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首页> 外文期刊>Applied Microbiology and Biotechnology >Expression of chondroitin-4-O-sulfotransferase in Escherichia coli and Pichia pastoris
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Expression of chondroitin-4-O-sulfotransferase in Escherichia coli and Pichia pastoris

机译：软骨素-4-O-磺膦酸酯在大肠杆菌和毕赤酵母中的表达

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Chondroitin sulfates are linear sulfated polysaccharides called glycosaminoglycans. They are important nutraceutical and pharmaceutical products that are biosynthesized through the action of chondroitin sulfotransferases on either an unsulfated chondroitin or a dermatan polysaccharide precursor. While the enzymes involved in the biosynthesis of chondroitin sulfates are well known, the cloning end expression of these membrane-bound Golgi enzymes continue to pose challenges. The major chondroitin-4-sulfotransferase, Homo sapiens C4ST-1, had been previously cloned and expressed from mammalian CHO, COS-7, and HEK 293 cells, and its activity was shown to require glycosylation. In the current study, a C4ST-1 construct was designed and expressed in both Escherichia coli and Pichia pastoris in its non-glycosylated and glycosylated forms. Both constructs showed similar activity albeit different kinetic parameters when acting on a microbially prepared unsulfated chondroitin substrate. Moreover, the glycosylated form of C4ST-1 showed lower stability than the non-glycosylated form.
机译：硫酸软骨素是称为糖胺聚糖的直链多糖。它们是通过软骨素磺脲类酶的作用在未卷发的软骨素或皮肤多糖前体上生物合成的重要营养保健品和药物产品。虽然涉及硫酸软骨素的生物合成的酶是众所周知的，但是这些膜结合的高尔基酶的克隆结束表达仍然存在挑战。主要克隆和表达来自哺乳动物Cho，COS-7和HEK 293细胞的主要软骨素-4-磺膦酸酶，Homo Sapiens C4st-1，并显示其活性要求糖基化。在目前的研究中，在其非糖基化和糖基化的形式中设计并表达了C4ST-1构建体和表达。两种构建体显示出类似的活性，尽管在微生物制备的未制备的未卷发的软骨素基质上作用时不同的动力学参数。此外，C 4 St-1的糖基化形式显示比非糖基化形式较低的稳定性。

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