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 K-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 7.0 kDa by SDS-PAGE, and showed a lower glycosylation than the natural bovine CMP

Production of human caseinomacropeptide in recombinant Saccharomyces cerevisiae and Pichia pastoris

Caseinomacropeptide is a polypeptide of 64 amino acid residues (106-169) derived from the C-terminal part of the mammalian milk k-casein. This macropeptide has various biological activities and is used as a functional food ingredient as well as a pharmaceutical compound. The gene encoding the human caseinomacropeptide (hCMP) was synthesized and expressed with an alpha-factor secretion signal in the two yeast strains, Saccharomyces cerevisiae and Pichia pastoris. The complete polypeptide of the recombinant hCMP was produced and secreted in a culture medium by both the strains, but the highest production was observed in S. cerevisiae with a galactose-inducible promoter. In a fed-batch bioreactor culture, 2.5 g/l of the recombinant hCMP was obtained from the S. cerevisiae at 97 h

Antiobesity effect of recombinant human caseinomacropeptide in Sprague-Dawley rat

Caseinomacropepticle (CMP) is a glycopepticle of 64 amino acid residues derived from the C-terminal of mammalian milk kappa-casein. Recently, human CMP (hCMP) was produced from the recombinant yeast Saccharomyces cerevisiae. In this study, the antiobesity activity of the recombinant hCMP (rhCMP) was investigated in vivo using Sprague-Dawley rats. The rhCMP did not affect the rats fed with a normal fat diet (fat content, 5.0%), but decreased the body weight gain of the rats fed with a high fat diet (fat content, 20%) by up to 19%, Autopsies revealed that the weights of the liver, kidney and adipose tissues decreased when the rats were given the rhCMP, which also reduced the lipid concentrations in the plasma and liver, but enhanced the fecal excretion of lipids. These results suggest that rhCMP prevent the accumulation of lipid by stimulating its fecal excretion, so could be used as a food supplement to alleviate the obesity problem caused by a high fat diet