Mathematical modelling and kinetic study for CD production catalysed by Toruzyme® and CGTase from Bacillus firmus strain 37

  • 230 Accesses


A new mathematical model was developed for the kinetics of α-, β- and γ-cyclodextrin production, expanding an existing model that only included the production of β- and γ-cyclodextrins, because a detailed kinetic modelling of the reactions involved allows the manipulation of the process yields. The kinetic behaviour of the commercial enzyme Toruzyme® was studied with maltodextrin as substrate at different concentrations and for CGTase from Bacillus firmus strain 37 at a concentration of 100 g L−1. The mathematical model showed a proper fit to the experimental data, within the 24-h period studied, confirming that the considered hypotheses represent the kinetic behaviour of the enzymes in the reaction medium. The kinetic parameters generated by the model allowed reproducing previous observed qualitative tendencies as it can be seen that changing experimental conditions in the reaction process such as enzyme and substrate concentrations results in large changes in the enzyme kinetics and using high substrate concentrations does not guarantee the highest conversion rates due to enzyme inhibition and reverse reactions. In addition, this new mathematical model complements previous qualitative observations enabling the manipulation of the direct and reverse reactions catalysed by the enzyme by adjusting the reaction conditions, to target quantitative results of increased productivity and better efficiency in the production of a desired cyclodextrin.

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

Access options

Buy single article

Instant unlimited access to the full article PDF.

US$ 39.95

Price includes VAT for USA

Subscribe to journal

Immediate online access to all issues from 2019. Subscription will auto renew annually.

US$ 199

This is the net price. Taxes to be calculated in checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5


  1. 1.

    French D (1957) The Schardinger dextrins. Adv Carbohydr Chem Biochem 12:189–260

  2. 2.

    Lo PK, Tan CY, Hassan O, Ahmad A, Mahadi NM, Illias RM (2009) Improvement of excretory overexpression for Bacillus sp. G1 cyclodextrin glucanotransferase (CGTase) in recombinant Escherichia coli though medium optimization. J Biotechnol 8:184–193

  3. 3.

    Szejtli J (1988) Cyclodextrin technology, Davies. Kluwer Academic Publishers, Dordrecht

  4. 4.

    Van Der Veen BA, Van Alebeek GJWM, Uitdehaag JCM, Dijkstra BW, Dijkhuizen L (2000) The three transglycosylation reactions catalysed by cyclodextrin glycosyltransferase from Bacillus circulans (strain 251) proceed via different kinetic mechanisms. Eur J Biochem 267:658–665

  5. 5.

    Muria SR, Cheirsilp B, Kitcha S (2011) Effect of substrate concentration and temperature on the kinetics and thermal stability of cyclodextrin glycosyltransferase for the production of β-cyclodextrin: experimental results vs. mathematical model. Process Biochem 46:1399–1404

  6. 6.

    Zheng M, Endo T, Zimmermann W (2002) Enzymatic synthesis and analysis of large-ring cyclodextrins. Aust J Chem 55:39–48

  7. 7.

    Han R, Li J, Shin H, Chen RR, Du G, Liu L, Chen J (2014) Recent advances in discovery, heterologous expression, and molecular engineering of cyclodextrin glycosyltransferase for versatile applications. Biotechnol Adv 32:415–428

  8. 8.

    Ban X, Gu X, Li C, Huang M, Cheng L, Hong Y, Zhaofeng L (2015) Mutations at calcium binding site III in cyclodextrin glycosyltransferase improve β-cyclodextrin specificity. Int J Biol Macromol 76:224–229

  9. 9.

    Moriwaki C, Costa GL, Pazzetto R, Zanin GM, Moraes FF, Portilho M et al (2007) Production and characterization of a new cyclodextrin glycosyltransferase from Bacillus firmus isolated from Brazilian soil. Process Biochem 42:1384–1390

  10. 10.

    Moriwaki C, Mangolim CS, Ruiz GB, Morais GR, Baesso ML, Matioli G (2014) Biosynthesis of CGTase by immobilized alkalophilic bacilli and crystallization of beta-cyclodextrin: effective techniques to investigate cell immobilization and the production of cyclodextrins. Biochem Eng J 83:22–32

  11. 11.

    Matioli G, Moraes FF, Zanin GM (2000) Ciclodextrinas e suas aplicações em: alimentos, fármacos, cosméticos, agricultura, biotecnologia, química analítica e produtos gerais. Eduem-Editora da Universidade Estadual de Maringá, Maringá

  12. 12.

    Zehentgruber D, Lundemo P, Svensson D, Adlercreutz P (2011) Substrate complexation and aggregation influence the cyclodextrin glycosyltransferase (CGTase) catalysed synthesis of alkyl glycosides. J Biotechnol 155:232–235

  13. 13.

    Sanjari S, Vahabzadeh F (2014) Experimental studies and kinetic modelling of cyclodextrin glucanotransferase production from starch with the use of Bacillus sp. DSM 2523. Starch/Stärke 66:606–614

  14. 14.

    Gastón JAR, Szerman N, Costa H, Krymkiewicz N, Ferrarotti SA (2009) Cyclodextrin glycosyltransferase from Bacillus circulans DF 9R: activity and kinetic studies. Enzyme Microb Technol 45:36–41

  15. 15.

    Zhekova BY, Stanchev VS (2011) Reaction conditions for maximal cyclodextrin production by cyclodextrin glucanotransferase from Bacillus megaterium. Pol J Microbiol 60:113–118

  16. 16.

    Souza M (2003) Modelagem cinética da produção de ciclodextrinas. Dissertação (Mestrado em Engenharia Química)–Curso de Pós-Graduação em Engenharia Química. Universidade Estadual de Maringá, Maringá

  17. 17.

