Skip to main content
SpringerLink
Log in

Effect of organic solvents on the yield and specificity of cyclodextrins by recombinant cyclodextrin glucanotransferase (CGTase) from Anaerobranca gottschalkii

  • Original Article
  • Published:
Journal of Inclusion Phenomena and Macrocyclic Chemistry Aims and scope Submit manuscript

Abstract

Cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19) is an enzyme that degrades starch and starch related glucans into cyclodextrins (CDs) by intramolecular transglycosylation reaction. The biochemical activity of recombinant CGTase from Anaerobranca gottschalkii for the yield and product specificity of cyclodextrins was investigated in the presence of organic solvents. Compared with the control of starch bioconversion, addition of various organic solvents generally increased the total CD and product specificity by affecting product inhibition and/or intermolecular transglycosylation reaction. The highest conversion (45 %) of starch to CDs was obtained in the presence of ethanol, while the simultaneous addition of two organic solvents, decanol-ethanol, comparatively showed a reduced total yield of 39 %. Despite this, the highest product ratio of 91 % α-CD, and 64 % β-CD was obtained in the presence of decanol and cyclohexane respectively. The effect of organic solvents on the yield and specificity of CD was attributed mainly to their effect on product inhibition and transglycosylation reaction. Although the use of two organic solvents showed almost a significant increase in total yield of CDs, it resulted in a comparatively lower specific product yield compared to their respective individual effect. Generally, normal enzyme activity was favoured at higher temperature of 65 °C, but the addition of organic solvents, in most cases, was found to decrease the bioconversion. Thus, the preferred optimal condition was reduced to 40 °C, where the maximal conversion of starch to CDs in general and α-CD in particular was achieved.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Li, Z., et al.: Extracellular expression and biochemical characterization of α-cyclodextrin glycosyltransferase from Paenibacillus macerans. Carbohydr. Res. 345, 886–892 (2010)

    Article  CAS  Google Scholar 

  2. Kuo, C–.C., et al.: Production of cyclodextrin glucanotransferase from an alkalophilic Bacillus sp. by pH-stat fed-batch fermentation. Biotechnol. Lett. 31, 1723–1727 (2009)

    Article  CAS  Google Scholar 

  3. Valle, E.M.M.D.: Cyclodextrins and their uses: a review. Process. Biochem. 39, 1033–1046 (2004)

    Article  Google Scholar 

  4. Moriwaki, C., et al.: A novel cyclodextrin glycosyltransferase from Bacillus sphaericus strain 41: production, characterization and catalytic properties. Biochem. Eng. J. 48, 124–131 (2009)

    Article  CAS  Google Scholar 

  5. Charoensakdi, R., et al.: A recombinant cyclodextrin glycosyltransferase cloned from Paenibacillus sp. strain RB01 showed improved catalytic activity in coupling reaction between cyclodextrins and disaccharides. J. Incl. Phenom. Macrocycl. Chem. 57, 53–59 (2007)

    Article  CAS  Google Scholar 

  6. Tardioli, P.W., Zanin, G.M., Moraes, FFd: Characterization of thermoanaerobacter cyclomaltodextrin glucanotransferase immobilized on glyoxyl-agarose. Enzyme. Microb. Technol. 39, 1270–1278 (2006)

    Article  CAS  Google Scholar 

  7. Blackwood, A.D., Bucke, C.: Addition of polar organic solvents can improve the product selectivity of cyclodextrin glycosyltransferase. Solvent effects on CGTase. Enzyme. Microb. Technol. 27, 704–708 (2000)

    Article  CAS  Google Scholar 

  8. Biwer, A., Antranikian, G., Heinzle, E.: Enzymatic production of cyclodextrins. Appl. Microbiol. Biotechnol. 59, 609–617 (2002)

