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

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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.

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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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Authors and Affiliations

  1. Tianjin Institute of Forestry and Pomology, Tianjin Academy of Agricultural Science, Tianjin, 300384, China

    Jianfeng Zheng, Ying Chen, Liwei Yang & Jun Zhang

  2. College of Life Science, Nankai University, Tianjin, 300071, China

    Jianfeng Zheng & Mingchun Li

Authors
  1. Jianfeng Zheng
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  2. Ying Chen
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  3. Liwei Yang
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  4. Mingchun Li
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  5. Jun Zhang
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Correspondence to Jun Zhang.

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Zheng, J., Chen, Y., Yang, L. et al. Preparation of Cross-Linked Enzyme Aggregates of Trehalose Synthase via Co-aggregation with Polyethyleneimine. Appl Biochem Biotechnol 174, 2067–2078 (2014). https://doi.org/10.1007/s12010-014-1104-y

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  • DOI: https://doi.org/10.1007/s12010-014-1104-y

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