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Fast protein liquid chromatography purification of hydrophobic fraction of bovine milk proteose-peptone and characterization by bidimensional electrophoresis

  • Jean-Michel Girardet (a1), Abderrahmane Mati (a1), Tibogo Sanogo (a1), Luc Etienne (a1) and Guy Linden (a1)...


Bovine milk Hydrophobic fraction of proteose-peptone was prepared by hydrophobic interaction fast protein liquid chromatography. This method has several advantages such as high rapidity, simple good reproducibility and less denaturation. The proteose-peptone was eluted from a TSK-Phenyl-5PW column with a 1 M-0 M ionic strength gradient of NaH2PO4, pH 6·8, using a 6 ml/min flow rate for 56 min. The quantity of protein injected was 62·5 mg; however, it could be increased up to 100 mg. The elution order was β-CN-4P < BSA (1·6% of total N) < β-CN-5P < β-CN-1P. The hydrophobic fraction was obtained in pure water at the end of the gradient (17·3% of total N). A proteose-peptone cartograph was achieved by bidimensional electrophoresis. This hydrophobic fraction represented three principal zones of Mr 30000–28000, 19000 and 11000, which were respectively composed of 13, 4 and 2 principal spots distributed between 4·9 and 6·1 isoelectric points (IP). These spots corresponded to glycoproteins. ·7, 5·0 and 5·1 IP which migrated to Mr 18000 while β-CN-1P was identified as Mr 9000 in two spots of 5·1 and 5·3 IP.



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