Evaluation of the biomethanation potential of enriched methanogenic cultures on gelatin

Abstract

Purpose

Biomethane is an environment-friendly, economic, and alternative energy resource for a clean and green future. In the present study, we have evaluated the biomethanation potential of acetate-utilizing methanogenic culture (AUMC) and gelatin-enriched mixed culture (GEMC) with Clostridium acetobutylicum NCIM 2841 (GEMC-CA.) on gelatin as a sole carbon and nitrogen source.

Methods

We conducted experiments for examining the specific-methanogenic activity of these cultures in the metabolic assay media containing 1% gelatin. The produced methane and consumed gelatin were quantified by standard experimental methods. Exchange metabolites produced by these cultures were qualitatively analyzed by mass spectrometry.

Results

Results of our study show that the growth-associated amino acid catabolism partially or completely supported the methanogenesis of these defined cultures. AUMC and GEMC found to be suitable for enhanced methanogenic activity on gelatin but a rapid degradation of amino acid was attributed by GEMC-CA. The ammonia released from these cultures was directly proportional to gelatin degradation. Mass spectral data analysis identified some key exchange metabolites from acidogenic culture and methanogenic culture for confirming the growth-associated methanogenesis.

Conclusion

The biomethanation potential of these cultures on gelatin is coupled with the Stickland reactions-directed methanogenesis in a syntrophic manner. The present study provides the importance for the development of a starter culture for the biomethanation of protein-based industrial wastes in effective ways.

Keywords

Amino acid catabolism / Methanogenesis / Methanogenic culture / Gelatin / Anaerobic digestion / Defined culture

References

[1] Balan V. Current challenges in commercially producing biofuels from lignocellulosic biomass. ISRN Biotechnol, 2014

[2] Chaney AL, Marbach P. Modified reagents for determination of urea and ammonia. Modified reagents for determination of urea and ammonia. Clin Chem, 1962, 8: 130-132.

[3] Chellapandi P, Uma L. Co-digestion of ossein factory waste for methane production in batch. Elixer Biotechnol, 2012, 42: 6383-6385.

[4] Chellapandi P, Uma L. Evaluation of methanogenic activity of biogas plant slurry on ossein factory wastes. J Environ Sci Health, 2012, 54: 10-13.

[5] Chellapandi P, Prabaharan D, Uma L. A preliminary study on co-digestion of ossein factory waste for methane production. Eur Asia J BioSci, 2008, 2: 110-114.

[6] Chellapandi P, Prabaharan D, Uma L. Evaluation of methanogenic activity of biogas plant slurry for monitoring anaerobic co-digestion of ossein factory waste and cyanobacterial biomass. Appl Biochem Biotechnol, 2010, 162: 534-535.

[7] Eastoe JE. The amino acid composition of mammalian collagen and gelatin. Biochem J, 1995, 61: 589-600.

[8] Fonknechten N, Chaussonnerie S, Tricot S, Lajus A, Andreesen JR, Perchat N, Pelletier E, Gouyvenoux M, Barbe MV, Salanoubat M, Le Paslier D, Weissenbach J, Cohen GN, Kreimeyer A. Clostridium sticklandii, a specialist in amino acid degradation: revisiting its metabolism through its genome sequence. BMC Genom, 2010, 11: 555.

[9] Halket JM, Waterman D, Przyborowska AM, Patel RKP, Fraser PD, Bramley PM. Chemical derivatization and mass spectral libraries in metabolic profiling by GC/MS and LC/MS/MS. J Exp Bot, 2005, 56: 219-243.

[10] Hartwich K, Poehlein A, Daniel R. The purine-utilizing bacterium Clostridium acidurici 9a: a genome-guided metabolic reconsideration. PLoS ONE, 2012, 7: e51662.

[11] Jain MK, Zeikus JG. Bioconversion of gelatin to methane by a co-culture of Clostridium collagenovorans and Methanosarcina barkeri. Appl Environ Microbiol, 1989, 55: 366-371.

