Alcohol oxidase from Pichia-type yeasts
First Claim
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1. A method comprising:
- preparing an aqueous fluid containing a suspension of cells of a methanol utilizing microorganism of genus Pichia, said aqueous fluid having a cell density effective for crystallization of alcohol oxidase in a recovery range solution under suitable conditions of pH and ionic strength;
homogenizing said fluid containing a suspension of cells to produce a homogenate; and
removing suspended solids from the homogenate, thereby producing a solution containing soluble alcohol oxidase.
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Abstract
A new alcohol oxidase is isolated from Pichia-type microorganisms in soluble or crystalline form. The crystalline novel enzyme is isolated by preparing an aqueous fluid containing cells of a Pichia-type microorganism, homogenizing the fluid and separating solids therefrom to produce an alcohol oxidase solution, adjusting the solution to have an ionic strength in the range of 0.05 to 0.01 in ionic strength to form a recovery range solution thereby causing the crystalline alcohol oxidase to form. The new enzyme is used to determine alcohol concentrations in fluid samples in which conditions are compatible with the enzyme'"'"'s activity.
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16 Claims
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1. A method comprising:
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preparing an aqueous fluid containing a suspension of cells of a methanol utilizing microorganism of genus Pichia, said aqueous fluid having a cell density effective for crystallization of alcohol oxidase in a recovery range solution under suitable conditions of pH and ionic strength; homogenizing said fluid containing a suspension of cells to produce a homogenate; and removing suspended solids from the homogenate, thereby producing a solution containing soluble alcohol oxidase. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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Specification