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Efficient production of (R)-(−)-mandelic acid using glutaraldehyde cross-linked Escherichia coli cells expressing Alcaligenes sp. nitrilase

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Abstract

Recombinant Escherichia coli cells expressing Alcaligenes sp. nitrilase were simply immobilized by direct cross-linking using glutaraldehyde. About 85 % of the total nitrilase activity was recovered under the optimal cross-linking conditions. The thermal stabilities of the cross-linked cells measured at 30, 40 and 50 °C were 4.5-, 5.3-, and 5.1-fold those of the free cells, respectively. The concentration of (R)-(−)-mandelic acid reached 280 mM after merely 2 h transformation with the immobilized cells using 300 mM mandelonitrile as substrate, affording an extremely high productivity of 510.7 g L−1 d−1. In addition, operational stability of the immobilized cells was obviously superior to that of free cells, without significant activity loss after 15 cycles of batch reactions or 8 cycles of repeated fed-batch reactions. Therefore, the easy preparation and robust characteristics of the immobilized biocatalyst make it a very promising biocatalyst for high-performance and low-cost production of optically pure (R)-(−)-mandelic acid.

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Acknowledgments

This work was financially supported by the National Science Foundation of China (Nos. 21276082 & 31200050), Ministry of Science and Technology, P. R. China (Nos. 2011CB710800, 2011AA02A210 & 2012AA022201), Shanghai Commission of Science and Technology (No. 11431921600) and the Fundamental Research Funds for the Central Universities (No. 222201314016).

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Authors and Affiliations

  1. Laboratory of Biocatalysis and Synthetic Biotechnology, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China

    Zhi-Jun Zhang, Jiang Pan, Chun-Xiu Li, Hui-Lei Yu, Gao-Wei Zheng, Xin Ju & Jian-He Xu

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  1. Zhi-Jun Zhang
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  2. Jiang Pan
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  3. Chun-Xiu Li
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Correspondence to Jian-He Xu.

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Zhang, ZJ., Pan, J., Li, CX. et al. Efficient production of (R)-(−)-mandelic acid using glutaraldehyde cross-linked Escherichia coli cells expressing Alcaligenes sp. nitrilase. Bioprocess Biosyst Eng 37, 1241–1248 (2014). https://doi.org/10.1007/s00449-013-1096-y

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