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Co-cultured methanogen improved the metabolism in the hydrogenosome of anaerobic fungus as revealed by gas chromatography-mass spectrometry analysis

  • Li, Yuqi (Laboratory of Gastrointestinal Microbiology, National Center for International Research on Animal Gut Nutrition, Nanjing Agricultural University) ;
  • Sun, Meizhou (Laboratory of Gastrointestinal Microbiology, National Center for International Research on Animal Gut Nutrition, Nanjing Agricultural University) ;
  • Li, Yuanfei (Laboratory of Gastrointestinal Microbiology, National Center for International Research on Animal Gut Nutrition, Nanjing Agricultural University) ;
  • Cheng, Yanfen (Laboratory of Gastrointestinal Microbiology, National Center for International Research on Animal Gut Nutrition, Nanjing Agricultural University) ;
  • Zhu, Weiyun (Laboratory of Gastrointestinal Microbiology, National Center for International Research on Animal Gut Nutrition, Nanjing Agricultural University)
  • Received : 2019.08.15
  • Accepted : 2020.01.06
  • Published : 2020.12.01

Abstract

Objective: The purpose of this study was to reveal the metabolic shift in the fungus cocultured with the methanogen (Methanobrevibacter thaueri). Methods: Gas chromatography-mass spectrometry was used to investigate the metabolites in anaerobic fungal (Pecoramyces sp. F1) cells and the supernatant. Results: A total of 104 and 102 metabolites were detected in the fungal cells and the supernatant, respectively. The partial least squares-discriminant analysis showed that the metabolite profiles in both the fungal cell and the supernatant were distinctly shifted when co-cultured with methanogen. Statistically, 16 and 30 metabolites were significantly (p<0.05) affected in the fungal cell and the supernatant, respectively by the co-cultured methanogen. Metabolic pathway analysis showed that co-culturing with methanogen reduced the production of lactate from pyruvate in the cytosol and increased metabolism in the hydrogenosomes of the anaerobic fungus. Citrate was accumulated in the cytosol of the fungus co-cultured with the methanogen. Conclusion: The co-culture of the anaerobic fungus and the methanogen is a good model for studying the microbial interaction between H2-producing and H2-utilizing microorganisms. However, metabolism in hydrogenosome needs to be further studied to gain better insight in the hydrogen transfer among microorganisms.

