N-acetyltransferase co-expression increases α-glucosidase expression level in Pichia pastoris.

Pichia pastoris is subjected to strong oxidative stress in the methanol induction phase. The oxidative stress inflicts severe injury to yeast cells, which causes cell death and reduces protein expression ability. N-acetyltransferase in Saccharomyces cerevisiae can protect yeast cells from damage caused by decreasing reactive oxygen species (ROS) in oxidative pressure environments such as ethanol treatment, freeze-thawing, or heat shock. In this study, N-acetyltransferase from P. pastoris (PpMpr1) was overexpressed for the first time to improve the anti-oxidative stress ability to protect cells from strong ROS damage during the methanol induction phase. Cell viability of the PpMpr1 overexpression strain increased significantly, while biomass was increased by 22.7% at high dissolved oxygen (DO). At the same time, the heterologous α-glucosidase (AGL) expression level at 25% DO was increased by 21.5%. The AGL degradation was greatly relieved in the fermentation supernatant of the PpMpr1 overexpression strain. This study shows that PpMpr1 has a great potential for improvement of anti-oxidative stress ability in P. pastoris and provides a promising recombinant microorganism for industrial production of proteins.

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