeric sugars are required. Here, we show that natural activated zeolites (i.e. trace metal activated zeolites) represent eminently suitable mineral microhabitats and potential carriers for immobilisation of microorganisms responsible for anaerobic hydrolysis of biopolymers stabilising related bacterial and methanogenic communities. A strategy for comprehensive analysis of immobilised anaerobic populations was developed that includes the visualisation of biofilm formation via scanning electron microscopy and confocal laser scanning microscopy, community and fingerprint analysis as well as enzyme activity and identification analyses. Using SDS polyacrylamide gel electrophoresis, hydrolytical active protein bands were traced by congo red staining. Liquid chromatography/mass spectroscopy revealed cellulolytical endo- and exoglucanase (exocellobiohydrolase) as well as hemicellulolytical xylanase/mannase after proteolytic digestion. Relations to hydrolytic/fermentative zeolite colonisers were obtained by using single-strand conformation polymorphism analysis (SSCP) based on amplification of bacterial and archaeal 16S rRNA fragments. Thereby, dominant colonisers were affiliated to the genera Clostridium, Pseudomonas and Methanoculleus. The specific immobilisation on natural zeolites with functional microbes already colonising naturally during the fermentation offers a strategy to systematically supply the biogas formation process responsive to population dynamics and process requirements.
KeywordsBiogas Zeolites Hemicellulases Cellulases Microbial community Grass silage
This study was performed within the Austrian Centre of Industrial Biotechnology ACIB and has been kindly supported by IPUS GmbH (Austria), the Federal Ministry of Economy, Family and Youth (BMWFJ), the Federal Ministry of Traffic, Innovation and Technology (bmvit), the Styrian Business Promotion Agency SFG, the Standortagentur Tirol and ZIT–Technology Agency of the City of Vienna through the COMET-Funding Program managed by the Austrian Research Promotion Agency FFG. Many thanks go to contributors of the collaborating organisations and institutes (FELMI-ZFE, ZMF and LfL).
- Amann RI, Binder BJ, Olson RJ, Chisholm SW, Devereux R, Stahl DA (1990) Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial populations. Appl Environ Microbiol 56(6):1919–1925Google Scholar
- Bischofsberger W, Böhnke B, Seyfried CF (2004) Mikrobiologische Grundlagen des anaeroben Abbaus – Überblick und Organismen. In: Dauber S (ed) Anaerobtechnik. Springer, Berlin, Germany, pp 23–49Google Scholar
- Bolstad AI, Jensen HB, Bakken V (1996) Taxonomy, biology and periodontal aspects of Fusobacterium nucleatum. Clin Microbiol Rev 9:55–71Google Scholar
- Bougrier C, Dognin D, Laroche C, Rivero JC (2011) Effects of additives solutions of trace elements on anaerobic digestion of maize silage. 1st International Conference on Biogas Microbiology. Leipzig, Germany, September 14–16, Proceedings P-10, p 75Google Scholar
- Holper J, Lesjak M, Heinzel U, Boos B (2005) Zeolith in der Biogasgewinnung. Sonn. 05450052.5 [EP 1 577 269 A1]. European Patent, Styria, AustriaGoogle Scholar
- Karita S, Sakka K, Ohmiya K (1997) Cellulosomes, cellulase complexes, of anaerobic microbes: their structure models and functions. In: Onodera R, Itabashi H, Ushida K, Yano H, Sasaki Y (eds) Rumen microbes and digestive physiology in ruminants. Japan Science Society Tokyo/Karger, Basel, pp 47–57Google Scholar
- Lieber A, Kiesel B, Babel W (2002) Microbial diversity analysis of soil by SSCP fingerprinting technique using TGGE Maxi System. In: Merbach W, Hütsch BW, Augustin J (eds) Teubner. Ökophysiologie des Wurzelraumes, Stuttgart, pp 61–65Google Scholar
- Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin-Phenol reagents. J Biol Chem 193:265–275Google Scholar
- Maestrojuán GM, Boone DR, Xun L, Mah RA, 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(2):117–122CrossRefGoogle Scholar
- Potivichayanon S, Sungmon T, Chaikongmao W, Kamvanin S (2011) Enhancement of biogas production from bakery waste by Pseudomonas aeruginosa. World Acad Sci Eng Technol 56:529–532Google Scholar
- Schwieger F, Tebbe C (1998) A new approach to utilize PCR-SSCP for 16S rRNA gene-based microbial community analysis. Appl Environ Microbiol 64(12):4870–4876Google Scholar
- Stahl DA, Amann R (1996) Development and application of nucleic acid probes. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematics. Wiley, Chichester, pp 205–248Google Scholar
- Takashima M, Speece RE (1989) Mineral nutrient requirements for high-rate methane fermentation of acetate at low SRT. J Wat Poll Cont Fed 61:1645–1650Google Scholar
- Van Gylswyk NO, van der Toorn JJTK (1986) Enumeration of Bacteroides succinogenes in the rumen of sheep fed maize-straw diets. FEBS Lett 38(4):205–209Google Scholar
- Zheng D, Alm EW, Stahl DA, Raskin L (1996) Characterization of universal small-unit rRNA hybridization probes for quantitative molecular ecology studies. Appl Environ Microbiol 62:4504–4513Google Scholar