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

Abstract

Abstract 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. rCalB-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.