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US 8,510,053 B2Granted
Method for modifying a property of a protein
AssigneeAjinomoto Co., Inc.
InventorsYosuke Nishio, Eiichiro Kimura, Yoshihiro Usuda, Kazuho Ikeo, Yoji Nakamura, Takashi Gojobori, Yutaka Kawarabayashi, Yumi Hino, Eiichi Hori, Jun Yamazaki
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Abstract
The present invention relates to a method for modifying a property of a protein. The present invention also relates to a method for producing a protein which has a modified property, and a method for producing a microorganism which has a modified property. The present invention is useful in the field of microbial industrial production and the like.
Claims9
A method for modifying the thermostability of a protein, comprising
- (a) selecting 1000 or more genes from the genome of a first microorganism, and selecting 1000 or more genes from the genome of a second microorganism, wherein the genes from the first microorganism are orthologs to the genes from the second microorganism, and wherein the second microorganism is closely related to the first microorganism, but grows differently under an optimum growth condition wherein the optimum growth condition is the optimum growth temperature when compared with the first microorganism, and
- (b) comparing an amino acid sequence encoded by one of the said 1000 or more genes from the first microorganism to an amino acid sequence encoded by the orthologous gene from the second microorganism,
- (c) detecting substitutions between the amino acid sequence encoded by one of the said 1000 or more genes from the first microorganism and the amino acid sequence encoded by the orthologous gene from the second microorganism for each pair of orthologous genes,
- (d) compiling the detected amino acid substitutions for each amino acid substitution type,
- (e) calculating the frequency of each amino acid substitution type, wherein for each detected amino acid substitution type, a correction is made by subtracting the total number of substitution types which occur from the first microorganism to the second microorganism from the total number of the same substitution type which occurs in the reverse direction, or from the second microorganism to the first microorganism,
- (f) identifying and labelling the amino acid substitutions which occur at a high frequency, and
- (g) modifying the thermostability of said protein by introducing one or more mutations into a gene encoding said protein so that the amino acid substitutions identified in (f) occur.
4 more elements
A method for producing a protein having a modified property comprising:
- (a) selecting 1000 or more genes from the genome of a first microorganism, and selecting 1000 or more genes from the genome of a second microorganism, wherein the genes from the first microorganism are orthologs to the genes from the second microorganism, and wherein the second microorganism is closely related to the first microorganism, but grows differently under at least one optimum growth condition when compared with the first microorganism,
- (b) comparing an amino acid sequence encoded by a gene from the first microorganism to an amino acid sequence encoded by the orthologous gene from the second microorganism,
- (c) detecting substitutions between the amino acid sequence encoded by a gene from the first microorganism and the amino acid sequence encoded by a gene from the second microorganism for each pair of orthologous genes,
- (d) compiling the detected amino acid substitutions for each amino acid substitution type,
- (e) calculating the frequency of each amino acid substitution type, wherein for each detected amino acid substitution type, a correction is made by subtracting the total number of substitution types which occur from the first microorganism to the second microorganism from the total number of the same substitution type which occurs in the reverse direction, or from the second microorganism to the first microorganism,
- (f) identifying and labelling the amino acid substitutions which occur at a high frequency as amino acid substitutions which are involved in said optimum growth condition, and
- (g) introducing one or more of the amino acid substitutions identified in (f) into the gene encoding the protein to modify a property of the protein,
- (h) introducing said gene obtained in (g) into a suitable host for gene expression to express the protein having a modified property.
5 more elements
A method for producing a microorganism having a modified property comprising:
- (a) selecting 1000 or more genes from the genome of a first microorganism, and selecting 1000 or more genes from the genome of a second microorganism, wherein the genes from the first microorganism are orthologs to the genes from the second microorganism, and wherein the second microorganism is closely related to the first microorganism, but grows differently under at least one optimum growth condition when compared with the first microorganism,
- (b) comparing an amino acid sequence encoded by a gene from the first microorganism to an amino acid sequence encoded by the orthologous gene from the second microorganism,
- (c) detecting substitutions between the amino acid sequence encoded by a gene from the first microorganism and the amino acid sequence encoded by a gene from the second microorganism for each pair of orthologous genes,
- (d) compiling the detected amino acid substitutions for each amino acid substitution type,
- (e) calculating the frequency of each amino acid substitution type, wherein for each detected amino acid substitution type, a correction is made by subtracting the total number of substitution types which occur from the first microorganism to the second microorganism from the total number of the same substitution type which occurs in the reverse direction, or from the second microorganism to the first microorganism,
- (f) identifying and labelling the amino acid substitutions which occur at a high frequency as amino acid substitutions which are involved in said optimum growth condition, and
- (g) introducing one or more of the amino acid substitutions identified in (f) into a chromosomal DNA of a microorganism to modify a property of said microorganism.
4 more elements
Dependencies are inferred from the claim text.