Dna encoding a protein having d-lactate dehydrogenase activity and uses thereof
First Claim
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1. A polynucleotide as described in any of the following (a) to (g):
- (a) a polynucleotide comprising the nucleotide sequence as shown in SEQ ID NO;
1;
(b) a polynucleotide that hybridizes under stringent conditions with a probe comprising all or part of the nucleotide sequence as shown in SEQ ID NO;
1 or a complementary strand thereof and that encodes a protein having D-lactate dehydrogenase activity;
(c) a polynucleotide that encodes a protein consisting of the amino acid sequence as shown in SEQ ID NO;
2;
(d) a polynucleotide that encodes a protein consisting of an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO;
2 by substitution, deletion, insertion, or addition of one or several amino acid residues and having D-lactate dehydrogenase activity;
(e) a polynucleotide that encodes a protein having 70% or higher homology to the amino acid sequence as shown in SEQ ID NO;
2 and having D-lactate dehydrogenase activity;
(f) a polynucleotide that encodes a protein having an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO;
4 by substitution of one or more amino acid residues selected from the list of amino acid residue substitutions shown in Table 1 and having D-lactate dehydrogenase activity;
TABLE 1 List of amino acid residue substitutions Substitution typePosition of substitutionAmino acid substituent 140Valine (Val)2112Isoleucine (Ile)3131Histidine (His)4139Isoleucine (Ile)5181Glutamic acid (Glu)6266Glycine (Gly)7267Leucine (Leu)8268Phenylalanine (Phe)9269Asparagine (Asn)10270Glutamic acid (Glu)11271Aspartic acid (Asp)12272Tryptophan (Trp)13273Serine (Ser)14274Glycine (Gly)15276Glutamic acid (Glu)16277Phenylalanine (Phe)17287Serine (Ser)18292Leucine (Leu)19293Valine (Val)
wherein positions of substitution are indicated as the positions from methionine, which corresponds to the initiation codon;
or (g) a polynucleotide that encodes a protein having an amino acid sequence containing at least amino acid residues 78 and 79, 152 to 175, 235, and 296 of the amino acid sequence as shown in SEQ ID NO;
2 and having D-lactate dehydrogenase activity.
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Abstract
This invention provides a polynucleotide that encodes a protein having lactate dehydrogenase activity and such protein that can be used for producing D-lactic acid. This polynucleotide has the nucleotide sequence as shown in SEQ ID NO: 1 (a), and it hybridizes under stringent conditions with a probe comprising all or part of the nucleotide sequence as shown in SEQ ID NO: 1 or a complementary strand thereof and encodes a protein having D-lactate dehydrogenase activity (b).
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Citations
15 Claims
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1. A polynucleotide as described in any of the following (a) to (g):
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(a) a polynucleotide comprising the nucleotide sequence as shown in SEQ ID NO;
1;
(b) a polynucleotide that hybridizes under stringent conditions with a probe comprising all or part of the nucleotide sequence as shown in SEQ ID NO;
1 or a complementary strand thereof and that encodes a protein having D-lactate dehydrogenase activity;
(c) a polynucleotide that encodes a protein consisting of the amino acid sequence as shown in SEQ ID NO;
2;
(d) a polynucleotide that encodes a protein consisting of an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO;
2 by substitution, deletion, insertion, or addition of one or several amino acid residues and having D-lactate dehydrogenase activity;
(e) a polynucleotide that encodes a protein having 70% or higher homology to the amino acid sequence as shown in SEQ ID NO;
2 and having D-lactate dehydrogenase activity;
(f) a polynucleotide that encodes a protein having an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO;
4 by substitution of one or more amino acid residues selected from the list of amino acid residue substitutions shown in Table 1 and having D-lactate dehydrogenase activity;
TABLE 1 List of amino acid residue substitutions Substitution type Position of substitution Amino acid substituent 1 40 Valine (Val) 2 112 Isoleucine (Ile) 3 131 Histidine (His) 4 139 Isoleucine (Ile) 5 181 Glutamic acid (Glu) 6 266 Glycine (Gly) 7 267 Leucine (Leu) 8 268 Phenylalanine (Phe) 9 269 Asparagine (Asn) 10 270 Glutamic acid (Glu) 11 271 Aspartic acid (Asp) 12 272 Tryptophan (Trp) 13 273 Serine (Ser) 14 274 Glycine (Gly) 15 276 Glutamic acid (Glu) 16 277 Phenylalanine (Phe) 17 287 Serine (Ser) 18 292 Leucine (Leu) 19 293 Valine (Val) wherein positions of substitution are indicated as the positions from methionine, which corresponds to the initiation codon;
or(g) a polynucleotide that encodes a protein having an amino acid sequence containing at least amino acid residues 78 and 79, 152 to 175, 235, and 296 of the amino acid sequence as shown in SEQ ID NO;
2 and having D-lactate dehydrogenase activity. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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2. A protein as described in any of the following (h) to (1):
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(h) a protein that consists of the amino acid sequence as shown in SEQ ID NO;
2 of the Sequence Listing;
(i) a protein that consists of an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO;
2 of the Sequence Listing by substitution, deletion, insertion, or addition of one or several amino acid residues and that has D-lactate dehydrogenase activity;
(j) a protein that has 70% or higher homology to the amino acid sequence as shown in SEQ ID NO;
2 and that has D-lactate dehydrogenase activity;
(k) a protein that has an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO;
4 by substitution of one or more amino acid residues selected from the list of amino acid residue substitutions shown in Table 1 and that has D-lactate dehydrogenase activity;
or(l) a protein that has an amino acid sequence containing at least amino acid residues 78 and 79, 152 to 175, 235, and 296 of the amino acid sequence as shown in SEQ ID NO;
2 and that has D-lactate dehydrogenase activity.
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Specification