Genomic and metabolic insights into solvent production by Thermoanaerobacterium thermosaccharolyticum GSU5

Document Type : Research Paper

Authors

Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, IQUIBICEN-CONICET, Buenos Aires, Argentina.

Abstract

Thermoanaerobacterium thermosaccharolyticum GSU5 was isolated from animal dung collected in a pasture plain in Buenos Aires, Argentina. This thermophilic and anaerobic microorganism was able to produce butanol and ethanol, but not acetone, using sugars such as xylose, arabinose, glucose, galactose, fructose, sucrose, and cellobiose. Key metabolic enzymes leading to solvent production were identified in its genome. A detailed analysis of the solvent and organic acid biosynthetic pathway genes of sequenced strains revealed new insights into the unique metabolic features of this species. Genes required for the synthesis of acetone are absent in the genomes of all sequenced Thermoanaerobacterium, suggesting that it is a general trait of the genus. Strains able to produce butanol synthesize butyrate through the one step pathway catalyzed by the butyryl-CoA:acetate-CoA transferase (But). The large range of fermentable substrates and the ability to produce both ethanol and butanol without acetone makes this species an interesting candidate for second generation biofuel production.

Graphical Abstract

Genomic and metabolic insights into solvent production by Thermoanaerobacterium thermosaccharolyticum GSU5

Highlights

  • Comparative genome analysis of Thermoanaerobacterium thermosaccharolyticum GSU5 was performed.
  • Ethanol and butanol production from different carbon sources without acetone was achieved.
  • Solventogenic and acidogenic pathways in Thermoanaerobacterium were elucidated.

