Protection of protein secondary structure by saccharides of different molecular weights during freeze-drying.
The protective effects of saccharides with various molecular weights (glucose, maltose, maltotriose, maltotetraose, maltopentaose, maltoheptaose, dextran 1060, dextran 4900, and dextran 10200) against lyophilization-induced structural perturbation of model proteins (BSA, ovalbumin) were studied. Fourier transform infrared (FT-IR) analysis of the proteins in initial solutions and freeze-dried solids indicated that maltose conferred the greatest protection against secondary structure change. The structure-stabilizing effect of maltooligosaccharides decreased in increasing the number of saccharide units. Larger molecules of dextran also showed a smaller structure-stabilizing effect. Increasing the effective saccharide molecular size by a borate-saccharide complexation reduced the protein structure-stabilizing effect of all of the saccharides except glucose. The results indicate that the larger saccharide molecules, and/or the complex formation with borate ion, reduce the free and accessible hydroxyl groups to interact with and stabilize the protein structure by a water-substitution mechanism.
Izutsu
Ken-ichi
K
National Institute of Health Sciences, Tokyo, Japan. izutsu@nihs.go.jp
Aoyagi
Nobuo
N
Kojima
Shigeo
S
D002152:Calorimetry, Differential Scanning; D004355:Drug Stability; D005612:Freeze Drying; D008970:Molecular Weight; D009005:Monosaccharides / Q000737:chemistry*; D009844:Oligosaccharides / Q000737:chemistry*; D010047:Ovalbumin / Q000737:chemistry; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared
D009005:Monosaccharides; D009844:Oligosaccharides; D011506:Proteins; D012710:Serum Albumin, Bovine; D010047:Ovalbumin