Purification and characterization of human caseinomacropeptide produced by a recombinant Saccharomyces cerevisiae.
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.
Kim
Yu-Jin
YJ
Department of Chemical Engineering, Institute for Environmental Technology and Industry, Pusan National University, 30 Jangjeon-dong, Geumjeong-gu, Busan 609-735, Republic of Korea.
Park
Sunghoon
S
Oh
You-Kwan
YK
Kang
Whankoo
W
Kim
Hee Sook
HS
Lee
Eun Yeol
EY
D000818:Animals; D002364:Caseins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D006031:Glycosylation; D006801:Humans; D030781:Organisms, Genetically Modified; D010446:Peptide Fragments / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors
D002364:Caseins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C065989:caseinomacropeptide