Articles & Issues

Language: English

Conflict of Interest: In relation to this article, we declare that there is no conflict of interest.

Publication history: Received November 23, 2008
Accepted January 25, 2009

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/bync/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

All issues

Improvement of the catalytic performance of immobilized penicillin acylase through assembly of macromolecular reagents in nanopore to create a crowding environment

Cheng Zhou^{1 2} Anming Wang^{1 2} Zhiqiang Du^{1 2} Shemin Zhu^{1 3} Shubao Shen^{1 2†}

¹National Engineering Research Center for Biotechnology, Nanjing University of Technology, Nanjing 210009, P. R. China ²College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, P. R. China ³College of Materials Science and Engineering, Nanjing University of Technology, Nanjing 210009, P. R. China

zsbshen@hotmail.com

Korean Journal of Chemical Engineering, July 2009, 26(4), 1065-1069(5), 10.1007/s11814-009-0177-8

Download PDF

Abstract

Macromolecular reagents were co-assembled with penicillin acylase (PA) and immobilized in mesocellular siliceous foams (MCFs) to resemble living cells. Types and concentrations of macromolecules were studied. The catalytic characteristic and stability of PA preparations were also investigated. PA assembled with dextran 10 k in MCFs showed maximum specific activity, 1.32-fold of that of the solely immobilized PA. The optimum pH of dextran and BSA derivatives_x000D_ shifted to neutrality, and the optimum temperature increased by 10 ℃. Also, Km of BSA derivative of PA declined 56.7% compared to solely immobilized PA, while the Kcat/Km of PA assembled with BSA was enhanced to 147%. After incubation at 50 ℃ for 6 h, residual activity of PA assembled with BSA exhibited 53.0%. The ficoll derivative showed 82.8% of its initial activity at 4 ℃ after 8-week storage. The results indicated that macromolecular reagents assembled with PA in MCFs could dramatically improve the catalytic performance and stability of im- mobilized enzyme.

Keywords

Penicillin Acylase Immobilization Covalent Mesocellular Siliceous Foams Macromolecular Crowding

References

Chandel AK, Rao LV, Narasu ML, Singh OV, Enzyme. Microb. Technol., 42, 199 (2008)
Sheldon RA, Adv. Synth. Catal., 349, 1289 (2007)
Kallenberg AI, Van Rantwijk F, Sheldon RA, Adv. Synth. Catal., 347, 905 (2005)
Wang HW, Kim IH, Park CS, Lee JH, Korean J. Chem. Eng., 25(4), 801 (2008)
Montes T, Grazu V, Manso I, Galan B, Lopez-Gallego F, Gonzalez R, Hermoso JA, Garcia JL, Guisan JM, Fernandez-Lafuente R, Adv. Synth. Catal., 349, 459 (2007)
Wang AM, Zhou C, Wang H, Shen SB, Xue JY, Ouyang PK, Chin. J. Chem. Eng., 15(6), 788 (2007)
Park K, Kim H, Maken S, Kim Y, Min B, Park J, Korean J. Chem. Eng., 22(3), 412 (2005)
Van Roon JL, SchroenCGPH, Tramper J, Beeftink HH, Biotechnol. Adv., 25, 137 (2007)
Minton AP, J. Cell. Sci., 119, 2863 (2006)
Zhou HX, Rivas G, Minton AP, Annu. Rev. Biophys., 37, 375 (2008)
Ellis RJ, Trends Biochem. Sci., 26, 597 (2001)
Kazan D, Erarslan A, J. Chem. Technol. Biotechnol., 74(12), 1157 (1999)
Mislovicova D, Masarova J, Bucko M, Gemeiner P, Enzyme Microb. Technol., 39(4), 579 (2006)
Schmidt-Winkel P, Lukens WW, Zhao DY, Yang PD, Chmelka BF, Stucky GD, J. Am. Chem. Soc., 121(1), 254 (1999)
Wang AM, Wang H, Zhu SM, Zhou C, Du ZQ, Shen SB, Bioprocess. Biosyst. Eng., 31, 509 (2008)
Wang AM, Liu MQ, Wang H, Zhou C, Du ZQ, Zhu SM, Shen SB, J. Biosci. Bioeng., 106, In press (2008)
Kim H, Jung JC, Yeom SH, Lee KY, Yi J, Song IK, Mater. Res. Bull., 42, 2132 (2007)
Brandford MMA, Anal. Biochem., 72, 248 (1976)
Shewale JG, Kumar KK, Ambekar GR, Biotechnol. Technique., 1, 69 (1987)
Cheung MS, Thirumalai D, J. Mol. Biol., 357, 632 (2006)
Zhou HX, Ken AD, Biochem., 40, 11289 (2001)
Minton AP, Curr. Opin. Struct. Biol., 10, 34 (2000)
Ellis RJ, Curr. Opin. Struct. Biol., 11, 114 (2001)
Koshland DE, Bioorg. Chem., 30, 211 (2002)
Jiang M, Guo ZH, J. Am. Chem. Soc., 129(4), 730 (2007)
Minton AP, J. Biol. Chem., 276, 10577 (2001)
Bernardino SMSA, Gallegos JFM, Santhagunam A, Barros DPC, Fernandes P, Fonseca LP, J. Biotechnol., 131, S97 (2007)