Novel thermostable gluconate dehydratase and use thereof
AssigneesPosco, Postech Foundation
InventorsSeonghun Kim, Sun Bok Lee
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Abstract
The present invention relates to a novel thermostable gluconate dehydratase from the thermoacidophilic archaeon Sulfolobus solfataricus, a coding sequence, and an expression system. The gluconate dehydratase has a molecular weight of about 320,000 to 380,000 daltons as the native protein, and about 40,000 to 50,000 daltons as the monomer protein, and catalyzes the dehydration reaction of aldonic acids to 2-keto-3-deoxy derivatives at temperatures of less than 120° C. The gluconate dehydratase can be produced from native or recombinant host cells and thereby used in the pharmaceutical, agricultural, and other industries.
Claims18
An isolated or purified polynucleotide encoding a gluconate dehydratase, wherein the gluconate dehydratase comprises a polynucleotide having at least a 50% identity to a nucleic acid sequence encoding a polypeptide comprising amino acid sequences of SEQ ID NO:2, or a polynucleotide complementary to the polynucleotide having at least a 50% identity to a polynucleotide encoding an polypeptide comprising amino acid sequences of SEQ ID NO:2.
A polypeptide comprising an amino acid sequence which is at least 50% identical to amino acid sequences of SEQ ID NO:2, wherein the polypeptide catalyzes dehydration of aldonic acid to 2-Keto-3-deoxy aldonic acid.
A method of purifying a gluconate dehydratase, comprising:
- conducting chromatography of a culture solution or a cell from a gluconate dehydratase producing microorganism through a column packed with resin attached to one or more kinds of functional groups selected from the group consisting of carboxy, carboxymethyl, sulpho, sulphomethyl, sulphoprophyl, aminoethyl, diethylaminoethyl, trimethylaminomethyl, triethylaminoethyl, dimethyl-2-hydroxyethylaminomethyl, diethyl-2-hydroxypropylaminoethyl, phospho, alkyl and hydroxylapatite, and
- wherein the matrix of the resin is selected from the group consisting of agarose, cellulose, dextran, polyacrylate, and polystyrene.
A method for producing a 2-keto-3-deoxy aldonic acid from aldonic acid, comprising:
- contacting the gluconate dehydratase to aldonic acid in water or an aqueous solvent at a temperature from 0° C. to 120° C. and a pH of 1.5 to 12, wherein the blend ratio of a gluconate dehydratase to aldonic acid is 1 ug:
- 0.01 to 1 mol.
Dependencies are inferred from the claim text.