    Article  CAS  Google Scholar 

  9. Doukyu, N., Ogino, H.: Organic solvent-tolerant enzymes. Biochem. Eng. J. 48, 270–282 (2010)

    Article  CAS  Google Scholar 

  10. Kaulpiboon, J., Hansakul, P.: Comparative studies on the synthesis of cyclodextrin from two bacterial CGTases in the presence of organic solvents. Thammasat. Int. J. Sc. Tech. 12(2), 10–17 (2007)

    Google Scholar 

  11. Qi, Q., Zimmermann, W.: Cyclodextrin glucanotransferase: from gene to applications. Appl. Microbiol. Biotechnol. 66, 475–485 (2005)

    Article  CAS  Google Scholar 

  12. Thiemann, V., et al.: Characterisation of a thermoalkali-stable cyclodextrin glycosyltransferase from the anaerobic thermoalkaliphilic bacterium Anaerobranca gottschalkii. Arch. Microbiol. 182, 226–235 (2004)

    Article  CAS  Google Scholar 

  13. Ding, R., et al.: Enhanced secretion of recombinant α-cyclodextrin glucosyltransferase from E. coli by medium additives. Process. Biochem. 45, 880–886 (2010)

    Article  CAS  Google Scholar 

  14. Rahman, K., et al.: Molecular cloning of a cyclodextrin glucanotransferase gene from alkalophilic Bacillus sp. TS1-1 and characterization of the recombinant enzyme. Enzyme. Microb. Technol. 39, 74–84 (2006)

    Article  CAS  Google Scholar 

  15. Cao, X., et al.: A novel cyclodextrin glycosyltransferase from an alkalophilic Bacillus species: purification and characterization. Food. Res. Int. 38, 309–314 (2005)

    Article  CAS  Google Scholar 

  16. Hannig, G., Makrides, S.C.: Strategies for optimizing heterologous protein expression in Escherichia coli. Trends. Biotechnol. 16(2), 54–60 (1998)

    Article  CAS  Google Scholar 

  17. Kelly, R.M., Dijkhuizen, L., Leemhuis, H.: The evolution of cyclodextrin glucanotransferase product specificity. Appl. Microbiol. Biotechnol. 84, 119–133 (2009)

    Article  CAS  Google Scholar 

  18. Astray, G., et al.: A review on the use of cyclodextrins in foods. Food. Hydrocolloid. 23, 1631–1640 (2009)

    Article  CAS  Google Scholar 

  19. Bruce, L.Z., Henrik, K.N., Robert, L.S.: Thermostable enzymes for industrial applications. J. Ind. Microbiol. 8, 71–82 (1991)

    Article  Google Scholar 

  20. Giordano, F., Novak, C., Moyano, J.R.: Thermal analysis of cyclodextrins and their inclusion compounds. Thermochim. Acta. 380, 123–151 (2001)

    Article  CAS  Google Scholar 

  21. Goh, K.M., et al.: The effects of reaction conditions on the production of γ-cyclodextrin from tapioca starch by using a novel recombinant engineered CGTase. J. Mol. Catal. B Enzym. 49, 118–126 (2007)

    Article  CAS  Google Scholar 

  22. Hamilton, L.M., Kelly, C.T., Fogarty, W.M.: Review: cyclodextrins and their interaction with amylolytic enzymes. Enzyme. Microb. Technol. 26, 561–567 (2000)

    Article  CAS  Google Scholar 

  23. Hedges, A.R.: Industrial applications of cyclodextrins. Chem. Rev. 98, 2035–2044 (1998)

    Article  CAS  Google Scholar 

  24. Jana, S., Deb, J.K.: Strategies for efficient production of heterologous proteins in Escherichia coli. Appl. Microbiol. Biotechnol. 67, 289–298 (2005)

    Article  CAS  Google Scholar 

  25. Kim, M.H., Sohn, C.B., Oh, T.K.: Cloning and sequencing of a cyclodextrin glycosyltransferase gene from Brevibacillus brevis CD162 and its expression in Escherichia coli. FEMS. Microbiol. Lett. 164, 411–418 (1998)

