Modified amadoriase and method for producing the same, agent for improving surfactant resistance of amadoriase and composition for measuring HbA1c using the same

    公开(公告)号:US11549134B2

    公开(公告)日:2023-01-10

    申请号:US16832712

    申请日:2020-03-27

    IPC分类号: C12Q1/26 C12N9/06 G01N33/72

    摘要: Provided is a composition by which glycated hemoglobin can be measured even in the presence of a stronger surfactant than a conventional case. Also provided is a buffer and/or stabilizer which maintains the residual activity of an amadoriase or lowers a reduction of residual activity. The present invention provides a composition for use in measuring glycated hemoglobin containing an amadoriase having substitution of one or more amino acid residues at a position(s) corresponding to an amino acid(s) selected from the group consisting of position 262, position 257, position 249, position 253, position 337, position 340, position 232, position 129, position 132, position 133, position 44, position 256, position 231 and position 81 of an amadoriase derived from the genus Coniochaeta and represented by SEQ ID No: 1 or 3, and having residual activity even in the presence of a surfactant. The present invention also provides a composition and kit for use in measuring glycated hemoglobin, comprising a specific stabilizer and/or a buffer. The present invention can provide an enzyme and a composition for use in measuring glycated hemoglobin, excellent in storage stability even if they are exposed to a surfactant.

    Amadoriase having enhanced dehydrogenase activity

    公开(公告)号:US11499143B2

    公开(公告)日:2022-11-15

    申请号:US15521104

    申请日:2015-10-23

    摘要: Provided are an amadoriase that is less likely to be influenced by oxygen concentration and a method and a reagent kit for measurement of HbA1c using such amadoriase. Provided are an amadoriase that is obtained via substitution of one or more amino acid residues at a position or positions corresponding to the position(s) selected from the group consisting of positions 280, 267, 269, 54, and 241 of the amadoriase derived from the genus Coniochaeta, a method for measurement of HbA1c, a reagent kit for measurement, and a sensor using such amadoriase. The modified amadoriase according to the invention has a lowered oxidase activity and an enhanced dehydrogenase activity, and this enables the use of an electron mediator, and this reduces the influence of oxygen concentration. Thus, HbA1c can be measured with high sensitivity.

    AMADORIASE HAVING ALTERED SUBSTRATE SPECIFICITY
    3.
    发明申请
    AMADORIASE HAVING ALTERED SUBSTRATE SPECIFICITY 有权
    具有改变基质特性的改性剂

    公开(公告)号:US20150247129A1

    公开(公告)日:2015-09-03

    申请号:US14715739

    申请日:2015-05-19

    IPC分类号: C12N9/06 C12Q1/26

    CPC分类号: C12N9/0032 C12N1/00 C12N15/63

    摘要: This invention provides an amadoriase having high substrate specificity to fructosyl valyl histidine. Such amadoriase comprises substitution of one or more amino acid residues at positions corresponding to amino acids selected from the group consisting of position 98, position 259, position 154, position 125, position 261, position 263, position 106, position 103, position 355, position 96, position 66, position 67, position 70, position 100, position 110, position 113, position 114, and position 156 in the amadoriase derived from the genus Coniochaeta. This invention enables accurate measurement of α-fructosyl valyl histidine derived from the β-chain amino terminus in glycated hemoglobin in the presence of ε-fructosyl lysine.

    摘要翻译: 本发明提供对果糖基缬氨酰组氨酸具有高底物特异性的阿马道里酶。 这样的阿马道里酶包含一个或多个氨基酸残基的取代,该位置对应于选自第98位,第259位,第154位,第125位,第261位,第263位,第106位,第103位,第355位的氨基酸。 位于第59位,位置66,位置67,位置70,位置100,位置110,位置113,位置114和位置156在源自孔雀属属的阿马多利亚酶。 本发明能够在糖果糖基赖氨酸的存在下,精确测定衍生自糖基化血红蛋白中的“氨基末端”的α-果糖基缬氨酰基组氨酸。

