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US 10,717,971 B2Granted
Protein deamidase
AssigneeAjinomoto Co., Inc.
InventorsNoriko Yokoyama, Mayu Nakano, Shuhei Hashiro, Masako Hiraga, Kazutaka Shimbo, Ayaka Tokunaga
Life of this patent
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Abstract
A novel protein deamidase having an activity of directly acting on a side chain amide group of an asparagine residue in a protein to form a side chain carboxyl group and release ammonia, a microorganism that produces the same, a gene encoding the same, a method for producing the same, and use of the same are provided. A bacterium classified into the class Actinobacteria is cultured to generate protein deamidase, and the enzyme is collected from culture.
Claims12
An isolated polynucleotide encoding a protein having an activity for catalyzing a reaction of deamidating an asparagine residue in a protein,
- wherein the protein having the activity is selected from the group consisting of the following (a), (b), and (c):
- (a) a protein comprising the amino acid sequence of SEQ ID NO:
- 2, 5, 7, 8, or 9, the amino acid sequence of positions 240 to 1355 of SEQ ID NO:
- 2, the amino acid sequence of positions 181 to 1180 or 67 to 1180 of SEQ ID NO:
- 5, the amino acid sequence of positions 193 to 1172 or 70 to 1172 of SEQ ID NO:
- 7, or the amino acid sequence of positions 146 to 989 or 21 to 989 of SEQ ID NO:
- 8;
- (b) a protein comprising the amino acid sequence of SEQ ID NO:
- 2, 5, 7, 8, or 9, the amino acid sequence of positions 240 to 1355 of SEQ ID NO:
- 2, the amino acid sequence of positions 181 to 1180 or 67 to 1180 of SEQ ID NO:
- 5, the amino acid sequence of positions 193 to 1172 or 70 to 1172 of SEQ ID NO:
- 7, or the amino acid sequence of positions 146 to 989 or 21 to 989 of SEQ ID NO:
- 8 but including substitution, deletion, insertion, or addition of one or several amino acid residues;
- and
- (c) a protein comprising an amino acid sequence showing an identity of 90% or higher to the amino acid sequence of SEQ ID NO:
- 2, 5, 7, 8, or 9, the amino acid sequence of positions 240 to 1355 of SEQ ID NO:
- 2, the amino acid sequence of positions 181 to 1180 or 67 to 1180 of SEQ ID NO:
- 5, the amino acid sequence of positions 193 to 1172 or 70 to 1172 of SEQ ID NO:
- 7, or the amino acid sequence of positions 146 to 989 or 21 to 989 of SEQ ID NO:
- 8.
17 more elements
A recombinant vector containing a polynucleotide encoding a protein having an activity for catalyzing a reaction of deamidating an asparagine residue in a protein,
- wherein the protein having the activity is selected from the group consisting of the following (a), (b), and (c):
- (a) a protein comprising the amino acid sequence of SEQ ID NO:
- 2, 5, 7, 8, or 9, the amino acid sequence of positions 240 to 1355 of SEQ ID NO:
- 2, the amino acid sequence of positions 181 to 1180 or 67 to 1180 of SEQ ID NO:
- 5, the amino acid sequence of positions 193 to 1172 or 70 to 1172 of SEQ ID NO:
- 7, or the amino acid sequence of positions 146 to 989 or 21 to 989 of SEQ ID NO:
- 8;
- (b) a protein comprising the amino acid sequence of SEQ ID NO:
- 2, 5, 7, 8, or 9, the amino acid sequence of positions 240 to 1355 of SEQ ID NO:
- 2, the amino acid sequence of positions 181 to 1180 or 67 to 1180 of SEQ ID NO:
- 5, the amino acid sequence of positions 193 to 1172 or 70 to 1172 of SEQ ID NO:
- 7, or the amino acid sequence of positions 146 to 989 or 21 to 989 of SEQ ID NO:
- 8 but including substitution, deletion, insertion, or addition of one or several amino acid residues;
- and
- (c) a protein comprising an amino acid sequence showing an identity of 90% or higher to the amino acid sequence of SEQ ID NO:
- 2, 5, 7, 8, or 9, the amino acid sequence of positions 240 to 1355 of SEQ ID NO:
- 2, the amino acid sequence of positions 181 to 1180 or 67 to 1180 of SEQ ID NO:
- 5, the amino acid sequence of positions 193 to 1172 or 70 to 1172 of SEQ ID NO:
- 7, or the amino acid sequence of positions 146 to 989 or 21 to 989 of SEQ ID NO:
- 8.
17 more elements
A method for producing a protein having an activity for catalyzing a reaction of deamidating an asparagine residue in a protein, the method comprising:
- culturing a microorganism capable of producing the protein having the activity in a medium such that the protein having the activity is generated;
- and
- collecting the protein having the activity from the culture broth,
- wherein the protein having the activity is selected from the group consisting of the following (a), (b), and (c):
- (a) a protein comprising the amino acid sequence of SEQ ID NO:
- 2, 5, 7, 8, or 9, the amino acid sequence of positions 240 to 1355 of SEQ ID NO:
- 2, the amino acid sequence of positions 181 to 1180 or 67 to 1180 of SEQ ID NO:
- 5, the amino acid sequence of positions 193 to 1172 or 70 to 1172 of SEQ ID NO:
- 7, or the amino acid sequence of positions 146 to 989 or 21 to 989 of SEQ ID NO:
- 8;
- (b) a protein comprising the amino acid sequence of SEQ ID NO:
- 2, 5, 7, 8, or 9, the amino acid sequence of positions 240 to 1355 of SEQ ID NO:
- 2, the amino acid sequence of positions 181 to 1180 or 67 to 1180 of SEQ ID NO:
- 5, the amino acid sequence of positions 193 to 1172 or 70 to 1172 of SEQ ID NO:
- 7, or the amino acid sequence of positions 146 to 989 or 21 to 989 of SEQ ID NO:
- 8 but including substitution, deletion, insertion, or addition of one or several amino acid residues;
- and
- (c) a protein comprising an amino acid sequence showing an identity of 90% or higher to the amino acid sequence of SEQ ID NO:
- 2, 5, 7, 8, or 9, the amino acid sequence of positions 240 to 1355 of SEQ ID NO:
- 2, the amino acid sequence of positions 181 to 1180 or 67 to 1180 of SEQ ID NO:
- 5, the amino acid sequence of positions 193 to 1172 or 70 to 1172 of SEQ ID NO:
- 7, or the amino acid sequence of positions 146 to 989 or 21 to 989 of SEQ ID NO:
- 8.
20 more elements
Dependencies are inferred from the claim text.