    Article  CAS  Google Scholar 

  26. Leemhuis, H., Kelly, R.M., Dijkhuizen, L.: Engineering of cyclodextrin glucanotransferases and the impact for biotechnological applications. Appl. Microbiol. Biotechnol. 85, 823–835 (2010)

    Article  CAS  Google Scholar 

  27. Li, Z.-F., et al.: Calcium Leads to further increase in glycine-enhanced extracellular secretion of recombinant α-cyclodextrin glycosyltransferase in Escherichia coli. J. Agric. Food. Chem. 57, 6231–6237 (2009)

    Article  CAS  Google Scholar 

  28. Loftsson, T., Duchêne, D.: Cyclodextrins and their pharmaceutical applications. Int. J. Pharm. 329, 1–11 (2007)

    Article  CAS  Google Scholar 

  29. Ong, R.M., et al.: Cloning, extracellular expression and characterizationof a predominant α-CGTase from Bacillus sp. G1 in E. coli. J. Ind. Microbiol. Biotechnol. 35, 1705–1714 (2008)

    Article  CAS  Google Scholar 

  30. Pishtiyski, I., Zhekova, B.: Effect of different substrates and their preliminary treatment on cyclodextrin production. World. J. Microbiol. Biotechnol. 22, 109–114 (2006)

    Article  CAS  Google Scholar 

  31. Szejtli, J.z.: Introduction and general overview of cyclodextrin chemistry. Chem. Rev. 98, 1743–1753 (1998)

    Article  CAS  Google Scholar 

  32. Tonkova, A.: Bacterial cyclodextrin glucanotransferase. Enzyme. Microb. Technol. 22, 678–686 (1998)

    Article  CAS  Google Scholar 

  33. Veen, BAvd, et al.: The three transglycosylation reactions catalyzed by cyclodextrin glycosyltransferase from Bacillus circulans (strain 251) proceed via different kinetic mechanisms. Eur. J. Biochem. 267, 658–665 (2000)

    Article  Google Scholar 

  34. Veen, BAvd, et al.: The role of arginine 47 in the cyclization and coupling reactions of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 Implications for product inhibition and product specificity. Eur. J. Biochem. 267, 3432–3441 (2000)

    Article  Google Scholar 

  35. Veen, BAvd, et al.: Engineering of cyclodextrin glycosyltransferase reaction and product specificity. Biochim. Biophys. Acta. 1543(2), 336–360 (2000)

    Article  Google Scholar 

  36. Villalonga, R., Cao, R., Fragoso, A.: Supramolecular chemistry of cyclodextrins in enzyme technology. Chem. Rev. 107, 3088–3116 (2007)

    Article  CAS  Google Scholar 

  37. Zhekova, B., et al.: Approaches for yield increase of β -cyclodextrin formed by cyclodextrin glucanotransferase from Bacillus megaterium. World. J. Microbiol. Biotechnol. 25, 1043–1049 (2009)

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported financially by the National Natural Science Foundation of China (30970057) and (31100048), the Key Technologies R & D Program of Jiangsu Province, China (BE2011711), the Key Program of National Natural Science Foundation of China (20836003), and the 111 Project (No. 111-2-06).

Author information

Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, People’s Republic of China

    Berhane T. Tesfai, Sheng Chen, Jian Chen & Jing Wu

  2. School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, People’s Republic of China

    Berhane T. Tesfai, Dan Wu, Sheng Chen, Jian Chen & Jing Wu

Authors
  1. Berhane T. Tesfai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dan Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sheng Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jian Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jing Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jing Wu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tesfai, B.T., Wu, D., Chen, S. et al. Effect of organic solvents on the yield and specificity of cyclodextrins by recombinant cyclodextrin glucanotransferase (CGTase) from Anaerobranca gottschalkii . J Incl Phenom Macrocycl Chem 77, 147–153 (2013). https://doi.org/10.1007/s10847-012-0225-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10847-012-0225-6

Keywords

Search

Navigation