Method for production of polypeptide

Abstract

The present invention provides a method capable of producing a natural or recombinant protein in high yield. The present invention relates to a method of producing a polypeptide, comprising culturing a cell which strongly expresses cysteine sulfinic acid decarboxylase and has a transferred DNA encoding a desired polypeptide and thereby allowing the cell to produce the polypeptide. Hamster cysteine sulfinic acid decarboxylase, a DNA encoding the same, a recombinant vector and a transformed cell are also provided.

1 . A method for producing a polypeptide, in which cells capable of high-yield production of β-alanine and into which DNA encoding a desired polypeptide has been transferred are cultured to produce the desired polypeptide. 
     
     
         2 . The production method of  claim 1 , wherein the cells capable of high-yield production of β-alanine are cells into which DNA encoding a protein having enzyme activity for synthesizing β-alanine from aspartic acid has been transferred. 
     
     
         3 . The production method of  claim 1 , wherein the cells capable of high-yield production of β-alanine are cells into which DNA encoding cysteine sulfinic acid decarboxylase, glutamate decarboxylase, or a mutant thereof has been transferred. 
     
     
         4 . The production method of  claim 1 , wherein the cells capable of high-yield production of β-alanine are cells in which the ratio of the intracellular concentration of β-alanine on the 6 th  or subsequent days after the initiation of the culture to that at the initiation of the culture is 2 or greater. 
     
     
         5 . The production method of  claim 1 , wherein the cells capable of high-yield production of β-alanine are capable of increasing the β-alanine concentration in the culture medium on 6 th  or subsequent days after the initiation of the culture by 50 μM or more. 
     
     
         6 . A cultured cell, in which the ratio of the intracellular concentration of β-alanine on the 6 th  or subsequent days after the initiation of the culture to that at the initiation of the culture is 2 or greater. 
     
     
         7 . A cultured cell, which is capable of increasing the β-alanine concentration in the culture medium on the 6 th  or subsequent days after the initiation of the culture by 50 μM or more. 
     
     
         8 . A method for producing a polypeptide, in which cells that strongly express cysteine sulfinic acid decarboxylase and into which DNA encoding a desired polypeptide has been transferred are cultured to produce the desired polypeptide. 
     
     
         9 . The production method of  claim 8 , wherein the cells that strongly express cysteine sulfinic acid decarboxylase are cells into which DNA encoding cysteine sulfinic acid decarboxylase has been transferred. 
     
     
         10 . The production method of  claim 8 , wherein the cells that strongly express cysteine sulfinic acid decarboxylase further express a taurine transporter strongly. 
     
     
         11 . The production method of  claim 10 , wherein the cells that strongly express a taurine transporter are cells into which DNA encoding a taurine transporter has been transferred. 
     
     
         12 . The method of  claim 9 , wherein the cell is Chinese hamster ovary cells. 
     
     
         13 . The method of  claim 8 , wherein the desired polypeptide is an antibody. 
     
     
         14 . The method of  claim 9 , wherein the DNA encoding the cysteine sulfinic acid decarboxylase is any one of the following (a) to (e):
 (a) a DNA encoding a polypeptide having the amino acid sequence as shown in SEQ ID NO: 2, 4, 6 or 8;   (b) a DNA encoding a polypeptide which has an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO: 2, 4, 6 or 8 by substitution, deletion, addition and/or insertion of one or more amino acid residues and yet has cysteine sulfinic acid decarboxylase activity;   (c) a DNA encoding a polypeptide having 70% or more amino acid sequence homology with the amino acid sequence as shown in SEQ ID NO: 2, 4, 6 or 8 and yet having cysteine sulfinic acid decarboxylase activity;   (d) a DNA having the nucleotide sequence as shown in SEQ ID NO: 1, 3, 5 or 7;   (e) a DNA which hybridizes to a DNA complementary to a DNA having the nucleotide sequence as shown in SEQ ID NO: 1, 3, 5 or 7 under stringent conditions and yet encodes a polypeptide having cysteine sulfinic acid decarboxylase activity.   
     
     
         15 . A DNA encoding cysteine sulfinic acid decarboxylase, which is any one of the following (a) to (e), provided that DNA having the nucleotide sequence as shown in SEQ ID NO: 3, DNA having the nucleotide sequence as shown in SEQ ID NO: 5, and DNA having the nucleotide sequence as shown in SEQ ID NO: 7 are excluded:
 (a) a DNA encoding a polypeptide having the amino acid sequence as shown in SEQ ID NO: 2;   (b) a DNA encoding a polypeptide which has an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO: 2 by substitution, deletion, addition and/or insertion of one or more amino acid residues and yet has cysteine sulfinic acid decarboxylase activity;   (c) a DNA encoding a polypeptide having 97% or more amino acid sequence homology with the amino acid sequence as shown in SEQ ID NO: 2 and yet having cysteine sulfinic acid decarboxylase activity;   (d) a DNA having the nucleotide sequence as shown in SEQ ID NO: 1;   (e) a DNA which hybridizes to a DNA complementary to a DNA having the nucleotide sequence as shown in SEQ ID NO: 1 under stringent conditions and yet encodes a polypeptide having cysteine sulfinic acid decarboxylase activity.   
     
     
         16 . A polypeptide which is any one of the following (A) to (E), provided that a polypeptide having the amino acid sequence as shown in SEQ ID NO: 4, a polypeptide having the amino acid sequence as shown in SEQ ID NO: 6, and a polypeptide having the amino acid sequence as shown in SEQ ID NO: 8 are excluded:
 (A) a polypeptide having the amino acid sequence as shown in SEQ ID NO: 2;   (B) a polypeptide which has an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO: 2 by substitution, deletion, addition and/or insertion of one or more amino acid residues and yet has cysteine sulfinic acid decarboxylase activity;   (C) a polypeptide having 97% or more amino acid sequence homology with the amino acid sequence as shown in SEQ ID NO: 2 and yet having cysteine sulfinic acid decarboxylase activity;   (D) a polypeptide encoded by a DNA having the nucleotide sequence as shown in SEQ ID NO: 1;   (E) a polypeptide encoded by a DNA which hybridizes to a DNA complementary to a DNA having the nucleotide sequence as shown in SEQ ID NO: 1 under stringent conditions and yet encodes a polypeptide having cysteine sulfinic acid decarboxylase activity.   
     
     
         17 . A recombinant vector comprising the DNA of  claim 15 . 
     
     
         18 . A cell into which the DNA of  claim 15  has been transferred. 
     
     
         19 . A cell which has a transferred DNA encoding cysteine sulfinic acid decarboxylase and a transferred DNA encoding a desired polypeptide. 
     
     
         20 . The cell of  claim 19 , which further has a transferred DNA encoding a taurine transporter. 
     
     
         21 . A cell which has a transferred DNA encoding cysteine sulfinic acid decarboxylase and a transferred DNA encoding a taurine transporter. 
     
     
         22 . A cell capable of high-yield production of β-alanine which can be obtained by transferring a DNA encoding cysteine sulfinic acid decarboxylase. 
     
     
         23 . The cell of  claim 22 , which further has a transferred DNA encoding a desired polypeptide.