An integrated purification and immobilization strategy for ELP-fusion β-glucosidase by its thermosensitivity and hydrophobicity

Abstract

In the field of biocatalysis, enzymes play a crucial role. However, they are faced with many challenges in practical applications, such as poor operational stability, high production costs and difficulties in recycling. Therefore, the efficient separation, purification and immobilization of enzymes are key to realize their industrial application. In this experiment, Elastin-like polypeptides (ELPs) tags with different sequence lengths, (VPGVG)n = 30, 40, 50, were designed to be fused with β-glucosidase (Glu) for expression. The temperature-responsive properties of ELPs have been utilized to achieve efficient separation and purification of recombinant enzymes, significantly improving the purification efficiency. Furthermore, the immobilization of Glu was achieved by enhancing the adsorption force between the recombinant enzyme and the carrier material based on the hydrophobicity of the ELPs. Compared to the free Glu, the immobilized Glu exhibited excellent thermal stability, pH stability, reusability and storage stability. In the catalytic hydrolysis of carboxymethyl cellulose (CMC), the glycosylation rate was increased by 27.26–28.05 % due to the synergistic action of immobilized Glu and cellulase. Therefore, this study developed an integrated method combining separation, purification with immobilization based on recombinant enzymes with ELPs, aiming to enhance the industrial application of immobilized enzymes.