Study On Modification Of 5-hydroxyme-Thylfurfural Oxidase And Catalase By Heterodimerizing Leucine Zipper

5-Hydroxymethylfurfural oxidase can convert 5-hydroxymethylfurfural to 2,5-furandicarboxylic acid by multi-step catalytic oxidation,which has great potential for application.The catalytic process of 5-hydroxymethylfurfural oxidase requires the cofactor flavin adenine dinucleotide,which catalyzes the production of two intermediates and a harmful by-product hydrogen peroxide.By introducing a catalase capable of eliminating by-products to construct a double enzyme catalytic system,the overall catalytic efficiency and enzyme stability can be effectively improved.This study constructed a pET28a-e-HMFO-ELP6 and pET28a-ELP6-CAT-r expression vector modified with a heterodimeric affinity peptide and a hexaploid elastin-like peptide.The self-assembly polymerization of free double enzyme e-HMFO-ELP6&ELP6-CAT-r was carried out in a solution environment by affinity peptide-mediated self-assembly.The catalytic efficiency of the catalytic oxidation of HMF showed that the substrate conversion rate of the two enzyme system e-HMFO-ELP6&ELP6-CAT-r exhibited a higher catalysis efficiency.In summary,the self-assembled double enzyme system constructed by the subject is a two-enzyme catalytic system in which HMFO is mainly responsible for catalytic oxidation of HMF and CAT as auxiliary elimination of by-product inhibition.The self-assembly property makes the two enzyme system have an improvement in catalytic activity and stability.