    Souza M, Faria SHB, Zanin GM, Moraes FF (2013) Kinetics of the simultaneous production of β- and γ-cyclodextrins catalysed by CGTase from alkalophilic Bacillus sp. Acta Sci Technol 35(4):687–693

  18. 18.

    Di Maggio J, Ricci JCD, Diaz MS (2010) Parameter estimation in kinetic models for large scale metabolic networks with advanced mathematical programming techniques. 20th European Symposium on Computer Aided Chemical Engineering 28:355–360

  19. 19.

    Fenelon VC, Aguiar MFA, Miyoshi JH, Martinez CO, Matioli G (2015) Ultrafiltration system for cyclodextrin production in repetitive batches by CGTase from Bacillus firmus strain 37. Bioprocess Biosyst Eng 38:1291–1301

  20. 20.

    Matioli G (1997) Seleção de microrganismo e caracterização de sua enzima ciclodextrina glicosiltransferase. Tese (Doutorado em Ciências–Bioquímica)—Curso de Pós-Graduação em Bioquímica. Universidade Federal do Paraná, Curitiba

  21. 21.

    Matioli G, Zanin GM, Moraes FF (2001) Characterization of cyclodextrin glycosyltransferase from Bacillus firmus strain no. 37. Appl Biochem Biotechnol 91–93:643–654

  22. 22.

    Calsavara LPV, Cunha ARD, Balbino TA, Zanin GM, Moraes FF (2011) Production of cyclodextrins from corn starch granules in a sequential batch mode and in the presence of ethanol. Appl Biochem Biotechnol 165:1485–1493

  23. 23.

    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

  24. 24.

    Moriwaki C, Mazzer C, Pazzetto R, Matioli G (2009) Produção, purificação e aumento da performance de ciclodextrina glicosiltransferases para produção de ciclodextrinas. Quím Nova 32:2360–2366

  25. 25.

    Segel IH (1975) Enzyme kinetics. Wiley, California

  26. 26.

    Zhekova B, Pishtiyski I, Stanchev V (2008) Investigation on cyclodextrin production with cyclodextrin glucanotransferase from Bacillus megaterium. Food Technol Biotechnol 46:328–334

  27. 27.

    Goh PH, Illias RMd, Goh KM (2012) Domain replacement to elucidate the role of B domain in CGTase thermostability and activity. Proc Biochem 47:2123–2130

  28. 28.

    Tardioli PW, Zanin GM, Moraes FF (2000) Production of cyclodextrins in a fluidized-bed reactor using cyclodextrin-glycosyl-transferase. Appl Biochem Biotechnol 84–86:1003–1019

  29. 29.

    Rakmai J, Cheirsilp B (2016) Continuous production of β-cyclodextrin by cyclodextrin glycosyltransferase immobilized in mixed gel beads: comparative study in continuous stirred tank reactor and packed bed reactor. Biochem Eng J 105:107–113

  30. 30.

    Mimi Sakinah AM, Ismail AF, Illias RMd, Zularisam AW, Hassan O, Matsuura T (2014) Effect of substrate and enzyme concentration on cyclodextrin production in a hollow fibre membrane reactor system. Sep Purif Technol 124:61–67

  31. 31.

    Wu D, Chen S, Wang N, Chen J, Wu J (2012) Gamma-cyclodextrin production using cyclodextrin glycosyltransferase from Bacillus clarkii 7364. Appl Biochem Biotechnol 167:1954–1962

  32. 32.

    Wind RD, Uitdehaag JCM, Buitelaar RM, Dijkstra BW, Dijkhuizeni L (1998) Engineering of cyclodextrin product specificity and pH Optima of the thermostable cyclodextrin glycosyltransferase from Thermoanaerobacterium thermosulfurigenes EM1. J Biol Chem 273:5771–5779

  33. 33.

    Mäkelä MJ, Korpela TK (1988) Determination of catalytic activity of cyclomaltodextrin glucanotransferase by maltotriose-methylorange assay. J Biochem Biophys Methods 15:307–318

  34. 34.

    Ozbek B, Yuceer S (2001) α-Amylase inactivation during wheat starch hydrolysis process. Process Biochem 37:87–95

  35. 35.

    Elbaz AF, Sobhi A, El Mekawy A (2015) Purification and characterization of cyclodextrin β-glucanotransferase from novel alkalophilic bacilli. Bioprocess Biosyst Eng 38:767–776

  36. 36.

    Rather MY, Ara KZG, Karlsson EN, Adlercreutz P (2015) Characterization of cyclodextrin glycosyltransferases (CGTases) and their application for synthesis of alkyl glycosides with oligomeric head group. Process Biochem 50:722–728

  37. 37.

    Matioli G, Zanin GM, Moraes FF (2002) Influence of substrate and product concentrations on the production of cyclodextrins by CGTase of Bacillus firmus, strain no. 37. Appl Biochem Biotechnol 98–100:947–961

Download references


This paper was supported by the Federal Institute of Paraná-Paranavaí, the State University of Maringá and the financial support of Novozymes and Amido Mundo Novo.

Author information

Correspondence to Keren Hapuque Pinheiro.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests being compliance with ethical standards.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Pinheiro, K.H., do Nascimento, L.B., Fenelon, V.C. et al. Mathematical modelling and kinetic study for CD production catalysed by Toruzyme® and CGTase from Bacillus firmus strain 37. Bioprocess Biosyst Eng 40, 1305–1316 (2017) doi:10.1007/s00449-017-1789-8

Download citation


  • Cyclomaltodextrin glucanotransferase (CGTase)
  • Modelling
  • Kinetics