Kim Yu-Jin, Park Sunghoon, Oh You-Kwan, Kang Whankoo, Kim Hee Sook, Lee Eun Yeol
Department of Chemical Engineering, Institute for Environmental Technology and Industry, Pusan National University, 30 Jangjeon-dong, Geumjeong-gu, Busan 609-735, Republic of Korea.
Protein Expr Purif. 2005 Jun;41(2):441-6. doi: 10.1016/j.pep.2005.02.021.
Caseinomacropeptide (CMP) is a biologically active polypeptide derived from the C-terminal of milk kappa-casein. CMP is heterogeneous since it is modified differently by glycosylation and phosphorylation after translation. Recently, recombinant human CMP (hCMP) has been produced as a secretory product in yeast. The present study aimed at the purification and characterization of recombinant hCMP. By sequential molecular cut-off ultrafiltration and anion-exchange chromatography, the recombinant hCMP in the culture broth could be purified to an HPLC purity over 94%. The authenticity of the purified hCMP was confirmed by sequence analysis of N-terminal amino acids. The recombinant hCMP was estimated to be 7.0kDa by SDS-PAGE, and showed a lower glycosylation than the natural bovine CMP.
酪蛋白巨肽（CMP）是一种源自牛奶κ-酪蛋白C末端的生物活性多肽。由于CMP在翻译后经糖基化和磷酸化修饰不同，所以它具有异质性。最近，重组人CMP（hCMP）已在酵母中作为分泌产物生产出来。本研究旨在对重组hCMP进行纯化和表征。通过连续的分子截留超滤和阴离子交换色谱法，可将培养液中的重组hCMP纯化至HPLC纯度超过94%。通过N端氨基酸序列分析证实了纯化的hCMP的真实性。经SDS-PAGE测定，重组hCMP的分子量约为7.0kDa，其糖基化程度低于天然牛CMP。
