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Amino acid-mediated aldolase immobilisation for enhanced catalysis and thermostability

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Abstract

To improve the properties of the immobilised 2-deoxy-d-ribose-5-phosphate aldolase (DERA), unreacted functional groups on support surface were blocked with amino acids. The relative activities of the immobilised enzyme were 144.7 and 141.9% when the post-immobilisation modification was done with Arg and Phe, respectively. The residual activity of immobilised DERA after heating at 60 °C for 120 min was 65.1% when Phe and Val were used as the blocking amino acids, a 2.0- and 2.87-fold increase over that of the immobilised (no post-immobilisation blocking) and free DERA. Immobilised DERA maintained maximal activity in 2-deoxyribose-5-phosphate (DR5P) synthesis up to 600 mM of acetaldehyde, which was much higher than the amount of acetaldehyde tolerated by free enzyme (300 mM). This superior resistance to high acetaldehyde concentrations would accelerate the DR5P reaction by shifting the reaction equilibrium towards the product. The results from this study suggest that the novel immobilised DERA may be useful for industrial applications.

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Acknowledgments

This work was supported financially by the National Natural Science Foundation of China (20906016, 21076053), the Technology Research and Development Program of Hangzhou (20090331N03), the Technology Research and Development Program of Zhejiang Province (2009C31086), the Special Foundation for New Researchers in Hangzhou Normal University (YS05203141) and the Foundation for Innovation of Science and Technology and Cultural Originality in Hangzhou Normal University (YS03201012).

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Authors and Affiliations

  1. Research Center for Biomedicine and Health, Hangzhou Normal University, No. 1378, West Wenyi Road, Hangzhou, 311112, People’s Republic of China

    Anming Wang, Weifang Gao, Fangkai Zhang, Feifei Chen, Fangchuan Du & Xiaopu Yin

  2. College of Biological and Environmental Sciences, Hangzhou Normal University, Hangzhou, 310036, People’s Republic of China

    Fangkai Zhang & Fangchuan Du

  3. College of Materials, Chemistry and Chemical Engineering, Hangzhou Normal University, No. 222, Wenyi Road, Hangzhou, 310036, People’s Republic of China

    Feifei Chen

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  1. Anming Wang
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  2. Weifang Gao
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  3. Fangkai Zhang
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Correspondence to Anming Wang.

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Wang, A., Gao, W., Zhang, F. et al. Amino acid-mediated aldolase immobilisation for enhanced catalysis and thermostability. Bioprocess Biosyst Eng 35, 857–863 (2012). https://doi.org/10.1007/s00449-011-0670-4

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  • DOI: https://doi.org/10.1007/s00449-011-0670-4

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