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Enhanced biochemical characteristics of β-glucosidase via adsorption and cross-linked enzyme aggregate for rapid cellobiose hydrolysis

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Abstract

With proper design, immobilization can be useful tool to improve the stability of enzymes, and in certain cases even their activity, selectivity, productivity and economic viability. An immobilized β-glucosidase (BGL, EC 3.2.1.21) through matrix adsorption and cross-linked enzyme aggregate (ad-CLEA) technology is presented in this work. After adsorption and precipitation, BGL was immobilized to poly(glycidyl methacrylate-co-ethylenedimethacrylate) (PGMA/EDMA) microparticles using glutaraldehyde as the cross-linker. Immobilized BGL exhibits lower apparent Km but much higher Vmax than that of the soluble enzyme, suggesting greater enzyme–substrate affinity and rapid velocity. Besides, ad-CLEA-BGL presents better thermostability retaining activity nearly 70% for 3 h and approximately 50% for 5 h at 70 °C, high operational reusability remaining more than 90% activity after nine uses and excellent storage stability holding about 95% activity after 45 days. Furthermore, the cellobiose is completely hydrolyzed within 1 h with ad-CLEA-BGL, which is significantly more efficient than soluble enzyme (about 4 h). Therefore, BGL was successfully immobilized on PGMA/EDMA microparticles with an ad-CLEA technology and the immobilization greatly enhances the biochemical characteristics. This work indicates promising application for ad-CLEA-BGL in utilizing agricultural remnants, bio-converting cellobiose to fermentable reducing sugar and ethanol production.

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Acknowledgements

Xu Deng and Tian He contributed equally to this work and should be considered co-first authors. This work was financially supported by National Natural Science Foundation of China (no. 21775100).

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Authors and Affiliations

  1. Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province and Key Laboratory of Medicinal Resources and Natural Pharmaceutical Chemistry of the Ministry of Education, Shaanxi Normal University, Xi’an, 710062, China

    Xu Deng, Tian He, Jun Li, Hui-Ling Duan & Zhi-Qi Zhang

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  1. Xu Deng
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  2. Tian He
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Correspondence to Zhi-Qi Zhang.

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Xu Deng declares that she has no conflict of interest. Tian He declares that he has no conflict of interest. Jun Li declares that she has no conflict of interest. Hui-Ling Duan declares that she has no conflict of interest. Zhi-Qi Zhang declares that he has no conflict of interest.

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Deng, X., He, T., Li, J. et al. Enhanced biochemical characteristics of β-glucosidase via adsorption and cross-linked enzyme aggregate for rapid cellobiose hydrolysis. Bioprocess Biosyst Eng 43, 2209–2217 (2020). https://doi.org/10.1007/s00449-020-02406-5

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