DNA for encoding D-hydantoin hydrolases, DNA for encoding N-carbamyl-D-amino acid hydrolases, recombinant DNA containing the genes, cells transformed with the recombinant DNA, methods for producing proteins utilizing the transformed cells and methods for producing D-amino acids

A recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein

• the DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-hydantoinase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 1 in the Sequence Listing;
• and
• (b) is a base sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 1 in the Sequence Listing under a stringent condition for encoding.

A cell transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein

• the DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-hydantoinase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 1 in the Sequence Listing;
• and
• (b) is a base sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 1 in the Sequence Listing under a stringent condition for encoding.

A method for producing a protein having D-hydantoinase activity, the method comprising:

• incubating a cell in a culture medium;
• and
• allowing a protein having D-hydantoinase activity to be accumulated in one or both of the culture medium and the cell, wherein
• the cell to be incubated being transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein
• the DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-hydantoinase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 1 in the Sequence Listing;
• and
• (b) is a base sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 1 in the Sequence Listing under a stringent condition for encoding.

A method for producing a protein having D-hydantoinase activity, the method comprising:

• incubating a cell in a culture medium;
• and
• allowing a protein having D-hydantoinase activity to be accumulated in one or both of the culture medium and the cell, wherein
• the cell to be incubated being transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein
• the DNA having an amino acid sequence represented by (c) or (d) and that encodes a protein having D-hydantoinase activity, wherein
• (c) is the amino acid sequence represented by Seq. ID No. 2 in the Sequence Listing;
• (d) is an amino acid sequence resulting from the substitution, deletion, insertion, addition or inversion of one or several amino acid residues in the amino acid sequence represented by Seq. ID No. 2 in the Sequence Listing.

A method for producing N-carbamyl-D-amino acid, the method comprising:

• producing a protein having D-hydantoinase activity by
• incubating a cell in a culture medium;
• and
• allowing a protein having D-hydantoinase activity to be accumulated in one or both of the culture medium and the cell;
• and
• producing N-carbamyl-D-amino acid by making the protein having D-hydantoinase activity react with a 5-substituted hydantoin, wherein
• the cell to be incubated being transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein
• the DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-hydantoinase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 1 in the Sequence Listing;
• and
• (b) is a base sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 1 in the Sequence Listing under a stringent condition for encoding.

A method for producing N-carbamyl-D-amino acid, the method comprising:

• producing a protein having D-hydantoinase activity by
• incubating a cell in a culture medium;
• and
• allowing a protein having D-hydantoinase activity to be accumulated in one or both of the culture medium and the cell;
• and
• producing N-carbamyl-D-amino acid by making the protein having D-hydantoinase activity react with a 5-substituted hydantoin, wherein
• the cell to be incubated being transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein
• the DNA having an amino acid sequence represented by (c) or (d) and that encodes a protein having D-hydantoinase activity, wherein
• (c) is the amino acid sequence represented by Seq. ID No. 2 in the Sequence Listing;
• (d) is an amino acid sequence resulting from the substitution, deletion, insertion, addition or inversion of one or several amino acid residues in the amino acid sequence represented by Seq. ID No. 2 in the Sequence Listing.

A method for producing D-amino acid comprising:

• producing a protein having D-hydantoinase activity by
• incubating a cell in a culture medium;
• and
• allowing a protein having D-hydantoinase activity to be accumulated in one or both of the culture medium and the cell;
• and
• producing D-amino acid by making the protein having D-hydantoinase activity and an enzyme hydrolyzing an N-carbamyl-D-amino acid or a material containing the enzyme react with a 5-substituted hydantoin, wherein
• the cell to be incubated being transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein
• the DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-hydantoinase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 1 in the Sequence Listing;
• and
• (b) is a base sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 1 in the Sequence Listing under a stringent condition for encoding.

A method for producing D-amino acid comprising:

• producing a protein having D-hydantoinase activity by
• incubating a cell in a culture medium;
• and
• allowing a protein having D-hydantoinase activity to be accumulated in one or both of the culture medium and the cell;
• and
• producing D-amino acid by making the protein having D-hydantoinase activity and an enzyme hydrolyzing an N-carbamyl-D-amino acid or a material containing the enzyme react with a 5-substituted hydantoin, wherein
• the cell to be incubated being transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein
• the DNA having an amino acid sequence represented by (c) or (d) and that encodes a protein having D-hydantoinase activity, wherein
• (c) is the amino acid sequence represented by Seq. ID No. 2 in the Sequence Listing;
• (d) is an amino acid sequence resulting from the substitution, deletion, insertion, addition or inversion of one or several amino acid residues in the amino acid sequence represented by Seq. ID No. 2 in the Sequence Listing.

A recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein

• the DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-carbamylase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 3 in the Sequence Listing;
• and
• (b) is a base sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 3 in the Sequence Listing under a stringent condition.

A recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein

• the DNA having an amino acid sequence represented by (c) or (d) and that encodes a protein having D-carbamylase activity, wherein
• (c) is the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing;
• and
• (d) is an amino acid sequence resulting from the substitution, deletion, insertion, addition or inversion of one or several amino acid residues in the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing.

A cell transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein

• the DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-carbamylase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 3 in the Sequence Listing;
• and
• (b) is abase sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 3 in the Sequence Listing under a stringent condition.

A cell transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein

• the DNA having an amino acid sequence represented by (c) or (d) and that encodes a protein having D-carbamylase activity, wherein
• (c) is the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing;
• and
• (d) is an amino acid sequence resulting from the substitution, deletion, insertion, addition or inversion of one or several amino acid residues in the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing.

A method for producing a protein having D-carbamylase activity, the method comprising:

• incubating a cell in a culture medium;
• and
• allowing a protein having D-carbamylase activity to be accumulated in one or both of the culture medium and the cell, wherein
• the cell to be incubated being transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein
• the DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-carbamylase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 3 in the Sequence Listing;
• and
• (b) is a base sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 3 in the Sequence Listing under a stringent condition.

A method for producing a protein having D-carbamylase activity, the method comprising:

• incubating a cell in a culture medium;
• and
• allowing a protein having D-carbamylase activity to be accumulated in one or both of the culture medium and the cell, wherein
• the cell to be incubated being transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein
• the DNA having an amino acid sequence represented by (c) or (d) and that encodes a protein having D-carbamylase activity, wherein
• (c) is the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing;
• and
• (d) is an amino acid sequence resulting from the substitution, deletion, insertion, addition or inversion of one or several amino acid residues in the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing.

A method for producing D-amino acid, the method comprising:

• producing a protein having D-carbamylase activity by
• incubating a cell in a culture medium;
• and
• allowing a protein having D-carbamylase activity to be accumulated in one or both of the culture medium and the cell;
• and
• producing D-amino acid by making the protein having D-carbamylase activity react with an N-carbamylamino acid, wherein
• the cell to be incubated being transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein
• the DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-carbamylase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 3 in the Sequence Listing;
• and
• (b) is a base sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 3 in the Sequence Listing under a stringent condition.

A method for producing D-amino acid, the method comprising:

• producing a protein having D-carbamylase activity by
• incubating a cell in a culture medium;
• and
• allowing a protein having D-carbamylase activity to be accumulated in one or both of the culture medium and the cell;
• and
• producing D-amino acid by making the protein having D-carbamylase activity react with an N-carbamylamino acid, wherein
• the cell to be incubated being transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein
• the DNA having an amino acid sequence represented by (c) or (d) and that encodes a protein having D-carbamylase activity, wherein
• (c) is the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing;
• and
• (d) is an amino acid sequence resulting from the substitution, deletion, insertion, addition or inversion of one or several amino acid residues in the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing.

A method for producing D-amino acid, the method comprising:

• producing a protein having D-carbamylase activity by
• incubating a cell in a culture medium;
• and
• allowing a protein having D-carbamylase activity to be accumulated in one or both of the culture medium and the cell;
• and
• producing D-amino acid by making the protein having D-carbamylase activity and an enzyme hydrolyzing a 5-substituted hydantoin or a material containing the enzyme react with a 5-substituted hydantoin, wherein
• the cell to be incubated being transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein
• the DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-carbamylase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 3 in the Sequence Listing;
• and
• (b) is a base sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 3 in the Sequence Listing under a stringent condition.

A method for producing D-amino acid, the method comprising:

• producing a protein having D-carbamylase activity by
• incubating a cell in a culture medium;
• and
• allowing a protein having D-carbamylase activity to be accumulated in one or both of the culture medium and the cell;
• and
• producing D-amino acid by making the protein having D-carbamylase activity and an enzyme hydrolyzing a 5-substituted hydantoin or a material containing the enzyme react with a 5-substituted hydantoin, wherein
• the cell to be incubated being transformed with a recombinant DNA resulting from a connection of a DNA with a vector DNA, wherein
• the DNA having an amino acid sequence represented by (c) or (d) and that encodes a protein having D-carbamylase activity, wherein
• (c) is the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing;
• and
• (d) is an amino acid sequence resulting from the substitution, deletion, insertion, addition or inversion of one or several amino acid residues in the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing.

A method for producing D-amino acid, the method comprising:

• producing a protein having D-hydantoinase activity by
• incubating a first cell in a first culture medium;
• and
• allowing a protein having D-hydantoinase activity to be accumulated in the first culture medium and/or the first cell, wherein
• the first cell to be incubated being transformed with a first recombinant DNA resulting from a connection of a first DNA with a first vector DNA, wherein
• the first DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-hydantoinase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 1 in the Sequence Listing;
• and
• (b) is a base sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 1 in the Sequence Listing under a stringent condition for encoding;
• producing a protein having D-carbamylase activity by
• incubating a second cell in a second culture medium;
• and
• allowing a protein having D-carbamylase activity to be accumulated in the second culture medium and/or the second cell, wherein
• the second cell to be incubated being transformed with a second recombinant DNA resulting from a connection of a second DNA with a second vector DNA, wherein
• the second DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-carbamylase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 3 in the Sequence Listing;
• and
• (b) is a base sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 3 in the Sequence Listing under a stringent condition;
• and
• producing D-amino acid by making the protein having D-hydantoinase activity and the protein having D-carbamylase activity react with a 5-substituted hydantoin.

