Oriented multivalent silaffin-affinity immobilization of recombinant lipase on diatom surface: Reliable loading and high performance of biocatalyst.
Academic Article
Overview
abstract
Microbial lipases are widely used biocatalysts; however, their functional surface immobilization should be designed for successful industrial applications. One of the unmet challenges is to develop a practical surface immobilization to achieve both high stability and activity of lipases upon the large loading. Herein, we present a silaffin-based multivalent design as a simple and oriented approach for Bacillus subtilis lipase A (LipA) immobilization on economic diatom biosilica matrix to yield highly-stable activity with reliable loading. Specifically, silaffin peptides Sil3H, Sil3K, and Sil3R, as monovalent or divalent genetic fusion tags, selectively immobilized LipA on biosilica surfaces. Sil3K peptide fusion to LipA termini most efficiently produced high catalytic activity upon immobilization. The activity was 70-fold greater than that of immobilized wild-type LipA. Compared to single fusion, the double Sil3K fusion displayed 1.7 higher enzymatic loading combined with high catalytic performances of LipA on biosilica surfaces. The multivalent immobilized LipA was distributed uniformly on biosilica surfaces. The biocatalyst was stable over a wide pH range with 98% retention activity after 10 reuses. The stabilized lipase fusion was compatible with laundry detergents, making it an attractive biocatalyst for detergent formulations. These findings demonstrate that multivalent surface immobilization is a plausible method for developing high-performance biocatalysts suitable for industrial biotechnological applications.
