Human pS2/trefoil factor 1:: Production and characterization in Pichia pastoris
Human pS2/trefoil factor 1:: Production and characterization in Pichia pastoris
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DOI：
10.1006/prep.2000.1352
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发表时间：
2001-02-01
影响因子：
1.6
通讯作者：
Rio, MC
中科院分区：
文献类型：
--
作者：
Kannan, R;Tomasetto, C;Rio, MC
重组蛋白人三叶因子1 (hTFF1)，以前称为hpS2，首次在毕赤酵母中以酵母为基础表达。在甘油醛-3-磷酸脱氢酶启动子的控制下，hTFF1在P， pastoris胞外培养基中大量分泌。含hTFF1的发酵液经切向流过滤浓缩，再经阴离子交换层析和阳离子交换层析纯化，制备高效液相层析纯化。Western blot分析发现hTFF1是完整的。进一步分析显示主要存在hTFF1肽的单体形式。最后，在体外，重组hTFF1被证明可以减少HCT116癌细胞的增殖，(C) 2001学术出版社。
The recombinant protein human trefoil factor 1 (hTFF1), formerly called hpS2, has been produced for the first time in a yeast-based expression in Pichia pastoris. hTFF1 was secreted in large amounts in the extracellular medium of P, pastoris under the control of the glyceraldehyde-3-phosphate dehydrogenase promoter. The fermentation broth containing hTFF1 was concentrated by tangential flow filtration prior to purification by anion- and cation-exchange chromatography, followed by preparative high-performance liquid chromatography. The resulting hTFF1 was found to be intact by Western blot analysis. Further analysis revealed mainly the presence of the monomeric form of the hTFF1 peptide. Finally, in vitro, the recombinant hTFF1 was shown to decrease proliferation of the HCT116 cancer cells, (C) 2001 Academic Press.