Keywords

References

[1] Altaras, N.E., Etzel, M.R., Cameron, D.C., 2001. Conversion of sugars to 1,2-propanediol by Thermoanaerobacterium thermosaccharolyticum HG-8. Biotechnol. Progr. 17(1), 52-56.
[2] Amador-Noguez, D., Brasg, I.A., Feng, X.J., Roquet, N., Rabinowitz, J.D., 2011. Metabolome remodeling during the acidogenic-solventogenic transition in Clostridium acetobutylicum. Appl. Environ. Microbiol. 77(22), 7984-7997.
[3] Amiri, H., Azarbaijani, R., Yeganeh, L.P., Fazeli, A.S., Tabatabaei, M., Salekdeh, G.H., Karimi, K., 2016. Nesterenkonia sp. strain F, a halophilic bacterium producing acetone, butanol and ethanol under aerobic conditions. Sci. Rep. 6(1), 1-10.
[4] Auch, A.F., Klenk, H.P., Göker, M., 2010. Standard operating procedure for calculating genome-to-genome distances based on high-scoring segment pairs. Stand. Genomic Sci. 2(1), 142-148.
[5] Berezina, O.V., Brandt, A., Yarotsky, S., Schwarz, W.H., Zverlov, V.V., 2009. Isolation of a new butanol-producing Clostridium strain: high level of hemicellulosic activity and structure of solventogenesis genes of a new Clostridium saccharobutylicum isolate. Syst. Appl. Microbiol. 32(7), 449-459.
[6] Bhandiwad, A., Guseva, A., Lynd, L.R., 2013. Metabolic engineering of Thermoanaerobacterium thermosaccharolyticum for increased n- butanol production for n -butanol production. Adv. Microbiol. 3, 46-51.
[7] Bhandiwad, A., Shaw, A.J., Guss, A., Guseva, A., Bahl, H., Lynd, L.R., 2014. Metabolic engineering of Thermoanaerobacterium saccharolyticum for n-butanol production. Metab. Eng. 21, 17-25.
[8] Biswas, R., Huntemann, M., Clum, A., Pillay, M., Palaniappan, K., Varghese, N., Gussc, A.M., 2018. Complete genome sequence of Thermoanaerobacterium sp. strain RBIITD, a butyrate- and butanol-producing thermophile. Genome Announc. 6(2), e01411-17.
[9] Cameron, D.C., Cooney, C.L., 1986. A novel ferementation: the production of R(-)-1,2-propanediol and acetol by Clostridium Thermosaccharolyticum. Nat. Biotechnol. 4(7), 651-654.
[10] Cao, G., Ren, N., Wang, A., Lee, D.J., Guo, W., Liu, B., Feng, Y., Zhao, Q., 2009. Acid hydrolysis of corn stover for biohydrogen production using Thermoanaerobacterium thermosaccharolyticum W16. Int. J. Hydrogen Energy. 34(17), 7182-7188.
[11] Cao, G.L., Zhao, L., Wang, A.J., Wang, Z.Y., Ren, N.Q., 2014. Single-step bioconversion of lignocellulose to hydrogen using novel moderately thermophilic bacteria. Biotechnol. Biofuels. 7(1), 82.
[12] Debarbouille, M., Gardan, R., Arnaud, M., Rapoport, G., 1999. Role of bkdR, a transcriptional activator of the SigL-dependent isoleucine and valine degradation pathway in Bacillus subtilis. J. Bacteriol. 181(7), 2059-2066.
[13] Demain, A.L., Newcomb, M., Wu, J.D., 2005. Cellulase, clostridia, and ethanol. Microbiol. Mol. Biol. Rev. 69(1), 124-154.
[14] Dürre, P., 2007. Biobutanol: an attractive biofuel. Biotechnol. J. Healthcare Nutr. Technol. 2(12), 1525-1534.
[15] Freier-Schröder, D., Wiegel, J., Gottschalk, G., 1989. Butanol formation by Clostridium thermosaccharolyticum at neutral pH. Biotechnol. Lett. 11(11), 831-836.
[16] Hill, J., Nelson, E., Tilman, D., Polasky, S., Tiffany, D., 2006. Environmental, economic, and energetic costs and benefits of biodiesel and ethanol biofuels. Proc. Natl. Acad. Sci. 103(30), 11206-11210.
[17] Hungate, R.E., 1950. The anaerobic mesophilic cellulolytic bacteria. Bacteriol. Rev. 14(1), 1.
[18] Jiang, Y., Guo, D., Lu, J., Dürre, P., Dong, W., Yan, W., Zhang, W., Ma, J., Jiang, M., Xin, F., 2018. Consolidated bioprocessing of butanol production from xylan by a thermophilic and butanologenic Thermoanaerobacterium sp. M5. Biotechnol Biofuels. 11(1), 1-14.
[19] Jones, D.T., Woods, D.R., 1986. Acetone-butanol fermentation revisited. Microbiol. Rev. 50(4), 484-524.
[20] Khamtib, S., Reungsang, A., 2012. Biohydrogen production from xylose by Thermoanaerobacterium thermosaccharolyticum KKU19 isolated from hot spring sediment. Int. J. Hydrogen Energy. 37(17), 12219-12228.
