The Pichia pastoris strain P52-5 was inoculated at an initial $OD_{600}$ of 1 into 250-mL Erlenmeyer flasks containing 50 mL of YNG and YNM media, respectively. After cultivation at 30 °C and 220 rpm for 96 h, cultures were harvested and centrifuged at 5,000 × g for 5 min. The resulting supernatants were collected and submitted to Shanghai Luming Biotechnology Co., Ltd. (Shanghai, China) for quality control (QC) and Pro DIA quantitative proteomic identification. The total protein expression levels were used to normalize the protein abundance across the three biological triplicates