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An efficient immobilizing technique of penicillin acylase with combining mesocellular silica foams support and p-benzoquinone cross linker

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Abstract

To improve the performance of covalently immobilized penicillin acylase (PA), the immobilization was carried out in mesocellular silica foams (MCFs) using p-benzoquinone as cross linker. The characterizations of the immobilized enzyme were studied carefully. The results showed that the relative activity of the immobilized PA was increased to 145% of that of free enzyme. The activity was 3.7 folds of that of PA on the silica nanoparticles. The enzyme in MCFs presented a turnover equal to that of free enzyme. It was also found that the optimum pH of the immobilized PA shifted to pH 7.5 and the optimum reaction temperature rose from 45 to 50 °C. Furthermore, the stability of PA was ameliorated greatly after immobilization. Fourier transform infrared spectroscopy showed no major secondary structural change for PA confined in MCFs. The proposed covalent immobilizing technique would rank among the potential strategies for efficient immobilization of PA.

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References

  1. Kallenberg AI, Rantwijk FV, Sheldon RA (2005) Immobilization of Penicillin G Acylase: The Key to Optimum Performance. Adv Synth Catal 347:905–926

    Article  CAS  Google Scholar 

  2. Sio CF, Quax WJ (2004) Improved β-lactam acylases and their use as industrial biocatalysts. Curr Opin Biotechnol 15:349–355

    Article  CAS  Google Scholar 

  3. Gómez L, Ramírez HL, Neira-Carrillo A, Villalonga R (2006) Polyelectrolyte complex formation mediated immobilization of chitosan-invertase neoglycoconjugate on pectin-coated chitin. Bioprocess Biosyst Eng 28:387–395

    Article  Google Scholar 

  4. Benkhelifa H, Bengoa C, Larre C, Guibal E, Popineau Y, Legrand J (2005) Casein hydrolysis by immobilized enzymes in a torus reactor. Process Biochem 40:461–467

    Article  CAS  Google Scholar 

  5. Tümtürk H, Sahin F, Demirel G (2007) A new method for immobilization of acetylcholinesterase. Bioprocess Biosyst Eng 30:141–145

    Article  Google Scholar 

  6. Katchalski-Katzir E, Kraemer DM (2000) Eupergit C, a carrier for immobilization of enzymes of industrial potential. J Mol Catal B: Enzym 10:157–176

    Article  CAS  Google Scholar 

  7. Wang Q, Gao Q, Shi J (2004) Enhanced catalytic activity of hemoglobin in organic solvents by layered titanate immobilization. J Am Chem Soc 126:14346–14347

    Article  CAS  Google Scholar 

  8. Maria Chong AS, Zhao XS (2004) Design of large-pore mesoporous materials for immobilization of penicillin G acylase biocatalyst. Catal Today 93–95:293–299

    Article  Google Scholar 

  9. Basso A, De Martin L, Ebert C et al (2003) Organically modified xerogels as novel tailor-made supports for covalent immobilization of enzymes (penicillin G acylase). Tetrahedron Lett 44:5889–5891

    Article  CAS  Google Scholar 

  10. Sheldon RA (2007) Enzyme immobilization: the quest for optimum performance. Adv Synth Catal 349:1289–1307

    Article  CAS  Google Scholar 

  11. Zhou HX, Dill KA (2001) Stabilization of proteins in confined spaces. Biochemistry 40:11289–11293

    Article  CAS  Google Scholar 

  12. Minton AP (2001) The influence of macromolecular crowding and macromolecular confinement on biochemical reactions in physiological media. J Biol Chem 276:10577–10580

    Article  CAS  Google Scholar 

  13. Lei C, Shin Y, Magnuson JK, Fryxell G (2006) Characterization of functionalized nanoporous supports for protein confinement. Nanotechnology 17:5531–5538

    Article  CAS  Google Scholar 

  14. Zong J, Chen YW, Shen SB (2006) Complex biocatalyst assembled by enzyme and inorganic/ metal nanoparticles. J Chem Ind Eng (China) 57:1777–1781

    Google Scholar 

  15. Xue P, Lu GZ, Liu WY (2006) Poly (GMA/MA/MBAA) copolymer beads: a highly efficient support immobilizing penicillin G acylase. Chin Chem Lett 17:129–132

    CAS  Google Scholar 

  16. Cheung MS, Thirumalai D (2006) Nanopore–protein interactions dramatically alter stability and yield of the native state in restricted spaces. J Mol Biol 357:632–643

    Article  CAS  Google Scholar 

  17. Cheung MS, Klimov D, Thirumalai D (2005) Molecular crowding enhances native state stability and refolding rates of globular proteins. Proc Natl Acad Sci USA 102:4753–4758

    Article  CAS  Google Scholar 

  18. Schmidt-Winkel P, Lukens WW, Zhao D et al (1999) Mesocellular siliceous foams with uniformly sized cells and windows. J Am Chem Soc 121:254–255

