Abstract

A mild and green immobilization strategy was proposed based on catechol compounds and elastin-like peptides-SpyCatcher (EC), where ECs could specifically bind to the target enzyme chimera (xylanase-SpyTag-lichenase, XL) and offer suitable arm length between the chimera and carriers. The EC-functionalized carriers (Poly(vinylidene fluoride), PVDF) exhibited excellent carrier stability. After incubation at 4 ℃ for 12 days, the retention rate of EC was as high as 92.3%. When applied for covalently immobilizing XL from the cell crude lysates, it achieved the enzyme load of 34.1μg/cm2. The immobilized XL showed enhanced affinity towards the substrates and excellent thermostability, where the half-lives of xylanase and lichenase in XL at 50 ℃ were significantly improved up to 632.8% and 1266.1%, respectively. The versatility of this strategy was also proved by the carriers of nylon and gauze, which also showed comparable kinetic parameters with those immobilized on PVDF. The encouraging results pave a sustainable and eco-friendly way for purification and immobilization of target enzymes in a single step, which offers a potentially renewable avenue for biocatalysis and other related biotechnological processes.