Boosting sugarcane trash decomposition: synergistic action and proximity effect of xylanase and feruloyl esterase co-displayed on the cell surface of Pichia pastoris (Komagataella phaffi)
Abstract
Yeast surface display technology offers a promising avenue for enhancing lignocellulosic biomass degradation and bioconversion processes. In this study, we investigated the synergistic action of cell-surface-displayed xylanase (XYN) and feruloyl esterase (FAE) using Pichia pastoris (Komagataella phaffi) and also explored the impact of the proximity effect when both enzymes are co-displayed on the same cell surface. We engineered three P. pastoris strains: X-Pichia displaying XYN, F-Pichia displaying FAE, and X/F-Pichia co-displaying both enzymes. Immunofluorescence confirmed successful cell-surface display. In hydrolysis experiments using acid-pretreated sugarcane trash, a clear synergy emerged. The mixing of X-Pichia and F-Pichia (“X-Pichia + F-Pichia”) exhibited an approximately 1.2 times higher reducing sugar yield compared to X-Pichia alone. Moreover, X/F-Pichia surpassed this mixture due to the proximity effect, yielding roughly 1.5 times more reducing sugar than X-Pichia alone. Additionally, X/F-Pichia produced about 1.1 times more ferulic acid than F-Pichia alone or “X-Pichia + F-Pichia.” These results underscore the potential of co-displaying multiple enzymes on the P. pastoris cell surface to significantly enhance biomass degradation. The enzyme co-display system with synergy and proximity effects holds promise for efficient lignocellulosic biomass utilization and sustainable bioprocessing. This work contributes to the development of more efficient and sustainable bioconversion processes for lignocellulosic biomass.
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