In this project, our aim was to utilize the powerful synthetic ability of yeast cells to efficiently secrete the lipase CALB. In the yeast cell factory, the transcriptional intensity of the promoter and the secretion ability of the signal peptide are crucial for the production and secretion of the target protein. Through literature review, we identified a series of promising promoters (AOX1, AOX713, FDH1, and FLD1) and signal peptides (α-factor SP, 0030-α-factor SP, SWP1-α-factor SP, KRE1-α-factor SP). We conducted combination tests on these parts and created a standardized complete combination, forming an efficient lipase (CALB) secretion system and successfully verifying its high secretion capability.

This collection is not merely a simple listing of independent components, but represents a modular toolkit designed through systems engineering. All these parts are in accordance with assembly compatibility and form a logically rigorous system: the promoters regulate the transcriptional intensity, the signal peptides guide the secretion efficiency, and the lipase genes provide the functional core.

We conducted complete construction and functional tests on all these parts and made detailed records (for details, please click on the part number 01f3fd18-99aa-47b8-b43f-f57e74945b15 ). This collection provides a complete secretion toolkit for the synthetic biology community that can be used for efficient secretion of exogenous proteins in Pichia pastoris. Other teams can select promoter-signal peptide combinations according to our functional test sections to meet different protein secretion requirements as needed.

Table 1 Part collection of in this project.