دورية أكاديمية
Expression of heparan sulfate sulfotransferases in Kluyveromyces lactis and preparation of 3'-phosphoadenosine-5'-phosphosulfate
| العنوان: | Expression of heparan sulfate sulfotransferases in Kluyveromyces lactis and preparation of 3'-phosphoadenosine-5'-phosphosulfate |
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| المؤلفون: | Zhou, X., Chandarajoti, K., Pham, T. Q., Liu, R., Liu, J. |
| المصدر: | Glycobiology, 21(6) |
| سنة النشر: | 2011 |
| المجموعة: | Carolina Digital Repository (UNC - University of North Carolina) |
| مصطلحات موضوعية: | Gene Expression, Sulfotransferases, heparitin sulfotransferase, Carbohydrate Conformation, Kluyveromyces, Phosphoadenosine Phosphosulfate, Polysaccharides |
| الوصف: | Heparan sulfate (HS) belongs to a major class of glycans that perform central physiological functions. Heparin is a specialized form of HS and is a clinically used anticoagulant drug. Heparin is a natural product isolated from pig intestine. There is a strong demand to replace natural heparin with a synthetic counterpart. Although a chemoenzymatic approach has been employed to prepare synthetic heparin, the scale of the synthesis is limited by the availability of sulfotransferases and the cofactor, 3′-phosphoadenosine-5′-phosphosulfate (PAPS). Here, we present a novel method to produce secreted forms of sulfotransferases in the yeast cells, Kluyveromyces lactis. Five sulfotransferases including N-sulfotransferase, 2-O-sulfotransferase, 3-O-sulfotransferase 1 and 6-O-sulfotransferases 1 and 3 were expressed using this method. Unlike bacterial-expressed sulfotransferases, the yeast proteins can be directly used to modify polysaccharides without laborious purification. The yeast-expressed sulfotransferases also tend to have higher specific activity and thermostability. Furthermore, we demonstrated the possibility for the gram-scale synthesis of PAPS from adenosine 5'-triphosphate at only 1/5000th of the price purchased from a commercial source. Our results pave the way to conduct the enzymatic synthesis of heparin in large quantities. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| العلاقة: | https://doi.org/10.17615/gfxh-k889Test; https://cdr.lib.unc.edu/downloads/8049gc76z?file=thumbnailTest; https://cdr.lib.unc.edu/downloads/8049gc76zTest |
| DOI: | 10.17615/gfxh-k889 |
| الإتاحة: | https://doi.org/10.17615/gfxh-k889Test https://cdr.lib.unc.edu/downloads/8049gc76z?file=thumbnailTest https://cdr.lib.unc.edu/downloads/8049gc76zTest |
| حقوق: | http://rightsstatements.org/vocab/InC/1.0Test/ |
| رقم الانضمام: | edsbas.C281E4A9 |
| قاعدة البيانات: | BASE |
| DOI: | 10.17615/gfxh-k889 |
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