Yeast surface display technology,which anchors target proteins on the cell surface,has been widely applied in the fields of vaccine preparation,drug screening,environmental management,and strain selection. In pursuit of high-efficiency production of therapeutic proteins and performing protein engineering in K. lactis,the study designed and implemented a surface display system utilizing Sed1 from Saccharomyces cerevisiae as an anchor and His Tag as a detection target. The high-throughput characterization technology that combined indirect immunofluorescence labeling and whole-cell flow cytometry was employed to comprehensively evaluate the performance of the system. Laser scanning confocal microscopy imaging further confirmed the successful display of Human serum albumin as a model therapeutic protein. Following optimization of detection conditions,a remarkable maximum display efficiency of 89.8% was achieved. This study was the first time to establish and systematically optimize a yeast surface display system in K. lactis and laid the foundation for conducting protein engineering in K. lactis.
Kluyveromyces lactisyeast surface displayoptimizationHuman serum albumin