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Protein-Coated Microcrystals from Candida rugosa Lipase: Its Immobilization, Characterization, and Application in Resolution of Racemic Ibuprofen

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Abstract

In this study, an economical heterogeneous biocatalyst, protein-coated microcrystals (PCMCs), was prepared from a commercial Candida rugosa lipase (CRL) and used for catalyzing esterification of (R, S)-ibuprofen enantiomers with isooctanol in isooctane. The main variables controlling the process (precipitating solvents, pH, saturated K2SO4 solution, and water content) were optimized via single-factorial experiments. Under optimum conditions, the enantiomeric excess of active S(+)-ibuprofen and total conversion rate were 97.34 and 49.83 %, respectively, and the corresponding enzyme (PCMC-CRL) activity attained 387.29 μmol/min/g protein, a 5.78-fold enhancement over the free lipase powder. Additionally, the thermostability, organic-solvent tolerance, and operational stability of PCMC-CRL were greatly improved as compared to the free enzyme. Fourier transform infrared (FTIR) spectroscopy was employed to reveal the correlation between conformation and enzyme activity enhancement. Moreover, the PCMC-CRL retained most of its original activity following use in more than 15 successive batches, suggesting that it exhibits adequate operational stability. These results indicate that PCMC-CRL is of great potential use in industrial applications.

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Acknowledgments

This work is financially supported by the National Natural Science Foundation of China (nos. 31170078, 31070089, and J1103514), the National High Technology Research and Development Program of China (nos. 2011AA02A204 and 2014AA093510), the Innovation Foundation of Shenzhen Government (no. JCYJ20120831111657864), the Innovation Foundation of HUST (nos. 2011TS100, 2014QN119, and 2014NY007), and the Fundamental Research Funds for the Central Universities HUST (no. 2172012SHYJ004). Many thanks to the Analytical and Testing Center of HUST for their valuable assistance with SEM and FT-IR measurements.

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  1. Key Laboratory of Molecular Biophysics, The Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, China

    Shuangshuang Huang, Xiang Li, Li Xu, Caixia Ke, Rui Zhang & Yunjun Yan

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  1. Shuangshuang Huang
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  2. Xiang Li
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Correspondence to Li Xu or Yunjun Yan.

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Huang, S., Li, X., Xu, L. et al. Protein-Coated Microcrystals from Candida rugosa Lipase: Its Immobilization, Characterization, and Application in Resolution of Racemic Ibuprofen. Appl Biochem Biotechnol 177, 36–47 (2015). https://doi.org/10.1007/s12010-015-1725-9

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