Split intein can efficiently ligate peptide segments via a splicing reaction in a traceless manner and therefore has attracted great attention. Based on the structural characteristics and splicing reaction process of split intein, this paper comprehensively reviewed the recent progresses on the performance optimization and expanded applications of split intein, and revealed its great potential in the field of chemical protein synthesis as an increasingly sophisticated protein engineering technology. Finally, the challenges presented in the split intein- mediated protein trans-splicing and potential solutions in the future research were briefly discussed.