Room‐Temperature Synthesis of Trypsin‐Inorganic Hybrid Nanocomposites for Fast and Efficient Protein Digestion

Abstract

AbstractFast and high‐efficiency protein digestion is essential for high‐throughput proteome analysis. Herein, a new kind of trypsin‐inorganic hybrid nanocomposites were room‐temperature synthesized based on self‐assembly stragety, where trypsin and nickel phosphate (Ni3(PO4)2) was used as organic and inorganic ligands, respectively. The general applicability of the synthetic method was also confirmed by using a series of other metal phosphates including Mg2+, Ba2+, Zn2+, and Co2+. The as‐prepared trypsin‐inorganic hybrid nanocomposites with flower‐like morphology showed higher enzymatic acticity (206 % enhancement) and faster catalytic kinetics than the free trypsin. Taking the merits togethers as mentioned above, the hybrid nanocomposites could be employed as an immmobilized enzyme reactor (IMER) for fast and high‐efficiency protein digestion. The peptide mass fingerprint analysis of the model proteins demonstrated that the coverage rate of the sequences treated by IMER could bear comparsion with that of the free one, but the digestion time was shorten from 12 h to 1 min. In addition, the IMER showed a good applicability for complex human serum sample, showing a promising potential in proteomics research.