Facile immobilization of Bacillus licheniformis γ-glutamyltranspeptidase onto graphene oxide nanosheets and its application to the biocatalytic synthesis of γ-l-glutamyl peptides

Abstract

For the practical application of Bacillus licheniformis γ-glutamyltranspeptidase (BlGGT), we illustrated a simple and efficient approach to fabricate a biocatalytic system by immobilizing the enzyme onto graphene oxide (GO) nanosheets via both non-covalent (GO-BlGGT) and covalent (GO/GA-BlGGT) bonds. The enzyme-loading capacity for the prepared GO/GA nanomaterial was 3.47 mg/mg support, corresponding to 68.7% recovery of the initial activity. Native and enzyme-bound layered GOs were characterized by X-ray diffraction, followed by Raman and Fouier transform infrared spectroscopy, elemental analysis and thermogram analysis. As compared to the free form of BlGGT, the immobilized enzymes exhibited significantly higher activity, possibly due to the beneficial effect of the layered GO carrier. The kinetic behaviors of GO-BlGGT and GO/GA-BlGGT were mostly consistent with those of free enzyme. The covalently immobilized enzyme had a comparable stability respective to free enzyme during a storage period of 30 days and could be recycled nine times with 45.3% retention of the initial activity. Besides, the biocatalytic synthesis of γ-l-glutamyl-phenylalanine and γ-l-glutamyl-leucine by immobilized enzymes resulted in the product yield of more than 31%. Taken together, these results suggest that the facile strategy is an economical means of depositing bioactive enzymes upon GO nanosheets for BlGGT-mediated biocatalysis.

Keywords: Bacillus licheniformis; Graphene oxide; Immobilization; γ-Glutamyltranspeptidase; γ-l-Glutamyl peptide.