Journal of Lipid Research, Vol 17, 307-313, Copyright © 1976 by Lipid Research, Inc.
Affinity chromatography of lipase with hydrophobic ligands coupled to cyanogen bromide-activated agarose
Y Kosugi and H Suzuki
The behavior of lipase produced by Pseudomans mephitica var. lipopytica
toward hydrophobic residues coupled to spacer gels that were prepared by
coupling a primary amine to CNBr-activated agarose, was studied. The lipase
adsorbed on the ligand of a long unbranched aliphatic chain, a benzene
ring, or deoxycholic acid was only slightly or not all eluted at pH 5 or pH
11 by buffers containing 1 M NcC1. The lipase was eluted by liquid
containing a surfactant or an organic solvent miscible with water,
indicating greater involvement of hydrophobic forces. The adsorption of
propane, cyclopentane, cyclohexane, cycloheptane, or chrysene appears to be
achieved through electrostatic forces, inasmuch as desorption was caused by
buffer containing 1 M NaC1 at pH 11. The amount of lipase adsorbed on these
hydrophobic ligands was about the same as that adsorbed on the ligands
belonging to the first group. Since little lipase wad adsorbed on
cyclopropane, cycloctane, pyridine, methane, n-pentane, or branched
aliphatic chains, these ligands appear to impose steric hindrance on the
adsorption of lipase, or they may be too small to fit into the hydrophobic
sites of lipase.
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