Abstract
Cell autolysis plays important physiological roles in the life cycle of clostridial cells. Understanding the genetic basis of the autolysis phenomenon of pathogenic Clostridium or solvent producing Clostridium cells might provide new insights into this important species. Genes that might be involved in autolysis of Clostridium acetobutylicum, a model clostridial species, were investigated in this study. Twelve putative autolysin genes were predicted in C. acetobutylicum DSM 1731 genome through bioinformatics analysis. Of these 12 genes, gene SMB_G3117 was selected for testing the in tracellular autolysin activity, growth profile, viable cell numbers, and cellular morphology. We found that overexpression of SMB_G3117 gene led to earlier ceased growth, significantly increased number of dead cells, and clear electrolucent cavities, while disruption of SMB_G3117 gene exhibited remarkably reduced intracellular autolysin activity. These results indicate that SMB_G3117 is a novel gene involved in cellular autolysis of C. acetobutylicum.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Allcock, E.R., Reid, S.J., Jones, D.T., and Woods, D.R. (1981). Autolytic activity and an autolysis-deficient mutant of Clostridium acetobutylicum. Appl Environ Microbiol 42, 929–935.
Andreesen, J., Bahl, H., and Gottschalk, G. (1989). Introduction to the physiology and biochemistry of the genus Clostridium. Clostridia. 27–62. Edited by Minton, N. and Clarke, D. New York and London: Plenum Press.
Bao, G., Wang, R., Zhu, Y., Dong, H., Mao, S., Zhang, Y., Chen, Z., Li, Y., and Ma, Y. (2011). Complete genome sequence of Clostridium acetobutylicum DSM 1731, a solvent-producing strain with multireplicon genome architecture. J Bacteriol 193, 5007–5008.
Barber, J.M., Robb, F.T., Webster, J.R., and Woods, D.R. (1979). Bacteriocin production by Clostridium acetobutylicum in an industrial fermentation process. Appl Environ Microbiol 37, 433–437.
Blackman, S.A., Smith, T.J., and Foster, S.J. (1998). The role of autolysins during vegetative growth of Bacillus subtilis 168. Microbiol-UK 144, 73–82.
ChapotChartier, M.P. (1996). Autolysins of lactic acid bacteria. Lait 76, 91–109.
Croux, C., Canard, B., Goma, G., and Soucaille, P. (1992a). Autolysis of Clostrtdium acetobutylicum ATCC 824. J Gen Microbiol 138, 861–869.
Croux, C., Canard, B., Goma, G., and Soucaille, P. (1992b). Purification and characterization of an extracellular muramidase of Clostridium acetobutylicum ATCC 824 that acts on non-N-acetylated peptidoglycan. Appl Environ Microbiol 58, 1075–1081.
Croux, C., and Garcia, J.L. (1992). Reconstruction and expression of the autolytic gene from Clostridium acetobutylicum ATCC 824 in Escherichia coli. FEMS Microbiol Lett 74, 13–20.
Dong, H., Tao, W., Zhang, Y., and Li, Y. (2012). Development of an anhydrotetracycline-inducible gene expression system for solvent-producing Clostridium acetobutylicum: A useful tool for strain engineering. Metab Eng 14, 59–67.
Dong, H., Zhang, Y., Dai, Z., and Li, Y. (2010). Engineering Clostridium strain to accept unmethylated DNA. PLoS ONE 5, e9038.
Eltsov, M., and Zuber, B. (2006). Transmission electron microscopy of the bacterial nucleoid. J Struct Biol 156, 246–254.
Foster, S.J. (1994). The role and regulation of cell-wall structural dynamics during differentiation of endospore-forming bacteria. J Appl Bacteriol 76, S25–39.
Foster, S.J. (1995). Molecular characterization and functional analysis of the major autolysin of Staphylococcus aureus 8325/4. J Bacteriol 177, 5723–5725.
Garcia, J.L., Garcia, E., Sanchezpuelles, J.M., and Lopez, R. (1988). Identification of a lytic enzyme of Clostridium acetobutylicum that degrades choline-containing pneumococcal cell walls. Fems MicrobiolLett 52, 133–137.
Hartmanis, M.G., and Gatenbeck, S. (1984). Intermediary metabolism in Clostridium acetobutylicum: levels of enzymes involved in the formation of acetate and butyrate. Appl Environ Microbiol 47, 1277–1283.
Heap, J.T., Kuehne, S.A., Ehsaan, M., Cartman, S.T., Cooksley, C.M., Scott, J.C., and Minton, N.P. (2010). The ClosTron: mutagenesis in Clostridium refined and streamlined. J Microbiol Methods 80, 49–55.