    Article  CAS  Google Scholar 

  19. Tian B, Liu X, Yu C et al (2002) Microwave assisted template removal of siliceous porous materials. Chem Commun (Camb) 7:1186–1187

    Article  Google Scholar 

  20. Stöber W, Fink A, Bohn E (1968) Controlled growth of monodisperse silica spheres in the micron size range. J Colloid Interface Sci 26:62–69

    Article  Google Scholar 

  21. Bradford MMA (1976) Rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–250

    Article  CAS  Google Scholar 

  22. Shewale JG, Kumar KK, Ambekar GR (1987) Evaluation of determination of 6-aminopenicillanic acid by p-dimethylaminobenzaldehyde. Biotechnol Tech 1:69–72

    Article  CAS  Google Scholar 

  23. Van Langen LM, Janssen MHA, Oosthoek NHP et al (2002) Active site titration as a tool for the evaluation of immobilization procedures of penicillin acylase. Biotech Bioeng 79:223–227

    Article  Google Scholar 

  24. Pchelintsev NA, Youshko MI, Ŝvedas VK (2006) A new method for spectrophotometric assay of activity of cross-linked penicillin acylase aggregates. Biochemistry (Mosc) 71:315–319

    Article  CAS  Google Scholar 

  25. Griebenow K, Klibanov AM (1995) Lyophilization-Induced Reversible Changes in the Secondary Structure of Proteins. Proc Natl Acad Sci USA 92:10969–10976

    Article  CAS  Google Scholar 

  26. Van Roon JL, Schroën CGPH Tramper J, Beeftink HH (2007) Biocatalysts: measurement, modelling and design of heterogeneity. Biotechnol Adv 25:137–147

    Article  Google Scholar 

  27. Atkins PW, De Paula J (2002) Atkins’ physical chemistry. 7th edn. Oxford University Press, NY

    Google Scholar 

  28. Chen B, Miller EM, Gross RA et al (2007) Effects of macroporous resin size on Candida antarctica lipase B adsorption, fraction of active molecules, and catalytic activity for polyester synthesis. Langmuir 23:1381–1387

    Article  CAS  Google Scholar 

  29. Norouzian D, Javadpour S, Moazami N, Akbarzadeha A et al (2002) Immobilization of whole cell penicillin G acylase in open pore gelatin matrix. Enzyme Microb Technol 30:26–29

    Article  CAS  Google Scholar 

  30. Kazan D, Ertan H, Erarslan A (1997) Stabilization of Escherichia coli penicillin G acylase against thermal inactivation by cross-linking with dextran dialdehyde polymers. Appl Microbiol Biotechnol 48:191–197

    Article  CAS  Google Scholar 

  31. Duggleby HJ, Tolley SP, Hill CP, Dodson EJ, Dodson G, Moody PCE (1995) Penicillin acylase has a single amino-acid catalytic centre. Nature 373:264–265

    Article  CAS  Google Scholar 

  32. Dong AC, Huang P, Caughey WS (1990) Proteins secondary structures in water from second-derivative amide-I infrared spectra. Biochemistry 29:3303–3308

    Article  CAS  Google Scholar 

  33. Asuri P, Bale SS, Karajanagi SS et al (2006) The protein–nanomaterial interface. Curr Opin Biotechnol 17:562–568

    Article  CAS  Google Scholar 

  34. Mcnay JL, Fernandez EJ (1999) How does a protein unfold on a reversed phase liquid chromatography surface. J Chromatogr A 849:135–148

    Article  CAS  Google Scholar 

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Acknowledgments

This work was financially supported by the National High Technology Research and Development Program of China (863 Program, No.2006AA02Z211), National Natural Science Foundation of China (20376034), Natural Science Foundation of Jiangsu Province of China (BK2006181) and Foundation of Jiangsu Province of China for College Postgraduate Students in Innovation Engineering (2007). The research was performed at the National Engineering Research Center for Biotechnology sponsored by the Minister of Science and Technology of PR China.

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Authors and Affiliations

  1. National Engineering Research Center for Biotechnology, College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology, No.5 Xinmofan Road, Nanjing, 210009, People’s Republic of China

    Anming Wang, Hua Wang, Cheng Zhou, Zhiqiang Du & Shubao Shen

  2. College of Materials Science and Engineering, Nanjing University of Technology, Nanjing, 210009, People’s Republic of China

    Shemin Zhu

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  1. Anming Wang
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  2. Hua Wang
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  3. Shemin Zhu
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  4. Cheng Zhou
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Correspondence to Shubao Shen.

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Wang, A., Wang, H., Zhu, S. et al. An efficient immobilizing technique of penicillin acylase with combining mesocellular silica foams support and p-benzoquinone cross linker. Bioprocess Biosyst Eng 31, 509–517 (2008). https://doi.org/10.1007/s00449-007-0189-x

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