Abstract
As a promising alternative biofuel, biobutanol can be produced through acetone/butanol/ethanol (ABE) fermentation. Currently, ABE fermentation is still a small-scale industry due to its low production and high input cost. Moreover, butanol toxicity to the Clostridium fermentation host limits the accumulation of butanol in the fermentation broth. The wild-type Clostridium acetobutylicum D64 can only produce about 13 g butanol/L and tolerates less than 2% (v/v) butanol. To improve the tolerance of C. acetobutylicum D64 for enhancing the production of butanol, nitrogen ion beam implantation was employed and finally five mutants with enhanced butanol tolerance were obtained. Among these, the most butanol tolerant mutant C. acetobutylicum NT642 can tolerate above 3% (v/v) butanol while the wide-type strain can only withstand 2% (v/v). In batch fermentation, the production of butanol and ABE yield of C. acetobutylicum NT642 was 15.4 g/L and 22.3 g/L, respectively, which were both higher than those of its parental strain and the other mutants using corn or cassava as substrate. Enhancing butanol tolerance is a great precondition for obtaining a hyper-yield producer. Nitrogen ion beam implantation could be a promising biotechnology to improve butanol tolerance and production of the host strain C. acetobutylicum.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Annous, B.A. and Blaschek, B.H. 1991. Isolation and characterization of Clostridium acetobutylicum mutants with enhanced amylolytic activity. Appl. Environ. Microbiol.7, 2544–2548.
Bowles, L.K. and Ellefson, W.L. 1985. Effects of butanol on Clostridium acetobutylicum. Appl. Environ. Microbiol.50, 1165–1170.
Chen, C.K. and Blaschek, B.H. 1999. Acetate enhances solvent production and prevents degeneration in Clostridium beijerinckii BA101. Appl. Microbiol. Biotechnol.52, 170–173.
Connor, M.R. and Liao, J.C. 2009. Microbial production of advanced transportation fuels in non-natural hosts. Curr. Opin. Biotechnol.20, 307–315.
Dunlop, M.J., Dossani, Z.Y., Szmidt, H.L., Chu, H.C., Lee, T.S., Keasling, J.D., Hadi, M.Z., and Mukhopadhyay, A. 2011. Engineering microbial biofuel tolerance and export using efflux pumps. Mol. Syst. Biol.7, 487–493.
Feng, H., Yu, Z. and Chu, P. 2006. Ion implantation of organisms. Mater. Sci. Eng. R.54, 49–120.
Formanek, J., Mackie, R., and Blaschek, H.P. 1997. Enhanced butanol production by Clostridium beijerinckii BA101 grown in semidefined P2 medium containing 6 percent maltodextrin or glucose. Appl. Environ. Microbiol.63, 2306–2310.
Guo, T., Tang, Y., Zhang, Q.Y., Du, T.F., Liang, D.F., Jiang, M., and Ouyang, P.K. 2011. Clostridium beijerinckii mutant with high inhibitor tolerance obtained by low-energy ion implantation. J. Ind. Microbiol. Biotechnol.39, 401–407.
**, X.Q., Zhu, H., Wu, X.M., Zhang, G.D., and He, B.F. 2008. A rapid screening method of producing strain in acetone-butanol fermentation. Chin. J. Process Eng.8, 1185–1189.
Kataoka, N., Tajima, T., Kato, J., Rachadech, W., and Vangnai, A.S. 2011. Development of butanol-tolerant Bacillus subtilis strain GRSW2-B1 as a potential bioproduction host. AMB Express1, 10–21.
Lin, Y.L. and Blaschek, H.P. 1983. Butanol production by a butanol-tolerant strain of Clostridium acetobutylicum in extruded corn broth. Appl. Environ. Microbiol.45, 966–973.
Liu, S.Q., Bischoff, K.M., Leathers, T.D., Qureshi, N., Rich, J.O., and Hughes, S.R. 2012. Adaptation of lactic acid bacteria to butanol. Biocatal. Agri. Biotechnol.1, 57–61.
Maddox, I.S. 1980. Production of n-butanol from whey filtrate using Clostridium acetobutylicum NCIB 2951. Biotechnol. Lett.2, 101–105.
Makarova, K., Slesarev, A., Wolf, Y., Sorokin, A., Mirkin, B., Koonin, E., Pavlov, A., Pavlova, N., Karamychev, V., Polouchine, N., andet al. 2006. Comparative genomics of the lactic acid bacteria. Proc. Natl. Acad. Sci. USA103, 15611–15616.
Mao, S.M., Luo, Y.M., Zhang, T.R., Li, J.S., Bao, G.H., Zhu, Y., Chen, Z.G., Zhang, Y.P., Li, Y., and Ma, Y.H. 2010. Proteome reference map and comparative proteomic analysis between a wild type Clostridium acetobutylicum DSM 1731 and its mutant with enhanced butanol tolerance and butanol yield. J. Proteome Res.9, 3046–3049.
Mutschlechner, O., Swoboda, H., and Gapes, J.R. 2000. Continuous two-stage ABE-fermentation using Clostridium beijerinckii NRRL B592 operating with a growth rate in the first stage vessel close to its maximal value. J. Mol. Microbiol. Biotechnol.2, 101–105.
Papoutsakis, E.T. 2008. Engineering solventogenic Clostridia. Curr. Opin. Biotechnol.19, 420–429.
Soucaille, P., Joliff, G., Izard, A., and Goma, G. 1987. Butanol tolerance and autobacteriocin production by Clostridium acetobutylicum. Curr. Microbiol.14, 295–295.
Tomas, C.A., Beamish, J., and Papoutsakis, E.T. 2004. Transcriptional analysis of butanol stress and tolerance in Clostridium acetobutylicum. J. Bacteriol.186, 2006–2018.
Winkler, J., Rehmann, M., and Kao, K.C. 2010. Novel Escherichia coli hybrids with enhanced butanol tolerance. Biotechnol. Lett.32, 915–920.
Xu, T.T., Bai, Z.Z., Wang, L.J., and He, B.F. 2008. Breeding of d(−)-lactic acid high producing strain by low-energy ion implantation and preliminary analysis of related metabolism. Appl. Biochem. Biotechnol.160, 314–321.
Yu, Z.L., Deng, J.G., He, J.J., and Huo, Y.P. 1991. Mutation breeding by ion implantation. Nucl. Instrum. Methods Phys. Res. Sect. B.59, 705–708.
Author information
Authors and Affiliations
Corresponding author
Additional information
Supplemental material for this article may be found at http://www.springer.com/content/120956.
Electronic supplementary material
Rights and permissions
About this article
Cite this article
Liu, XB., Gu, QY., Yu, XB. et al. Enhancement of butanol tolerance and butanol yield in Clostridium acetobutylicum mutant NT642 obtained by nitrogen ion beam implantation. J Microbiol. 50, 1024–1028 (2012). https://doi.org/10.1007/s12275-012-2289-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12275-012-2289-9