Heidrich, C., Templin, M.F., Ursinus, A., Merdanovic, M., Berger, J., Schwarz, H., de Pedro, M.A., and Holtje, J.V. (2001). Involvement of N-acetylmuramyl-L-alanine amidases in cell separation and antibiotic-induced autolysis of Escherichia coli. Mol Microbiol 41, 167–178.
Hirsch, A., and Grinsted, E. (1954). Methods for the growth and enumeration of anaerobic spore-formers from cheese, with observations on the effect of nisin. J. Dairy Res. 21, 101–110.
Holtje, J.V. (1995). From growth to autolysis: the murein hydrolases in Escherichia coli. Arch Microbiol 164, 243–254.
Jayaswal, R.K., Lee, Y.I., and Wilkinson, B.J. (1990). Cloning and expression of a Staphylococcus aureus gene encoding a peptidoglycan hydrolase activity. J Bacteriol 172, 5783–5788.
Jones, S.W., Paredes, C.J., Tracy, B., Cheng, N., Sillers, R., Senger, R.S., and Papoutsakis, E.T. (2008). The transcriptional program underlying the physiology of clostridial sporulation. Genome Biol 9 R114.
Ju, C.X., Gu, H.W., and Lu, C.P. (2012). Characterization and functional analysis of atl, a novel gene encoding autolysin in Streptococcus suis. J Bacteriol 194, 1464–1473.
Kuehne, S.A., Heap, J.T., Cooksley, C.M., Cartman, S.T., and Minton, N.P. (2011). ClosTron-mediated engineering of Clostridium. In Strain Engineering (Springer), pp. 389–407.
Lee, S.Y., Park, J.H., Jang, S.H., Nielsen, L.K., Kim, J., and Jung, K.S. (2008). Fermentative butanol production by Clostridia. Biotechnol Bioeng 101, 209–228.
Lutke-Eversloh, T., and Bahl, H. (2011). Metabolic engineering of Clostridium acetobutylicum: recent advances to improve butanol production. Curr Opin Biotechnol 22, 634–647.
Mermelstein, L.D., Welker, N.E., Bennett, G.N., and Papoutsakis, E.T. (1992). Expression of cloned homologous fermentative genes in Clostridium acetobutylicum ATCC 824. Nat Biotech 10, 190–195.
Nolling, J., Breton, G., Omelchenko, M.V., Makarova, K.S., Zeng, Q.D., Gibson, R., Lee, H.M., Dubois, J., Qiu, D.Y., Hitti, J., et al. (2001). Genome sequence and comparative analysis of the solvent-producing bacterium Clostridium acetobutylicum. J Bacteriol 183, 4823–4838.
Rashid, M.H., Kuroda, A., and Sekiguchi, J. (1993). Bacillus subtilis mutant deficient in the major autolytic amidase and glucosaminidase is impaired in motility. FEMS Microbiol Lett 112, 135–140.
Rehner, S.A., and Samuels, G.J. (1994). Taxonomy and phylogeny of gliocladium analyzed from nuclear large subunit ribosomal DNA-sequences. Mycol Res 98, 625–634.
Shockman, G.D., and Holtje, J.-V. (1994). Microbial peptidoglycan (murein) hydrolases. In Bacterial Cell Wall, J.-M. Ghuysen, and R. Hakenbeck, eds. (Amsterdam, Elsevier), pp. 131–166.
Smith, T.J., Blackman, S.A., and Foster, S.J. (2000). Autolysins of Bacillus subtilis: multiple enzymes with multiple functions. Microbiol-UK 146, 249–262.
Tamura, H., Yamada, A., and Kato, H. (2012). Identification and characterization of an autolysin gene, atlA, from Streptococcus criceti. J Microbiol 50, 777–784.
Webster, J.R., Reid, S.J., Jones, D.T., and Woods, D.R. (1981). Purification and characterization of an autolysin from Clostridium acetobutylicum. Appl Environ Microbiol 41, 371–374.
Yoshino, S., Ogata, S., and Hayashida, S. (1982). Some properties of autolysin of Clostridium saccharoperbutylacetonicum. Agric Biol Chem 46, 1243–1248.
Zhang, Y.H., Zhang, Y.P., Zhu, Y., Mao, S.M., and Li, Y. (2010). Proteomic analyses to reveal the protective role of glutathione inresistance of Lactococcus lactis to osmotic stress. Appl Environ Microbiol 76, 3177–3186.
Zingaro, K.A., and Terry Papoutsakis, E. (2013). GroESL overexpression imparts Escherichia coli tolerance to i-, n-, and 2-butanol, 1,2,4-butanetriol and ethanol with complex and unpredictable patterns. Metab Eng 15, 196–205.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yang, L., Bao, G., Zhu, Y. et al. Discovery of a novel gene involved in autolysis of Clostridium cells. Protein Cell 4, 467–474 (2013). https://doi.org/10.1007/s13238-013-3025-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13238-013-3025-x