Fructosyl peptidyl oxidase
First Claim
Patent Images
1. A device for assaying fructosyl valine, fructosyl valyl histidine, a fructosyl hexapeptide or HbA1c in a sample, comprising an electron transfer mediator and one of:
- a fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and a Km value toward fructosyl valyl histidine of 1 mM or less;
a fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and a Vmax/Km value toward fructosyl valyl histidine of 10 U/mg·
mM or more; and
a fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and residual activity of 50% or more when heat-treated at 50°
C. for 10 minutes.
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Abstract
In one form, a fructosyl peptidyl oxidase derived from a budding yeast Phaeosphaeria nodorum for assaying a glycated protein in a sample is provided. The fructosyl peptidyl oxidase has higher activity toward fructosyl valine as well as fructosyl valyl histidine, and may be useful in assaying HbA1c with higher sensitivity and specificity. Still, other forms include unique methods, techniques, systems and devices involving a fructosyl peptidyl oxidase.
10 Citations
16 Claims
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1. A device for assaying fructosyl valine, fructosyl valyl histidine, a fructosyl hexapeptide or HbA1c in a sample, comprising an electron transfer mediator and one of:
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a fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and a Km value toward fructosyl valyl histidine of 1 mM or less;a fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and a Vmax/Km value toward fructosyl valyl histidine of 10 U/mg·
mM or more; anda fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and residual activity of 50% or more when heat-treated at 50°
C. for 10 minutes. - View Dependent Claims (2, 3, 4)
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5. A kit for assaying fructosyl valine, fructosyl valyl histidine, a fructosyl hexapeptide or HbA1c in a sample, comprising an electron transfer mediator and one of:
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a fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and a Km value toward fructosyl valyl histidine of 1 mM or less;a fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and a Vmax/Km value toward fructosyl valyl histidine of 10 U/mg·
mM or more; anda fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and residual activity of 50% or more when heat-treated at 50°
C. for 10 minutes. - View Dependent Claims (6, 7, 8)
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9. An enzyme electrode, comprising one of:
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a fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and a Km value toward fructosyl valyl histidine of 1 mM or less;a fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and a Vmax/Km value toward fructosyl valyl histidine of 10 U/mg·
mM or more; anda fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and residual activity of 50% or more when heat-treated at 50°
C. for 10 minutes; andwherein the fructosyl peptidyl oxidase is immobilized on the electrode. - View Dependent Claims (10, 11, 12)
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13. An enzyme sensor for assaying fructosyl valine, fructosyl valyl histidine, a fructosyl hexapeptide or HbA1c, comprising a working electrode including one of:
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a fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and a Km value toward fructosyl valyl histidine of 1 mM or less;a fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and a Vmax/Km value toward fructosyl valyl histidine of 10 U/mg·
mM or more; anda fructosyl peptidyl oxidase comprising an amino acid sequence having at least 90% identity with the amino acid sequence set forth in SEQ ID NO;
1 and residual activity of 50% or more when heat-treated at 50°
C. for 10 minutes; andwherein the fructosyl peptidyl oxidase is immobilized on the electrode. - View Dependent Claims (14, 15, 16)
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Specification