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Journal of Biological Physics

, Volume 43, Issue 3, pp 445–459 | Cite as

Optimization of monomethoxy polyethyleneglycol-modified oxalate decarboxylase by response surface methodology

  • Han Long
  • XingHua Cai
  • Hui Yang
  • JunBin He
  • Jia Wu
  • RiHui LinEmail author
Original Paper

Abstract

In order to improve the stability of oxalate decarboxylase (Oxdc), response surface methodology (RSM), based on a four-factor three-level Box-Behnken central composite design was used to optimize the reaction conditions of oxalate decarboxylase (Oxdc) modified with monomethoxy polyethyleneglycol (mPEG5000). Four independent variables such as the ratio of mPEG-aldehyde to Oxdc, reaction time, temperature, and reaction pH were investigated in this work. The structure of modified Oxdc was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier transform infrared (FTIR) spectroscopy, the stability of the modified Oxdc was also investigated. The optimal conditions were as follows: the mole ratio of mPEG-aldehyde to Oxdc of 1:47.6, time of 13.1 h, temperature at 29.9 °C, and the reaction pH of 5.3. Under optimal conditions, experimental modified rate (MR = 73.69%) and recovery rate (RR = 67.58%) were matched well with the predicted value (MR = 75.11%) and (RR = 69.17%). SDS-PAGE and FTIR analysis showed that mPEG was covalently bound to the Oxdc. Compared with native Oxdc, the modified Oxdc (mPEG-Oxdc) showed higher thermal stability and better tolerance to trypsin or different pH treatment. This work will provide a further theoretical reference for enzyme modification and conditional optimization.

Keywords

Oxalate decarboxylase Monomethoxy polyethyleneglycol Response surface method (RSM) 

Notes

Acknowledgements

This research was supported by Guangxi Higher Education Institutes Talent Highland Innovation Team Scheme (GJR201147-12), Guangxi Natural Science Foundation Project (2014GXNSFAA118045). The authors also would like to thank the teacher ChengYu Wei for the help of information collection and equipment provided.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

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Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  • Han Long
    • 1
  • XingHua Cai
    • 1
  • Hui Yang
    • 2
  • JunBin He
    • 1
  • Jia Wu
    • 1
  • RiHui Lin
    • 1
    • 2
    • 3
    Email author
  1. 1.Guangxi Colleges and Universities Key Laboratory of Utilization of Microbial and Botanical Resources, School of Marine Sciences and BiotechnologyGuangxi University for NationalitiesNanningPeople’s Republic of China
  2. 2.Key Laboratory of New Techniques for Chemical and Biological Conversion Process, School of Chemistry and Chemical EngineeringGuangxi University for NationalitiesNanningPeople’s Republic of China
  3. 3.Guangxi Key Laboratory Cultivation Base for Polysaccharide Materials and their ModificationGuangxi University for NationalitiesNanningChina

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