Multi-layered And Site-specific Protein Immobilization On Cellulose Surface

In early stage,immobilized enzyme technology is mainly focused on single enzyme immobilization,and in recent years more and more researchers began to study the technology of multi enzyme co-immobilization.However,no matter the immobilization of single enzyme or multi enzyme,there is a small amount of immobilized enzyme and the activity center of the immobilized enzyme was hidden and so on.Therefore,The purpose of this research is to find a new and efficient immobilization technology to solve the above problems,and to provide theoretical basis and technical support for the immobilization of industrial enzyme preparation.In this paper,the cellulose was selected as the immobilized carrier and the green fluorescent protein as the immobilized target protein.With the aid of gene recombination technology,a complete set of self loading and multi-layer orientation immobilization system on protein was constructed.And the scaffold protein was introduced into the system as a connecting bridge between the immobilized carrier and the immobilized target protein.This paper has two innovative points,the first point is that each scaffold protein carrier surface can not only be fixed at least two times of the target protein but also can adjust the length of the scaffold protein to indirect regulation immobilized target protein quantity;the second poin is that the scaffold protein and the immobilized target protein can be specifically targeted,thus avoiding the destruction of the active functional structure of the immobilized target protein.The scaffold protein first through its cellulose binding domain connect to the carrer,then every bit on a scaffold protein fix green fluorescent protein(GFP)with anchoring domain and binding domain of affinity adsorption,and finally to achieve the carrer and the protein scaffold as well as the GFP directed self-assembly.The experimental results showed that the maximum GFP loading capacity reached 0.508 umol/g cellulose;in addition,the number of GFP was 3 times of the single layer fixed method,which shows that the efficiency of the fixed target protein was increased by 300%.