One-step separation and immobilization of his-tagged enzyme directly from cell lysis solution by biomimetic mineralization approach

We report a facile approach to fabricate biocatalyst, which is the BglucLH-Cu-3(PO4)(2) nanoflowers (BglucH-NF) possessed microscale flower-like structure assembled with nanoscale petals, by integrating the separation and immobilization of his-tagged enzyme directly from cell lysis solution in a single biomineralization step without the preparation of support. The recombinant beta-glucosidase-linker-His (BglucLH) being rich in imidazole group was constructed to enhance the specific binding of enzyme with copper ions. We have demonstrated that the activity recovery and separation efficiency of BglucLH-NF were improved by 253.2 % and 290.1 % in comparison with Bgluc-Cu-3(PO4)(2) nanoflowers (Bgluc-NF), indicating the positive reinforcement of his-tag for improving separation and immobilization efficiency. The BglucLH-NF could maintain 73.2 +/- 2.77 % of the initial activity after repeating 15 cycles, exhibiting excellent reusability. The integrated separation and immobilization of histagged enzyme directly from cell lysis solution by biomimetic mineralization approach displays great potential in the convenient fabrication of biocatalyst with high enzyme loading, stability, and recyclability.

Automated summary

This study presents a simple method to separate and immobilize histagged enzymes directly from cell lysis solution through a single biomineralization step, without the need for support preparation. The results demonstrate that this approach significantly enhances enzyme activity recovery and separation efficiency, while also exhibiting excellent reusability.