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Purification and Characterization of a GH11 Xylanase from Biobutanol-Producing Clostridium beijerinckii G117

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Abstract

Most biobutanol-producing Clostridium strains are unable to ferment polysaccharides such as cellulose and xylan due to the lack of hydrolyzing enzymes. In this study, we show that Clostridium beijerinckii G117, a newly isolated biobutanol-producing strain, expresses xylanase enzyme in the presence of 1 % beechwood xylan. The xylanase activity in the medium containing actively growing culture and 1 % of beechwood xylan can reach up to 2.66 U/ml after 14 h of fermentation. Using salting-out and size-exclusion chromatography, we purify the crude xylanase by 8.7-fold from the supernatant with a yield of 32.2 %. This purified xylanase has a molecular weight of 22.6 kDa, making it one of the smallest reported clostridial xylanases. Conserved domain analysis reveals that the xylanase belongs to glycoside hydrolase family 11 (GH11) but lacks a carbohydrate binding domain. When beechwood xylan is used as substrate for the xylanase, majority of the products are xylo-oligosaccharide (~98 %), suggesting that this is an endo-1,4-β-xylanase.

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References

  1. Jin, C., Yao, M., Liu, H., Lee, C. F., & Ji, J. (2011). Renewable and Sustainable Energy Reviews, 15, 4080–4106.

    Article  CAS  Google Scholar 

  2. Ezeji, T. C., Qureshi, N., & Blaschek, H. P. (2007). Current Opinion in Biotechnology, 18, 220–227.

    Article  CAS  Google Scholar 

  3. Sun, Y., & Cheng, J. (2002). Bioresource Technology, 83, 1–11.

    Article  CAS  Google Scholar 

  4. Gírio, F. M., Fonseca, C., Carvalheiro, F., Duarte, L. C., Marques, S., & Bogel-Łukasik, R. (2010). Bioresource Technology, 101, 4775–4800.

    Article  Google Scholar 

  5. Jeffries, T. W. (2006). Current Opinion in Biotechnology, 17, 320–326.

    Article  CAS  Google Scholar 

  6. Menon, V., & Rao, M. (2012). Progress in Energy and Combustion Science, 38, 522–550.

    Article  CAS  Google Scholar 

  7. Bobleter, O. (1994). Progress in Polymer Science, 19, 797–841.

    Article  CAS  Google Scholar 

  8. Biely, P. (1985). Trends in Biotechnology, 3, 286–290.

    Article  CAS  Google Scholar 

  9. Paes, G., Berrin, J. G., & Beaugrand, J. (2012). Biotechnology Advances, 30, 564–592.

    Article  CAS  Google Scholar 

  10. Lee, S. F., Forsberg, C. W., & Gibbins, L. N. (1985). Applied and Environmental Microbiology, 50, 1068–1076.

    CAS  Google Scholar 

  11. Rajagopalan, G., Yew, K. W., He, J., & Yang, K. L. (2012). BioEnergy Research, 6, 448–457.

    Article  Google Scholar 

  12. Chua, T. K., Liang, D. W., Qi, C., Yang, K. L., & He, J. (2013). Bioresource Technology, 135, 372–378.

    Article  CAS  Google Scholar 

  13. Wu, Y. R., Li, Y., Yang, K. L., & He, J. (2012). Journal of Bacteriology, 194, 5470–5471.

    Article  CAS  Google Scholar 

  14. He, J., Ritalahti, K. M., Yang, K.-L., Koenigsberg, S. S., & Loffler, F. E. (2003). Nature, 424, 62–65.

    Article  CAS  Google Scholar 

  15. Miller, G. L. (1959). Analytical Chemistry, 31, 426–428.

    Article  CAS  Google Scholar 

  16. Sa-Pereira, P., Paveia, H., Costa-Ferreira, M., & Aires-Barros, M. R. (2003). Molecular Biotechnology, 24, 257–281.

    Article  CAS  Google Scholar 

  17. Marichamy, S., & Mattiasson, B. (2005). Enzyme and Microbial Technology, 37, 497–504.

    Article  CAS  Google Scholar 

  18. Palop, M. L., Vallés, S., Piñaga, F., & Flors, A. (1991). Journal of Chemical Technology and Biotechnology, 51, 105–114.

    Article  CAS  Google Scholar 

  19. Kosugi, A., Murashima, K., & Doi, R. H. (2001). Journal of Bacteriology, 183, 7037–7043.

    Article  CAS  Google Scholar 

  20. Murty, M. V. S., & Chandra, T. S. (1991). Enzyme and Microbial Technology, 13, 430–435.

    Article  CAS  Google Scholar 

  21. Rani, D. S., & Nand, K. (2000). Process Biochemistry, 36, 355–362.

    Article  CAS  Google Scholar 

  22. Lemmel, S. A., Datta, R., & Frankiewicz, J. R. (1986). Enzyme and Microbial Technology, 8, 217–221.

    Article  CAS  Google Scholar 

  23. Akila, G., & Chandra, T. S. (2003). FEMS Microbiology Letters, 219, 63–67.

    Article  CAS  Google Scholar 

  24. Ali, M. K., Rudolph, F. B., & Bennett, G. N. (2005). Journal of Industrial Microbiology and Biotechnology, 32, 12–18.