[21] Lamed, R., Zeikus, J.G., 1980. Ethanol production by thermophilic bacteria: relationship between fermentation product yields of and catabolic enzyme activities in Clostridium thermocellum and Thermoanaerobium brockii. J. Bacteriol. 144(2), 569-578.
[22] Li, T., Zhang, C., Yang, K.L., He, J., 2018. Unique genetic cassettes in a Thermoanaerobacterium contribute to simultaneous conversion of cellulose and monosugars into butanol. Sci Adv. 4(3), e1701475.
[23] Lynd, L.R., Weimer, P.J., Van Zyl, W.H., Pretorius, I.S., 2002. Microbial cellulose utilization: fundamentals and biotechnology. Microbiol. Mol. Biol. Rev. 66(3), 506-577.
[24] Mendez, B.S., Pettinari, M.J., Ivanier, S.E., Ramos, C.A., Siñeriz, F., 1991. Clostridium thermopapyrolyticum sp. nov., a cellulolytic thermophile. Int. J. Syst. Evol. Microbiol. 41(2), 281-283.
[25] Onyenwoke, R.U., Wiegel, J., 2015. Thermoanaerobacter. Bergey's Manual of Systematics of Archaea and Bacteria, pp.1-29.
[26] Overbeek, R., Olson, R., Pusch, G.D., Olsen, G.J., Davis, J.J., Disz, T., Edwards, R.A., Gerdes, S., Parrello, B., Shukla, M., Vonstein, V., 2014. The SEED and the rapid annotation of microbial genomes using subsystems technology (RAST). Nucleic Acids Res. 42(D1), D206-D214.
[27] Panitz, J.C., Zverlov, V.V., Pham, V.T.T., Stürzl, S., Schieder, D., Schwarz, W.H., 2014. Isolation of a solventogenic Clostridium sp. strain: fermentation of glycerol to n-butanol, analysis of the bcs operon region and its potential regulatory elements. Syst. Appl. Microbiol. 37(1), 1-9.
[28] Richter, M., Rosselló-Móra, R., Oliver Glöckner, F., Peplies, J., 2016. JSpeciesWS: a web server for prokaryotic species circumscription based on pairwise genome comparison. Bioinformatics. 32(6), 929-931.
[29] Romano, I., Dipasquale, L., Orlando, P., Lama, L., d’Ippolito, G., Pascual, J., Gambacorta, A, 2010. Thermoanaerobacterium thermostercus sp. nov., a new anaerobic thermophilic hydrogen-producing bacterium from buffalo-dung. Extremophiles. 14(2), 233-240.
[30] Shaw, A.J., Podkaminer, K.K., Desai, S.G., Bardsley, J.S., Rogers, S.R., Thorne, P.G., Hogsett D.A., Lynd, L.R, 2008. Metabolic engineering of a thermophilic bacterium to produce ethanol at high yield. Proc. Natl. Acad. Sci. U.S.A. 105(37), 13769-13774.
[31] Shaw, J.A., Covalla, S.F., Miller, B.B., Firliet, B.T., Hogsett, D.A., Herring, C.D, 2012. Urease expression in a Thermoanaerobacterium saccharolyticum ethanologen allows high titer ethanol production. Metab. Eng. 14(5), 528-532.
[32] Sompong, O., Khongkliang, P., Mamimin, C., Singkhala, A., Prasertsan, P., Birkeland, N.K., 2017. Draft genome sequence of Thermoanaerobacterium sp. strain PSU-2 isolated from thermophilic hydrogen producing reactor. Genomics Data. 12, 49-51.
[33] Sompong, O., Prasertsan, P., Karakashev, D., Angelidaki, I., 2008. Thermophilic fermentative hydrogen production by the newly isolated Thermoanaerobacterium thermosaccharolyticum PSU-2. Int. J. Hydrogen Energy. 33(4), 1204-1214.
[34] Tian, L., Papanek, B., Olson, D.G., Rydzak, T., Holwerda, E.K., Zheng, T., Zhou, J., Maloney, M., Jiang, N., Giannone, R.J., Hettich, R.L., 2016. Simultaneous achievement of high ethanol yield and titer in Clostridium thermocellum. Biotechnol. Biofuels. 9(1), 116.
[35] Vazquez, G.J., Pettinari, M.J., Méndez, B.S., 2001. Phosphotransbutyrylase expression in Bacillus megaterium. Curr. Microbiol. 42(5), 345-349.
[36] Vital, M., Howe, A.C., Tiedje, J.M., 2014. Revealing the bacterial butyrate synthesis pathways by analyzing (meta)genomic data. MBio. 5(2), e00889-14.
[37] Wang, S., Huang, H., Moll, J., Thauer, R.K., 2010. NADP+ reduction with reduced ferredoxin and NADP+ reduction with NADH are coupled via an electron-bifurcating enzyme complex in Clostridium kluyveri. J. Bacteriol. 192(19), 5115-5123.
[38] Wietzke, M., Bahl, H., 2012. The redox-sensing protein Rex, a transcriptional regulator of solventogenesis in Clostridium acetobutylicum. Appl. Microbiol. Biotechnol. 96(3), 749-761.
[39] Yamada, R., Hasunuma, T., Kondo, A., 2013. Endowing non-cellulolytic microorganisms with cellulolytic activity aiming for consolidated bioprocessing. Biotechnol. Adv. 31(6), 754-763.

Files

Cited by

Share

How to cite

Statistics