Unit for Molecular Glycobiology, Department for Molecular Biomedical Research, VIB, Ghent, Belgium.
Microb Cell Fact. 2010 Nov 23;9:93. doi: 10.1186/1475-2859-9-93.
Yeast expression systems with altered N-glycosylation are now available to produce glycoproteins with homogenous, defined N-glycans. However, data on the behaviour of these strains in high cell density cultivation are scarce.
Here, we report on cultivations under controlled specific growth rate of a GlycoSwitch-Man5 Pichia pastoris strain producing Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) at high levels (hundreds of milligrams per liter). We demonstrate that homogenous Man5GlcNAc2 N-glycosylation of the secreted proteins is achieved at all specific growth rates tested.
Together, these data illustrate that the GlycoSwitch-Man5 P. pastoris is a robust production strain for homogenously N-glycosylated proteins.
目前已有经过修饰的酵母表达系统可用于生产具有均一、定义明确的 N-聚糖的糖蛋白。然而，关于这些菌株在高密度培养中的行为的数据却很少。
在这里，我们报告了在控制特定生长速率下，一株产生粒细胞-巨噬细胞集落刺激因子（GM-CSF）的 GlycoSwitch-Man5 毕赤酵母菌株的培养情况，该菌株的产量很高（每升数百毫克）。我们证明，在所有测试的特定生长速率下，分泌蛋白的均一 Man5GlcNAc2 N-糖基化都能实现。
综上所述，这些数据表明 GlycoSwitch-Man5 毕赤酵母是一种生产均一 N-糖基化蛋白的稳健生产菌株。