Keywords

References

  1. Orpin CG. Studies on the rumen flagellate Neocallimastix frontalis. J Gen Microbiol 1975;91:249-62. https://doi.org/10.1099/00221287-91-2-249
  2. Li Y, Li Y, Jin W, et al. Combined genomic, transcriptomic, proteomic, and physiological characterization of the growth of Pecoramyces sp. F1 in monoculture and co-culture with a syntrophic methanogen. Front Microbiol 2019;10:435. https://doi.org/10.3389/fmicb.2019.00435
  3. Edwards JE, Forster RJ, Callaghan TM, et al. PCR and omics based techniques to study the diversity, ecology and biology of anaerobic fungi: insights, challenges and opportunities. Front Microbiol 2017;8:1657. https://doi.org/10.3389/fmicb. 2017.01657
  4. Teunissen MJ, Kets EPW, Op den Camp HJM, Huis in't Veld JHJ, Vogels GD. Effect of coculture of anaerobic fungi isolated from ruminants and non-ruminants with methanogenic bacteria on cellulolytic and xylanolytic enzyme activities. Arch Microbiol 1992;157:176-82. https://doi.org/10.1007/BF00245287
  5. Bauchop T, Mountfort DO. Cellulose fermentation by a rumen anaerobic fungus in both the absence and the presence of rumen methanogens. Appl Environ Microbiol 1981;42:1103-10. https://doi.org/10.1128/AEM.42.6.1103-1110.1981
  6. Jin W, Cheng YF, Mao SY, Zhu WY. Isolation of natural cultures of anaerobic fungi and indigenously associated methanogens from herbivores and their bioconversion of lignocellulosic materials to methane. Bioresour Technol 2011; 102:7925-31. https://doi.org/10.1016/j.biortech.2011.06.026
  7. Joblin KN, Naylor GE, Williams AG. Effect of Methanobrevibacter smithii on xylanolytic activity of anaerobic ruminal fungi. Appl Environ Microbiol 1990;56:2287-95. https://doi.org/10.1128/AEM.56.8.2287-2295.1990
  8. Cheng YF, Edwards JE, Allison GG, Zhu WY, Theodorou MK. Diversity and activity of enriched ruminal cultures of anaerobic fungi and methanogens grown together on lignocellulose in consecutive batch culture. Bioresour Technol 2009;100:4821-8. https://doi.org/10.1016/j.biortech.2009.04.031
  9. Barichievich EM, Calza RE. Supernatant protein and cellulase activities of the anaerobic ruminal fungus Neocallimastix frontalis EB188. Appl Environ Microbiol 1990;56:43-8. https://doi.org/10.1128/AEM.56.1.43-48.1990
  10. Haitjema CH, Solomon KV, Henske JK, Theodorou MK, O'Malley MA. Anaerobic gut fungi: Advances in isolation, culture, and cellulolytic enzyme discovery for biofuel production. Biotechnol Bioeng 2014;111:1471-82. https://doi.org/10.1002/bit.25264
  11. Marvin-Sikkema FD, Gomes TMP, Grivet JP, Gottschal JC, Prins RA. Characterization of hydrogenosomes and their role in glucose metabolism of Neocallimastix sp. L2. Arch Microbiol 1993;160:388-96. https://doi.org/10.1007/BF00252226
  12. Ma B, Liu J, Zhang Q, et al. Metabolomic profiles delineate signature metabolic shifts during estrogen deficiency-induced bone loss in rat by GC-TOF/MS. PLOS One 2013;8:e54965. https://doi.org/10.1371/journal.pone.0054965
  13. Ma B, Li X, Zhang Q, et al. Metabonomic profiling in studying anti-osteoporosis effects of strontium fructose 1,6-diphosphate on estrogen deficiency-induced osteoporosis in rats by GC/TOF-MS. Eur J Pharmacol 2013;718:524-32. https://doi.org/10.1016/j.ejphar.2013.06.030
  14. Thevenot EA, Roux A, Xu Y, Ezan E, Junot C. Analysis of the human adult urinary metabolome variations with age, body mass index, and gender by implementing a comprehensive workflow for univariate and OPLS statistical analyses. J Proteome Res 2015;14:3322-35. https://doi.org/10.1021/acs.jproteome.5b00354
  15. Li Y, Jin W, Cheng Y, Zhu W. Effect of the associated methanogen Methanobrevibacter thaueri on the dynamic profile of end and intermediate metabolites of anaerobic fungus Piromyces sp. F1. Curr Microbiol 2016;73:434-41. https://doi.org/10.1007/s00284-016-1078-9
  16. Nakashimada Y, Srinivasan K, Murakami M, Nishio N. Direct conversion of cellulose to methane by anaerobic fungus Neocallimastix frontalis and defined methanogens. Biotechnol Lett 2000;22:223-7. https://doi.org/10.1023/A:1005666428494
  17. Leis S, Dresch P, Peintner U, et al. Finding a robust strain for biomethanation: anaerobic fungi (Neocallimastigomycota) from the Alpine ibex (Capra ibex) and their associated methanogens. Anaerobe 2014;29:34-43. https://doi.org/10.1016/j.anaerobe.2013.12.002
  18. Wei YQ, Yang HJ, Luan Y, Long RJ, Wu YJ, Wang ZY. Isolation, identification and fibrolytic characteristics of rumen fungi grown with indigenous methanogen from yaks (Bos grunniens) grazing on the Qinghai-Tibetan Plateau. J Appl Microbiol 2016;120:571-87. https://doi.org/10.1111/jam.13035
  19. Wei YQ, Long RJ, Yang H, et al. Fiber degradation potential of natural co-cultures of Neocallimastix frontalis and Methanobrevibacter ruminantium isolated from yaks (Bos grunniens) grazing on the Qinghai Tibetan Plateau. Anaerobe 2016;39: 158-64. https://doi.org/10.1016/j.anaerobe.2016.03.005
  20. Sun M, Jin W, Li Y, Mao S, Cheng Y, Zhu W. Isolation and identification of cellulolytic anaerobic fungi and their associated methanogens from Holstein cow. Acta Microbiologica Sinica 2014;54:563-71. https://doi.org/10.13343/j.cnki.wsxb.2014.05.011
  21. Li YF, Jin W, Mu CL, Cheng YF, Zhu WY. Indigenously associated methanogens intensified the metabolism in hydrogenosomes of anaerobic fungi with xylose as substrate. J Basic Microbiol 2017;57:933-40. https://doi.org/10.1002/jobm.201700132
  22. Cheng YF, Jin W, Mao SY, Zhu WY. Production of citrate by anaerobic fungi in the presence of co-culture methanogens as revealed by 1H NMR spectrometry. Asian-Australas J Anim Sci 2013;26:1416-23. https://doi.org/10.5713/ajas.2013.13134
  23. Kown M, Song JY, Ha JK, Park HS, Chang JS. Analysis of functional genes in carbohydrate metabolic pathway of anaerobic rumen fungus Neocallimastix frontalis PMA02. Asian-Australas J Anim Sci 2009;22:1555-65. https://doi.org/10.5713/ajas.2009.80371
  24. Hackstein JHP, Akhmanova A, Boxma B, Harhangi HR, Voncken FGJ. Hydrogenosomes: eukaryotic adaptations to anaerobic environments. Trends Microbiol 1999;7:441-7. https://doi.org/10.1016/S0966-842X(99)01613-3
  25. Akhmanova A, Voncken FGJ, Hosea KM, et al. A hydrogenosome with pyruvate formate-lyase: Anaerobic chytrid fungi use an alternative route for pyruvate catabolism. Mol Microbiol 1999;32:1103-14. https://doi.org/10.1046/j.1365-2958.1999.01434.x

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