[12] Liu WY, Liao B. Anaerobic co-digestion of vegetable and fruit market waste in LBR+ CSTR two-stage process for waste reduction and biogas production. Appl Biochem Biotechnol, 2018

[13] Masion E, Amine J, Marczak R. Influence of amino acid supplements on the metabolism of Clostridium acetobutylicum. FEMS Microbiol Ecol, 1987, 43: 269-274.

[14] Moench TT, Zeikus JG. Nutritional growth requirements for Butyribacterium methylotrophicum on single carbon substrates and glucose. Curr Opin Microbiol, 1983, 9: 151-154.

[15] Moore S, Stein WH. A modified ninhydrin reagent for the photometric determination of amino acids and related compounds. J Biol Chem, 1954, 211: 907-913.

[16] Nagase M, Matsuo T. Interactions between amino-acid degrading bacteria and methanogenic bacteria in anaerobic digestion. Biotechnol Bioeng, 1982, 24: 2227-2239.

[17] Nisman B. The Stickland reaction. Bacteriol Rev, 1954, 18: 16-42.

[18] Noble JE, Bailey MJA. Quantitation of protein. Methods Enzymol, 2009, 463: 73-95.

[19] Ollivier B, Smiti N, Mah RA, Garcia JL. Thermophilic methanogenesis from gelatin by a mixed defined bacterial culture. Appl Microbiol Biotechnol, 1986, 24: 79-83.

[20] Orlygsson J, Houwen FP, Svensson BH. Influence of hydrogenothrophic methane formation on the thermophilic anaerobic degradation of protein and amino acids. FEMS Microbiol Ecol, 1994, 13: 327-334.

[21] Orlygsson J, Houwen FP, Svensson BH. Thermophilic anaerobic amino acid degradation: deamination rates and end-product formation. Appl Microbiol Biotechnol, 1995, 43: 235-241.

[22] Patel CN, Chellapandi P. Anaerobic digestion of cotton seed cake using developed mixed consortia. Electron J Environ Agric Food Chem, 2009, 8: 333-344.

[23] Perisin MA, Sund CJ. Human gut microbe co-cultures have greater potential than monocultures for food waste remediation to commodity chemicals. Sci Rep, 2018, 8: 15594.

[24] Ramasamy IR, Pullammanmappallil PC. Protein degradation during anaerobic wastewater treatment: derivation of stoichiometry. Biodegradation, 2001, 12: 247-257.

[25] Sangavai C, Chellapandi P. Amino acid catabolism-directed biofuel production in Clostridium sticklandii: an insight into model-driven systems engineering. Biotechnol Rep, 2017, 16: 32-43.

[26] Sasaki K, Morita M, Sasaki D, Nagaoka J, Matsumoto N, Ohmura N, Shinozaki H. Syntrophic degradation of proteinaceous materials by the thermophilic strains Coprothermobacter proteolyticus and Methanothermobacter thermautotrophicus. J Biosci Bioeng, 2011, 112: 469-472.

[27] Stickland LH. Studies in the metabolism of the strict anaerobes (genus Clostridium): the chemical reactions by which Clostridium sporogenes obtains its energy. Biochem J, 1934, 28: 1746-1759.

[28] Tang Y, Shigematsu T, Morimura S, Kida KJ. Microbial community analysis of mesophilic anaerobic protein degradation process using bovine serum albumin (BSA)-fed continuous cultivation. J Biosci Bioeng, 2005, 99: 150-164.

[29] Vieira AM, Bergamasco R, Gimenes ML, Nakamura CV, Dias B, Filho P. Microbial populations of up-flow anaerobic sludge blanket reactor treating waste water from a gelatin industry. Environ Technol, 2001, 22: 1477-1485.

[30] Vlysidis A, Binns M, Webb C, Theodoropoulos C. A techno-economic analysis of biodiesel biorefineries: assessment of integrated designs for the co-production of fuels and chemicals. Energy, 2011, 36: 4671-4683.

[31] Wolfe RS. Techniques for cultivating methanogens. Methods Enzymol, 2011, 494: 1-21.

[32] Wu J, Cao Z, Hu Y, Wang X, Wang G, Zuo J, Wang K, Qian Y. Microbial insight into a pilot-scale enhanced two-stage high-solid anaerobic digestion system treating waste activated sludge. Int J Environ Res Public Health, 2017, 14: 1483.