Efficient recovery of beta-mannanase fused with LysM3014 domain in Escherichia coli by glass bead based-cell disruption method

Abstract

Toward the development of whole cell biocatalysts via non-GMO strategy, in this study, three E. coli cell disruptions, including sonication, freezing - thawing, and bead-shaking have been evaluated to recover efficiently fusion protein LysM3014-ManB containing a beta-mannanase (ManB) from Bacillus licheniformis fused with a single LysM3014 domain derived from a putative extracellular transglycosylase Lp_3014 of Lactobacillus plantarum WCFS1. The results showed that the highest volumetric activity of beta-mannanase (~46,651 U/l fermentation) was obtained when shaking the cells with glass beads. Moreover, the hybrid enzymatic activity of LysM3014-ManB increased approximately 1.4 folds (~66,000 U/l fermentation) under the optimum conditions of glass bead-based cell disruption with falcon 50 ml, glass bead ratio of 1.0 g/ml and disruption time of 8 min. The study indicates that shaking the cells with glass beads is likely to be the simplest and the most efficient cell disruption method in comparison with other disruption methods. Hybrid enzymes can be used to further study the immobilisation of ManB on the surface of L.
plantarum through an anchor containing a LysM domain.