Saccharomyces cerevisiae is a promising host for heterologous protein production due to its safety, genetic tractability, and ability to perform complex post-translational modifications. Recent advances in synthetic biology and metabolic engineering have significantly improved the efficiency of protein production in yeast. This review summarizes the key strategies for enhancing heterologous protein production in S. cerevisiae, including hyperexpression systems, secretion engineering, glycosylation pathway engineering, and systems metabolic engineering. These strategies aim to increase protein yield, improve secretion efficiency, and optimize metabolic pathways for sustainable production. Codon optimization, increased gene copy number, and transcriptional regulation are among the most effective methods for enhancing protein expression. Promoter engineering, including the use of synthetic promoters and intron-containing promoters, has also been employed to improve gene expression levels. Terminator engineering is crucial for controlling mRNA stability and protein expression. Protein secretion engineering involves modifying secretion signals, ER folding, and vesicle trafficking to enhance secretion efficiency. Glycosylation pathway engineering is used to modify protein glycosylation patterns, which can improve protein activity and stability. Systems metabolic engineering aims to optimize metabolic pathways for efficient protein production, reducing metabolic burden and oxidative stress. The integration of artificial intelligence and machine learning has further enhanced the ability to design and optimize genetic elements and metabolic pathways. Overall, these strategies have significantly improved the efficiency and sustainability of heterologous protein production in S. cerevisiae, making it a promising platform for future biomanufacturing.Saccharomyces cerevisiae is a promising host for heterologous protein production due to its safety, genetic tractability, and ability to perform complex post-translational modifications. Recent advances in synthetic biology and metabolic engineering have significantly improved the efficiency of protein production in yeast. This review summarizes the key strategies for enhancing heterologous protein production in S. cerevisiae, including hyperexpression systems, secretion engineering, glycosylation pathway engineering, and systems metabolic engineering. These strategies aim to increase protein yield, improve secretion efficiency, and optimize metabolic pathways for sustainable production. Codon optimization, increased gene copy number, and transcriptional regulation are among the most effective methods for enhancing protein expression. Promoter engineering, including the use of synthetic promoters and intron-containing promoters, has also been employed to improve gene expression levels. Terminator engineering is crucial for controlling mRNA stability and protein expression. Protein secretion engineering involves modifying secretion signals, ER folding, and vesicle trafficking to enhance secretion efficiency. Glycosylation pathway engineering is used to modify protein glycosylation patterns, which can improve protein activity and stability. Systems metabolic engineering aims to optimize metabolic pathways for efficient protein production, reducing metabolic burden and oxidative stress. The integration of artificial intelligence and machine learning has further enhanced the ability to design and optimize genetic elements and metabolic pathways. Overall, these strategies have significantly improved the efficiency and sustainability of heterologous protein production in S. cerevisiae, making it a promising platform for future biomanufacturing.