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US 7,510,861 B2Granted
Gluconate dehydratase
AssigneeMitsui Chemicals, Inc.
InventorsHitoki Miyake, Toshifumi Yamaki, Toshihiro Oikawa, Takeshi Nakamura, Hiroki Ishibashi, Yasushi Fukuiri, Atsushi Sakuma, Hironori Komatsu, Tomoyuki Ando, Kazuhiko Togashi, Hideki Umetani
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Abstract
A novel gluconate dehydratase derived from Achromobacter xylosoxidans and a gene encoding the gluconate dehydratase are provided. By reacting the gluconate dehydratase or a transformed cell containing the gene with an aldonic acid, the corresponding 2-keto-3-deoxyaldonic acid can be efficiently produced.
Claims12
A process for producing a 2-deoxyaldonic acid, comprising the steps of:
- reacting in a water based medium an aldonic acid or salt thereof with a gluconate dehydratase capable of dehydrating D-gluconic acid to produce 2-keto-3-deoxy-D-gluconic acid, thereby converting said aldonic acid or salt thereof into a 2-keto-3-deoxyaldonic acid;
- and
- reacting in a water based medium said 2-keto-3-deoxyaldonic acid with an oxidizing agent, thereby decarboxylating said 2-keto-3-deoxyaldonic acid and reducing the carbon number of said 2-keto-3-deoxyaldonic acid by 1 to produce said 2-deoxyaldonic acid;
- wherein:
- said gluconate dehydratase maintains at least 95% of the enzyme activity thereof after being allowed to stand in 30 mM tris(hydroxymethyl) aminomethane buffer solution with a pH of about 8.5 containing 1 mM of sodium D-gluconate and 1 mM of magnesium chloride at 55° C. for 2 hours;
- and
- said gluconate dehydratase comprises the amino acid sequence shown in SEQ ID NO:2.
4 more elements
A process for producing a 2-deoxyaldose, comprising the steps of:
- reacting in a water-based medium an aldonic acid or salt thereof with a gluconate dehydratase capable of dehydrating D-gluconic acid to produce 2-keto-3-deoxy-D-gluconic acid, thereby converting said aldonic acid or salt thereof into a 2-keto-3-deoxyaldonic acid;
- reacting in a water-based medium said 2-keto-3-deoxyaldonic acid with a reducing agent to produce a 2-hydroxy-3-deoxyaldonic acid;
- and
- reacting in a water-based medium said 2-hydroxy-3-deoxyaldonic acid with an oxidizing agent, thereby decarboxylating said 2-hydroxy-3-deoxyaldonic acid and reducing the carbon number of said 2-hydroxy-3-deoxyaldonic acid by 1 to produce said 2-deoxyaldose;
- wherein:
- said gluconate dehydratase maintains at least 95% of the enzyme activity thereof after being allowed to stand in 30 mM tris(hydroxymethyl) aminomethane buffer solution with a pH of about 8.5 containing 1 mM of sodium D-gluconate and 1 mM of magnesium chloride at 55° C. for 2 hours;
- and
- said gluconate dehydratase comprises the amino acid sequence shown in SEQ ID NO:2.
5 more elements
A process for converting an aldonic acid into a corresponding 2-keto-3-deoxyaldonic acid, comprising the step of:
- reacting in a water based medium an aldonic acid or salt thereof with (a) a transformed cell prepared by transforming a host cell with a plasmid containing a gene encoding a gluconate dehydratase capable of dehydrating D-gluconic acid to produce 2-keto-3-deoxy-D-gluconic acid, or (b) processed products from the transformed cell;
- wherein:
- said gluconate dehydratase maintains at least 95% of the enzyme activity thereof after being allowed to stand in 30 mM tris(hydroxymethyl) aminomethane buffer solution with a pH of about 8.5 containing 1 mM of sodium D-gluconate and 1 mM of magnesium chloride at 55° C. for 2 hours;
- and
- said gluconate dehydratase comprises the amino acid sequence shown in SEQ ID NO:2.
2 more elements
A process for producing a 2-deoxyaldonic acid, comprising the steps of:
- reacting in a water based medium an aldonic acid or salt thereof with (a) a transformed cell prepared by transforming a host cell with a plasmid containing a gene encoding a gluconate dehydratase capable of dehydrating D-gluconic acid to produce 2-keto-3-deoxy-D-gluconic acid, or (b) processed products from the transformed cell, thereby converting said aldonic acid or salt thereof into a 2-keto-3-deoxyaldonic acid;
- and
- reacting in a water based medium said 2-keto-3-deoxyaldonic acid with an oxidizing agent, thereby decarboxylating said 2-keto-3-deoxyaldonic acid and reducing the carbon number of said 2-keto-3-deoxyaldonic acid by 1 to produce said 2-deoxyaldonic acid;
- wherein:
- said gluconate dehydratase maintains at least 95% of the enzyme activity thereof after being allowed to stand in 30 mM tris(hydroxymethyl) aminomethane buffer solution with a pH of about 8.5 containing 1 mM of sodium D-gluconate and 1 mM of magnesium chloride at 55° C. for 2 hours;
- and
- said gluconate dehydratase comprises the amino acid sequence shown in SEQ ID NO:2.
4 more elements
A process for producing a 2-deoxyaldose, comprising the steps of:
- reacting in a water based medium an aldonic acid or salt thereof with (a) a transformed cell prepared by transforming a host cell with a plasmid containing a gene encoding a gluconate dehydratase capable of dehydrating D-gluconic acid to produce 2-keto-3-deoxy-D-gluconic acid, or (b) processed products from the transformed cell, thereby converting said aldonic acid or salt thereof into a 2-keto-3-deoxyaldonic acid;
- reacting in a water based medium said 2-keto-3-deoxyaldonic acid with a reducing agent to produce a 2-hydroxy-3-deoxyaldonic acid;
- and
- reacting in a water based medium said 2-hydroxy-3-deoxyaldonic acid with an oxidizing agent, thereby producing thereby decarboxylating said 2-hydroxy-3-deoxyaldonic acid and reducing the carbon number of said 2-hydroxy-3-deoxyaldonic acid by 1 to produce said 2-deoxyaldose;
- wherein:
- said gluconate dehydratase maintains at least 95% of the enzyme activity thereof after being allowed to stand in 30 mM tris(hydroxymethyl) aminomethane buffer solution with a pH of about 8.5 containing 1 mM of sodium D-gluconate and 1 mM of magnesium chloride at 55° C. for 2 hours;
- and
- said gluconate dehydratase comprises the amino acid sequence shown in SEQ ID NO:2.
5 more elements
A process for converting an aldonic acid into a corresponding 2-keto-3-deoxyaldonic acid, comprising the step of:
- reacting in a water based medium an aldonic acid or salt thereof with a gluconate dehydratase capable of dehydrating D-gluconic acid to produce 2-keto-3-deoxy-D-gluconic acid;
- wherein:
- said gluconate dehydratase maintains at least 95% of the enzyme activity thereof after being allowed to stand in 30 mM tris(hydroxymethyl) aminomethane buffer solution with a pH of about 8.5 containing 1 mM of sodium D-gluconate and 1 mM of magnesium chloride at 55° C. for 2 hours;
- and
- said gluconate dehydratase comprises the amino acid sequence shown in SEQ ID NO:2.
2 more elements
Dependencies are inferred from the claim text.