Method for purifying cholesterol esterase
AssigneeEastman Kodak Company
InventorsTheodore Walter Esders, James Robert Schaeffer, Harry Wayne Harris
Life of this patent
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Abstract
Cholesterol esterase is purified by removal of the separable lipase in an enzyme preparation containing both cholesterol esterase and lipase activities by treatment using a hydrophobic chromatography column comprising carboxylated polysaccharide material having lower alkylamine with less than 8 carbon atoms attached thereto.
Claims10
A method for purifying a microbial enzyme preparation having separable cholesterol esterase and lipase activities, said method comprising:
- treating the enzyme preparation by hydrophobic chromatography using carboxylated polysaccharide material having attached thereto lower alkylamine with less than 8 carbon atoms to remove the separable lipase from the enzyme preparation to yield a purified cholesterol esterase preparation.
A method for purifying a microbial cholesterol esterase preparation having separable lipase activity, said method comprising:
- a. treating the cholesterol esterase preparation by hydrophobic chromatography using a column comprising carboxylated polysaccharide material having attached thereto lower alkylamine with less than 8 carbon atoms;
- and
- b. eluting cholesterol esterase separately from the separable lipase activity using a stepwise salt gradient.
A method for purifying a microbial cholesterol esterase preparation by removing separable lipase activity, said method comprising:
- a. applying the cholesterol esterase preparation to a hydrophobic chromatography column comprising a carbodiimide reaction product of (1) alkylamine having less than 8 carbon atoms and (2) cyanogen bromide-activated agarose having free carboxyl-containing group attached thereto;
- and
- b. eluting the cholesterol esterase with a stepwise salt gradient to remove separable lipase activity thereby obtaining a purified cholesterol esterase preparation.
A method for removing separable lipase from an enzyme preparation having cholesterol esterase activity obtained by growing Candida rugosa, said method comprising:
- a. applying the enzyme preparation to a hydrophobic chromatography column comprising a reaction product of (1) butylamine and (2) cyanogen bromide-activated agarose having aminohexanoic acid attached thereto;
- and
- b. eluting the cholesterol esterase activity using a stepwise salt gradient to obtain a purified enzyme preparation having cholesterol esterase activity with separable lipase removed.
Dependencies are inferred from the claim text.