    Amadoriase having enhanced anionic surfactant tolerance

    公开(公告)号:US11198852B2

    公开(公告)日:2021-12-14

    申请号:US15523806

    申请日:2015-11-06

    摘要: The present invention provides a composition that enables measurement of glycated hemoglobin in the presence of a stronger surfactant than conventional surfactants. To this end, the present invention provides an amadoriase in which one or more amino acids have been substituted at positions corresponding to positions selected from the group consisting of positions 80, 71, 175, 172, 279, 12, 9, 77, 30, 28, 13, 3, 4, 286, 204, 338, 44, 340, and 194 of the amadoriase derived from the genus Coniochaeta having the amino acid sequence as shown in SEQ ID NO: 1 as well as a composition for measurement of glycated hemoglobin comprising an amadoriase that retains activity in the presence of an anionic surfactant. The present invention can provide an enzyme and a composition for measurement of glycated hemoglobin that sufficiently remain stable even when exposed to anionic surfactants.

    Cytochrome-fused glucose dehydrogenase and glucose measurement method

    公开(公告)号:US11208466B2

    公开(公告)日:2021-12-28

    申请号:US15778726

    申请日:2016-11-30

    摘要: A cytochrome b-glucose dehydrogenase fusion protein having modified electron transfer properties, and a glucose measurement method and measuring kit using the cytochrome b-glucose dehydrogenase fusion protein are provided. Provided are a cytochrome b-glucose dehydrogenase fusion protein in which glucose dehydrogenase having homology with SEQ ID NO: 1 or SEQ ID NO: 4 and cytochrome b are linked together, as well as a glucose measurement method, a measurement reagent kit and a sensor using the cytochrome b-glucose dehydrogenase fusion protein. The cytochrome b-glucose dehydrogenase fusion protein of the present invention has modified electron transfer properties, and can be used for measuring glucose in the presence of a free-form mediator in reduced concentration or in the absence of a free-form mediator, and can be used, for example, in continuous glucose monitoring.

    HbA1c dehydrogenase
    9.
    发明授权

    公开(公告)号:US11111517B2

    公开(公告)日:2021-09-07

    申请号:US16094924

    申请日:2017-04-21

    摘要: This invention provides an HbA1c dehydrogenase that is capable of directly acting on hemoglobin A1c and is less likely to be influenced by oxygen concentration and a method for measurement and a kit of assay reagents using such HbA1c dehydrogenase. The HbA1c dehydrogenase having dehydrogenase activity and capable of directly acting on HbA1c is obtained by substitution of one or more amino acid residues at positions corresponding to positions 280, 269, 54, 241, and 267 of the amadoriase that is capable of directly acting on hemoglobin A1c and is derived from, for example, the genus Coniochaeta. This invention also provides a method for measurement of HbA1c, a kit of assay reagents, and a sensor using such HbA1c dehydrogenase. Such HbA1c dehydrogenase is capable of directly acting on hemoglobin A1c and has lowered oxidase activity and/or enhanced dehydrogenase activity. This not only eliminates the need for treatment of hemoglobin A1c with a protease but also enables the use of an electron mediator in the measurement of HbA1c, thereby reducing effects due to oxygen concentration, and enables HbA1c measured with high sensitivity.

    Hemoglobin A1c measurement method and measurement kit

    公开(公告)号:US11078517B2

    公开(公告)日:2021-08-03

    申请号:US15031037

    申请日:2014-10-24

    摘要: This invention provides an amadoriase that acts on the β-chain of hemoglobin A1c (HbA1c) and generates hydrogen peroxide, a method for measurement of HbA1c using such amadoriase, and a reagent kit for measurement of HbA1c using such amadoriase. The method for measurement of HbA1c involves the use of an amadoriase that has specific activity of 0.1 U/mg or greater to αF6P and oxidizes the HbA1c β-chain amino terminus so as to generate hydrogen peroxide, and the reagent kit for measurement of HbA1c comprises such amadoriase. The method and the kit for measurement of HbA1c enable quantification of HbA1c to be performed rapidly, simply, and accurately.