Advertisement

Biotechnology Letters

, Volume 36, Issue 8, pp 1661–1667 | Cite as

Impact of the configuration of a chiral, activating carrier on the enantioselectivity of entrapped lipase from Candida rugosa in cyclohexane

  • Jan Tobis
  • Joerg C. Tiller
Original Research Paper

Abstract

Lipase from Candida rugosa was loaded into an amphiphilic polymer co-network (APCN) composed of the chiral poly[(R)-N-(1-hydroxybutan-2-yl) acrylamide] [P-(R)-HBA] and P-(S)-HBA, respectively, linked by poly(dimethylsiloxane). The nanophase-separated amphiphilic morphology affords a 38,000-fold activation of the enzyme in the esterification of 1-phenylethanol with vinyl acetate. Further, the enantioselectivity of the entrapped lipase was influenced by the configuration of the chiral, hydrophilic polymer matrix. While the APCN with the (S)-configuration of the APCN affords 5.4 faster conversion of the (R)-phenylethanol compared to the respective (S)-enantiomer, the (R)-APCN allows an only a 2.8 faster conversion of the (R)-enantiomer of the alcohol. Permeation-experiments reveal that the enantioselectivity of the reaction is at least partially caused by specific interactions between the substrates and the APCN.

Keywords

Amphiphilic polymer co-networks Biocatalysis Enantioselectivity Immobilization Lipase Non-aqueous media Organic solvents 

References

  1. Adlercreutz P (2013) Immobilisation and application of lipases in organic media. Chem Soc Rev 42(15):6406–6436PubMedCrossRefGoogle Scholar
  2. Broos J, Kakodinskaya IK, Engbersen JFJ, Verboom W, Reinhoudt DN (1995) Large activation of serine proteases by pretreatment with crown ethers. J Chem Soc Chem Commun 2:255–256CrossRefGoogle Scholar
  3. Bruns N, Tiller JC (2005) Amphiphilic network as nanoreactor for enzymes in organic solvents. Nano Lett 5:45–48PubMedCrossRefGoogle Scholar
  4. Bruns N, Tiller JC (2006) Nanophasic amphiphilic co-networks with a fluorophilic phase. Macromolecules 39:4386–4394CrossRefGoogle Scholar
  5. Bruns N, Bannwarth W, Tiller JC (2008) Amphiphilic co-networks as activating carriers for the enhancement of enzymatic activity in supercritical CO2. Biotechnol Bioeng 101(1):19–26PubMedCrossRefGoogle Scholar
  6. Carrea G, Riva S (2000) Properties and synthetic applications of enzymes in organic solvents. Angew Chem Int Ed Engl 39:2226–2254PubMedCrossRefGoogle Scholar
  7. Dech S, Cramer T, Ladisch R, Bruns N, Tiller JC (2011) Solid–solid interface adsorption of proteins and enzymes in nanophase-separated amphiphilic co-networks. Biomacromolecules 12:1594–1601PubMedCrossRefGoogle Scholar
  8. Erdodi G, Kennedy JP (2006) Amphiphilic co-networks: definition, synthesis, applications. Prog Polym Sci 31:1–18CrossRefGoogle Scholar
  9. Fitzpatrick PA, Klibanov AM (1991) How can the solvent affect enzyme enantioselectivity. J Am Chem Soc 113:3166–3171CrossRefGoogle Scholar
  10. Godoy CA, Romero O, delas Rivas B, Mateo C, Fernandez-Lorente G, Guisan JM, Palomo JM (2013) Changes on enantioselectivity of a genetically modified thermophilic lipase by site-directed oriented immobilization. J Mol Catal B 87:121–127CrossRefGoogle Scholar
  11. Khmelnitsky YL, Welch SH, Clark DS, Dordick JS (1994) Salts dramatically enhance activity of enzymes suspended in organic solvents. J Am Chem Soc 116:2647–2648CrossRefGoogle Scholar
  12. Mallin H, Menyes U, Vorhaben T, Hoehne M, Bornscheuer UT (2013) Immobilization of two (R)-amine transaminases on an optimized chitosan support for the enzymatic synthesis of optically pure amines. ChemCatChem 5:588–593CrossRefGoogle Scholar
  13. Natalia D, Greiner L, Leitner W, Ansorge-Schumacher MB (2012) Stability, activity, and selectivity of benzaldehyde lyase in supercritical fluids. J Supercrit Fluids 62:173–177CrossRefGoogle Scholar
  14. Paradkar VM, Dordick JS (1994) Aqueous-like activity of alpha-chymotrypsin dissolved in nearly anhydrous organic solvents. J Am Chem Soc 116:5009–5010CrossRefGoogle Scholar
  15. Reetz MT (1997) Entrapment of biocatalysts in hydrophobic sol–gel materials for use in organic chemistry. Adv Mater 9(12):943–954CrossRefGoogle Scholar
  16. Reetz MT, Wilensek S, Zha D, Jaeger K-E (2001) Directed evolution of an enantioselective enzyme through combinatorial multiple-cassette mutagenesis. Angew Chem Int Ed Engl 40:3589–3591PubMedCrossRefGoogle Scholar
  17. Schoenfeld I, Dech S, Ryabenky B, Daniel B, Glowacki B, Ladisch R, Tiller JC (2013) Investigations on diffusion limitations of biocatalyzed reactions in amphiphilic polymer co-networks in organic solvents. Biotechnol Bioeng 110:2333–2342PubMedCrossRefGoogle Scholar
  18. Tobis J, Thomann Y, Tiller JC (2010) Synthesis and characterization of chiral and thermo responsive amphiphilic co-networks. Polymer 51:35–45CrossRefGoogle Scholar
  19. Tobis J, Boch L, Thomann Y, Tiller JC (2011) Amphiphilic polymer co-networks as chiral separation membranes. J Membr Sci 372:219–227CrossRefGoogle Scholar
  20. Wang P, Sergeeva MV, Lim L, Dordick JS (1997) Biocatalytic plastics as active and stable materials for biotransformations. Nat Biotechnol 15(8):789–793PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  1. 1.Freiburg Material Research Center and Institute for Macromolecular ChemistryUniversity of FreiburgFreiburgGermany
  2. 2.Department of Bio- and Chemical EngineeringTU DortmundDortmundGermany

Personalised recommendations