Engineering actively magnetic crosslinked inclusion bodies of Candida antarctica lipase B: An efficient and stable biocatalyst for enzyme-catalyzed reactions

In our previous study, the soluble expression of recombinant Candida antarctica lipase B (rCalB) in E. coli is achieved through fusion with polyamino acid tails. The present study showed that even in the form of inclusion bodies (rCalB-IBs), rCalB-IBs were still active towards enzyme-catalyzed reactions. rCa1B-IBs could be directly crosslinked by the oxidized dextran in the presence of Fe3O4 nanoparticles to obtain magnetic aggregates (rCalB-mclIBs), and an activity recovery of 113.9 % +/- 2.2 % could be achieved under optimized conditions. The obtained rCalB-mclIBs exhibited excellent thermal and storage stabilities, and a good tolerance in changing pH environmental. rCalB-mclIBs could be effectively recycled and could retain 85.4 % +/- 2.5 % of its initial activity, and an e.e.(p) of 84.4 % +/- 2.2 % in (R, S)-NEMPA-ME hydrolysis was observed after the 10th cycle. After the application of rCalB-mclIBs into a promiscuous carbon-carbon bond-forming reaction, the MBH and the Aldol products could be obtained simultaneously. This study provides a simple and inexpensive strategy to highlight the application potential of rCalB-IBs as a biocatalyst.

Automated summary

This study demonstrates that recombinant Candida antarctica lipase B expressed in the form of inclusion bodies remains active and can be crosslinked with oxidized dextran to form magnetic aggregates with high activity recovery. These aggregates show excellent thermal stability, storage stability, and pH tolerance, while also exhibiting the ability to be recycled effectively and retain high activity levels. Additionally, they can be used in carbon-carbon bond-forming reactions to produce multiple products simultaneously, showcasing their potential as a versatile biocatalyst.