JP2015171359A5 - - Google Patents

Claims (11)

At least 90% identity with the amino acid sequence shown by SEQ ID NO: 2, 9, 10, 11, 12, 13 or 14 and the amino acid sequence shown by SEQ ID NO: 2, 9, 10, 11, 12, 13 or 14 In an amino acid sequence or an amino acid sequence in which one or several amino acids are deleted, substituted or added in the amino acid sequence shown in SEQ ID NO: 2, 9, 10, 11, 12, 13 or 14 ,
At least one amino acid residue of amino acids at positions corresponding to P147, L214, L216, Q421, H423 or F447 of the amino acid sequence represented by SEQ ID NO: 2 is substituted with alanine ;
Compared to the pyranose oxidase before modification, the ratio (PDH / POD) of pyranose dehydrogenase activity (PDH specific activity) per protein amount and pyranose oxidase activity (POD specific activity) per protein amount is increased. A modified pyranose oxidase. The amino acid sequence shown in SEQ ID NO: 2, the amino acid sequence having at least 90% identity with the amino acid sequence shown in SEQ ID NO: 2, or one or several amino acids in the amino acid sequence shown in SEQ ID NO: 2 are deleted or substituted Or in the added amino acid sequence:
At least one amino acid residue of amino acids at positions corresponding to P147, L214, L216, Q421, H423 or F447 of the amino acid sequence represented by SEQ ID NO: 2 is substituted with alanine ;
Compared to the pyranose oxidase before modification, the ratio (PDH / POD) of pyranose dehydrogenase activity (PDH specific activity) per protein amount and pyranose oxidase activity (POD specific activity) per protein amount is increased. A modified pyranose oxidase. The amino acid at the position corresponding to P147 is proline, the amino acid at the position corresponding to L214 is leucine or isoleucine, the amino acid at the position corresponding to L216 is leucine, and the amino acid at the position corresponding to Q421 is glutamine, The modified pyranose oxidase according to claim 1 or 2, wherein the amino acid at the position corresponding to H423 is histidine, and the amino acid at the position corresponding to F447 is phenylalanine. The modified pyranose oxidation according to any one of claims 1 to 3 , wherein PDH / POD is at least twice when PDH / POD of the enzyme having the amino acid sequence represented by SEQ ID NO: 2 is made 1 time. enzyme. A polynucleotide encoding the modified pyranose oxidase according to any one of claims 1 to 4 . A recombinant vector comprising the polynucleotide according to claim 5 . A transformed cell transformed with the vector according to claim 6 . A method for producing a modified pyranose oxidase, comprising culturing the transformed cell of claim 7 and collecting the modified pyranose oxidase from the obtained culture. The pyranose measuring method using the modified pyranose oxidase of any one of Claims 1-4 . A reagent composition for measuring pyranose comprising the modified pyranose oxidase according to any one of claims 1 to 4 . A biosensor for measuring pyranose, comprising the modified pyranose oxidase according to any one of claims 1 to 4 .