A method for producing D-amino acid, the method comprising:

• producing a protein having D-hydantoinase activity by
• incubating a first cell in a first culture medium;
• and
• allowing a protein having D-hydantoinase activity to be accumulated in the first culture medium and/or the first cell, wherein
• the first cell to be incubated being transformed with a first recombinant DNA resulting from a connection of a first DNA with a first vector DNA, wherein
• the first DNA having abase sequence represented by (a) or (b) and that encodes a protein having D-hydantoinase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 1 in the Sequence Listing;
• and
• (b) is a base sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 1 in the Sequence Listing under a stringent condition for encoding;
• producing a protein having D-carbamylase activity by
• incubating a second cell in a second culture medium;
• and
• allowing a protein having D-carbamylase activity to be accumulated in the second culture medium and/or the second cell, wherein
• the second cell to be incubated being transformed with a second recombinant DNA resulting from a connection of a second DNA with a second vector DNA, wherein
• the second DNA having an amino acid sequence represented by (c) or (d) and that encodes a protein having D-carbamylase activity, wherein
• (c) is the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing;
• and
• (d) is an amino acid sequence resulting from the substitution, deletion, insertion, addition or inversion of one or several amino acid residues in the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing;
• and
• producing D-amino acid by making the protein having D-hydantoinase activity and the protein having D-carbamylase activity react with a 5-substituted hydantoin.

A method for producing D-amino acid, the method comprising:

• producing a protein having D-hydantoinase activity by
• incubating a first cell in a first culture medium;
• and
• allowing a protein having D-hydantoinase activity to be accumulated in the first culture medium and/or the first cell, wherein
• the first cell to be incubated being transformed with a first recombinant DNA resulting from a connection of a first DNA with a first vector DNA, wherein
• the first DNA having an amino acid sequence represented by (c) or (d) and that encodes a protein having D-hydantoinase activity, wherein
• (c) is the amino acid sequence represented by Seq. ID No. 2 in the Sequence Listing;
• (d) is an amino acid sequence resulting from the substitution, deletion, insertion, addition or inversion of one or several amino acid residues in the amino acid sequence represented by Seq. ID No. 2 in the Sequence Listing;
• producing a protein having D-carbamylase activity by
• incubating a second cell in a second culture medium;
• and
• allowing a protein having D-carbamylase activity to be accumulated in the second culture medium and/or the second cell, wherein
• the second cell to be incubated being transformed with a second recombinant DNA resulting from a connection of a second DNA with a second vector DNA, wherein
• the second DNA DNA having a base sequence represented by (a) or (b) and that encodes a protein having D-carbamylase activity, wherein
• (a) is the base sequence represented by Seq. ID No. 3 in the Sequence Listing;
• and
• (b) is abase sequence hybridizing with a complementary base sequence of the base sequence represented by Seq. ID No. 3 in the Sequence Listing under a stringent condition;
• and
• producing D-amino acid by making the protein having D-hydantoinase activity and the protein having D-carbamylase activity react with a 5-substituted hydantoin.

A method for producing D-amino acid, the method comprising:

• producing a protein having D-hydantoinase activity by
• incubating a first cell in a first culture medium;
• and
• allowing a protein having D-hydantoinase activity to be accumulated in the first culture medium and/or the first cell, wherein
• the first cell to be incubated being transformed with a first recombinant DNA resulting from a connection of a first DNA with a first vector DNA, wherein
• the first DNA having an amino acid sequence represented by (c) or (d) and that encodes a protein having D-hydantoinase activity, wherein
• (c) is the amino acid sequence represented by Seq. ID No. 2 in the Sequence Listing;
• (d) is an amino acid sequence resulting from the substitution, deletion, insertion, addition or inversion of one or several amino acid residues in the amino acid sequence represented by Seq. ID No. 2 in the Sequence Listing;
• producing a protein having D-carbamylase activity by
• incubating a second cell in a second culture medium;
• and
• allowing a protein having D-carbamylase activity to be accumulated in the second culture medium and/or the second cell, wherein
• the second cell to be incubated being transformed with a second recombinant DNA resulting from a connection of a second DNA with a second vector DNA, wherein
• the second DNA having an amino acid sequence represented by (c) or (d) and that encodes a protein having D-carbamylase activity, wherein
• (c) is the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing;
• and
• (d) is an amino acid sequence resulting from the substitution, deletion, insertion, addition or inversion of one or several amino acid residues in the amino acid sequence represented by Seq. ID No. 4 in the Sequence Listing;
• and
• producing D-amino acid by making the protein having D-hydantoinase activity and the protein having D-carbamylase activity react with a 5-substituted hydantoin.