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The invention discloses a method for enhancing the synthetic ability of yeast total protein and dipeptide tripeptide. Relates to the technical field of synthetic biology and metabolic engineering. According to the invention, the saccharomyces cerevisiae engineering strain with high protein synthesis capability and high endogenous dipeptide and tripeptide accumulation level is successfully constructed. The metabonomics analysis reveals the unique peptide fragment accumulation spectrum, especially the generation of a large amount of lysine-rich tripeptide at the C terminal, and proposes a potential mechanism of metabolic pathway'bottleneck 'caused by peptidase activity down-regulation in combination with proteome data. The recombinant strain has important application potential in the fields of single-cell protein, functional peptide products and the like.