Abstract:

The synthetic Tbfmoc-Met-OH is used to replace Fmoc-Met-OH as a building block for solid phase peptide synthesis of ubiquitin. Based on the significant differences in gel filtration and UV absorbance between the Tbfmocubiquitin and the impurities, Tbfmocubiquitin can be separated from the impurities using gel filtration easily and efficiently. After removing the Tbfmoc group under mild condition (pH=8.5), the ubiquitin product is obtained with high purity, which is confirmed by amino acid analysis, MS and HPLC measurements. The strategy used in the paper can overcome the Tbfmoc-Cl’s weak points, including long reaction time, low coupling efficiency, more by-product producing and product purification difficulty, and the established new method can be used for the synthesis, separation and purification of ubiquitin easily and efficiently.

Key words: Tbfmoc-Met-OH; ubiquitin; solid phase peptide synthesis