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Applied Biochemistry and Biotechnology

, Volume 174, Issue 6, pp 2067–2078 | Cite as

Preparation of Cross-Linked Enzyme Aggregates of Trehalose Synthase via Co-aggregation with Polyethyleneimine

  • Jianfeng Zheng
  • Ying Chen
  • Liwei Yang
  • Mingchun Li
  • Jun ZhangEmail author
Article

Abstract

Trehalose synthase (TreS) from Meiothermus ruber was co-aggregated with polyethyleneimine (PEI) and precipitated with polyethylene glycol (PEG), followed by cross-linking with glutaraldehyde to obtain TreS-polyethyleneimine cross-linked enzyme aggregates (termed as CLEAs-PEI-PEG). The TreS solution at 0.5 mg mL−1 protein concentration, with PEI at a mass ratio of 1:0.8 (enzyme/PEI, w/w) and 25 % (w/v) PEG concentration were found to be most adequate for the co-aggregation of TreS. CLEAs-PEI-PEG was most active with glutaraldehyde at a mass ratio of 1:0.5 (enzyme/glutaraldehyde, w/w) to cross-link the co-aggregates. The CLEAs-PEI-PEG prepared in this work had an optimum pH of 6.5 and optimum temperature of 60 °C. For lower concentrations of enzyme, using PEI could enhance the cross-linking efficiency of TreS. The thermal stability and pH tolerance of CLEAs-PEI-PEG were significantly improved. Scanning electron microscopy revealed that the main structure of CLEAs-PEI-PEG showed scaffolding morphology which was constituted by structured ball-like particles with a size of 1–2.5 μm in diameter.

Keywords

Trehalose synthase Polyethyleneimine Co-aggregation Cross-linking 

Notes

Acknowledgments

The current work was supported by the National Natural Science Foundation of China (no. 21076162) and Tianjin Natural Science Foundation (no. 10JCYBJC05000, no. 10JCYBJC09600).

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Jianfeng Zheng
    • 1
    • 2
  • Ying Chen
    • 1
  • Liwei Yang
    • 1
  • Mingchun Li
    • 2
  • Jun Zhang
    • 1
    Email author
  1. 1.Tianjin Institute of Forestry and PomologyTianjin Academy of Agricultural ScienceTianjinChina
  2. 2.College of Life ScienceNankai UniversityTianjinChina

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