    Article  CAS  Google Scholar 

  25. Subramaniyan, S., & Prema, P. (2002). Critical Reviews in Biotechnology, 22, 33–64.

    Article  CAS  Google Scholar 

  26. Stoppok, W., Rapp, P., & Wagner, F. (1982). Applied and Environmental Microbiology, 44, 44–53.

    CAS  Google Scholar 

  27. Han, S. O., Yukawa, H., Inui, M., & Doi, R. H. (2003). Journal of Bacteriology, 185, 6067–6075.

    Article  CAS  Google Scholar 

  28. Narayanaswamy, N., Dheeran, P., Verma, S., & Kumar, S. (2013). Biological Pretreatment of Lignocellulosic Biomass for Enzymatic Saccharification. In Z. Fang (Ed.), Pretreatment Techniques for Biofuels and Biorefineries (pp. p. 3–p. 34). Berlin Heidelberg: Springer.

    Chapter  Google Scholar 

  29. Ali, M. K., Rudolph, F. B., & Bennett, G. N. (2004). Journal of Industrial Microbiology and Biotechnology, 31, 229–234.

    Article  CAS  Google Scholar 

  30. Murty, M. V., & Chandra, T. S. (1992). Antonie van Leeuwenhoek, 61, 35–41.

    Article  CAS  Google Scholar 

  31. Wang, Y., Li, X., Mao, Y., & Blaschek, H. P. (2012). BMC Genomics, 13, 102.

    Article  CAS  Google Scholar 

  32. Biely, P., Vrsanska, M., Tenkanen, M., & Kluepfel, D. (1997). Journal of Biotechnology, 57, 151–166.

    Article  CAS  Google Scholar 

  33. Swaroopa Rani, D., & Nand, K. (2001). Anaerobe, 7, 45–53.

    Article  CAS  Google Scholar 

  34. Sakka, K., Kojima, Y., Kondo, T., Karita, S., Shimada, K., & Ohmiya, K. (1994). Bioscience, Biotechnology, and Biochemistry, 58, 1496–1499.

    Article  CAS  Google Scholar 

  35. Hayashi, H., Takehara, M., Hattori, T., Kimura, T., Karita, S., Sakka, K., & Ohmiya, K. (1999). Applied Microbiology and Biotechnology, 51, 348–357.

    Article  CAS  Google Scholar 

  36. Bérenger, J.-F., Frixon, C., Bigliardi, J., & Creuzet, N. (1985). Canadian Journal of Microbiology, 31, 635–643.

    Article  Google Scholar 

  37. Bastawde, K. B. (1992). World Journal of Microbiology and Biotechnology, 8, 353–368.

    Article  CAS  Google Scholar 

  38. Ali, M. K., Hayashi, H., Karita, S., Goto, M., Kimura, T., Sakka, K., & Ohmiya, K. (2001). Bioscience, Biotechnology, and Biochemistry, 65, 41–47.

    Article  CAS  Google Scholar 

  39. Nagarajan, D. R., Rajagopalan, G., & Krishnan, C. (2006). Applied Microbiology and Biotechnology, 73, 591–597.

    Article  CAS  Google Scholar 

  40. Knob, A., & Carmona, E. C. (2010). Applied Biochemistry and Biotechnology, 162, 429–443.

    Article  CAS  Google Scholar 

  41. Sunna, A., & Antranikian, G. (1997). Critical Reviews in Biotechnology, 17, 39–67.

    Article  CAS  Google Scholar 

  42. Lee, S. F., Forsberg, C. W., & Rattray, J. B. (1987). Applied and Environmental Microbiology, 53, 644–650.

    CAS  Google Scholar 

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Acknowledgments

This work was supported by the Singapore National Research Foundation (NRF) under grant NRF 2009 NRF-CRP 001-039. The contributions of Dr. Wu Yirui to this work are greatly appreciated.

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Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, National University of Singapore, Block E5 #02-09, 4 Engineering Drive 4, Singapore, 117585, Singapore

    Choong Hey Ng & Kun-Lin Yang

  2. Department of Civil and Environmental Engineering, National University of Singapore, Block E1A #07-03, 1 Engineering Drive 2, Singapore, 117576, Singapore

    Jianzhong He

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  1. Choong Hey Ng
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  2. Jianzhong He
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Correspondence to Kun-Lin Yang.

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Table S1

Table summarizing the purification steps of G117 xylanase from the crude enzyme to the final purified product. (DOCX 12 kb)

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Ng, C.H., He, J. & Yang, KL. Purification and Characterization of a GH11 Xylanase from Biobutanol-Producing Clostridium beijerinckii G117. Appl Biochem Biotechnol 175, 2832–2844 (2015). https://doi.org/10.1007/s12010-014-1470-5

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  • DOI: https://doi.org/10.1007/s12010-014-1470-5

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